Diversity and distribution of heterotrophic flagellates in seawater of the Powell Basin, Antarctic Peninsula

Heterotrophic flagellates are essential components of the marine microbial food web. However, how the changes in flagellate populations reflect environmental changes in marine ecosystems is still unclear, especially in polar regions. In this study, we used pyrosequencing to examine the community structure of heterotrophic flagellates (HFs) in the Powell Basin’s surface waters of the northern Antarctic Peninsula. OTUs (operational taxonomic units) of different taxa and the correlations between community structure and environmental factors were analysed. Eight taxa of HFs were selected for the principal analysis: Telonemia, Picozoa, Rhizaria, Amoebozoa, Apusomonas, Centrohelida, Choanomonada and marine stramenopiles (MASTs). The HFs were defined as heterotrophic picoflagellates (HPFs; <3 μm) and heterotrophic nanoflagellates (HNFs; >3 μm, <20 μm), which had similar dominant phyla (MASTs and Telonemia). However, their taxonomic composition differed. Environmental factors exerted similar effects on the community structure of both HPFs and HNPs. Compared with the correlation between HPF and environmental factors, the correlation between HNF and environmental factors was stronger. Salinity, bacterial biomass and the biological interactions amongst dominant taxa were the main variables to influence the diversity and community structure of HFs.

Diel population dynamics and mortality of Prochlorococcus in the North Pacific Subtropical Gyre

Marine ecosystem models often consider temporal dynamics on the order of months to years, and spatial dynamics over regional and global scales as a means to understand the ecology, evolution, and biogeochemical impacts of marine life. Large-scale dynamics are themselves driven over diel scales as a result of light-driven forcing, feedback, and interactions. Motivated by high-frequency measurements taken by Lagrangian sampling in the North Pacific Subtropical Gyre, we develop a hierarchical set of multitrophic community ecology models to investigate and understand daily ecological dynamics in the near-surface ocean including impacts of light-driven growth, infection, grazing, and phytoplankton size structure. Using these models, we investigate the relative impacts of viral-induced and grazing mortality for Prochlorococcus; and more broadly compare in silico dynamics with in situ observations. Via model-data fitting, we show that a multi-trophic model with size structure can jointly explain diel changes in cyanobacterial abundances, cyanobacterial size structure, viral abundance, viral infection rates, and grazer abundances. In doing so, we find that a significant component (between 5% to 55%) of estimated Prochlorococcus mortality is not attributed to either viral lysis (by T4- or T7-like cyanophage) or grazing by heterotrophic nanoflagellates. Instead, model-data integration suggests a heightened ecological relevance of other mortality mechanisms -- including grazing by other predators, particle aggregation, and stress-induced loss mechanisms. Altogether, linking mechanistic multitrophic models with high-resolution measurements provides a route for understanding of diel origins of large-scale marine microbial community and ecosystem dynamics.

Community Assembly Processes as a Mechanistic Explanation of the Predator-Prey Diversity Relationship in Marine Microbes

Predator and prey α-diversities are often positively associated; yet, understandings of the underlying mechanisms require manipulative experiments and thus remain unclear. We attempt to address this issue by deciphering how α-diversity of predator and prey influences each other’s community assembly processes, which subsequently determine their α-diversity. The occurrence of assembly processes was indicated by the mean pairwise taxonomic index within a community (αMPTI), assuming assembly processes left traceable imprints on species’ phylogeny. Specifically, αMPTI quantifies deviations of observed phylogenetic distances from that of random, so that it can be used to hint at the occurrence of non-random/deterministic assembly processes. Larger αMPTI of a community implies the occurrence of weaker homogenizing deterministic assembly processes, which suggests that this community might be comprised of less similar species and thus has higher α-diversity. We hypothesize that higher predator and prey α-diversity would be positively associated with each other’s αMPTI, which would then be positively associated with their α-diversity. To test the hypothesis, we calculated Shannon diversity and αMPTI for heterotrophic nanoflagellates (HNF; predator) and bacteria (prey) communities in the East China Sea (ECS). The HNF Shannon diversity was found to be positively associated with αMPTI of bacteria, which was then positively associated with bacterial Shannon diversity. In contrast, bacterial Shannon diversity did not correlate with HNF’s αMPTI. We argue that top-down control is one of the explanations to the positive α-diversity association among trophic levels in microbes of the ECS.

Mechanisms and fluid dynamics of foraging in heterotrophic nanoflagellates

ABSTRACTHeterotrophic nanoflagellates are the main consumers of bacteria and picophytoplankton in the ocean. In their micro-scale world, viscosity impedes predator-prey contact, and the mechanisms that allow flagellates to daily clear a volume of water for prey corresponding to 106 times their own volume is unclear. It is also unclear what limits observed maximum ingestion rates of about 104 bacterial prey per day. We used high-speed video-microscopy to describe feeding flows, flagellum kinematics, and prey searching, capture, and handling in four species with different foraging strategies. In three species, prey-handling times limit ingestion rates and account well for their reported maximum values. Similarly, observed feeding flows match reported clearance rates. Simple point-force models allowed us to estimate the forces required to generate the feeding flows, between 4-13 pN, and consistent with the force produced by the hairy (hispid) flagellum, as estimated using resistive force theory. Hispid flagella can produce a force that is much higher than the force produced by a naked flagellum with similar kinematics, and the hairy flagellum is therefore key to foraging in most nanoflagellates. Our findings provide a mechanistic underpinning of observed functional responses of prey ingestion rates in nanoflagellates.

Elevated Contribution of Low Nucleic Acid Prokaryotes and Viral Lysis to the Prokaryotic Community Along the Nutrient Gradient From an Estuary to Open Ocean Transect

Prokaryotes represent the largest living biomass reservoir in aquatic environments and play a crucial role in the global ocean. However, the factors that shape the abundance and potential growth rate of the ecologically distinct prokaryotic subgroups [i.e., high nucleic acid (HNA) and low nucleic acid (LNA) cells] along varying trophic conditions in the ocean remain poorly understood. This study conducted a series of modified dilution experiments to investigate how the abundance and potential growth rate of HNA and LNA prokaryotes and their regulating factors (i.e., protozoan grazing and viral lysis) change along a cross-shore nutrient gradient in the northern South China Sea. The results showed that the abundance of both HNA and LNA cells was significantly positively correlated with the abundance of heterotrophic nanoflagellates and viruses, whereas only HNA abundance exhibited a significant positive correlation with nutrient level. With a decreasing nutrient concentration, the potential growth rate of the HNA subgroup declined significantly, while that of the LNA subgroup was significantly enhanced, leading to an elevated relative potential growth rate of the LNA to HNA subgroup under decreasing nutrient levels. Furthermore, our data revealed different regulatory roles of protozoan grazing and viral lysis on the HNA and LNA subgroups, with HNA suffering higher mortality pressure from grazing than from lysis in contrast to LNA, which experienced equivalent pressures. As the nutrient levels declined, the relative contribution of lysis to the mortality of the HNA subgroup increased significantly, in contrast to the insignificant change in that of the LNA subgroup. Our results indicated the elevated role of LNA cells in the prokaryotic community and the enhanced viral lysis pressure on the total prokaryotes under oligotrophic conditions. This implies a weakened efficiency of carbon cycling within the microbial loop and enhanced viral lysis to shunt more carbon and energy flow in the future ocean, in which oligotrophication will be strengthened due to global warming.

Contribution of large bacteria to bacterial biomass in a deep freshwater lake (Lake Biwa, Japan)

Despite the importance of bacterial cell volume in microbial ecology in aquatic environments, literature regarding the effects of seasonal and spatial variations on bacterial cell volume remains scarce. We used transmission electron microscopy to examine seasonal and spatial variations in bacterial cell size for 18 mo in 2 layers (epilimnion 0.5 m and hypolimnion 60 m) of Lake Biwa, Japan, a large and deep freshwater lake. During the stratified period, we found that the bacterial cell volume in the hypolimnion ranged from 0.017 to 0.12 µm3 (median), whereas that in the epilimnion was less variable (0.016 to 0.033 µm3, median) and much lower than that in the hypolimnion. Additionally, in the hypolimnion, cell volume during the stratified period was greater than that during the mixing period (up to 5.7-fold). These differences in cell volume resulted in comparable bacterial biomass in the hypolimnion and epilimnion, despite the fact that there was lower bacterial abundance in the hypolimnion than in the epilimnion. We also found that the biomass of larger bacteria, which are not likely to be grazed by heterotrophic nanoflagellates, increased in the hypolimnion during the stratified period. Our data suggest that estimation of carbon flux (e.g. bacterial productivity) needs to be interpreted cautiously when cell volume is used as a constant parametric value. In deep freshwater lakes, a difference in cell volume with seasonal and spatial variation may largely affect estimations.

Ocean acidification reduces growth and grazing impact of Antarctic heterotrophic nanoflagellates

High-latitude oceans have been identified as particularly vulnerable to ocean acidification if anthropogenic CO2 emissions continue. Marine microbes are an essential part of the marine food web and are a critical link in biogeochemical processes in the ocean, such as the cycling of nutrients and carbon. Despite this, the response of Antarctic marine microbial communities to ocean acidification is poorly understood. We investigated the effect of increasing fCO2 on the growth of heterotrophic nanoflagellates (HNFs), nano- and picophytoplankton, and prokaryotes (heterotrophic Bacteria and Archaea) in a natural coastal Antarctic marine microbial community from Prydz Bay, East Antarctica. At CO2 levels ≥634 µatm, HNF abundance was reduced, coinciding with increased abundance of picophytoplankton and prokaryotes. This increase in picophytoplankton and prokaryote abundance was likely due to a reduction in top-down control of grazing HNFs. Nanophytoplankton abundance was elevated in the 634 µatm treatment, suggesting that moderate increases in CO2 may stimulate growth. The taxonomic and morphological differences in CO2 tolerance we observed are likely to favour dominance of microbial communities by prokaryotes, nanophytoplankton, and picophytoplankton. Such changes in predator–prey interactions with ocean acidification could have a significant effect on the food web and biogeochemistry in the Southern Ocean, intensifying organic-matter recycling in surface waters; reducing vertical carbon flux; and reducing the quality, quantity, and availability of food for higher trophic levels.

Cascading effects in freshwater microbial food webs by predatory Cercozoa, Katablepharidacea and ciliates feeding on aplastidic bacterivorous cryptophytes

ABSTRACT Heterotrophic nanoflagellates (HNF) are considered as major planktonic bacterivores, however, larger HNF taxa can also be important predators of eukaryotes. To examine this trophic cascading, natural protistan communities from a freshwater reservoir were released from grazing pressure by zooplankton via filtration through 10- and 5-µm filters, yielding microbial food webs of different complexity. Protistan growth was stimulated by amendments of five Limnohabitans strains, thus yielding five prey-specific treatments distinctly modulating protistan communities in 10- versus 5-µm fractions. HNF dynamics was tracked by applying five eukaryotic fluorescence in situ hybridization probes covering 55–90% of total flagellates. During the first experimental part, mainly small bacterivorous Cryptophyceae prevailed, with significantly higher abundances in 5-µm treatments. Larger predatory flagellates affiliating with Katablepharidacea and one Cercozoan lineage (increasing to up to 28% of total HNF) proliferated towards the experimental endpoint, having obviously small phagocytized HNF in their food vacuoles. These predatory flagellates reached higher abundances in 10-µm treatments, where small ciliate predators and flagellate hunters also (Urotricha spp., Balanion planctonicum) dominated the ciliate assemblage. Overall, our study reports pronounced cascading effects from bacteria to bacterivorous HNF, predatory HNF and ciliates in highly treatment-specific fashions, defined by both prey-food characteristics and feeding modes of predominating protists.

Multivariate control of heterotrophic bacterial abundance and zooplankton grazing in Labrador fjords (northeastern Canada)

This study was conducted in 4 Labrador fjords (Nachvak, Saglek, Okak, and Anaktalak) during the summers of 2007 and 2013, early fall 2010, and late fall 2009. Our results show that water temperature combined with the availability of nutrients and organic substrates are the main abiotic factors controlling the abundance of heterotrophic bacteria in Labrador fjords. Bacterivory also played a crucial role, with heterotrophic bacteria exerting a significant bottom-up control on the abundance of heterotrophic nanoflagellates (r = 0.35, p < 0.05) and ciliates (r = 0.70, p < 0.01). During summer 2013, the intrinsic phytoplankton growth rate varied between <0 and 0.64 d-1, with a mean value of 0.36 d-1. The herbivory rate was highly variable, ranging from 0.01 to 0.86 d-1, with a mean value of 0.31 d-1. Grazing mortality was 6-fold higher than phytoplankton growth rate. Mean phytoplankton growth and herbivory rates in Labrador fjords were comparable to the Barents and Bering seas. The intrinsic growth rate of total heterotrophic bacteria ranged between <0 and 0.68 d-1, with a mean value of 0.30 d-1. Bacterivory varied from 0.01 to 0.95 d-1, with a mean of 0.30 d-1. Mortality due to grazing was up to 2.3 times higher than total bacterial growth rate. This study improves our understanding of the factors influencing the dynamics of heterotrophic bacteria and indicates that herbivory and bacterivory exert substantial control on microbial communities in Labrador fjords.

Weekly variations of viruses and heterotrophic nanoflagellates and their potential impact on bacterioplankton in shallow waters of the central Red Sea

ABSTRACT Bacterioplankton play a pivotal role in marine ecosystems. However, their temporal dynamics and underlying control mechanisms are poorly understood in tropical regions such as the Red Sea. Here, we assessed the impact of bottom-up (resource availability) and top-down (viruses and heterotrophic nanoflagellates) controls on bacterioplankton abundances by weekly sampling a coastal central Red Sea site in 2017. We monitored microbial abundances by flow cytometry together with a set of environmental variables including temperature, salinity, dissolved organic and inorganic nutrients and chlorophyll a. We distinguished five groups of heterotrophic bacteria depending on their physiological properties relative nucleic acid content, membrane integrity and cell-specific respiratory activity, two groups of Synechococcus cyanobacteria and three groups of viruses. Viruses controlled heterotrophic bacteria for most of the year, as supported by a negative correlation between their respective abundances and a positive one between bacterial mortality rates and mean viral abundances. On the contrary, heterotrophic nanoflagellates abundance covaried with that of heterotrophic bacteria. Heterotrophic nanoflagellates showed preference for larger bacteria from both the high and low nucleic acid content groups. Our results demonstrate that top-down control is fundamental in keeping heterotrophic bacterioplankton abundances low (&lt; 5 × 10 5 cells mL−1) in Red Sea coastal waters.