Investigation of relationships between marine diatoms and the bacterium Listeria monocytogenes

Relationships between marine diatoms and the bacterium Listeria monocytogenes have been studied by routine algological methods and high-resolution video-enhanced differential interference contrast light microscopy. The study showed that the relationship between the listeria and the benthic diatom Navicula sp. has a parasitic character, whereas the relationship between the listeria and the planktonic diatom Phaeodactylum tricornutum is protocooperative.     

Quantifying Translocation of Listeria monocytogenes in Rats by Using Urinary Nitric Oxide-Derived Metabolites

The urinary nitric oxide metabolites NO₂⁻ and NO₃⁻ (summed as NO_x) are a noninvasive, quantitative biomarker of translocation of salmonella from the intestinal lumen to systemic organs. Listeria monocytogenes is a food-borne gram-positive pathogen that can also cross the intestinal epithelium. In this study, we tested the efficacy of urinary NO_x as a marker of listeria translocation. Rats (eight per group) were orally infected with increasing doses of L. monocytogenes; control rats received heat-killed listeria. The kinetics of urinary NO_x and population levels of listeria in feces were determined for 7 days. Another group of rats was killed 1 day after infection to verify translocation by culturing viable listeria from systemic organs. Oral administration of increasing doses of L. monocytogenes resulted in a time- and dose-dependent increase in urinary NO_x excretion. Translocation was a prerequisite for inducing a NO_x response, since heat-killed L. monocytogenes did not elevate NO_x excretion in urine. Fecal counts of listeria also showed dose and time dependency. Moreover, the number of viable L. monocytogenes cells in mesenteric lymph nodes also increased in a dose-dependent manner and correlated with urinary NO_x. In conclusion, urinary NO_x is a quantitative, noninvasive biomarker of listeria translocation.     

Diatoms in northeast Pacific surface sediments as paleoceanographic proxies

Fossil diatom total abundances (# valves/g) in 54 surface-sediment samples from the northeast (NE) Pacific Ocean reflect the position of high primary production associated with coastal upwelling and that possible biases associated with dilution or dissolution are small. Diatom species assemblages, defined by Q-mode factor analysis in 30 samples with abundant diatoms, are related to modern oceanographic properties. Five statistical assemblages, given by five specific diatom species and/or groups, are related to upwelling (Chaetoceros spores), subtropical (Thalassionema nitzschioides), subarctic (Rhizosolenia hebetata), transitional (Neodenticula seminae) and freshwater (freshwater diatoms) ecological environments. These factors are significantly correlated with primary productivity, temperature, nutrient concentrations and salinity, although the strongest relationship is that between diatom assemblages and productivity. However, it is not possible to distinguish between coastal and open-ocean (curl-driven) upwelling based on Chaetoceros spores relative percentages by themselves or on the floral factors.     

Morphological and physiological alterations in the diatom Gomphonema pseudoaugur due to heavy metal stress

Periphytic diatom communities were analyzed from several heavy metal contaminated water bodies of Haryana, India. Among the analyzed sites, site HO3 (Saraswati Dham, Kurukshetra, Pehowa) showed significant response in the periphytic diatom community in terms of community shift (dominance of Gomphonema pseudoaugur) and lower biodiversity indices (species richness and Shannon index). Gomphonema pseudoaugur responded more specifically through induction of lipid bodies and occurrence of deformities in diatom frustules. PCA analysis showed that site HO3 is contaminated with heavy metals, especially Pb and Se. Pearson's correlation analysis showed a positive and statistically strong relationship between induction of lipid bodies and deformities with heavy metals (Pb and Se). Finally, from the present study, we concluded that heavy metal stress induces increased lipid body (LBs) size and deformities in the diatom species Gomphonema pseudoaugur, which could be a valuable indicator species for biomonitoring and a consideration in biofuel production.     

The in vitro inhibition of growth of intracellular Listeria monocytogenes by lymphocyte products

The listeria-inhibiting activity of culture supernatants from listeria-immune and nonimmune lymphocytes was assessed on listeria-infected macrophage cultures. It was found that supernatant from listeria-immune lymphocyte cultures stimulated in vitro with antigen was markedly inhibitory to the multiplication of intracellular listeria. Some inhibitory activity was also evident in supernatant from antigen-stimulated non-immune lymphocyte cultures. Supernatant from listeria-immune lymphocytes stimulated in vivo with antigen was capable of inhibiting listeria. Some inhibitory activity was still evident upon dilution of active supernatant at 1:100.     

Specificity of Acquired Resistance Produced by Immunization with Listeria monocytogenes and Listeria Fractions

Mice were immunized with 1.0 mg of an attenuated strain of Listeria monocytogenes to determine the period of protection afforded by this strain when the mice were challenged intravenously with 5 MLD of listeria. Protection appeared 2 days after immunization and was still apparent 4 weeks after immunization. If the challenge dose was decreased to 1 MLD, protection was apparent at 10 weeks. Mice immunized with a comparable dose of mycobacterial cells and challenged intravenously with 1 MLD of listeria showed no protection at 10 weeks. The magnitude of the immune response to listeria challenge was not increased in mice immunized with the same virulent strain as that used for challenge. It was also found that resistance to listeria challenge appeared early after listeria immunization if the immunizing dose was large. As the immunizing dose was decreased and the challenge dose increased, resistance appeared later. Listeria killed by heat or ultraviolet irradiation, living but nonmultiplying streptomycin-dependent listeria, or listeria ribosomal fraction gave no protection against listeria challenge. The magnitude of the immune responses after listeria immunization to listeria challenge and to mycobacteria challenge were compared. It was found that protection after listeria challenge was of longer duration. In addition, a 100-fold larger vaccinating dose was required to give comparable protection against tuberculous infection.     

Comparison of diatom community structure from epilithon and fish guts: implications for inferring past changes in water quality

The sampling skills of three common European fish species (Barbus barbus, Chondrostoma nasus and Squalius cephalus) were tested to assess their potential as biomonitors of past changes in river water quality through the analysis of diatoms in fish guts. The study was performed on three rivers with different chemical and physical conditions. Comparison of similarity indices revealed low diatom assemblage resemblance between epilithic samples and samples collected from fish guts at the same locality. In contrast, a mixed-effect linear model identified significant differences between locations through comparison of diatom-based water quality indices. Among fish species, diatom indices calculated from gut samples of B. barbus and C. nasus were in most cases not significantly different from those from epilithic samples while those from S. cephalus were often significantly different. The results of the study demonstrate that diatom analysis of fish guts provides a clear distinction between eutrophic and hypereutrophic rivers. Finer nutrient variations within hypereutrophic conditions were also found to be significant for some diatom indices. In this context, choice of appropriate diatom water quality index proved to be crucial in maximizing the sensitivity of the method.     

Diatoms and Other Epibionts Associated with Olive Ridley (Lepidochelys olivacea) Sea Turtles from the Pacific Coast of Costa Rica

Although the sea turtles have long been familiar and even iconic to marine biologists, many aspects of their ecology remain unaddressed. The present study is the first of the epizoic diatom community covering the olive ridley turtle’s (Lepidochelys olivacea) carapace and the first describing diatoms living on sea turtles in general, with the primary objective of providing detailed information on turtle epibiotic associations. Samples of turtle carapace including the associated diatom biofilm and epizoic macro-fauna were collected from Ostional beach (9° 59´ 23.7´´ N 85° 41´ 52.6´´ W), Costa Rica, during the arribada event in October 2013. A complex diatom community was present in every sample. In total, 11 macro-faunal and 21 diatom taxa were recorded. Amongst diatoms, the most numerous were erect (Achnanthes spp., Tripterion spp.) and motile (Haslea sp., Navicula spp., Nitzschia spp., Proschkinia sp.) forms, followed by adnate Amphora spp., while the most common macro-faunal species was Stomatolepas elegans (Cirripedia). Diatom densities ranged from 8179 ± 750 to 27685 ± 4885 cells mm^-2. Epizoic microalgae were either partly immersed or entirely encapsulated within an exopolymeric coat. The relatively low diatom species number, stable species composition and low inter-sample dissimilarities (14.4% on average) may indicate a mutualistic relationship between the epibiont and the basibiont. Dispersal of sea turtle diatoms is probably highly restricted and similar studies will help to understand both diatom diversity, evolution and biogeography, and sea turtle ecology and foraging strategies.     

Species diversity in postglacial diatom lake communities of Finland

Species diversity in three sections of Flandrian sediment from the Finnish Lake District was studied, together with orthodox diatom stratigraphy, to analyse the pattern of change in diatom communities over the postglacial time.The general agreement of the two approaches is clear, and in particular there is a positive direct relationship between the temporal development of these aquatic systems and changes in the diatom diversity.As indicated by both the quantitative inventory and the species diversity, these diatom communities consisted of few species immediately after glaciation at 10 000 radiocarbon years B.P. The minimum diversity at 9 000 B.P. coincides with the maximum of Melosira islandica ssp. helvetica. A trend to development of a larger number of equally abundant species, and higher total diversity followed isolation of the lake from the Baltic Sea at 8 000 B.P. This is consistent with earlier results of Duthie and Sreenivasa that the number of individuals and total diversity of the samples increase over the postglacial time.     

Aggregation and Sedimentation of Thalassiosira weissflogii (diatom) in a Warmer and More Acidified Future Ocean

Increasing Transparent Exopolymer Particle (TEP) formation during diatom blooms as a result of elevated temperature and pCO₂ have been suggested to result in enhanced aggregation and carbon flux, therewith potentially increasing the sequestration of carbon by the ocean. We present experimental results on TEP and aggregate formation by Thalassiosira weissflogii (diatom) in the presence or absence of bacteria under two temperature and three pCO₂ scenarios. During the aggregation phase of the experiment TEP formation was elevated at the higher temperature (20°C vs. 15°C), as predicted. However, in contrast to expectations based on the established relationship between TEP and aggregation, aggregation rates and sinking velocity of aggregates were depressed in warmer treatments, especially under ocean acidification conditions. If our experimental findings can be extrapolated to natural conditions, they would imply a reduction in carbon flux and potentially reduced carbon sequestration after diatom blooms in the future ocean.     

Diatom surface sediment assemblages around Iceland and their relationships to oceanic environmental variables

Canonical correspondence analysis of diatoms from surface sediment samples and oceanographic environmental variables shows that summer and winter sea-surface temperatures, water depth and winter sea-surface salinity are the main environmental factors affecting diatom distribution around Iceland. Of these, summer sea-surface temperature is the most important. Five diatom assemblages are distinguished and the distribution of these assemblages is clearly correlated with oceanic current patterns in the region. The sea-ice diatom assemblage is limited to the area where the East Greenland Current (Polar Water) has its strongest influence, and the cold diatom assemblage is basically controlled by the less cold East Icelandic Current (Modified Polar Water). The mixing diatom assemblage results from the interaction between the cold East Greenland and East Icelandic Currents and the warm Irminger Current. The warm diatom assemblage is located in the area dominated by the Irminger Current and may be used as an indicator of warm-water masses (Atlantic Water). The coastal diatom assemblage is the only one strongly influenced by both water depth and summer water temperatures.     

Living inside a sponge skeleton: the association of a sponge,a macroalga and a diatom

The relationships between sponges and macroalgae have been poorly investigated, especially in temperate waters. This work describes the consortium between the dictyoceratid sponge Dysidea pallescens and the red alga Acrochaetium spongicola permeating spongin fibres in the Mediterranean Sea; moreover, this is the first report of a diatom, Amphora pediculus, living also inside the spongin skeleton. The annual trend of the total algal biomass did not vary over time in relation to the temperature, irradiance and incorporation of foreign bodies. Our analyses, conducted by light and electron microscopy, suggest that both the macroalga and the diatom invade the skeleton of the sponge from the external environment, and that the benthic diatom, epiphyte of the macroalga, is passively carried inside the fibres through the growth of Acrochaetium spongicola. All the examined samples of D. pallescens showed that the macroalga permeated at least some fibres, while the presence of the diatom was occasional. The superficial layer of the sponge, thin and reticulate, likely allows the passage of the light and ensures the algal survival inside the sponge tissue. The high occurrence of the association with A. spongicola, together with the morphological adaptations of the sponge favouring the algal life, suggest that the relationship is mutualistic. The possible benefits for the involved partners are hypothesized. The taxonomy and ecology of endozoic Acrochaetiales are controversial due to the reduced size of thalli, the absence of peculiar diagnostic characters and unknown reproductive structures. Therefore, detailed studies on the relationships between the algae and their hosts will provide helpful information for the algal identification.     

Distribution patterns of diatom surface sediment assemblages in the Laptev Sea (Arctic Ocean)

The modern diatom distribution in the Laptev Sea, Arctic Ocean, was investigated in 89 surface sediment samples. Diatom concentrations are relatively low showing values between 0.01×10⁶ and 6.7×10⁶ valves per gram dry sediment. Based on a factor analysis using seventeen taxa or taxa groups five diatom surface sediment assemblages can be defined: the ice diatom assemblage of the central region of the Laptev Sea, the Chaetoceros assemblage of the eastern and southeastern shelf, the Thalassiosira antarctica assemblage of the continental slope and deep sea, the freshwater diatom assemblage in the vicinity of river mouths and deltas, and the Thalassiosira nordenskioeldii assemblage which shows a patchy occurrence on the central Laptev Sea shelf. The distribution pattern of diatom assemblages in surface sediment is significantly related to oceanographic conditions of surface water masses. The main factors controlling the distribution of diatoms in the Laptev Sea are the riverine freshwater input during the summer which strongly affects the salinity conditions, and the sea-ice extent. Furthermore, the composition of the Thalassiosira antarctica assemblage of the continental slope is largely influenced by dissolution and lateral transport processes.     

Effect of Immersion Solutions Containing Enterocin AS-48 on Listeria monocytogenes in Vegetable Foods

The effect of immersion solutions containing enterocin AS-48 alone or in combination with chemical preservatives on survival and proliferation of Listeria monocytogenes CECT 4032 inoculated on fresh alfalfa sprouts, soybean sprouts, and green asparagus was tested. Immersion treatments (5 min at room temperature) with AS-48 solutions (25 μg/ml) reduced listeria counts of artificially contaminated alfalfa and soybean sprouts by approximately 2.0 to 2.4 log CFU/g compared to a control immersion treatment in distilled water. The same bacteriocin immersion treatment applied on green asparagus had a very limited effect. During storage of vegetable samples treated with immersion solutions of 12.5 and 25 μg of AS-48/ml, viable listeria counts were reduced below detection limits at days 1 to 7 for alfalfa and soybean sprouts at 6 and 15°C, as well as green asparagus at 15°C. Only a limited inhibition of listeria proliferation was detected during storage of bacteriocin-treated alfalfa sprouts and green asparagus at 22°C. Treatment with solutions containing AS-48 plus lactic acid, sodium lactate, sodium nitrite, sodium nitrate, trisodium phosphate, trisodium trimetaphosphate, sodium thiosulphate, n-propyl p-hydroxybenzoate, p-hydoxybenzoic acid methyl ester, hexadecylpyridinium chloride, peracetic acid, or sodium hypochlorite reduced viable counts of listeria below detection limits (by approximately 2.6 to 2.7 log CFU/g) upon application of the immersion treatment and/or further storage for 24 h, depending of the chemical preservative concentration. Significant increases of antimicrobial activity were also detected for AS-48 plus potassium permanganate and in some combinations with acetic acid, citric acid, sodium propionate, and potassium sorbate.     

Diatoms are better indicators of urban stream conditions: A case study in Beijing,China

Urbanization dramatically affects hydrology, water quality and aquatic ecosystem composition. Here we characterized changes in diatom assemblages along an urban-to-rural gradient to assess impacts of urbanization on stream conditions in Beijing, China. Diatoms, water chemistry, and physical variables were measured at 22 urban (6 in upstream and 16 in downstream) and 7 rural reference stream sites during July and August of 2013. One-way ANOVA showed that water physical and chemical variables were significantly different (p < 0.05) between urban downstream and both reference and urban upstream sites, but not between reference and urban upstream sites (p > 0.05). Similarly, structural metrics, including species richness (S), Shannon diversity (H′), species evenness (J′) and Simpson diversity (D′), were significantly different (p < 0.05) between urban downstream and both reference and urban upstream sites, but not (p > 0.05) between reference and urban upstream sites. However, diatom assemblages were very different among all sites. Achnanthidium minutissima was a consistent dominant species in reference sites; Staurosira construens var. venter and Pseudostaurosira brevistriata were the dominant species in urban upstream sites; and Nitzschia palea was the dominant species in urban downstream sites. Clustering analyses based on the relative abundance of diatom species, showed all the samples fit into three groups: reference sites, urban upstream sites, and urban downstream sites. Canonical correspondence analysis (CCA) and Monte Carlo permutation tests showed that concentration of K⁺, EC, TN, Cl⁻ and pH were positively correlated with relative abundance of dominant diatom species in urban downstream samples; WT and F⁻ were correlated with reference and urban stream diatom composition. Our results demonstrate that the composition of diatom species was more sensitive to urbanization than the water physical and chemical parameters, and that diatom assemblage structure metrics more accurately assessed water quality. Some species, such as Amphora pediculus and Cocconeis placentula were among the dominant species in low nutrients stream sites; however, they were considered to be high nutrient indicators in some streams in USA. We suggest using caution in applying indicator indices based on species composition from other regions. It is necessary to build a complete set of diatom species data and their co-ordinate environment data for specific regions.     

Diatom communities along pH and hydrological gradients in three montane mires,central China

The distribution patterns of epiphytic diatom assemblages in three montane mires in central China were investigated to examine their relationships with selected environmental variables (pH and depth to water table, DWT). Two of the mires are considered to be in good ecological condition (Dajiuhu and Qizimeishan Mires) while Erxianyan Mire is extensively affected by acid deposition and human activities. A total of 206 taxa belonging to 56 genera were found in 44 Sphagnum samples. Multivariate analysis revealed that pH and DWT were significantly correlated with diatom distribution. In Erxianyan Mire, the characteristic taxa (Eunotia minor and Eunotia intermedia) had lower pH optima and may therefore be useful indicators of highly-acidic conditions. In Dajiuhu Mire, the dominant species had higher pH optima, and abundant xerotolerant taxa (Hantzschia amphioxys, Pinnularia borealis, Luticola mutica and Diadesmis contenta) were observed. In the partial canonical correspondence analyses with mire location as a covariable, the correlation between diatom data and pH was insignificant, likely because pH differences between mires were greater than those within mires. In contrast, diatom data were significantly correlated with DWT, suggesting that diatoms are good sensors of hydrological variability along the hollow to hummock gradient. Together, these data can expand current autecological information for these potential diatom indicator species, which is critical for refining our interpretations of bio-monitoring and palaeolimnological studies in montane mires.     

Diatom taphocoenoses in the coastal upwelling area off South West Africa

The diatom floral composition of 124 sediment samples from the South East Atlantic records the influence of coastal upwelling on sediment composition off South West Africa. Inner shelf samples between 19° and 24°S are rich in diatom valves and the patterns of diatom species distribution in these samples are related to the coastal upwelling process. Comparison with recent phytoplankton data shows that the sediment assemblages preserve many of the important species of the diatom biocoenoses, including Chaetoceros (resting spores), Delphineis karstenii, Thalassiosira eccentrica, and Thalassionema nitzschioides. Delphineis karstenii, a pioneer species in enriched coastal water, occurs nearshore and Chaetoreros resting spores are widespread, with highest relative abundance values in some more offshore samples. The abundance of the Thalassiosira eccentrica group and of Thalassionema nitzschioides in sediment samples in and near Walvis Bay reflects the recurrence of intense upwelling off this part of the coast. An abundance of large centric species has been reported in hydrological conditions characteristic of newly upwelled waters and, correspondingly, Actinocyclus octonarius and some large Coscinodiscus species occur in the sediments in nearshore patches or belts.     

Psammodiscus nov. gen. Based on Coscinodiscus nitidus

The epipsammic diatom Coscinodiscus nitidus differs from typicalCoscinodiscus species in areola arrangement and structure, closingmembrane structure, rimoportula arrangement, and habitat. Severalsimilarities exist between it and Rhaphoneis species but, whilethe relationship between these is close, the two cannot be consideredcongeneric. A new genus, Psammodiscus, is created to includeC. nitidus. Coscinodiscus nitidus, Rhaphoneis, Psammodiscus nov. gen., epipsammic diatom, diatom taxonomy     

Comparing diatom and Alexandrium catenella/tamarense blooms in Thau lagoon: Importance of dissolved organic nitrogen in seasonally N-limited systems

Diatom blooms in Thau lagoon are always related to rain events leading to inputs of inorganic nutrients such as phosphate, ammonium and nitrate through the watershed with time lags of about 1 week. In contrast, blooms of Alexandrium catenella/tamarense can occur following periods of 3 weeks without precipitation and no significant input of conventional nutrients such as nitrate and phosphate. Field results also indicate a significant drop (from 22–25 to 15–16 μM over 3 days) in dissolved organic nitrogen (DON) at the bloom peak, as well as a significant inverse relationship between A. catenella/tamarense cell density and DON concentrations that is not apparent for diatom blooms. Such dinoflagellate blooms are also associated with elevated (6–9 μM) ammonium concentrations, a curious feature also observed by other investigators, possibly the results of ammonium excretion by this organism during urea or other organic nitrogen assimilation.The potential use of DON by this organism represents short cuts in the nitrogen cycle between plants and nutrients and requires a new model for phytoplankton growth that is different from the classical diatom bloom model. In contrast to such diatom blooms that are due to conventional (nitrate, phosphate) nutrient pulses, Alexandrium catenella/tamarense blooms on the monthly time scale are due to organic nutrient enrichment, a feature that allows net growth rates of about 1.3 d⁻¹, a value higher than that generally attributed to such organisms.     

The harpacticoid copepod Tisbe holothuriae is resistant to the insidious effects of polyunsaturated aldehyde-producing diatoms

Numerous species of diatoms liberate oxylipins including polyunsaturated aldehydes (PUAs) in response to cellular damage such as may occur during grazing. PUAs are cyto- and genotoxic and negatively disrupt reproductive processes in copepods, their principal grazers, although experimental evidence would suggest that the grazer response may be species specific. The reproduction of the benthic harpacticoid copepod Tisbe holothuriae was compared over two generations. Copepods were reared using four diet treatments: PUA-producing diatom strains Skeletonema marinoi (Adriatic Sea Isolate FE6) and Melosira nummuloides (CCAP 1048/6); and non-PUA-producing diatom strains Phaeodactylum tricornutum (CCAP 1052/A) and S. marinoi (Seasalter (Walney) Ltd). Life tables were generated for each treatment using measured reproductive parameters and the net reproductive rate (R₀) calculated. No significant differences were observed between the individual reproductive parameters of T. holothuriae fed PUA-producing diatoms compared to those fed non-PUA-producing diatoms although diets of P. tricornutum resulted in some decreases in individual reproductive parameters in the second generation. There were no significant differences in the R₀ values between the four tested diets. These observations indicate that T. holothuriae exhibits a tolerance of known PUA-producing diatom diets that has not been similarly demonstrated in pelagic calanoid copepods. Harpacticoid copepods may have a greater capacity to detoxify diatom oxylipins than their planktonic calanoid counterparts.