Pulmonary surfactant (PS) reduces surface tension at the air-liquid interface in the alveolar epithelium of the lung, which is required for breathing and for the pulmonary maturity of the developing foetus. However, the origin of PS had never been thoroughly investigated, although it was assumed to be secreted from the foetal developing lung. Human amniotic membrane (hAM), particularly its epithelial cell layer, composes the amniotic sac enclosing the amniotic fluid. In this study, we therefore aimed to investigate a potential contribution of the cellular components of the hAM to pulmonary surfactant found in amniotic fluid. We identified that cells within the native membrane contain lamellar bodies and express all four surfactant proteins as well as ABCA3. Lipidomic profiling by nanoESI – MS/MS revealed the presence of the essential lipid species as found in PS. Also, the biophysical activity of conditioned cell culture supernatant obtained from hAM was tested with captive bubble surfactometry. hAM supernatant showed the ability to reduce surface tension, similar to human PS obtained from bronchoalveolar lavage. This means that hAM produces the essential PS-associated components and can therefore contribute as second potential source of PS in amniotic fluid aside from the foetal lung.

Mimiviridae is a group of viruses with large genomes and virions. Ecological relevance of Mimiviridae in marine environments has been increasingly recognized through the discoveries of novel isolates and metagenomic studies. To facilitate ecological profiling of Mimiviridae, we previously proposed a meta-barcoding approach based on 82 degenerate primer pairs (i.e., MEGAPRIMER) targeting the DNA polymerase gene of Mimiviridae. The method detected a larger number of operational taxonomic units (OTUs) in environmental samples than previous methods. However, it required large quantities of DNA and was laborious due to the use of individual primer pairs. Here, we examined coastal seawater samples using varying PCR conditions and purification protocols to streamline the MEGAPRIMER method. Mixing primer pairs in “cocktails” reduced the required amount of environmental DNA by 90%, while reproducing the results obtained by the original protocol. We compared the results obtained by the meta-barcoding approach with quantifications using qPCR for selected OTUs. This revealed possible amplification biases among different OTUs, but the frequency profiles for individual OTUs across multiple samples were similar to those obtained by qPCR. We anticipate that the newly developed MEGAPRIMER protocols will be useful for ecological investigation of Mimiviridae in a larger set of environmental samples.

Coastal seawater is the habitat of diverse microbial communities. These communities are affected by seasonal environmental changes and fluctuating nutrient availability, as well as competitive and cooperative interspecific interactions. The complex interplay of these factors affects elemental cycles and therefore, the environment, which in turn affects microbial communities. In this work, we investigated the seasonal dynamics of communities of eukaryotes, a major group of double-stranded DNA viruses infecting eukaryotes (i.e., Mimiviridae), as well as prokaryotes in the Uranouchi Inlet, Kochi, Japan. This inlet harbors several microalgae that recurrently form blooms throughout the year. We performed metabarcoding using ribosomal RNA genes and a Mimiviridae DNA polymerase gene as marker genes and counted cells of major algal species in 43 seawater samples collected during 20 months. Communities of eukaryotes, Mimiviridae, and prokaryotes characterized at the amplicon sequence variant (ASV) level showed similar seasonal cycles but differences in several aspects such as the recovery rate after a year. The ASV persistence level differed among eukaryote, Mimiviridae, and prokaryote communities. Mimiviridae ASVs were less persistent than were eukaryotic ASVs, and prokaryotic ASVs were the most persistent. These observations are discussed in the context of survival strategies of the major microbes in these three communities.

Coastal microbial communities are affected by seasonal environmental change, biotic interactions, and fluctuating nutrient availability. We investigated the seasonal dynamics of communities of eukaryotes, a major group of double-stranded DNA viruses that infect eukaryotes (order Imitervirales; phylum Nucleocytoviricota), and prokaryotes in the Uranouchi Inlet, Kochi, Japan. We performed metabarcoding using ribosomal RNA genes and viral polB genes as markers in 43 seawater samples collected over 20 months. Eukaryotes, prokaryotes, and Imitervirales communities characterized by the compositions of amplicon sequence variants (ASVs) showed synchronic seasonal cycles. However, the community dynamics showed intriguing differences in several aspects, such as the recovery rate after a year. We also showed that the differences in community dynamics were at least partially explained by differences in recurrence/persistence levels of individual ASVs among eukaryotes, prokaryotes, and Imitervirales. Prokaryotic ASVs were the most persistent, followed by eukaryotic ASVs and Imitervirales ASVs, which were the least persistent. We argue that the differences in the specificity of interactions (virus–eukaryote vs. prokaryote–eukaryote) as well as the niche breadth of community members were at the origin of the distinct community dynamics among eukaryotes, their viruses, and prokaryotes.

Viruses are important regulatory factors of the marine microbial community including microeukaryotes. However, little is known about their role in the northern Chukchi Sea in the Arctic basin, which has oligotrophic conditions in summer. To clarify the link between microbial eukaryotic communities and viruses as well as environmental conditions, we investigated the community structures of microeukaryotes (from 3 to144 μm and from 0.23 μm size bio‐particles collected from seawater) and Imitervirales (from 0.23 μm size bio‐particles collected from seawater), a dominant group of viruses infecting marine microeukaryotes. To the best of our knowledge, no study has investigated both Imitervirales and eukaryotic communities in the Arctic Ocean. Surface water samples were collected at 21 ocean stations located in the northeastern Chukchi Sea and an adjacent area outside the Beaufort Gyre (Adjacent Sea), and at two melt ponds on sea ice in the summer of 2018. At the ocean stations, nutrient concentrations were low in most of the locations, except the shelf in the adjacent sea. The community variations were significantly correlated between eukaryotes and Imitervirales, even within the northeastern Chukchi Sea characterized by relatively homogeneous environmental conditions. The association of the eukaryotic community with the viral community was stronger than that with geographical and physicochemical environmental factors. These results suggest that Imitervirales actively infect their hosts even in the cold and oligotrophic seawater in the Arctic Ocean.

Viruses are important regulatory factors of marine microbial community including microeukaryotes. However, little is known about their role in the northern Chukchi Sea of the Arctic basin, which remains oligotrophic conditions in summer. To elucidate linkages of microbial eukaryotic community with viruses as well as environmental variables, we investigated the community structures of microeukaryotes (3-144 μm and 0.2-3 μm size fractions) and Imitervirales (0.2-3 μm size fraction), a major group of viruses infecting marine microeukaryotes. Surface water samples were collected at 21 ocean stations located in the northeastern Chukchi Sea (NECS), an adjacent area outside the Beaufort Gyre (Adjacent Sea; AS), and two melt ponds on sea ice in the summer of 2018. At the ocean stations, nutrient concentrations were low in most of the locations expect at the shelf in the AS. The community variations were significantly correlated between eukaryotes and Imitervirales, even within the NECS characterized by relatively homogeneous environmental conditions. The association of the eukaryotic community with the viral community was stronger than that with geographical and physicochemical environmental factors. These results suggest that Imitervirales are actively infecting their hosts even in cold and oligotrophic sea water in the Arctic Ocean.