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.     

The potential use of nisin to control Listeria monocytogenes in poultry

Nisin (Nisaplin^TM) was added to scald water and used to treat Listeria monocytogenes which were either directly suspended in the water or attached to turkey skin. There was at least a 1 log decrease of listeria for the nisin treatments. This was followed by further reductions when the samples were stored under refrigeration. Heat exhibited a synergistic relationship with the nisin when the cells were directly suspended in the scald water. A 2 log decrease was observed immediately after this treatment and there was complete elimination of the listeria after 48 h of refrigeration. However, this relationship was not as obvious when the cells were first attached to the turkey skin. In these latter experiments, the nisin treatments resulted in reductions in the listeria levels but they were smaller than those observed when the cells were directly suspended in the scald water.     

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.     

Combined eco-morphological functional groups are reliable indicators of colonisation processes of benthic diatom assemblages in a lowland stream

Classifying benthic diatom taxa based on ecological and morphological features became increasingly important in recent years due to the demand of understanding the dynamics and functioning of diatom assemblages. The great potential in using these functional classifications in diatom ecology involves further refinement of current classification. In our experimental study, colonisation processes of diatom assemblages were studied in a typical small lowland stream, using both diatom guilds and cell size categories. We also tested newly proposed combined eco-morphological functional groups (ecological guilds combined with cell size categories) in the study of the colonisation process in benthic diatom assemblages. We hypothesised that (i) there is a decrease in the proportion of low profile guild, while an increase in that of high profile and motile guilds in time with the decreasing rate of physical disturbance; (ii) the presence of small size categories will be pronounced at the beginning of the colonisation processes, while proportion of larger size categories will be higher in the latter phases of colonisation; and (iii) the relationship between taxa and environmental factors are better reflected by the use of combined eco-morphological functional groups than by the sole analyses of rough guilds or cell size categories. The first hypothesis was not confirmed, and our second hypothesis was only partially confirmed by the results. We found that the relationship between environmental factors and guilds, as well as cell size categories was not appropriate to reveal the relationship between abiotic factors and taxa composition. In contrast we found that compositional changes in colonisation were appropriately reflected by the newly defined combined eco-morphological functional groups. In the combined eco-morphological functional groups, such kind of taxonomical and ecological features can be prevailed which are hidden in guilds or cell size categories separately. Thus these combined eco-morphological functional groups could help to come one step closer to develop a widely used ecological classification in diatom researches.     

Specificity of the Anamnestic Response Produced by Listeria monocytogenes or Mycobacterium tuberculosis to Challenge With Listeria monocytogenes

When mice immunized with Listeria monocytogenes were given a second injection of listeria, they showed an anamnestic immune response to intravenous challenge with listeria, as measured by enumeration of the viable infecting organisms in the spleens of the infected animals. This response was independent of the effects of the challenge dose. When mice immunized with living or heat-killed attenuated mycobacterial cells were boosted with living H37Ra, there was also an accelerated response to listeria challenge. The response was greater in the mice given the primary immunization with living cells than in those immunized with heat-killed cells. The response to listeria challenge in mice immunized and boosted with mycobacteria was of less magnitude than that in the mice immunized and boosted with listeria. Growth of listeria in the mice immunized and boosted with mycobacteria was retarded only during the first 2 days of the infection, whereas the infecting listeria in mice immunized and boosted with listeria were permanently inactivated. Mice immunized with mycobacterial ribosomal fraction and restimulated with living mycobacterial cells showed no accelerated response to listeria challenge. It is evident from these results that resistance to these organisms is specifically evoked, but that once evoked it is not completely nonspecific in action. Also, the resistance produced by the mycobacterial ribosomal fraction to challenge with mycobacteria is completely specific in action. Therefore, it has been shown that there are two mechanisms involved in acquired immunity to facultative, intracellular parasites. One is nonspecific and mediated by activated macrophages. The other is specific and mediated by a mechanism as yet unknown.     

Correlation of surface sediment diatoms with the present lake water pH in 28 Algoma lakes,Ontario, Canada

The surface sediment diatom analysis of 28 Algoma lakes (pH 4.40–8.13) indicates that even though each lake has a widely different aquatic environment and characteristic diatom assemblage, a definite relationship exists between the lake water pH and their diatom assemblages. In the acidic lakes acidobiontic and acidophilous diatom species predominate whereas in circumneutral and alkaline lakes circumneutral and alkaliphilous diatoms were most common. Cluster analysis of the pH indicator diatom assemblages grouped the study lakes into three distinct cluster groups. These groups also closely corresponded to lake water pH. On the basis of published ecological information as well as their presence in our study lakes, the pH indicator status of a number of diatom taxa have been discussed. A detailed listing of the diatom taxa identified and their pH indicator status is provided in order to facilitate their use in future diatom-inferred pH studies.     

A technique for the direct identification of haemolytic-pathogenic listeria on selective plating media

A technique based on the addition of a red cells top layer to a selective plating medium after listeria growth is proposed in order to detect directly the haemolytic activity of pathogenic listeria colonies. It was applied to different selective plating media (modified McBride agar, lithium chloride-phenylethanol-moxalactam, listeria selective medium–Oxford formulation, polymyxin-acriflavine-lithium chloride-ceftazidime-aesculin-mannitol and LSAMM). The haemolytic activity of listeria colonies was more easily detected with the top layer than when red cells were incorporated in the selective plating medium. The LSAMM was the best medium for the recovery and identification of Listeria monocytogenes colonies by this technique (three Listeria monocytogenes colonies were distinguished among 2520 Listeria innocua colonies in raw milk).     

Biogenic silica as an estimate of siliceous microfossil abundance in Great Lakes sediments

Biogenic silica concentration (BSi) in sediment cores from the Great Lakes is evaluated as an estimate of siliceous microfossil abundance. A significant linear relationship was found between measured BSi and diatom valve abundance for sediment cores from the Bay of Quinte, Lake Ontario, Lake Erie, Lake Michigan and Lake Superior and between measured BSi and diatom biovolume for Lake Erie, Lake Michigan, and Lake Superior but not for Lake Ontario. Diatom silica predicted from diatom species abundance and an estimated silica content per cell in the Lake Erie cores accounted for 117% and 103% of measured BSi, respectively. By contrast, predicted diatom silica could only account for 28% of measured BSi in the Lake Michigan core and only 25% in the Lake Superior core. A few large diatoms with a large silica content per cell comprised a major portion of predicted diatom silica in all cores. The discrepancy between chemically measured BSi and the silica predicted from diatoms in the Lake Michigan and Lake Superior cores was partially due to the inability of the regression model, used to estimate diatom silica content, to account for different degrees of silicification in the diatom asemblages from the more dissolved silica rich Lake Michigan and Lake Superior.     

Transfer functions for estimating paleoecological conditions (pH) from East African diatoms

Our purpose is to establish the quantitative relationship between recent diatom floras and ecological parameters, in order to extrapolate the results to the past. The parameter pH is here considered as an example. This work is based on the study of about I60 diatom samples from East Africa and of their corresponding biotopes. We propose some statistical methods to interpret the data. Correspondence analysis allows us to define the pH-indicator species. The regression calculations allow pH values to be calculated using the percentage of the diatom species in a sample.     

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     

Regulation of the immune response by macrophages

Regulation of the immune response by macrophages was studied with cellular resistance to Listeria monocytogenes as parameter. The use of agents which suppress macrophage activity during the induction-phase of immunity enabled the induction of protective immunity with killed listeria. Fractionation of the cell content of listeria yielded an RNA'se sensitive fraction which in a dose of 300 ng and in combination with the cationic surfactant dimethyl dioctadecyl ammonium bromide induced protective immunity against listeria.     

Influence of small-scale gold mining on French Guiana streams: Are diatom assemblages valid disturbance sensors?

The ongoing gold-rush in French Guiana could cause severe disturbance to ecosystems. Although illegal, small gold mining sites are rapidly expanding. Few studies have attempted to measure the consequences of the increased gold mining on the biota of small forest streams, and to date no study has dealt with primary producers. Here we measured the response of diatom assemblages to gold mining in ten sites differently affected by the mining activity (i.e., reference, formerly gold-mined and currently exploited). Our results showed that both taxonomic and functional structure of the diatom assemblages were influenced by the intensity of gold mining activity. A significant relationship between soil erosion and diatom motility ability has been demonstrated. These findings show that diatom assemblages are sensitive to gold mining disturbance and suggest that diatom communities may be used as sensors of the environmental stress caused by small-scale gold mining activities.     

Quantifying translocation of Listeria monocytogenes in rats by using urinary nitric oxide-derived metabolites

The urinary nitric oxide metabolites NO(2)(-) and NO(3)(-) (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.     

Biological characteristics of Listeria cultivating on the novel media for their accumulation and isolation

Biological characteristics of isolated strains of listeria cultivating on growth media developed for accumulation and isolation of these microorganisms. Stability of main cultural-morphologic characteristics, pathogenic enzyme synthesis, profile of plasmid DNA and specific bacteriophage lysing of listeria during their cultivation on novel media was shown.     

Incidence of Listeria in tropical fish and fishery products

Thirty-five tropical fish and fishery products were tested for the presence of listeria. Listeria innocua was detected in three of 10 fresh samples and five of 14 frozen samples. No listeria were detected in dried, salted fish. Listeria monocytogenes was not detected in any of the samples tested.     

CORRELATED EVOLUTION OF GENOME SIZE AND CELL VOLUME IN DIATOMS (BACILLARIOPHYCEAE)1

A correlation between genome size and cell volume has been observed across diverse assemblages of eukaryotes. We examined this relationship in diatoms (Bacillariophyceae), a phylum in which cell volume is of critical ecological and biogeochemical importance. In addition to testing whether there is a predictive relationship across extant species, we tested whether evolutionary divergences in genome size were correlated with evolutionary divergences in cell size (using independent contrasts). We estimated total DNA content for 16 diatom species using a flow cytometer and estimated cell volumes using critical dimensions with scaling equations. Our independent contrast analyses indicated a significant correlated evolution between genome size and cell volume. We then explored the evolutionary and ecological implications of this evolutionary relationship. Diatom cell volume is an important component of the global carbon cycle; therefore, understanding the mechanisms that drive diatom genome evolution has both evolutionary and ecological importance.     

Specificity of Acquired Resistance Produced by Immunization with Mycobacterial Cells and Mycobacterial Fractions

Mice were immunized intraperitoneally with 5.0 mg of living and 5.0 mg of heat-killed H37Ra cells of the attenuated strain Mycobacterium tuberculosis and challenged intraperitoneally with Listeria monocytogenes and Klebsiella pneumoniae. The period of protection provided by the living and heat-killed H37Ra cells against both heterologous infections was the same. When mice were immunized intraperitoneally with graded doses of living and heat-killed H37Ra and challenged intraperitoneally with listeria or klebsiella, the lowest immunizing dose providing protection against both klebsiella and listeria challenge was the same for living and heat-killed cells. Living and heat-killed cells also immunized equally effectively when the routes of immunization and challenge were different. Mice also were immunized intraperitoneally with mycobacterial ribosomal fraction, mycobacterial cell walls, and several nonspecific agents (Escherichia coli endotoxin, mineral oil emulsion, and Freund's incomplete adjuvant). The mice were challenged intraperitoneally with listeria or klebsiella at varying times after immunization. The mycobacterial components and all the nonspecific agents provided transitory protection lasting no longer than 4 days after immunization. Only the mycobacterial cell walls and the endotoxin provided protection against listeria challenge. It was concluded that the protection provided by the mycobacterial ribosomal fraction is specific for tuberculosis infection, since this fraction provided no protection against listeria infection and only transitory protection against klebsiella. It was also concluded that the mycobacterial component providing protection against heterologous infections is heat stable and probably is found in the cell wall.     

Relationship Between Periodic Resuspension Events and Planktonic Diatom Community Structure in Lake Michigan: A Field and Laboratory Investigation

Lake Michigan provides an ideal location for comparing episodic physical forcing events (storms) on phytoplankton processes and the more persistent seasonal variability of phytoplankton communities. This is due to the duration and extent of the highly turbid, recurrent coastal plume (RCP) in the lake during the winter/early spring. Although the RCP can coincide with the initiation of the basin wide spring diatom bloom, linkages between the duration and intensity of the plume and the prominent role of light availability in regulating Lake Michigan phytoplankton growth during the spring isothermal period have been postulated, but not verified. The concurrent physical and biological events provide a novel opportunity to examine phenomena associated with the RCP affecting the distribution and abundance of species in Lake Michigan's planktonic diatom flora. In this study, planktonic diatom assemblages from pre, post and active spring plume events were examined from stations along Lake Michigan's southern shoreline. Species abundance changed rapidly during storm events. Sediment resuspension via storm activity created a suboptimal growth environment. Post‐storm event diatom phytoplankton communities were floristically distinct from prestorm event communities, with resting cell‐forming taxa playing a significant role in these community‐restructuring periods. Laboratory simulations of resuspension events using Lake Michigan sediments were conducted under a variety of environmental conditions. Parameters varied included day length, temperature, and silica. The resulting assemblages were quantitatively counted. A statistically significant relationship was identified between day length and the vegetative growth of many resting cell‐forming diatom species. When day length was calculated for poststorm event field data, it revealed a high correlation between poststorm event communities and those predicted by the laboratory simulations. Timing of storm events and latitudinal position of the aquatic system are then important elements for consideration when predicting diatom phytoplankton community structure due to the relationship these factors have with day length.     

Relationship Between Surface Sediment Diatoms and Summer Water Quality in Shallow Lakes of the Middle and Lower Reaches of the Yangtze River

The relationship between surface sediment diatoms and summer water quality was investigated at 49 lakes in the middle and lower reaches of the Yangtze River. Lakes ranging from oligomesotrophic to hypereutrophic were examined, providing an obvious nutrient gradient. With the shift from mesotrophic to eutrophic levels, diatom multi-ecotypes dominated by epiphytic and facultative planktonic taxa were replaced by nutrient-tolerant planktonic taxa, such as Cyclotella meneghiniana Skvortzow, C. atomus Hustedt,Cyclostephanos Round, and Stephanodiscus Ehrenberg etc., reflecting the nutrient changes in the lake.The relationship between diatoms and summer water quality indices was explored further using numeric analysis. Canonical correspondence analysis (CCA) with forward selection and a Monte Carlo permutation test revealed that of all 25 summer water environmental variables, total phosphorus (TP), chlorophyll a (Chzl a), Secchi depth (SD), dissolved inorganic phosphorus, C1-, SO42-, Mg2 , CO32-, and water depth were significant variables (P＜0.05) in explaining diatom distributions. Of these, TP, Chl a, SD, and C1-, were the most important variables. The result of the correlation analysis also showed that a significant correlation exists among these variables, implying that these indices are either interconnected or independent in explaining the diatom data. For phosphorus-limited sites, TP was the most significant variable affecting the diatoms, also affecting changes in Chl a, SD, and iron concentrations. The independence of Chl a may be related to algal competition induced by lake eutrophication, resulting in the feedback to diatom community.In addition to TP, SD can be related to sediment disturbance by wave action and the growth of macrophytes in large shallow lakes. These relationships between diatom ecotypes and water quality provide the basis for a future quantitative reconstruction of historic lake nutrient evolution in the study area and will also provide a wealth of modern ecological knowledge that can be used to interpret fossil diatom records.     

Applicability of borrowed diatom-based water quality assessment indices in streams around São Carlos-SP,Brazil

Diatom-based indices are increasingly becoming important tools for the assessment of ecological conditions in lotic systems. The applicability of regional and foreign diatom-based water quality assessment indices to streams around São Carlos-SP, Brazil, is discussed. The relationship between measured water quality variables and diatom index scores was assessed. The indices, when compared to chemical analyses, proved useful in providing an indication of the quality of the investigated waters. Though all borrowed indices were applicable to the study area because many widely distributed diatom species have similar environmental tolerances to those recorded for these species elsewhere, ecological requirements of some diatom species from Brazil need to be clarified and incorporated in a diatom-based water quality assessment protocol unique to the region.