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1.
PLoS One ; 16(8): e0256639, 2021.
Article in English | MEDLINE | ID: mdl-34437618

ABSTRACT

Pontibacillus sp. ALD_SL1 and Psychroflexus sp. ALD_RP9 are two novel bacterial isolates from mangrove sediment and a moderately hypersaline pool on the Aldabra Atoll, Seychelles. The isolates represent two novel species were characterised physiologically and genomically. Pontibacillus sp. ALD_SL1 is a facultatively anaerobic yellow, motile, rod-shaped Gram-positive, which grows optimally at a NaCl concentration of 11%, pH 7 and 28°C. It is the third facultatively anaerobic member of the genus Pontibacillus. The organism gains energy through the fermentation of pyruvate to acetate and ethanol under anaerobic conditions. The genome is the first among Pontibacillus that harbours a megaplasmid. Psychroflexus sp. ALD_RP9 is an aerobic heterotroph, which can generate energy by employing bacteriorhodopsins. It forms Gram-negative, orange, non-motile rods. The strain grows optimally at NaCl concentrations of 10%, pH 6.5-8 and 20°C. The Psychroflexus isolate tolerated pH conditions up to 10.5, which is the highest pH tolerance currently recorded for the genus. Psychroflexus sp. ALD_RP9 taxonomically belongs to the clade with the smallest genomes. Both isolates show extensive adaptations to their saline environments yet utilise different mechanisms to ensure survival.


Subject(s)
Bacillaceae/isolation & purification , Flavobacteriaceae/isolation & purification , Geologic Sediments/microbiology , Bacillaceae/enzymology , Bacillaceae/growth & development , Bacillaceae/ultrastructure , Flavobacteriaceae/enzymology , Flavobacteriaceae/growth & development , Flavobacteriaceae/ultrastructure , Genome, Bacterial , Kinetics , Phylogeny , Seychelles , Water Microbiology
2.
Molecules ; 26(11)2021 May 24.
Article in English | MEDLINE | ID: mdl-34073991

ABSTRACT

Although axenic microbial cultures form the basis of many large successful industrial biotechnologies, the production of single commercial microbial strains for use in large environmental biotechnologies such as wastewater treatment has proved less successful. This study aimed to evaluate the potential of the co-culture of two halophilic bacteria, Marinirhabdus sp. and Marinobacter hydrocarbonoclasticus for enhanced protease activity. The co-culture was significantly more productive than monoculture (1.6-2.0 times more growth), with Marinobacter hydrocarbonoclasticus being predominant (64%). In terms of protease activity, enhanced total activity (1.8-2.4 times) was observed in the co-culture. Importantly, protease activity in the co-culture was found to remain active over a much broader range of environmental conditions (temperature 25 °C to 60 °C, pH 4-12, and 10-30% salinity, respectively). This study confirms that the co-culturing of halophilic bacteria represents an economical approach as it resulted in both increased biomass and protease production, the latter which showed activity over arange of environmental conditions.


Subject(s)
Flavobacteriaceae/enzymology , Marinobacter/enzymology , Peptide Hydrolases/biosynthesis , Coculture Techniques , Flavobacteriaceae/growth & development , Hydrogen-Ion Concentration , Marinobacter/growth & development , Salinity , Temperature
3.
Sci Rep ; 11(1): 6983, 2021 03 26.
Article in English | MEDLINE | ID: mdl-33772091

ABSTRACT

We investigated the dynamics of the bacterial composition and metabolic function within Akashiwo sanguinea bloom using a 100-L indoor microcosm and metagenomic next-generation sequencing. We found that the bacterial community was classified into three groups at 54% similarity. Group I was associated with "during the A. sanguinea bloom stage" and mainly consisted of Alphaproteobacteria, Flavobacteriia and Gammaproteobacteria. Meanwhile, groups II and III were associated with the "late bloom/decline stage to post-bloom stage" with decreased Flavobacteriia and Gammaproteobacteria in these stages. Upon the termination of the A. sanguinea bloom, the concentrations of inorganic nutrients (particularly PO43-, NH4+ and dissolved organic carbon) increased rapidly and then decreased. From the network analysis, we found that the A. sanguinea node is associated with certain bacteria. After the bloom, the specific increases in NH4+ and PO43- nodes are associated with other bacterial taxa. The changes in the functional groups of the bacterial community from chemoheterotrophy to nitrogen association metabolisms were consistent with the environmental impacts during and after A. sanguinea bloom. Consequently, certain bacterial communities and the environments dynamically changed during and after harmful algal blooms and a rapid turnover within the bacterial community and their function can respond to ecological interactions.


Subject(s)
Alphaproteobacteria/isolation & purification , Dinoflagellida/growth & development , Flavobacteriaceae/isolation & purification , Gammaproteobacteria/isolation & purification , Harmful Algal Bloom , Metagenome , Seawater/microbiology , Alphaproteobacteria/genetics , Alphaproteobacteria/growth & development , Carbon/analysis , Dinoflagellida/microbiology , Flavobacteriaceae/genetics , Flavobacteriaceae/growth & development , Gammaproteobacteria/genetics , Gammaproteobacteria/growth & development , High-Throughput Nucleotide Sequencing , Nitrogen/analysis , Phosphorus/analysis
4.
Mol Biochem Parasitol ; 237: 111279, 2020 05.
Article in English | MEDLINE | ID: mdl-32360511

ABSTRACT

Studies of Plasmodium sporozoites and liver stages require dissection of Anopheles mosquitoes to obtain sporozoites for experiments. Sporozoites from the rodent parasite P. yoelii are routinely used to infect hepatocytes for liver stage culture, but sometimes these cultures become contaminated. Using standard microbiological techniques, a single colony type of Gram-negative rod-shaped bacteria was isolated from contaminated cultures. Mass spectrometry and sequencing of the bacterial 16S ribosomal RNA gene identified the contaminant as Elizabethkingia spp. Based on sequence comparison and published studies of the Anopheles microbiome, the best match was E. anophelis. Culture contamination was not ameliorated by density gradient purification of sporozoites. However, the addition of vancomycin to the culture media consistently reduced contamination and improved culture outcomes as measured by liver stage parasite size. Thus, mosquito salivary gland-derived E. anophelis is identified a potential contaminant of Plasmodium liver stage cultures that can be mitigated by the addition of antibiotics.


Subject(s)
Anti-Bacterial Agents/pharmacology , Flavobacteriaceae/drug effects , Hepatocytes/drug effects , Plasmodium yoelii/growth & development , Sporozoites/growth & development , Vancomycin/pharmacology , Animals , Anopheles/microbiology , Anopheles/parasitology , Bacterial Typing Techniques , Cells, Cultured , Flavobacteriaceae/genetics , Flavobacteriaceae/growth & development , Flavobacteriaceae/pathogenicity , Hepatocytes/microbiology , Hepatocytes/parasitology , Malaria/parasitology , Microbial Sensitivity Tests , Microbiota/drug effects , Microbiota/genetics , Plasmodium yoelii/ultrastructure , RNA, Ribosomal, 16S/genetics , Rodentia/parasitology , Sporozoites/ultrastructure
5.
Chembiochem ; 21(9): 1279-1284, 2020 05 04.
Article in English | MEDLINE | ID: mdl-31845464

ABSTRACT

Anopheles mosquito microbiomes are intriguing ecological niches. Within the gut, microbes adapt to oxidative stress due to heme and iron after blood meals. Although metagenomic sequencing has illuminated spatial and temporal fluxes of microbiome populations, limited data exist on microbial growth dynamics. Here, we analyze growth interactions between a dominant microbiome species, Elizabethkingia anophelis, and other Anopheles-associated bacteria. We find E. anophelis inhibits a Pseudomonas sp. via an antimicrobial-independent mechanism and observe biliverdins, heme degradation products, upregulated in cocultures. Purification and characterization of E. anophelis HemS demonstrates heme degradation, and we observe hemS expression is upregulated when cocultured with Pseudomonas sp. This study reveals a competitive microbial interaction between mosquito-associated bacteria and characterizes the stimulation of heme degradation in E. anophelis when grown with Pseudomonas sp.


Subject(s)
Anopheles/microbiology , Bacterial Proteins/metabolism , Flavobacteriaceae/metabolism , Heme/metabolism , Microbiota , Virulence , Animals , Coculture Techniques , Flavobacteriaceae/growth & development , Genome, Bacterial , Phylogeny , Sequence Analysis, DNA
6.
Microbiologyopen ; 8(8): e00818, 2019 08.
Article in English | MEDLINE | ID: mdl-30809963

ABSTRACT

Algicidal bacteria are important players regulating the dynamic changes of plankton assemblages. Most studies on these bacteria have focused on the effect on single algal species in simple incubation experiments. Considering the complexity of species assemblages in the natural plankton, such incubations represent an oversimplification and do not allow making further reaching conclusions on ecological interactions. Here, we describe a series of co-incubation experiments with different level of complexity to elucidate the effect of the algicidal bacterium Kordia algicida on mixed cultures of a resistant and a susceptible diatom. The growth of the resistant diatom Chaetoceros didymus is nearly unaffected by K. algicida in monoculture, while cells of the susceptible diatom Skeletonema costatum are lysed within few hours. Growth of C. didymus is inhibited if mixed cultures of the two diatoms are infected with the bacterium. Incubations with filtrates of the infected cultures show that the effects are chemically mediated. In non-contact co-culturing we show that low concentrations of the lysed algae support the growth of C. didymus, while higher concentrations trigger population decline. Complex cascading effects of algicidal bacteria have thus to be taken into account if their ecological role is concerned.


Subject(s)
Antibiosis , Diatoms/growth & development , Flavobacteriaceae/growth & development , Plankton
7.
Appl Environ Microbiol ; 85(7)2019 04 01.
Article in English | MEDLINE | ID: mdl-30737345

ABSTRACT

Plankton communities consist of complex microbial consortia that change over time. These fluctuations can be only partially explained by limiting resources. Biotic factors such as herbivores and pathogens also contribute to the control of algal blooms. Here we address the effects of algicidal bacteria on a natural plankton community in an indoor enclosure experiment. The algicidal bacteria, introduced into plankton taken directly from the North Sea during a diatom bloom, caused the rapid decline of the bloom-forming Chaetoceros socialis within only 1 day. The haptophyte Phaeocystis, in contrast, is resistant to the lytic bacteria and could benefit from the removal of the competitor, as indicated by an onset of a bloom in the treated enclosures. This cascading effect caused by the bacterial pathogen accelerated the succession of Phaeocystis, which bloomed with a delay of only several weeks in the in situ waters at Helgoland Roads in the North Sea. The algicidal bacteria can thus modulate the community within the limits of the abiotic and biotic conditions of the local environment. Implications of our findings for plankton ecosystem functioning are discussed.IMPORTANCE Plankton communities change on a seasonal basis in temperate systems, with distinct succession patterns; this is mainly due to algal species that have their optimal timing relative to environmental conditions. We know that bacterial populations are also instrumental in the decay and termination of phytoplankton blooms. Here, we describe algicidal bacteria as modulators of this important species succession. Upon treatment of a natural plankton consortium with an algicidal bacterium, we observed a strong shift in the phytoplankton community structure, compared to controls, resulting in formation of a succeeding Phaeocystis bloom. Blooms of this alga have a substantial impact on global biogeochemical and ecological cycles, as they are responsible for a substantial proportion of primary production during spring in the North Sea. We propose that one of the key factors influencing such community shifts may be algicidal bacteria.


Subject(s)
Antibiosis , Flavobacteriaceae/growth & development , Flavobacteriaceae/physiology , Plankton/growth & development , Seawater/microbiology , Diatoms/drug effects , Diatoms/growth & development , Ecosystem , Eutrophication/drug effects , Marine Biology , North Sea , Pest Control, Biological , Phytoplankton/drug effects , Population Dynamics , Seasons
8.
Environ Microbiol ; 21(1): 374-388, 2019 01.
Article in English | MEDLINE | ID: mdl-30411473

ABSTRACT

Fungi living in sediments ('mycobenthos') are hypothesized to play a role in the degradation of organic matter deposited at the land-sea interface, but the environmental factors influencing the mycobenthos are poorly understood. We used mock community calibrated Illumina sequencing to show that the mycobenthos community structure in a coastal lagoon was significantly changed after exposure to a lignocellulose extract and subsequent development of benthic anoxia over a relatively short (10 h) incubation. Saprotrophic taxa dominated and were selected for under benthic anoxia, specifically Aquamyces (Chytridiomycota) and Orbilia (Ascomycota), implicating these genera as important benthic saprotrophs. Protein encoding genes involved in energy and biomass production from Fungi and the fungal-analogue group Labyrinthulomycetes had the highest increase in expression with the added organic matter compared with all other groups, indicating that lignocellulose stimulates metabolic activity in the mycobenthos. Flavobacteria dominated the active bacterial community that grew rapidly with the lignocellulose extract and crashed sharply upon O2 depletion. Our findings indicate that the diversity, activity and trophic potential of the mycobenthos changes rapidly in response to organic matter and decreasing O2 concentrations, which together with heterotrophic Flavobacteria, undergo 'boom and bust' dynamics during lignocellulose degradation in estuarine ecosystems.


Subject(s)
Ascomycota/growth & development , Chytridiomycota/growth & development , Humic Substances/microbiology , Lignin/metabolism , Mycobiome/physiology , Stramenopiles/growth & development , Anaerobiosis , Ascomycota/isolation & purification , Biomass , Chytridiomycota/isolation & purification , Ecosystem , Flavobacteriaceae/growth & development , Flavobacteriaceae/metabolism , Heterotrophic Processes , Oxygen/metabolism , Stramenopiles/metabolism
9.
FEMS Microbiol Ecol ; 94(12)2018 12 01.
Article in English | MEDLINE | ID: mdl-30239661

ABSTRACT

Recent studies have highlighted the potential role of microbiota in the biology of the Aedes albopictus mosquito vector. This species is highly anthropogenic and exhibits marked ecological plasticity, with a resulting high potential to colonize a wide range of habitats-including anthropized areas-under various climatic conditions. We put forward the hypothesis that climate and anthropogenic activities, such as the use of antibiotics in agriculture and human medicine, might affect the mosquito-associated bacterial community. We thus studied the additive impact of a temperature decrease and antibiotic ingestion on the temporal dynamics of Ae. albopictus survival and its associated bacterial communities. The results showed no effects of disturbances on mosquito survival. However, short-term temperature impacts on bacterial diversity were observed, while both the community structure and bacterial diversity were affected by early antibiotic ingestion. The genera Elizabethkingia, Chryseobacterium and Wolbachia, as well as an unclassified member of the Bacteroidales order were particularly affected. Antibiotics negatively impacted Elizabethkingia abundance, while Chryseobacterium was completely eliminated following both disturbances, to the benefit of Wolbachia and the unclassified Bacteroidales species. These results generated fresh insight into the effects of climate and anthropogenic activities such as the use of antibiotics on mosquito microbiota.


Subject(s)
Aedes/microbiology , Anti-Bacterial Agents/pharmacology , Dysbiosis/chemically induced , Microbiota/drug effects , Animals , Bacteroidetes/growth & development , Chryseobacterium/growth & development , Climate , Flavobacteriaceae/growth & development , Humans , Mosquito Vectors/microbiology , Temperature , Wolbachia/growth & development
10.
FEMS Microbiol Ecol ; 94(9)2018 09 01.
Article in English | MEDLINE | ID: mdl-30011002

ABSTRACT

To understand prokaryotic responses during a spring bloom in offshore shelf waters, prokaryotic parameters were measured daily at a station located in the middle of the East China Sea over a six-week period from March 25 to May 19. The site experienced a phytoplankton bloom in late April, triggering changes in prokaryotic abundance and production after a lag of approximately one week. Before the bloom, changes in prokaryotic composition were small. Both during the bloom and in the post-bloom period, successive changes among bacterial groups were apparent. A SAR11 group became more dominant during the bloom period, and diverse groups belonging to the Flavobacteriia occurred dominantly during both the bloom and post-bloom periods. However, bacterial community changes at the species level during the bloom and post-bloom periods occurred rapidly in a time scale of a few days. Especially, NS5, NS4 and Formosa bacteria belonging to Flavobacteriia and bacteria belonging to Halieaceae and Arenicellaceae families of Gammaproteobacteria showed a successive pattern with large short-term variation during the period. The changes in prokaryotic composition were found to be related to phytoplankton biomass and composition, as well as seawater temperature and variations in nutrients.


Subject(s)
Flavobacteriaceae/growth & development , Gammaproteobacteria/growth & development , Phytoplankton/growth & development , Seawater/microbiology , Biomass , China , Flavobacteriaceae/classification , Gammaproteobacteria/classification , Oceans and Seas , Seasons
11.
Curr Microbiol ; 75(9): 1126-1132, 2018 Sep.
Article in English | MEDLINE | ID: mdl-29761217

ABSTRACT

A Gram-stain-negative, aerobic, non-motile, and carrageenan-degrading bacterial strain, designated OISW-17T, was isolated from seawater around Oido, an island of South Korea, and its taxonomic position was determined using a polyphasic study. Optimal growth of the novel strain occurred at 20-25 °C and in the presence of 2.0% (w/v) NaCl. Phylogenetic trees using 16S rRNA gene sequences showed that strain OISW-17T forms a cluster with the type strains of Dokdonia species. The novel strain exhibited 16S rRNA gene sequence similarities of 95.7-96.3% to the type strains of Dokdonia species and of < 93.4% to the type strains of the other recognized species. Menaquinone-6 (MK-6) was found as the predominant menaquinone and iso-C15:0, iso-C15:1 G, and iso-C17:0 3-OH were the major fatty acids. Phosphatidylethanolamine, one unidentified lipid, and one unidentified aminolipid were major polar lipids detected in strain OISW-17T. Strain OISW-17T had DNA G+C content of 39.7 mol%. The differential phenotypic characteristics, along with the phylogenetic data, made strain OISW-17T to be distinct from recognized species of the genus Dokdonia. On the basis of the data given, strain OISW-17T represents a novel species of the genus Dokdonia, for which the name Dokdonia ponticola sp. nov. is proposed. The type strain of the novel species is OISW-17T (= KACC 19437T = KCTC 62187T = CGMCC 1.16527T).


Subject(s)
Carrageenan/metabolism , Flavobacteriaceae/classification , Flavobacteriaceae/metabolism , Phylogeny , Seawater/microbiology , DNA, Bacterial/genetics , Fatty Acids/analysis , Flavobacteriaceae/genetics , Flavobacteriaceae/growth & development , Phospholipids/analysis , RNA, Ribosomal, 16S/genetics , Republic of Korea , Sequence Analysis, DNA , Species Specificity , Vitamin K 2/analysis , Water Microbiology
12.
World J Microbiol Biotechnol ; 33(12): 215, 2017 Nov 27.
Article in English | MEDLINE | ID: mdl-29181599

ABSTRACT

Menaquinone (MK) has important applications in the pharmaceutical and food industries. To increase the production rate (QP) of MK-4, we developed a straightforward biotransformation method for MK-4 synthesis directly from its precursors 1,4-dihydroxy-2-naphthoate (DHNA) and farnesol using whole cells of genetically engineered Elizabethkingia meningoseptica. Results showed that MK-4 can be produced directly from farnesol and DHNA using both free and immobilized FM-D198 cells. MK-4 yield peaked at 29.85 ± 0.36 mg/L in the organic phase and 24.08 ± 0.33 mg/g DCW after 12 h of bioconversion using free cells in a two-phase conversion system. MK-4 yield reached 26.34 ± 1.35 mg/L and 17.44 ± 1.05 mg/g DCW after 8 h using immobilized cells. Although this yield was lower than that using free cells, immobilized cells can be re-used for MK-4 production via repeated-batch culture. After ten batch cultures, efficient MK-4 production was maintained at a yield of more than 20 mg/L. After optimizing the catalysis system, the MK-4 yield reached 26.91 ± 1.27 mg/L using the immobilized cells and had molar conversion rates of 58.56 and 76.90% for DHNA and farnesol, respectively.


Subject(s)
Farnesol/metabolism , Flavobacteriaceae/growth & development , Naphthols/metabolism , Vitamin K 2/metabolism , Batch Cell Culture Techniques , Biocatalysis , Biotransformation , Cell Culture Techniques , Cells, Immobilized/metabolism , Flavobacteriaceae/genetics , Flavobacteriaceae/metabolism , Genetic Engineering
13.
Antonie Van Leeuwenhoek ; 110(11): 1445-1452, 2017 Nov.
Article in English | MEDLINE | ID: mdl-28608316

ABSTRACT

A Gram-stain negative, non-flagellated, facultatively anaerobic, slightly halophilic bacterium, WNB302T, was isolated from a marine solar saltern in Wendeng, China (122°0'38.85″E, 36°57'56.49″N). Cells of strain WNB302T were 0.2-0.7 µm wide and 2.0-10.0 µm long, catalase- positive and oxidase-negative. Colonies were opaque, orange and approximately 1.0-2.0 mm in diameter after culture for 96 h on marine agar 2216. Growth occurs at 15-37 °C (optimum, 33-35 °C), pH 6.0-8.5 (optimum, pH 7.0-7.5), and with 0.5-7% NaCl (optimum, 2-3% NaCl). Phylogenetic analysis based on 16S rRNA gene sequences showed that strain WNB302T belongs to the genus Winogradskyella and is closely related to Winogradskyella exilis KCTC 32356T. Strain WNB302T exhibited 96.2 and 96.0% 16S rRNA gene sequence similarities with W. exilis KCTC 32356T and W. litoriviva KCTC 23972T. Average nucleotide identity (ANI) value based on draft genomes between strain WNB302T and Winogradskyella thalassocola KCTC 12221T showed a relatedness of 81.1% (with 93.7% of 16S rRNA gene sequence similarity). The major respiratory quinone of strain WNB302T was found to be MK-6, and the dominant fatty acids were found to be iso-C15:0 and iso-C15:1 G. The major polar lipids of strain WNB302T were three unidentified lipids (L1, L2 and L3), one unidentified aminolipids (AL1) and phosphatidylethanolamine (PE). The genomic DNA G+C content was 37.0 mol%. On the basis of the data presented, strain WNB302T is considered to represent a novel species of the genus Winogradskyella, for which the name Winogradskyella aurantia sp. nov. is proposed. The type strain is WNB302T (= KCTC 52614T = MCCC 1H00172T).


Subject(s)
Flavobacteriaceae/classification , Phylogeny , Seawater/microbiology , Bacterial Typing Techniques , Base Composition , Catalase/metabolism , China , DNA, Bacterial/genetics , Fatty Acids/analysis , Flavobacteriaceae/chemistry , Flavobacteriaceae/genetics , Flavobacteriaceae/growth & development , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Sequence Homology, Nucleic Acid
14.
Environ Microbiol ; 19(6): 2164-2181, 2017 06.
Article in English | MEDLINE | ID: mdl-28205313

ABSTRACT

Comprehension of the degradation of macroalgal polysaccharides suffers from the lack of genetic tools for model marine bacteria, despite their importance for coastal ecosystem functions. We developed such tools for Zobellia galactanivorans, an algae-associated flavobacterium that digests many polysaccharides, including alginate. These tools were used to investigate the biological role of AlyA1, the only Z. galactanivorans alginate lyase known to be secreted in soluble form and to have a recognizable carbohydrate-binding domain. A deletion mutant, ΔalyA1, grew as well as the wild type on soluble alginate but was deficient in soluble secreted alginate lyase activity and in digestion of and growth on alginate gels and algal tissues. Thus, AlyA1 appears to be essential for optimal attack of alginate in intact cell walls. alyA1 appears to have been recently acquired via horizontal transfer from marine Actinobacteria, conferring an adaptive advantage that might benefit other algae-associated bacteria by exposing new substrate niches. The genetic tools described here function in diverse members of the phylum Bacteroidetes and should facilitate analyses of polysaccharide degradation systems and many other processes in these common but understudied bacteria.


Subject(s)
Alginates/metabolism , Flavobacteriaceae/genetics , Flavobacteriaceae/metabolism , Phaeophyceae/microbiology , Polysaccharide-Lyases/genetics , Biomass , Cell Wall/metabolism , Flavobacteriaceae/enzymology , Flavobacteriaceae/growth & development , Glucuronic Acid/metabolism , Hexuronic Acids/metabolism , Polysaccharide-Lyases/metabolism , Sequence Deletion/genetics
16.
Arch Microbiol ; 197(9): 1095-9, 2015 Nov.
Article in English | MEDLINE | ID: mdl-26362038

ABSTRACT

A bacterial strain, JH03(T), was isolated from gravel adjacent to Geommeolle beach on Udo Island, South Korea. The cells were Gram-stain-negative, aerobic, non-motile and rod shaped. The ranges of temperature, pH and NaCl concentration for growth of the bacterium were 10-45 °C, pH 6.0-9.5 and 0.5-5.0 % (w/v), respectively. The major fatty acids of the bacterium were iso-C(15:0) (15.4 %), iso-C(15:1) G (14.1 %), iso-C(16:0) 3-OH (14.1 %), iso-C(17:0) 3-OH (11.5 %) and anteiso-C(15:0) (11.3 %). The major isoprenoid quinone was MK-6. The polar lipids included phosphatidylethanolamine, two unidentified amino lipids and three unidentified lipids. The DNA G+C content was 34.2 mol%. The phylogenetic analysis of the 16S rRNA gene sequences showed that strain JH03(T) was most closely related to Jejuia pallidilutea EM39(T) (96.5 % sequence similarity). Based on the polyphasic analysis, strain JH03(T) is a novel species of the genus Jejuia, for which the name Jejuia marina sp. nov. is proposed. The type strain is JH03(T) (= KCTC 42342(T) = JCM 30601(T)).


Subject(s)
Flavobacteriaceae/classification , Geologic Sediments/microbiology , Phylogeny , Bacterial Typing Techniques , Base Composition , Fatty Acids/analysis , Flavobacteriaceae/chemistry , Flavobacteriaceae/genetics , Flavobacteriaceae/growth & development , Hydrogen-Ion Concentration , RNA, Ribosomal, 16S/genetics , Republic of Korea , Salinity , Temperature
17.
Environ Microbiol ; 17(10): 3766-80, 2015 Oct.
Article in English | MEDLINE | ID: mdl-25684402

ABSTRACT

Rare microbial taxa are increasingly recognized to play key ecological roles, but knowledge of their spatio-temporal dynamics is lacking. In a time-series study in coastal waters, we detected 83 bacterial lineages with significant seasonality, including environmentally relevant taxa where little ecological information was available. For example, Verrucomicrobia had recurrent maxima in summer, while the Flavobacteria NS4, NS5 and NS2b clades had contrasting seasonal niches. Among the seasonal taxa, only 4 were abundant and persistent, 20 cycled between rare and abundant and, remarkably, most of them (59) were always rare (contributing < 1% of total reads). We thus demonstrate that seasonal patterns in marine bacterioplankton are largely driven by lineages that never sustain abundant populations. A fewer number of rare taxa (20) also produced episodic 'blooms', and these events were highly synchronized, mostly occurring on a single month. The recurrent seasonal growth and loss of rare bacteria opens new perspectives on the temporal dynamics of the rare biosphere, hitherto mainly characterized by dormancy and episodes of 'boom and bust', as envisioned by the seed-bank hypothesis. The predictable patterns of seasonal reoccurrence are relevant for understanding the ecology of rare bacteria, which may include key players for the functioning of marine ecosystems.


Subject(s)
Bacteria/growth & development , Plankton/growth & development , Seasons , Bacteria/classification , Bacteria/genetics , Ecology , Ecosystem , Environment , Flavobacteriaceae/growth & development , Oceans and Seas , Plankton/genetics , RNA, Ribosomal, 16S , Verrucomicrobia/growth & development
18.
Appl Environ Microbiol ; 81(6): 2233-43, 2015 Mar.
Article in English | MEDLINE | ID: mdl-25595771

ABSTRACT

Flavobacteria (members of the family Flavobacteriaceae) dominate the bacterial community in the Anopheles mosquito midgut. One such commensal, Elizabethkingia anophelis, is closely associated with Anopheles mosquitoes through transstadial persistence (i.e., from one life stage to the next); these and other properties favor its development for paratransgenic applications in control of malaria parasite transmission. However, the physiological requirements of E. anophelis have not been investigated, nor has its capacity to perpetuate despite digestion pressure in the gut been quantified. To this end, we first developed techniques for genetic manipulation of E. anophelis, including selectable markers, reporter systems (green fluorescent protein [GFP] and NanoLuc), and transposons that function in E. anophelis. A flavobacterial expression system based on the promoter PompA was integrated into the E. anophelis chromosome and showed strong promoter activity to drive GFP and NanoLuc reporter production. Introduced, GFP-tagged E. anophelis associated with mosquitoes at successive developmental stages and propagated in Anopheles gambiae and Anopheles stephensi but not in Aedes triseriatus mosquitoes. Feeding NanoLuc-tagged cells to A. gambiae and A. stephensi in the larval stage led to infection rates of 71% and 82%, respectively. In contrast, a very low infection rate (3%) was detected in Aedes triseriatus mosquitoes under the same conditions. Of the initial E. anophelis cells provided to larvae, 23%, 71%, and 85% were digested in A. stephensi, A. gambiae, and Aedes triseriatus, respectively, demonstrating that E. anophelis adapted to various mosquito midgut environments differently. Bacterial cell growth increased up to 3-fold when arginine was supplemented in the defined medium. Furthermore, the number of NanoLuc-tagged cells in A. stephensi significantly increased when arginine was added to a sugar diet, showing it to be an important amino acid for E. anophelis. Animal erythrocytes promoted E. anophelis growth in vivo and in vitro, indicating that this bacterium could obtain nutrients by participating in erythrocyte lysis in the mosquito midgut.


Subject(s)
Anopheles/microbiology , Flavobacteriaceae/growth & development , Flavobacteriaceae/genetics , Host-Pathogen Interactions , Aedes/microbiology , Animals , Gastrointestinal Tract/microbiology , Genes, Reporter , Genetics, Microbial/methods , Larva/microbiology , Luminescent Proteins/analysis , Luminescent Proteins/genetics , Molecular Biology/methods , Molecular Sequence Data , Sequence Analysis, DNA
19.
Environ Microbiol ; 17(10): 3597-609, 2015 Oct.
Article in English | MEDLINE | ID: mdl-24911121

ABSTRACT

Deep-sea coral reefs do not receive sunlight and depend on plankton. Little is known about the plankton composition at such reefs, even though they constitute habitats for many invertebrates and fish. We investigated plankton communities from three reefs at 260-350 m depth at hydrocarbon fields off the mid-Norwegian coast using a combination of cultivation and small subunit (SSU) rRNA gene and transcript sequencing. Eight months incubations of a reef water sample with minimal medium, supplemented with carbon dioxide and gaseous alkanes at in situ-like conditions, enabled isolation of mostly Alphaproteobacteria (Sulfitobacter, Loktanella), Gammaproteobacteria (Colwellia) and Flavobacteria (Polaribacter). The relative abundance of isolates in the original sample ranged from ∼ 0.01% to 0.80%. Comparisons of bacterial SSU sequences from filtered plankton of reef and non-reef control samples indicated high abundance and metabolic activity of primarily Alphaproteobacteria (SAR11 Ia), Gammaproteobacteria (ARCTIC96BD-19), but also of Deltaproteobacteria (Nitrospina, SAR324). Eukaryote SSU sequences indicated metabolically active microalgae and animals, including codfish, at the reef sites. The plankton community composition varied between reefs and differed between DNA and RNA assessments. Over 5000 operational taxonomic units were detected, some indicators of reef sites (e.g. Flavobacteria, Cercozoa, Demospongiae) and some more active at reef sites (e.g. Gammaproteobacteria, Ciliophora, Copepoda).


Subject(s)
Alphaproteobacteria/isolation & purification , Anthozoa/microbiology , Deltaproteobacteria/isolation & purification , Gammaproteobacteria/growth & development , Microbial Consortia/physiology , Plankton/growth & development , Alphaproteobacteria/genetics , Alphaproteobacteria/growth & development , Animals , Base Sequence , Coral Reefs , Deltaproteobacteria/genetics , Deltaproteobacteria/growth & development , Ecosystem , Flavobacteriaceae/genetics , Flavobacteriaceae/growth & development , Flavobacteriaceae/isolation & purification , Gammaproteobacteria/genetics , Gammaproteobacteria/isolation & purification , Molecular Sequence Data , Norway , Plankton/genetics , Seawater/microbiology
20.
Environ Microbiol ; 17(3): 781-95, 2015 Mar.
Article in English | MEDLINE | ID: mdl-24903166

ABSTRACT

We studied the seasonal growth potential of opportunistic bacterial populations in Lake Zurich (Switzerland) by a series of grazer-free dilution culture assays. Pronounced shifts in the composition of the bacterial assemblages were observed within one doubling of total cell numbers, from initially abundant Actinobacteria to other fast-growing microbial lineages. Small populations with growth potentials far above community average were detected throughout the year with striking seasonal differences in their respective taxonomic affiliations. Members of Cytophaga-Flavobacteria (CF) were disproportionally proliferating only during phytoplankton blooms in spring and summer, while Beta- and Gammaproteobacteria showed superior growth at all other occasions. Growth rates of Alphaproteobacteria and esp. Sphingomonadaceae were significantly correlated to water temperatures and were far above community average in summer. Within the genus Flavobacterium, two species-like populations showed a tendency for fast growth in most experiments, while four others were exclusively proliferating either during a spring or during a summer phytoplankton bloom. Their high growth potentials but low in situ abundances hint at a tight control by bacterivorous grazers and at a consequently accelerated carbon flux to higher trophic levels.


Subject(s)
Bacteria/classification , Bacteria/growth & development , Carbon Cycle , Lakes/microbiology , Phytoplankton/growth & development , Actinobacteria/growth & development , Alphaproteobacteria/growth & development , Betaproteobacteria/growth & development , Carbon/metabolism , Cytophaga/growth & development , Ecosystem , Flavobacteriaceae/growth & development , Seasons , Switzerland , Temperature
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