Association of Marine Archaea with the Digestive Tracts of Two Marine Fish Species

Recent studies have shown that archaea which were always thought to live under strict anoxic or extreme environmental conditions are also present in cold, oxygenated seawater, soils, the digestive tract of a holothurian deep-sea-deposit feeder, and a marine sponge. In this study, we show, by using PCR-mediated screening in other marine eukaryotes, that marine archaea are also present in the digestive tracts of flounder and grey mullet, two fish species common in the North Sea, in fecal samples of flounder, and in suspended particulate matter of the North Sea water column. No marine archaea could be detected in the digestive tracts of mussels or the fecal pellets of a copepod species. The archaeal 16S ribosomal DNA clone libraries of feces of flounder and the contents of the digestive tracts of grey mullet and flounder were dominated by group II marine archaea. The marine archaeal clones derived from flounder and grey mullet digestive tracts and feces formed a distinct cluster within the group II marine archaea, with 76.7 to 89.8% similarity to previously described group II clones. Fingerprinting of the archaeal community of flounder digestive tract contents and feces by terminal restriction fragment length polymorphism of archaeal 16S rRNA genes after restriction with HhaI showed a dominant fragment at 249 bp, which is likely to be derived from group II marine archaea. Clones of marine archaea that were closely related to the fish-associated marine archaea clones were obtained from suspended particulate matter of the water column at two stations in the North Sea. Terminal restriction fragment length polymorphism fingerprinting of the archaeal community present in suspended particulate matter showed the same fragment pattern as was found for the archaeal community of the flounder digestive tract contents and feces. These data demonstrate that marine archaea are present in the digestive tracts and feces of very common marine fish. It is possible that the marine archaea associated with the digestive tracts of marine fish are liberated into the water column through the feces and subsequently contribute to the marine archaeal community of suspended particulate matter.     

Estimation of the air/sea exchange of ammonia for the North Atlantic Basin

As gas phase atmospheric ammonia reacts with acidic aerosol particles it affects the chemical, physical, and optical properties of the particles. A knowledge of the source strengths of NH₃ is useful in determining the effect of NH₃ on aerosol properties on a regional basis. Here, an attempt is made to determine the direction and magnitude of the air/sea flux of ammonia for the North Atlantic Basin from both measured and modeled seawater and atmospheric ammonia concentrations. Previously reported measured seawater concentrations range from less than 30 to 4600 nM with the highest concentrations reported for the Caribbean Sea, the North Sea, and the Belgium coast. Measured atmospheric ammonia concentrations range from 2 to 500 nmol m^–3 with the largest values occurring over the Sargasso Sea, the Caribbean Sea, and the North Sea. For comparison to the measurements, seawater ammonia concentrations were calculated by the Hamburg Model of the Ocean Carbon Cycle (HAMOCC3). HAMOCC3 open ocean values agree well with the limited number of reported measured concentrations. Calculated coastal values are lower than those measured, however, due to the coarse resolution of the model. Atmospheric ammonia concentrations were calculated by the Acid Deposition Model of the Meteorological Synthesizing Center (MSC-W) and by the global 3-dimensional model Moguntia. The two models predict similar annually averaged values but are about an order of magnitude lower than the measured concentrations. Over the North Sea and the NE Atlantic, the direction and magnitude of the air/sea ammonia flux calculated from MSC-W and Moguntia agree within the uncertainty of the calculations. Flux estimates derived from measured data are larger in both the positive and negative direction than the model derived values. The discrepancies between the measured and modeled concentrations and fluxes may be a result of sampling artifacts, inadequate chemistry and transport schemes in the models, or the difficulty in comparing point measurements to time-averaged model values. Sensitivity tests were performed which indicate that, over the range of values expected for the North Atlantic, the accuracy of the calculated flux depends strongly on seawater and atmospheric ammonia concentrations. Clearly, simultaneous and accurate measurements of seawater and atmospheric ammonia concentrations are needed to reduce the uncertainty of the flux calculations, validate the model results, and characterize the role of oceanic ammonia emissions in aerosol processing and nitrogen cycling for the North Atlantic.     

A study of rotifer feeding and digestive processes using erythrocytes as microparticulate markers

Feeding and passage of nutrient particles through the digestive tract of the rotifer Brachionus plicatilis was examined using the following types of particles: yeast cells, mammalian (human, bovine, ovine) erythrocytes and erythrocyte-ghosts. The decrease of erythrocytes within the medium was found to follow first order kinetics. The rate constant was influenced by the density of rotifers, as well as by the initial density of particles in suspension at the beginning of the experiment. Passage of particles through the digestive tract depended on the supply of fresh particles taken up by the wheel organ. Passage time approximately doubled when the rotifers had no access to fresh particles. On provision of fresh particles, the contents of the digestive tract were expelled even if they included absorbable substances. The contents of the digestive tract of B. plicatilis can be quickly and easily exchanged by introducing a new type of particle to the medium. By replacing fluorescence-labelled erythrocyte ghosts with unlabelled ghosts, residual fluorescence indicating absorptive processes could be demonstrated.     

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.     

Bacteria in bivalve shellfish with special reference to the oyster

The bacterial flora of the Pacific oyster Crassostrea gigas, the sea mussel Perna viridis and the arkshell clam Scapharca cornea differed considerably from that of seawater in both numbers and generic composition. The numbers of heterotrophic bacteria in the bivalve shellfish, including the anaerobes and spore-forming bacteria, were greater than that in the surrounding water. Pseudomonas spp. were the dominant organisms, comprising over one third of the 321 strains characterized after isolation from the bivalves and seawater. Other bacteria isolated from the shellfish included Vibrio, Acinetobacter, and Aeromonas spp., whereas the seawater flora consisted mainly of coliform organisms, coryneform bacteria and Flavobacterium/Cytophaga spp. Bacteria associated with the deposit-feeding clams were higher in density and more distinct in generic composition as compared with those in the suspension-feeding oysters and mussels. Over 90% of the coliform and heterotrophic bacteria in oysters were found in organs associated with the digestive tract. Coliforms were mainly found in the stomach while heterotrophs were present in both stomach and the lower intestine. The results suggest that the stomach flora of oysters are mainly derived from the external environment and, through a process of selection and multiplication, that it may be gradually replaced by a more indigenous population which dominates the lower digestive tract.     

Cultivation of particle-associated heterotrophic bacteria during a spring phytoplankton bloom in the North Sea

Seawater contains free-living and particle-attached bacteria. Only a small fraction is cultivable on plates. As free-living and particle-associated bacteria differ in their physiological traits, their cultivability on plates may coincide with particle association. Using filtration and Imhoff sedimentation cones, particles were collected during a spring phytoplankton bloom off Helgoland (North Sea) in order to obtain particle-associated bacteria as inocula. Direct dilution plating resulted in 526 strains from 3 µm filtration retentates and 597 strains from settled particles. Motile Gammaproteobacteria from the genera Pseudoalteromonas, Shewanella, Psychrobacter, Vibrio and Colwellia, as well as particle-attached Flavobacteriia affiliating with the genera Tenacibaculum and Gramella, were frequently isolated. As a result, a diverse collection comprised of 266 strains was deposited. Two strains were most likely to represent novel genera and 78 strains were probably novel species. Recently, a high-throughput cultivation study from the same site using seawater as an inoculum had retrieved 271 operational phylogenetic units (OPUs) that represented 88% of the 4136 characterized strains at the species level. A comparison of 16S rRNA gene sequences revealed that the collection obtained matched 104 of the 271 seawater OPUs at the species level and an additional 113 at the genus level. This large overlap indicated a significant contribution of particle-associated bacteria to the cultivable microbiome from seawater. The presence of 49 genera not identified in the larger seawater study suggested that sample fractionation was an efficient strategy to cultivate rare members of the planktonic microbiome. The diverse collection of heterotrophic bacteria retrieved in this study will be a rich source for future studies on the biology of particle-associated bacteria.     

Bacteria in bivalve shellfish with special reference to the oyster

K ueh , C.S.W. & C han , K-Y. 1985. Bacteria in bivalve shellfish with special reference to the oyster. Journal of Applied Bacteriology , 59 , 41–47.
The bacterial flora of the Pacific oyster Crassostrea gigas , the sea mussel Perna viridis and the arkshell clam Scapharca cornea differed considerably from that of seawater in both numbers and generic composition. The numbers of heterotrophic bacteria in the bivalve shellfish, including the anaerobes and spore-forming bacteria, were greater than that in the surrounding water. Pseudomonas spp. were the dominant organisms, comprising over one third of the 321 strains characterized after isolation from the bivalves and seawater. Other bacteria isolated from the shellfish included Vibrio, Acinetobacter , and Aeromonas spp., whereas the seawater flora consisted mainly of coliform organisms, coryneform bacteria and Flavobacterium/ Cytophaga spp. Bacteria associated with the deposit-feeding clams were higher in density and more distinct in generic composition as compared with those in the suspension-feeding oysters and mussels. Over 90% of the coliform and heterotrophic bacteria in oysters were found in organs associated with the digestive tract. Coliforms were mainly found in the stomach while heterotrophs were present in both stomach and the lower intestine. The results suggest that the stomach flora of oysters are mainly derived from the external environment and, through a process of selection and multiplication, that it may be gradually replaced by a more indigenous population which dominates the lower digestive tract.     

Virus Decay and Its Causes in Coastal Waters

Recent evidence suggests that viruses play an influential role within the marine microbial food web. To understand this role, it is important to determine rates and mechanisms of virus removal and degradation. We used plaque assays to examine the decay of infectivity in lab-grown viruses seeded into natural seawater. The rates of loss of infectivity of native viruses from Santa Monica Bay and of nonnative viruses from the North Sea in the coastal seawater of Santa Monica Bay were determined. Viruses were seeded into fresh seawater that had been pretreated in various ways: filtration with a 0.2-(mu)m-pore-size filter to remove organisms, heat to denature enzymes, and dissolved organic matter enrichment to reconstitute enzyme activity. Seawater samples were then incubated in full sunlight, in the dark, or under glass to allow partitioning of causative agents of virus decay. Solar radiation always resulted in increased rates of loss of virus infectivity. Virus isolates which are native to Santa Monica Bay consistently degraded more slowly in full sunlight in untreated seawater (decay ranged from 4.1 to 7.2% h(sup-1)) than nonnative marine bacteriophages which were isolated from the North Sea (decay ranged from 6.6 to 11.1% h(sup-1)). All phages demonstrated susceptibility to degradation by heat-labile substances, as heat treatment reduced the decay rates to about 0.5 to 2.0% h(sup-1) in the dark. Filtration reduced decay rates by various amounts, averaging 20%. Heat-labile, high-molecular-weight dissolved material (>30 kDa, probably enzymes) appeared responsible for about 1/5 of the maximal decay. Solar radiation was responsible for about 1/3 to 2/3 of the maximal decay of nonnative viruses and about 1/4 to 1/3 of that of the native viruses, suggesting evolutionary adaptation to local light levels. Our results suggest that sunlight is an important contributing factor to virus decay but also point to the significance of particles and dissolved substances in seawater.     

Localization of norovirus and poliovirus in Pacific oysters

Aims:  To examine the uptake and tissue distribution of norovirus (NoV) and poliovirus (PV) experimentally bioaccumulated in feeding Pacific oysters ( Crassostrea gigas ).
Methods and Results:  Pacific oysters were allowed to bioaccumulated either PV or NoV under tidally synchronized feeding conditions in laboratory tanks. Oysters were then either fixed and paraffin wax embedded prior to localizing virus within tissues by immunohistochemistry (IHC), or they were dissected into digestive tract (stomach, intestine and digestive diverticula), gill and labial palp tissues, and the viral load determined by quantitative RT-PCR. Both PV and NoV immunoreactivities were predominantly found in the lumen and within cells of the digestive tract tissues; however, PV was also found within cells of nondigestive tract tissues, and in the gills and labial palp. Quantitative RT-PCR of tissue extracts corroborate the immunohistochemical data in that the major site for virus localization is the gut, but significant amounts of viral RNA were identified in the gills and labial palp.
Conclusions:  The human enteric viruses, PV and NoV, are readily bioaccumulated by feeding Pacific oysters and that some of the virus is internalized within cells of both digestive and nondigestive tissues.
Significance and Impact of the Study:  Oysters that have been virally contaminated even after depuration (cleaning) in uncontaminated seawater could pose a human health risk if consumed.     

Monoamine- and peptide-containing elements in the nemertine digestive tract

Detailed comparative-histological data on localization and morphological peculiarities of catecholamine-, serotonin-, neurotensin- and FMRFamide-containing elements in the nemertine digestive system are presented for the first time. Using fluorescent histochemistry and immunohistochemistry, pharynx, esophagus, and midgut were studied in five species from three genera of the White Sea nemertines. Described in various nemertine species are intra- and subepithelial cells of the open and closed type, containing biologically active substances. Their processes are distributed in basi- and subepithelial nerve plexuses and can make contact with lumen of the digestive tract. Species-specificity in localization of the cells containing certain biologically active substances is noticed both along the length of the tract and with respect to its epithelial layer. Peculiarities and common regularities in distribution are discussed, as well as a possible function of monoamine- and peptide-containing elements in the digestive tract of the studied nemertines and other invertebrates.     

Survival of foraminifera in the gut of holothuroids from Elba Island (Mediterranean Sea)

Detritivorous macroorganisms have important functions in nutrient remineralization and may exert significant predation pressure on meiofaunal communities. We have examined food excrements of detritus grazing holothuroids off Elba Island (Mediterranean Sea) to investigate whether ingested foraminifera survive the passage through the digestive tract. Our findings provide evidence that some benthic foraminifera pass through the complete digestive tract without being harmed by its predators. Sediment-feeding holothurians may thus affect microdistributional processes ultimately resulting in an allochthonous redistribution of foraminifera within the environment.     

Prevalence and microdiversity of Alteromonas macleodii-like microorganisms in different oceanic regions

The presence, prevalence and variability of microorganisms related to the species Alteromonas macleodii, a well known culturable gamma-Proteobacterium, has been studied in different seawater samples from diverse geographical locations, in both the Northern and Southern hemispheres, and tested with two molecular techniques (rRNA hybridization and gene cloning and sequencing). Results show that A. macleodii-like microorganisms are present in high proportions in North Atlantic and, especially, Mediterranean waters, being higher at deep samples and particle-associated fractions, in agreement with previous findings. In contrast, Southern samples (all from very cold areas near Antarctica) presented no significant hybridization signals. The analysis of the ribosomal ITS (16S-23S internal transcribed spacers) revealed that A. macleodii-like microorganisms from Mediterranean, North Atlantic, Caribbean and Red Sea waters differed in both size and sequence, mostly depending on their geographical origin, with Mediterranean and North Atlantic clones clustering into two main groups whereas Caribbean and Red Sea clones appeared separated.     

Retention of fluid and particles in the digestive tract of the llama (Lama guanacoe f. glama)

The retention times of fluid and particles of different lengths were measured in the digestive tract of the llama. PEG was used as a fluid marker; hay particles were labelled with cerium or samarium and the samples were determined by neutron activation analysis. In compartments 1 and 2 the retention time of fluid (9.7 hr) was significantly shorter than that of particles having a length of 0.2-1.0 cm (19.2 hr) and 2.5-4.0 cm (25.0 hr). In the intestine slightly significant differences in the retention times of fluid (20.1 hr) and particles (26.2 hr) were seen. In the whole digestive tract retention times were 36.2 hr for fluid, 52.0 hr for smaller particles and 59.9 hr for larger particles.     

NADPH-diaphorase activity in the digestive system of gastropod molluscs <Emphasis Type="Italic">Achatina fulica</Emphasis> and <Emphasis Type="Italic">Littorina littorea</Emphasis>

Distribution and morphological peculiarities of nitroxidergic elements throughout the entire length of digestive tract was studied for the first time in gastropod molluscs Littorina littorea (Prosobranchia) and Achatina fulica (Pulmonata) using histochemical detection of NADPH-diaphorase (NADPHd). NO-ergic cells and fibers were revealed in all parts of the mollusc digestive system beginning from esophagus. Intensive NADPHd activity is found in a great number of intraepithelial cells of the open type and their processes in the intraand subepithelial nerve plexuses, subepithelial neurons, granular connective tissue cells, and multiple nervous fibers distributed among muscular elements of digestive tract as well as those in nerves innervating the tract. NADPHd was also revealed in receptor cells in the oral area and in the A. fulica CNS ganglia innervating the digestive tract. A. fulica has a more complicated organization of A. fulica nitroxidergic system of the digestive tract. A system of glomerular structures formed by thin NADPHd-positive neural fibers coming from epithelium is found directly beneath the epithelium in esophagus, stomach, and midgut of the mollusc. More superficially under the main groups of muscular elements there are revealed small clusters of NADPHd-positive neurons that can be classified as primitive, non-structured microganglia. The distribution pattern and a possible functional role of nitroxidergic elements in digestive tract of molluscs as compared with other invertebrate and vertebrate animals are discussed.     

The utilization of fly ash by two tube-building polychaetes

Benthic tube-building polychaetes which utilize sediment particles have been found to incorporate fly ash in their tubes. Experiments were conducted on North Sea specimens of Myriochele oculata Zachs and Pectinaria koreni (Malmgren) in artificial sediments offering a range of fly ash concentrations. A marked selection for non-ash particles was shown by Myriochele oculata, and a marginal selection against ash by Pectinaria koreni. It is suggested that this selection is based on the inorganic nature of fresh ash particles, assuming initial collection for feeding: as ash particles at the sea bed develop an organic component, such selection will disappear.     

Evidence by electron micrographs for a high incidence of bacteriophage particles in the waters of Yaquina Bay, oregon: ecological and taxonomical implications.

A variety of viral particles, the majority of them clearly identifiable as bacteriophages, were found in the seawater of Yaquina Bay, Oregon. These phages were obtained as free particles from the seawater without employing specific hosts for enrichments or further purification in the laboratory. A variety of electron micrographs showing different morphologies of phages as well as phage-bacterium interactions found in the seawater are presented. In the area where the bay received organic enrichment from seafood processing plants, a minimum of 10(4) phage particles per ml was estimated. Since the technique used was designed to concentrate particles 0.2 micrometer in diameter or larger it is assumed that the actual number of phage particles is higher than 10(4) particles per ml. The implications of the presence of such phage concentrations in bays and estuaries with a certain level of eutrophication are of obvious importance in considering the microbial ecology of these environments.     

Role of bats in the ecology of Paracoccidioides brasiliensis: the survival of Paracoccidioides brasiliensis in the intestinal tract of frugivorous bat, Artibeus lituratus

The fruit-eating bat, Artibeus lituratus, was fed known quantities of viable yeast cells and mycelial particles of Paracoccidioides brasiliensis in an attempt to assess the role of this animal in the distribution of this agent in nature. Results of mycosal cultures of the stomach, upper intestine, lower intestine and rectum clearly showed that the fungal cells were unable to survive more than 8 hours in the digestive tract of the bat. The mycelial particles were more susceptible than the yeast and were killed before passing to the rectum. The fungus died rapidly in the voided fecal material. These findings indicate the improbability of isolating P. brasiliensis from the digestive tract of wild captured bats and show that A. lituratus probably plays no role in the distribution of this fungus in nature.     

Sulfate transport mechanisms in epithelial systems

A novel invertebrate gastrointestinal transport mechanism has been shown to couple chloride-sulfate exchange in an electrogenic fashion. In the lobster, Homarus americanus, the hepatopancreas, or digestive gland, exists as an outpocketing of the digestive tract, representing a single cell layer separating the gut lumen and an open circulatory system composed of hemolymph. Investigations utilizing independently prepared brush border and basolateral membrane vesicles revealed discrete antiport systems which possess the capacity to bring about a transcellular secretion of sulfate. The luminal antiport system functions as a high-affinity, one-to-one chloride-sulfate exchanger that is stimulated by an increase in luminal hydrogen ion concentration. Such a system would take advantage of the high chloride concentration of ingested seawater as well as the high proton concentrations generated during digestion, which further suggests a potential regulation by resident sodium-proton exchangers. Exchange of one chloride for one divalent sulfate ion provides the driving force for electrogenic vectorial translocation. The basolateral antiport system was found to be electroneutral in nature, responsive to gradients of the dicarboxylic anion oxalate while lacking in proton stimulation. No evidence of sodium-sulfate co-transport, commonly reported for the brush border of vertebrate renal and intestinal epithelia, was observed in either membrane preparation. The two antiporters together can account for the low hemolymph to seawater sulfate levels previously described in decapod crustaceans. A secretory pathway for sulfate based upon electrogenic chloride-antiport may appear among invertebrates partly in response to digestion taking place in a seawater environment. J. Exp. Zool. 289:245-253, 2001.     

Evidence by electron micrographs for a high incidence of bacteriophage particles in the waters of Yaquina Bay, oregon: ecological and taxonomical implications.

A variety of viral particles, the majority of them clearly identifiable as bacteriophages, were found in the seawater of Yaquina Bay, Oregon. These phages were obtained as free particles from the seawater without employing specific hosts for enrichments or further purification in the laboratory. A variety of electron micrographs showing different morphologies of phages as well as phage-bacterium interactions found in the seawater are presented. In the area where the bay received organic enrichment from seafood processing plants, a minimum of 10(4) phage particles per ml was estimated. Since the technique used was designed to concentrate particles 0.2 micrometer in diameter or larger it is assumed that the actual number of phage particles is higher than 10(4) particles per ml. The implications of the presence of such phage concentrations in bays and estuaries with a certain level of eutrophication are of obvious importance in considering the microbial ecology of these environments.     

Westward range expansion of the Lessepsian migrant Fistularia commersonii (Fistulariidae) in the Mediterranean Sea, with notes on its parasites

One adult female bluespotted cornetfish Fistularia commersonii was captured near the coast of Arbatax (Sardinia, Italy) in late October 2005. Macroscopic and microscopic analysis of the gonad revealed a postspawning ovary. Several body parts of the damselfish Chromis chromis were found in its stomach contents. Different developmental stages of 58 metazoan parasites belonging to Arthropoda, Nematoda and Platyhelminthes were detected in its mouth and digestive tract. The lepocreadiid Allolepidapedon fistulariae was reported for the first time from the Mediterranean and, with the exception of this species, all the parasites found were reported for the first time in this host. This new record of F. commersonii may confirm the rapid range expansion of this Lessepsian migrant in the western Mediterranean Sea, as well as the presence of its endoparasite A. fistulariae .