Campylobacter jejuni is a recognized and common gastrointestinal pathogen in most parts of the world. Human infections are often food borne, and the bacterium is frequent among poultry and other food animals. However, much less is known about the epidemiology of C. jejuni in the environment and what mechanisms the bacterium depends on to tolerate low pH. The sensitive nature of C. jejuni stands in contrast to the fact that it is difficult to eradicate from poultry production, and even more contradictory is the fact that the bacterium is able to survive the acidic passage through the human stomach. Here we expand the knowledge on C. jejuni acid tolerance by looking at protozoa as a potential epidemiological pathway of infection. Our results showed that when C. jejuni cells were coincubated with Acanthamoeba polyphaga in acidified phosphate-buffered saline (PBS) or tap water, the bacteria could tolerate pHs far below those in their normal range, even surviving at pH 4 for 20 h and at pH 2 for 5 h. Interestingly, moderately acidic conditions (pH 4 and 5) were shown to trigger C. jejuni motility as well as to increase adhesion/internalization of bacteria into A. polyphaga. Taken together, the results suggest that protozoa may act as protective hosts against harsh conditions and might be a potential risk factor for C. jejuni infections. These findings may be important for our understanding of C. jejuni passage through the gastrointestinal tract and for hygiene practices used in poultry settings.

The proposed plan for enrichment of the Sulu Sea, Philippines, a region of rich marine biodiversity, with thousands of tonnes of urea in order to stimulate algal blooms and sequester carbon is flawed for multiple reasons. Urea is preferentially used as a nitrogen source by some cyanobacteria and dinoflagellates, many of which are neutrally or positively buoyant. Biological pumps to the deep sea are classically leaky, and the inefficient burial of new biomass makes the estimation of a net loss of carbon from the atmosphere questionable at best. The potential for growth of toxic dinoflagellates is also high, as many grow well on urea and some even increase their toxicity when grown on urea. Many toxic dinoflagellates form cysts which can settle to the sediment and germinate in subsequent years, forming new blooms even without further fertilization. If large-scale blooms do occur, it is likely that they will contribute to hypoxia in the bottom waters upon decomposition. Lastly, urea production requires fossil fuel usage, further limiting the potential for net carbon sequestration. The environmental and economic impacts are potentially great and need to be rigorously assessed.

Some microalgae are able to kill or inhibit nutrient-competing microalgae, a process called allelopathy. Inhibiting or killing competitors enable these species to monopolize limiting resources, such as nitrogen and phosphorus. Prymnesium parvum is known to produce such allelopathic compounds, substances that seem identical to the ichthyotoxins identified from this species. Biotic and abiotic environmental factors influence not only growth rates but also toxin/allelopathic compounds production by P. parvum cells. Toxin production, as well as allelopathy, including grazer deterrence, increases dramatically in light, temperature, or nutrient stressed P. parvum cells. Correspondingly, toxicity and allelopathy may decrease, or cease completely, if cells are grown with high amounts of N and P in balanced proportions. However, even under nutrient (N and P) sufficient conditions, P. parvum is able to produce toxins/allelopathic compounds, with negative effects on other phytoplankton species or grazers, if cells densities of P. parvum are high relative to other species. This negative effect might shift the plankton community to more toxin resistant species. Filtrates from nutrient-deficient P. parvum cultures have almost the same strong negative effect on grazers and other phytoplankton species as when Prymnesium cells are grown together with the target organisms. Eutrophication, the increased input of N and P to aquatic ecosystems, besides increasing nutrient concentrations, is usually provoking unbalanced N:P condition for the optimal growth of phytoplankton, deviating from the Redfield ratio, i.e., the phytoplankton cellular nitrogen to phosphorus ratio, N:P = 16:1 (by atoms) or 7.2:1 (by weight). Eutrophication thus both enhances P. parvum growth and increases production of toxins and allelopathic compounds. Supplying N-deficient or P-deficient P. parvum cells with the deficient nutrient reduces toxicity to less than half within 24 h after additions. As P. parvum is mixotrophic, uptake of dissolved or particulate organic N (DON or PON) can also reduce toxicity and allelopathy in the same manner as addition of inorganic N to N-starved cells. In conclusion, P. parvum, by increasing its toxicity and allelopathic ability under poor environmental conditions, outcompetes the co-occurring phytoplankton species.

This paper describes the effects posed by stormwater runoff from an industrial log-yard on the microalgae Scenedesmus subspicatus. The effects of stormwater runoff sampled during two rain events were determined by exposing S. subspicatus cells to different concentrations (% v:v) of each sample. The effects were measured as the percentage change in growth rates in relation to a control culture after exposure times of 24, 48, 72 and 96h. The runoff from the first rain event had no negative effects to S.subspicatus, posing in most cases growth stimulation, whereas the runoff from the second rain event inhibited algae growth. Differences in runoff physico-chemical characteristics combined with the hydrological factors of each rain event explained these opposite effects. The hypothesis of toxic first flush phenomenon was confirmed in the second rain event on the basis of normalized inhibitory effects and runoff volume. It was found that 42, 51 and 50% of the inhibitory effects during exposures of 24, 48 and 72h are associated with the initial 4% of the total discharged volume. The fact that negative effects were observed in the two runoff events analyzed here, raises concern about the potential environmental threats posed by runoff originated from wood-based industrial areas during the entire hydrological year.

In industries based on dry processes, such as wood floor and wood furniture manufacture, wastewater is mainly generated after cleaning of surfaces, storage tanks and machinery. Owing to the small volumes, onsite treatment options and potential environmental risks posed to aquatic ecosystems due to discharge of these wastewaters are seldom investigated. In the present study, the effects of cleaning wastewater streams generated at two wood floor production lines on Desmodesmus subspicatus were investigated. The microalgae was exposed to different wastewater concentrations (100, 50, 25, 12.5 and 6.25% v:v) and the algae growth evaluation was based on in vivo chlorophyll fluorescence, cell density, cell size (number of cells/colony) and cell ratio (length/width). Inhibitory effects of the tested wastewaters on the microalgae were positively related to concentration and negatively related to exposure time. The EC50,24 h of blade cleaning wastewater (BCW) and floor cleaning wastewater (FCW) were 3.36 and 5.87% (v:v), respectively. No negative effect on cell colony formation was caused by BCW, whereas an increase of 90% unicellular cells was observed in FCW concentrations below 50% (v:v). At the lowest concentration (3.13% v:v) where no growth inhibition was observed, both wastewater streams caused changes in cell dimensions by increasing cell length and width. To conclude, wastewaters generated during cleaning procedures in the wood floor industries can have severe environmental impacts on aquatic organisms, even after high dilution. Therefore, these wastewaters must be treated before being discharged into water bodies.

Despite the paramount importance of bacteria for biogeochemical cycling of carbon and nutrients, little is known about the potential effects of climate change on these key organisms. The consequences of the projected climate change on bacterioplankton community dynamics were investigated in a Baltic Sea spring phytoplankton bloom mesocosm experiment by increasing temperature with 3°C and decreasing pH by approximately 0.4 units via CO₂ addition in a factorial design. Temperature was the major driver of differences in community composition during the experiment, as shown by denaturing gradient gel electrophoresis (DGGE) of amplified 16S rRNA gene fragments. Several bacterial phylotypes belonging to Betaproteobacteria were predominant at 3°C but were replaced by members of the Bacteriodetes in the 6°C mesocosms. Acidification alone had a limited impact on phylogenetic composition, but when combined with increased temperature, resulted in the proliferation of specific microbial phylotypes. Our results suggest that although temperature is an important driver in structuring bacterioplankton composition, evaluation of the combined effects of temperature and acidification is necessary to fully understand consequences of climate change for marine bacterioplankton, their implications for future spring bloom dynamics, and their role in ecosystem functioning.

We verified an active uptake of kleptoplastids in the toxic and bloom forming dinoflagellates genus Dinophysis from its preferred prey, the ciliate Myrionecta rubra, using a quantitative real-time PCR technique. During a 65 days starvation/feeding experiment with Dinophysis caudata, changes in plastid 16S rRNA, plastid autofluorescence and plastid/nuclear DNA ratio through the cell-cycle was followed with quantitative real-time PCR and flow-cytometry. During starvation, the cultures initially showed a rapid growth and an 3.5-fold increase of cells ml^-1, when at the same time, plastid DNA cell^-1 showed a 3.5-fold decrease, and phycoerythrin fluorescence cell^-1 a 3.6-fold decrease. The decrease in plastid DNA cell^-1day^-1 closely followed culture growth rate (r = 0.91, Pearson correlation), indicating that existing plastids were diluted within the growing population, and that no new plastids were synthesised by the cells. When starved cells were re-fed by the ciliate M. rubra on day 43-51 of the experiment, plastid DNA cell^-1 increased 7-fold up to 14 000 16S DNA copies per cell, thereby directly revealing the kleptoplastic behaviour. The implication is that not only availability of the prey M. rubra itself, but also the supply of suitable kleptoplastids might be an important controlling factor for Dinophysis spp. bloom formation and decline.

We verified an active uptake of kleptoplastids in the toxic and bloom-forming dinoflagellatesof the genus Dinophysis from its preferred prey, the ciliate Myrionecta rubra, using a quantitativereal-time PCR technique. During a 65 d starvation/feeding experiment with Dinophysis caudata,changes in plastid 16S rRNA, plastid autofluorescence and plastid/nuclear DNA ratio throughthe cell cycle were followed with quantitative real-time PCR and flow cytometry. During starvation,the cultures initially showed a rapid growth and a 3.5-fold increase of number of cells ml–1, while atthe same time, plastid DNA cell–1 showed a 3.5-fold decrease, and a 3.6-fold decrease in phycoerythrinfluorescence cell–1. The decrease in plastid DNA cell–1 d–1 closely followed culture growth rate(Pearson correlation, r = 0.91), indicating that existing plastids were diluted within the growing populationand that no new plastids were synthesised by the cells. When starved cells were re-fed by theciliate M. rubra on Days 43 to 51 of the experiment, plastid DNA cell–1 increased 7-fold up to 14 00016S DNA copies per cell, thereby directly revealing the kleptoplastic behaviour. The implication isthat not only availability of the prey M. rubra itself, but also the supply of suitable kleptoplastidsmight be an important controlling factor for Dinophysis spp. bloom formation and decline.

The raphidophyte Heterosigma akashiwo, which forms toxic blooms, causes major economical losses to the fish industry because of the fish kills involved. It is therefore important to be able to detect not only H. akashiwo but other toxic phytoplankton species as well, rapidly and accurately to reduce losses by fish kills. With this purpose, DNA sequences from H. akashiwo 18S and 28S rRNA gene regions were studied in silico to design species-specific probes to be used in a microarray format. Three strains of H. akashiwo (AC 265, AC 266 and GUMACC 120) were grown at optimal conditions and transferred into new environmental conditions changing either the light intensity, salinity, temperature or nutrient concentrations, to check if any of these environmental conditions induced changes in the cellular RNA concentration. The aim of this experiment was the calibration of several species-specific probes for the quantification of H. akashiwo. Differences on RNA content were not significant (p < 0.05) in any of the treatments, therefore the calibration curves were validated. The designed probes are reliable for the detection and quantification of H. akashiwo cells in natural waters. (C) 2013 Elsevier B.V. All rights reserved.

In the North Sea, Tripos and Dinophysis are commonly occurring mixotrophic planktonic dinoflagellate genera. In order to understand their bloom dynamics, an occurring bloom dominated by T. furca and D. norvegica was followed for several days. High cell abundances of these species were located to estimate: in situ growth rates from cell cycle analyses, depth distributions, growth rates sustained by photosynthesis, and parasite infection prevalence in all T. furca, T. fusus, D. norvegica and D. acuminata. Cell abundances were over 10000 cells L−1 for T. furca and up to 18000 cells L−1 for D. norvegica. Cells accumulated between 15-25 m depth and presented low specific in situ growth rates of 0.04-0.15 d−1 for T. furca and 0.02-0.16 d−1 for D. norvegica. Photosynthesis could sustain growth rates of 0.01-0.18 d−1 for T. furca and 0.02 to 0.14 d−1 for D. norvegica, suggesting that these species were relying mainly on photosynthesis. Parasite infections where generally low, with occasional high prevalence in D. norvegica (by Parvilucifera sp.) and T. fusus (by Amoebophrya sp.), while both parasites showed comparable prevalence in D. acuminata, which could offset in situ growth rates by parasite-induced host mortality. The restructuring effect of parasites on dinoflagellate blooms is often overlooked and this study elucidates their effect to cell abundances and their growth at the final stages of a bloom.

The response of natural Baltic Sea spring plankton communities topredicted higher temperature and increased seawater acidification was studied in an indoor-mesocosm experiment. Plankton communities collected in a coastal area of the Baltic Sea (Kalmar Sound) were incubated for 20 days in the following conditions of: normal pH and temperature (pH=7.52, 3°C; control condition); lower pH (pH=7.14,3°C); high temperature (pH=7.52, 6°C) and lower pH-high temperature,(pH=7.14, 6 °C). Results showed that phytoplankton biomass (expressed as chlorophyll a), growth rates, plankton cell densities and community composition were significantly influenced by higher temperature, lower pH and to a greater extent subjected to both factors in combination. At higher temperature, phytoplankton biomass, particulate organic carbon(POC) and growth rates in addition to copepod densities were significantly enhanced. Highest bacteriae and heterotrophic nanoflagellates densities were observed in the higher temperature and lower pH treatment. Furthermore, the highest total phytoplankton and plankton communities diversity were found in this treatment as well. Increase in temperature and acidification accelerated the spring bloom by ca. 1 day °C-1. The phytoplankton community shifted from a dominance of Skeletonema costatum, in favor of haptophytes and dinoflagellates; and from dominance of the ciliate Myrionecta rubra in favor of tintiniids and oligotrichids. Our results suggest that the concomitant increased temperature and acidification of the Baltic Sea will increase the spring bloom biomass, and induce an early appearance of phytoplankton species typical of summer, thus decreasing the dominance of diatoms during the spring bloom. Decreased ciliates and copepod abundances will probably lead to sinking of higher amount phytoplankton biomass to deep water layers, expanding the area of oxygen depletion. If that is the case, the extra-incorporated carbon will not be channeled up the food chain.

Infection of marine dinoflagellates by the parasitic dinoflagellate Amoebophrya spp. plays an important role in population dynamics and carbon flow in marine food webs. It has been extensively reported that Amoebophrya parasitoids occur in temperate coastal areas of the northern hemisphere; however, little is known about their distribution and importance in tropical areas and southern oceans. We used an rRNA-based, fluorescent in situ hybridization assay to detect Amoebophrya spp. infections during the decline of a late-summer dinoflagellate population dominated by Ceratium falcatiforme in a tropical coastal area of the southern Atlantic Ocean subjected to recurrent upwelling-downwelling cycles. Conditions during our survey were typical of downwelling when oligotrophic waters dominate the area. C. falcatiforme was the most infected host, with a prevalence averaging 2% over the study area at the beginning of sampling. At a fixed sampling station monitored over 4 wk, Amoebophrya prevalence escalated from 1 to 7% over a 6 d period, concomitant to a 94% decrease in host cell numbers. Infection by Amoebophrya was estimated to have killed ca. 11% of the host cell population within this period; thus, parasitism was not the main factor behind the C. falcatiforme population decline. Estimates based on biovolume calculations indicate that ca. 6.5% of the carbon in the decaying C. falcatiforme population was transformed into parasitoid dinospores, which became available to tintinnid ciliates that were abundant during our survey. Such a trophic link might be relevant in tropical oligotrophic waters when the system is characterized by a microbial food web structure.

Free-living, photosynthetic marine dinoflagellates are frequently infected by microparasites of the genus Amoebophrya. Attacks by Amoebophrya can contribute to the termination of dinoflagellate blooms and have been suggested to influence the geographical distribution of certain host species. Because infection terminates with the killing of the host (i.e. Amoebophrya behaves like a parasitoid), the interaction can be considered, from a modeling point of view, similar to the process of predation, with the difference that it takes a longer time for the parasitoid to kill the host as compared to typical predator-prey interactions. In the present work, we explored the population dynamics in Amoebophrya and their dinoflagellate hosts using the Rosenzweig-MacArthur modification of the traditional Lotka-Volterra predation model. The model was parameterized for 3 systems, Akashiwo sanguinea, Gymnodinium instriatum, and Karlodinium micrum, and their respective Amoebophrya parasitoids, using published experimental data. Parameter validation was possible for parasitoid search rate and mortality. The potential for host control by Amoebophrya and the probability for host extinction were studied with respect to carrying capacity, a parameter that is influenced by e. g. eutrophication. The model may be useful to predict conditions under which Amoebophrya can control populations of its dinoflagellate hosts.