Abstract
Bacteria are an active and diverse component of pelagic communities. The identification of main factors governing microbial diversity and spatial distribution requires advanced mathematical ...analyses. Here, the bacterial community composition was analysed, along with a depth profile, in the open Adriatic Sea using amplicon sequencing of bacterial 16S rRNA and the Neural gas algorithm. The performed analysis classified the sample into four best matching units representing heterogenic patterns of the bacterial community composition. The observed parameters were more differentiated by depth than by area, with temperature and identified salinity as important environmental variables. The highest diversity was observed at the deep chlorophyll maximum, while bacterial abundance and production peaked in the upper layers. The most of the identified genera belonged to Proteobacteria, with uncultured AEGEAN-169 and SAR116 lineages being dominant Alphaproteobacteria, and OM60 (NOR5) and SAR86 being dominant Gammaproteobacteria. Marine
Synechococcus
and
Cyanobium
-related species were predominant in the shallow layer, while
Prochlorococcus
MIT 9313 formed a higher portion below 50 m depth. Bacteroidota were represented mostly by uncultured lineages (NS4, NS5 and NS9 marine lineages). In contrast, Actinobacteriota were dominated by a candidatus genus
Ca.
Actinomarina. A large contribution of Nitrospinae was evident at the deepest investigated layer. Our results document that neural network analysis of environmental data may provide a novel insight into factors affecting picoplankton in the open sea environment.
By combining qualitative 16S metabarcoding and quantitative CARD-FISH methods with neural gas analysis, different patterns of the picoplankton community were revealed at finer taxonomic levels in ...response to changing environmental conditions in the Adriatic Sea. We present the results of a one-year study carried out in an oligotrophic environment where increased salinity was recently observed. We have shown that the initial state of community structure changes according to environmental conditions and is expressed as qualitative and quantitative changes. A general pattern of increasing diversity under harsh environmental conditions, particularly under the influence of increasing salinity at the expense of community abundance was observed. Considering the trend of changing seawater characteristics due to climate change, this study helps in understanding a possible structural change in the microbial community of the Adriatic Sea that could affect higher levels of the marine food web.
Human-induced climate change is expected to increase the frequency and severity of vegetation fires. The Mediterranean region is considered particularly prone to fire episodes in summer. It is well ...known that pyrogenic particles are an important source of external nutrients for the marine environment, especially in oligotrophic areas. In this study, the plankton components of the sea surface layers were integrated to evaluate, for the first time, their dynamics over six months and their response to fire events in a typical coastal area of the Adriatic Sea. Concentrations of nutrients and organic compounds, together with plankton communities were significantly higher in the sea surface microlayer (SML, < 1 mm thick), than in the underlying water from 1 m depth. The piconeuston community and chlorophyll a responded with extreme abundance and concentration to the most intense fire event that enriched the SML with NH4+. Phytoneuston abundance increased with a delay of 2 weeks, while diversity indices decreased slightly after the fire events. The large abundances of the studied piconeuston parameters could be explained by the high availability of organic compounds and the immediate availability of NH4+, while the phytoneuston community responded to an increased NO3‐ concentration, triggered by the fire events. We confirmed that fast-acting marine heterotrophs are important members of biogeochemical cycles associated with fire events and that, together with phytoplankton, they are unavoidable parameters to detect environmental changes.
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•Climate change is expected to increase frequency and severity of vegetation fires•Adriatic coastline is particularly vulnerable to fires in summer•Piconeuston and chlorophyll a responded immediately to NH4+ derived from fire event•Phytoneuston responded to fire events with a delay of 2 weeks
An assessment of the temperature increase effect on processes within the microbial food web provides a better insight into the carbon transfer and energy flow processes in marine environments in the ...global warming perspective. Modified laboratory dilution experiments that allow simultaneous estimates of protozoan grazing and viral lysis on picoplankton groups (bacteria, Prochlorococcus, Synechococcus and pico-eukaryotic algae) under in situ and 3°C above in situ temperatures were performed at seasonal scale. Picoplankton mortality due to grazing was generally higher than that caused by viral lysis, especially in the cold months. The largest part of HNF carbon demand was satisfied by grazing on bacteria throughout the year. Although ciliates satisfied their carbon demand predominantly through grazing on HNF and bacteria, the role of autotrophic picoplankton (APP) as their prey increased significantly in the cold months. Bacteria constituted the most important host for viruses throughout the year. However, during the warm months, APP groups were also significant hosts for viral infection. Under the warming condition the amount of picoplankton biomass transferred to protozoan grazers exceeded the lysed biomass, suggesting that global warming could further increase picoplankton carbon flow toward higher trophic levels in the Adriatic Sea.
Marine bacterioplankton represent a diverse assembly of species differing largely in their abundance, physiology, metabolic activity, and role in microbial food webs. To analyze their sensitivity to ...bottom-up and top-down controls, we performed a manipulation experiment where grazers were removed, with or without the addition of phosphate. Using amplicon-reads normalization by internal standard (ARNIS), we reconstructed growth curves for almost 300 individual phylotypes. Grazer removal caused a rapid growth of most bacterial groups, which grew at rates of 0.6 to 3.5 day−1, with the highest rates (>4 day−1) recorded among Rhodobacteraceae, Oceanospirillales, Alteromonadaceae, and Arcobacteraceae. Based on their growth response, the phylotypes were divided into three basic groups. Most of the phylotypes responded positively to both grazer removal as well as phosphate addition. The second group (containing, e.g., Rhodobacterales and Rhizobiales) responded to the grazer removal but not to the phosphate addition. Finally, some clades, such as SAR11 and Flavobacteriaceae, responded only to phosphate amendment but not to grazer removal. Our results show large differences in bacterial responses to experimental manipulations at the phylotype level and document different life strategies of marine bacterioplankton. In addition, growth curves of 130 phylogroups of aerobic anoxygenic phototrophs were reconstructed based on changes of the functional pufM gene. The use of functional genes together with rRNA genes may significantly expand the scientific potential of the ARNIS technique. IMPORTANCE Growth is one of the main manifestations of life. It is assumed generally that bacterial growth is constrained mostly by nutrient availability (bottom-up control) and grazing (top-down control). Since marine bacteria represent a very diverse assembly of species with different metabolic properties, their growth characteristics also largely differ accordingly. Currently, the growth of marine microorganisms is typically evaluated using microscopy in combination with fluorescence in situ hybridization (FISH). However, these laborious techniques are limited in their throughput and taxonomical resolution. Therefore, we combined a classical manipulation experiment with next-generation sequencing to resolve the growth dynamics of almost 300 bacterial phylogroups in the coastal Adriatic Sea. The analysis documented that most of the phylogroups responded positively to both grazer removal and phosphate addition. We observed significant differences in growth kinetics among closely related species, which could not be distinguished by the classical FISH technique.
•Responses of MFW structure to temperature change are reproducible in time.•Similar changes in MFW structure occurred regardless of the trophic status.•Importance of microbial heterotrophic ...activities increases with temperature.•Importance of autotrophic picoplankton increases with temperature.
Global and atmospheric climate change is altering the thermal conditions in the Adriatic Sea and, consequently, the marine ecosystem. Along the eastern Adriatic coast sea surface temperature (SST) increased by an average of 1.03 °C during the period from 1979 to 2015, while in the recent period, starting from 2008, a strong upward almost linear trend of 0.013 °C/month was noted. Being mainly oligotrophic, the middle Adriatic Sea is characterized by the important role played by the microbial food web in the production and transfer of biomass and energy towards higher trophic levels. It is very important to understand the effect of warming on microbial communities, since small temperature increases in surface seawater can greatly modify the microbial role in the global carbon cycle. In this study, the Self-Organizing Map (SOM) procedure was used to analyse the time series of a number of microbial parameters at two stations with different trophic status in the central Adriatic Sea. The results show that responses of the microbial food web (MFW) structure to temperature changes are reproducible in time. Furthermore, qualitatively similar changes in the structure of the MFW occurred regardless of the trophic status. The rise in temperature was associated with: (1) the increasing importance of microbial heterotrophic activities (increase bacterial growth and bacterial predator abundance, particularly heterotrophic nanoflagellates) and (2) the increasing importance of autotrophic picoplankton (APP) in the MFW.
The use of a suitable method for the enumeration of indicator microorganisms is of crucial importance for reliable monitoring and assessment of the quality of bathing waters. Among other ...characteristics, the method should be selective enough and ensure acceptable relative recovery of target microorganisms. This study presents the basic parameters, relative recovery and categorical performance characteristics of Tryptone Bile X-glucuronide (TBX) agar for
Escherichia coli
(
E. coli
) enumeration in bathing water samples using the membrane filtration method.
The results of the relative recovery study, in which TBX agar was compared against temperature-modified ISO 9308-1:
2014
, showed that in order to achieve a satisfactory relative recovery of
E. coli
with TBX agar at 44 ± 0.5 °C, the resuscitation period on a non-selective medium (Minerals Modified Glutamate Agar, MMGA) at 36 ± 2 °C is crucial. Incubation on a double-layer MMGA/TBX medium with a 6-h resuscitation period and alternating incubation on single-layer MMGA and TBX agar with a 4-h resuscitation period resulted in acceptable and very similar relative recovery. The achieved performance characteristics of the tested medium, double-layer MMGA/TBX agar, are acceptable. The selectivity was matrix-dependent and was 60.6% for inland and 69.9% for coastal waters. No significant effect of the resuscitation period on selectivity was recorded. Finally, the results showed that when the resuscitation period on a non-selective medium is included, TBX agar is a suitable medium for
E. coli
enumeration in bathing water samples using the membrane filtration method and that its use, theoretically, would not have negative effects on the assessment of bathing water quality.
Aerobic anoxygenic phototrophs are a newly discovered member of the bacterial community in the Adriatic Sea. During the last seven years, when we started to study these organisms, we have collected a ...considerable number of samples from different environments, from the coast, the estuary, and the open sea. Here we have compiled data from 34 georeferenced study sites from four studies that summarize all that is known about aerobic anoxygenic phototrophs and examine the spatial and vertical distribution and environmental factors affecting this community in the Adriatic Sea. We found horizontal and vertical influences on AAP distribution, mainly salinity, nitrates, chlorophyll
a
, ammonium, temperature, and soluble reactive phosphorus. Much is known about their ecology in the Adriatic, and with a new survey underway, we will expand our knowledge of their community composition and their role in carbon flux to higher trophic levels.
Aerobni anoksigeni fototrofi relativno su novi član bakterijske zajednice u Jadranskom moru. U posljednjih sedam godina, od kada smo započeli proučavati ove organizme, prikupili smo znatan broj uzoraka iz različitih okoliša, obalnog, estuarnog i područja otvorenog mora. U ovom radu objedinili smo podatke s 34 georeferencirane lokacije istraživanja iz četiri studije koje sažimaju sva saznanja o aerobnim anoksigenim fototrofima te istražili prostornu i vertikalnu raspodjelu, kao i okolišne čimbenike koji utječu na ovu zajednicu u Jadranskom moru. Utvrdili smo čimbenike koji utječu na horizontalnu i vertikalnu raspodjelu AAP, uglavnom salinitet, nitrate, klorofil a, amonijeve ione, temperaturu te topljivi reaktivni fosfor. Iako se dosta zna o njihovoj ekologiji u Jadranu, aktualnim istraživanjem proširit ćemo naše znanje o sastavu ove zajednice i njihovoj ulozi u protoku ugljika prema višim trofičkim razinama.
Temperature and phosphorus positively interacted in controlling picoplankton biomass production and its transfer towards higher trophic levels. Two complementary approaches (experimental and field ...study) indicated several coherent patterns: (1) the impact of temperature on heterotrophic bacteria was high at temperatures lower than 16°C and levelled off at higher temperatures, whereas this impact on autotrophic picoplankton was linear along the entire range of the investigated temperatures; (2) the addition of phosphorus increased the values of picoplankton production and grazing, but did not change the nature of their relationships with temperature substantially; (3) the picoplankton carbon flux towards higher trophic levels was larger during the warmer months (grazing by HNF dominated during the warmer period and by ciliates during the colder period) and also strengthened in conditions without phosphorus limitation; (4) the hypothesis that the available phosphorus can be better utilized at higher temperatures was confirmed for both autotrophic and heterotrophic picoplankton; (5) the hypothesis that the rise in temperature stimulates growth only in conditions of sufficient phosphorus was confirmed only for heterotrophic bacteria. Therefore, in the global warming scenario, an increase of the picoplankton carbon flux towards higher trophic levels can be expected in the Adriatic Sea, particularly under unlimited phosphorus conditions.
•Anomalous physical and microbial properties were detected in the middle Adriatic.•Local and remote drivers contributed to record-breaking salinities.•Picoplankton biomass and production sharply ...declined in high salinity conditions.•The documented conditions could be more frequent in the future climate.
Oceanographic measurements carried out in the middle Adriatic during summer 2017 revealed anomalous conditions in both physical and microbial properties. High salinities were observed throughout the entire water column, with an ‘inverse’ salinity profile in August and a maximum in the surface layer, recorded for the first time in the middle Adriatic. Surface salinity of 39.02 recorded in August was 2.5 standard deviations above the long-term average (1961–2016). The observed salinity distributions are the result of both local and remote drivers, whereby the North Ionian cyclonic gyre controlled by the Adriatic-Ionian Bimodal Oscillating System has been responsible for the overall above-average salinities since 2011. Yet, local factors present in 2017, such as strong evaporation caused by extremely high air temperatures, lack of precipitation and low river discharges, combined with a decrease in horizontal transport estimated from the Regional Ocean Modeling System simulations, contributed substantially to the observed surface salinity anomaly. The decrease in horizontal advection was conjoined with high values of repelling barriers in the fields of the finite-time Lyapunov exponent. Documented physical conditions were synchronized with considerably lower bacterial production and abundance of the most studied picoplankton groups in comparison to values during the last decade. The exception was the euryhaline organism Synechococcus, whose abundance was 88% higher than the average in the study area. Nutrient content and chlorophyll-a concentrations followed regular seasonal cycles during 2017, with typical low values pointing to salinity as a possible driver of the observed changes in the microbial food web. Following ongoing climate change and future projections, these documented anomalous physical and microbiological conditions may become more frequent in the Adriatic Sea.
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