New Method for Isolating Barophiles from Intestinal Contents of Deep-Sea Fishes Retrieved from the Abyssal Zone

We devised a new method (the dorayaki method) using marine agar under in situ pressures to isolate barophilic bacteria from the intestinal contents of three deep-sea fishes (two Coryphaenoides yaquinae samples and one Ilyophis sp. sample) retrieved from depths of 4,700 to 6,100 m in the Northwest Pacific Ocean. All 10 strains isolated from one sample (C. yaquinae) were obligately barophilic. One of the 10 strains did not grow at atmospheric pressure and 103.4 MPa but did grow well between 20.7 and 82.7 MPa, with optimal growth at 41.4 MPa. This method is useful for isolating psychrophilic and barophilic deep-sea bacteria.     

Effects of Hydrostatic Pressure on Catalysis by Epaxial Muscle Phosphofructokinase from an Abyssal Fish

SYNOPSIS: Phosphofructokinase (PFK) extracted from muscle ofabyssal Coryphaenoides fishes common in the deep waters aroundthe Galápagos Archipelago is extremely unstable upondecompression and extraction and can be recovered only in lowactivities. Preliminary studies indicate that the pressure responsesof the enzyme are complex: At low pressures, the enzyme is activated;at moderate pressures, activity passes through a pressure optimum;at high pressures, maximum catalytic activity is decelerated.At low pressures, the volume change of activation is about –11cm³/mole at 3°C and increases to about –46 cm³/moleat 28°C. The homologous enzyme from a surface species (Oligoplitesmundus) appears to be more pressure sensitive at low temperatures.     

Extrapolation from Individual-Level Responses to Population Growth Rate Using Population Modeling

Ecological risk assessments of chemicals are often based on simple measurements of toxicity in individuals. However, the protection goals are often set at the population and community levels. Population models may be a useful tool to extrapolate from individual-level measurements to population-level endpoints. In the present study, the population growth rate (λ) was calculated for three sets of full life-cycle data (Tetranychus urticae exposed to agrimek, and Daphnia pulex exposed to spinosad and diazinon). The results were compared to λ from population models, where survival and/or reproduction were adjusted according to 4 d of data from the same life-cycle data. This was done to determine whether truncated demographic data can give results similar to that obtained with full life-cycle data. The resulting correlations were strong when both effects on survival and reproduction were included in the model (p < .001, 0.93 < R₂ < 1.00). There were also strong correlations in several cases when only effects on survival or reproduction were considered, although the total risk to the population tended to be underestimated. The results of the present study show that population models can be useful to extrapolate truncated data on the individual level to more ecologically relevant population-level endpoints.     

Deep-Sea Bacteria: Growth and Utilization of Hydrocarbons at Ambient and In Situ Pressure

Microorganisms present in Atlantic Ocean sediment samples collected at a depth of 4,940 m were found to be capable of utilizing hydrocarbons under both ambient and in situ pressures. The rate of utilization under in situ pressure (500 atm) and ambient temperature (20 C) was found to be significantly less compared with hydrocarbon utilization examined under conditions of ambient temperature (20 C) and pressure (1 atm).     

Vertical Migrations of a Deep-Sea Fish and Its Prey

It has been speculated that some deep-sea fishes can display large vertical migrations and likely doing so to explore the full suite of benthopelagic food resources, especially the pelagic organisms of the deep scattering layer (DSL). This would help explain the success of fishes residing at seamounts and the increased biodiversity found in these features of the open ocean. We combined active plus passive acoustic telemetry of blackspot seabream with in situ environmental and biological (backscattering) data collection at a seamount to verify if its behaviour is dominated by vertical movements as a response to temporal changes in environmental conditions and pelagic prey availability. We found that seabream extensively migrate up and down the water column, that these patterns are cyclic both in short-term (tidal, diel) as well as long-term (seasonal) scales, and that they partially match the availability of potential DSL prey components. Furthermore, the emerging pattern points to a more complex spatial behaviour than previously anticipated, suggesting a seasonal switch in the diel behaviour mode (benthic vs. pelagic) of seabream, which may reflect an adaptation to differences in prey availability. This study is the first to document the fine scale three-dimensional behaviour of a deep-sea fish residing at seamounts.     

Effects of Growth Pressure and Temperature on Fatty Acid Composition of a Barotolerant Deep-Sea Bacterium

The effects of pressure and temperature on the fatty acid composition in a barotolerant deep-sea bacterium that had branched-chain fatty acids were examined. The major fatty acids of the strain at atmospheric pressure were iso-C_15:0, C_16:1, iso-C_17:0, and iso-C_17:1. As the growth pressure increased, the proportion of unsaturated fatty acid increased because of an increase in the proportion of iso-C_17:1. On the other hand, as the growth temperature decreased, the proportion of unsaturated fatty acid increased because of the increase in the proportion of C_16:1 and C_18:1.     

Peripheral Blood Parameters of Two Species of the Deep-Sea Fish Family Paralepididae

Biology Bulletin - The hematological and cytochemical parameters of the peripheral blood of the spotted barracudina Arctozenus risso and the sharpchin barracudina Paralepis coregonoides...     

Plant Growth Analysis: Growth and Yield Component Responses to Population Density in Forage Maize

Forage maize (Zea mays L.) was grown in monocultures at populationdensities ranging from 4·9 to 11·1 plants m^–2.Data for plant growth analysis were obtained from six harvestscarried out from 21 to 115 d after planting. Conventional plantgrowth analysis indicated that improvements in forage productivityper unit land area by high population density resulted directlyfrom increased plant presence. Reduction in dry weight per shootat high population density was associated with reduced unitleaf rate. Leaf area ratio was little affected, which may implythat competition for soil nutrients or oxygen was the chiefcause of plant interference. Yield component analysis demonstratedthe increasing importance of population density treatments asa source of variation as growth progressed. Direct relationshipsbetween variation in yield per plant and variation in two yieldcomponents, stem diameter and the inverse of leaf area ratio,were demonstrated. Both conventional plant growth analysis andyield component analysis indicated complex physiological andmorphological adjustments to species population density. Plant growth analysis, yield component analysis, Zea mays L     

Pressure Effects on Cholesterol and Lipid Synthesis by the Swimbladder of an Abyssal Coryphaenoides Species

SYNOPSIS. The swimbladder gas gland is recognized as a cholesterolsynthesis site in abyssal rattail fishes, Coryphaenoides sp.(from 2000 m depth), and Galápagos Islands surface fish,Orthopristis forbesi, Seriola mazatlana, and Sphoeroides annulatus.This relates to high levels of cholesterol in the gas gland(up to 21% of lipid) and high cholesterol levels in the fattyswimbladder interior (up to 49% of lipid). The gas gland hasmore protein (45.4%) than the internal fatty mass (18%). Lipidssynthesized include phospholipids and triglycerides in 2:1 ratioin the gas gland and 1:2 ratio in the liver. Deep fish havefatty livers (66%) compared to shallow fish (28%). Shallow fishincorporated five times as much acetate-l-¹⁴C into lipids asdid deep fish, and seven to eight times as much acetate-l-¹⁴Cinto cholesterol. Pressure facilitation of cholesterol synthesiswas observed in gas gland and liver of O. forbesi and Coryphaenoides,whereas total lipid synthesis was inhibited by higher pressures.Optimal acetate-l-¹⁴C incorporation into lipids occurred at5000 psi and 2°C in Coryphaenoides; it occurred at 14.7psi and 15° in O. forbesi. These conditions closely approximatethe environment of the fish.     

Trophosome of the Deep-Sea Tubeworm Riftia pachyptila Inhibits Bacterial Growth

The giant tubeworm Riftia pachyptila lives in symbiosis with the chemoautotrophic gammaproteobacterium Cand. Endoriftia persephone. Symbionts are released back into the environment upon host death in high-pressure experiments, while microbial fouling is not involved in trophosome degradation. Therefore, we examined the antimicrobial effect of the tubeworm’s trophosome and skin. The growth of all four tested Gram-positive, but only of one of the tested Gram-negative bacterial strains was inhibited by freshly fixed and degrading trophosome (incubated up to ten days at either warm or cold temperature), while no effect on Saccharomyces cerevisiae was observed. The skin did not show antimicrobial effects. A liquid chromatography-mass spectrometric analysis of the ethanol supernatant of fixed trophosomes lead to the tentative identification of the phospholipids 1-palmitoleyl-2-lyso-phosphatidylethanolamine, 2-palmitoleyl-1-lyso-phosphatidylethanolamine and the free fatty acids palmitoleic, palmitic and oleic acid, which are known to have an antimicrobial effect. As a result of tissue autolysis, the abundance of the free fatty acids increased with longer incubation time of trophosome samples. This correlated with an increasing growth inhibition of Bacillus subtilis and Listeria welshimeri, but not of the other bacterial strains. Therefore, the free fatty acids produced upon host degradation could be the cause of inhibition of at least these two bacterial strains.     

Population Structure and Phylogenetic Characterization of Marine Benthic Archaea in Deep-Sea Sediments

During the past few years Archaea have been recognized as a widespread and significant component of marine picoplankton assemblages and, more recently, the presence of novel archaeal phylogenetic lineages has been reported in coastal marine benthic environments. We investigated the relative abundance, vertical distribution, phylogenetic composition, and spatial variability of Archaea in deep-sea sediments collected from several stations in the Atlantic Ocean. Quantitative oligonucleotide hybridization experiments indicated that the relative abundance of archaeal 16S rRNA in deep-sea sediments (1500 m deep) ranged from about 2.5 to 8% of the total prokaryotic rRNA. Clone libraries of PCR-amplified archaeal rRNA genes (rDNA) were constructed from 10 depth intervals obtained from sediment cores collected at depths of 1,500, 2,600, and 4,500 m. Phylogenetic analysis of rDNA sequences revealed the presence of a complex archaeal population structure, whose members could be grouped into discrete phylogenetic lineages within the two kingdoms, Crenarchaeota and Euryarchaeota. Comparative denaturing gradient gel electrophoresis profile analysis of archaeal 16S rDNA V3 fragments revealed a significant depth-related variability in the composition of the archaeal population.     

Supernatant from Cultured Intestinal Cells Inhibits the Growth of Gram-Negative Bacteria

Bacterial growth chambers of Transwell units bearing intestinal epithelial monolayers (C2BBe) was consistently observed to be stagnant during the course of transmigration studies with Salmonella typhi. This limitation could not be explained by varying the bacterial load in the inoculum. Conditioned media produced by cultured C2BBe cells would not support bacterial growth. Growth support of the media was restored by heating to 95 C for 10 min. C2BBe conditioned media had bacteriostatic activity for a large variety of gram-negative, enteropathogenic bacteria but had no effect on gram-positive bacteria.     

Extrapolating Growth Reductions in Fish to Changes in Population Extinction Risks: Copper and Chinook Salmon

Fish commonly respond to stress, including stress from chemical exposures, with reduced growth. However, the relevance to wild populations of subtle and sometimes transitory growth reductions may not be obvious. At low-level, sustained exposures, Cu is one substance that commonly causes reduced growth but little mortality in laboratory toxicity tests with fish. To explore the relevance of growth reductions under laboratory conditions to wild populations, we (1) estimated growth effects of low-level Cu exposures to juvenile Chinook salmon (Oncorhynchus tshawytscha), (2) related growth effects to reduced survival in downriver Chinook salmon migrations, (3) estimated population demographics, (4) constructed a demographically structured matrix population model, and (5) projected the influence of Cu-reduced growth on population size, extinction risks, and recovery chances. Reduced juvenile growth from Cu in the range of chronic criteria concentrations was projected to cause disproportionate reductions in survival of migrating juveniles, with a 7.5% length reduction predicting about a 23% to 52% reduction in survival from a headwaters trap to the next census point located 640 km downstream. Projecting reduced juvenile growth out through six generations (～30 years) resulted in little increased extinction risk; however, population recovery times were delayed under scenarios where Cu-reduced growth was imposed.     

Two New Species of Abyssal Pogonophores from the Antarctic Ocean

Two new species of the genus Polybrachia Ivanov, Polybrachia macrolamellosa sp. n. and P. romanovi sp. n., found at two stations in the Atlantic sector of the Antarctic Ocean on cruises 11 and 43 of the R/V Akademik Kurchatov (1971 and 1985–1986) are described. The material was collected in the Scotia Sea (off northern Elephant Isle, at a depth of 3110 m) and in the South Sandwich Trench (8004–8116 m). The main distinctive features of the new species are the extremely large cuticular plaque (up to 88 µm in maximum diameter) on the trunk metameric papillae in P. macrolamellosa sp. n. and the very long cephalic lobe (L_cl/D_f is about 2) in P. romanovi sp. n.Original Russian Text Copyright ¢ 2005 by Biologiya Morya, Smirnov.     

Monounsaturated but Not Polyunsaturated Fatty Acids Are Required for Growth of the Deep-Sea Bacterium Photobacterium profundum SS9 at High Pressure and Low Temperature

There is considerable evidence correlating the production of increased proportions of membrane unsaturated fatty acids (UFAs) with bacterial growth at low temperatures or high pressures. In order to assess the importance of UFAs to microbial growth under these conditions, the effects of conditions altering UFA levels in the psychrotolerant piezophilic deep-sea bacterium Photobacterium profundum SS9 were investigated. The fatty acids produced by P. profundum SS9 grown at various temperatures and pressures were characterized, and differences in fatty acid composition as a function of phase growth, and between inner and outer membranes, were noted. P. profundum SS9 was found to exhibit enhanced proportions of both monounsaturated (MUFAs) and polyunsaturated (PUFAs) fatty acids when grown at a decreased temperature or elevated pressure. Treatment of cells with cerulenin inhibited MUFA but not PUFA synthesis and led to a decreased growth rate and yield at low temperature and high pressure. In addition, oleic acid-auxotrophic mutants were isolated. One of these mutants, strain EA3, was deficient in the production of MUFAs and was both low-temperature sensitive and high-pressure sensitive in the absence of exogenous 18:1 fatty acid. Another mutant, strain EA2, produced little MUFA but elevated levels of the PUFA species eicosapentaenoic acid (EPA; 20:5n-3). This mutant grew slowly but was not low-temperature sensitive or high-pressure sensitive. Finally, reverse genetics was employed to construct a mutant unable to produce EPA. This mutant, strain EA10, was also not low-temperature sensitive or high-pressure sensitive. The significance of these results to the understanding of the role of UFAs in growth under low-temperature or high-pressure conditions is discussed.     

Population Sizes and Distribution of Primates in the Lower Tana River Forests,Kenya

We studied the population size and distribution of diurnal primates in the lower Tana River forests, Kenya. They are the only remaining habitats for 2 threatened primates: the Tana River red colobus (Procolobus rufomitratus) and the Tana River crested mangabey (Cercocebus galeritus galeritus). We conducted censuses in 73 forest patches from January through March 2001. We estimate population size of the red colobus to be 788 individuals in 82 groups and that of the crested mangabeys to be 2,070 individuals in 59 groups. The data suggest that over a 7-year period (1994-2001), there was an 18% increase in the crested mangabey population and a 5% decline in red colobus numbers. Further, the red colobus range has expanded both north and south, whereas that of crested mangabeys has only expanded south. Fifty-six percent of crested mangabeys and 46% of red colobus groups were inside the Tana River Primate National Reserve (TRPNR). Other primates encountered included 170 groups of Sykes' monkeys (Cercopithecus mitis), 70 groups of yellow baboons (Papio cynocephalus) and 4 groups of grivets [Chlorocebus (Cercopithecus) aethiops]. Mean group densities of the 2 endangered primates and of baboons were higher inside than outside the TRPNR, reinforcing the importance of TRPNR for their conservation. An intervention program is required to stem further decline in the red colobus population and to protect small isolated groups in forest patches outside TRPNR.     

Distribution of Aeromonas Species in the Intestinal Tracts of River Fish

Aeromonas isolates were obtained from fish intestines, water, and sediments from an urban river and identified by the DNA-DNA microplate hybridization method. The isolates were Aeromonas veronii (22%), Aeromonas caviae (18%), Aeromonas hydrophila (13%), Aeromonas sobria (8%), Aeromonas jandaei (7%), and other Aeromonas spp. (33%). Aeromonas species occurred at high densities with high incidences, regardless of season. The results strongly suggest that aeromonads are indigenous in fish intestines, water, and sediments of rivers and have the potential to be predominant in aquatic environments.     

Species Distribution and Population Connectivity of Deep-Sea Mussels at Hydrocarbon Seeps in the Gulf of Mexico

Hydrocarbon seepage is widespread and patchy in the Gulf of Mexico, and six species of symbiont containing bathymodiolin mussels are found on active seeps over wide and overlapping depth and geographic ranges. We use mitochondrial genes to discriminate among the previously known and a newly discovered species and to assess the connectivity among populations of the same species in the northern Gulf of Mexico (GoM). Our results generally validate the morphologically based distribution of the three previously known GoM species of Bathymodiolus, although we found that approximately 10% of the morphologically based identifications were incorrect and this resulted in some inaccuracies with respect to their previously assigned depth and geographical distribution patterns. These data allowed us to confirm that sympatry of two species of Bathymodiolus within a single patch of mussels is common. A new species of bathymodiolin, Bathymodiolus sp. nov., closely related to B. heckerae was also discovered. The two species live at the same depths but have not been found in sympatry and both have small effective population sizes. We found evidence for genetic structure within populations of the three species of Bathymodiolinae for which we had samples from multiple sites and suggest limited connectivity for populations at some sites. Despite relatively small sample sizes, genetic diversity indices suggest the largest population sizes for B. childressi and Tamu fisheri and the smallest for B. heckerae and B. sp. nov. among the GoM bathymodiolins. Moreover, we detected an excess of rare variants indicating recent demographic changes and population expansions for the four species of bathymodiolins from the Gulf of Mexico.