Effects of exogenous addition of outer membrane vesicles on biofilm formation of Pseudoalteromonas marina and settlement of mussel (Mytilus coruscus)
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摘要: 为探究胞外囊泡（outer membrane vesicles，OMVs）对海假交替单胞菌（Pseudoalteromonas marina）生物被膜形成及厚壳贻贝（Mytilus coruscus）（壳长为0.88 mm±0.16 mm，壳高为0.55 mm±0.12 mm）稚贝附着的影响，从海假交替单胞菌中提取并纯化了OMVs，并添加到海假交替单胞菌中共同孵育形成生物被膜，在OMVs终质量浓度为0.1、1、10、20 μg/mL时检测形成的生物被膜对贻贝稚贝附着的诱导作用。结果表明：在海假交替单胞菌中成功提取到了OMVs，从菌体中提取获得的OMVs较上清液中更多；添加10 μg/mL的OMVs时，形成的生物被膜对贻贝稚贝的附着诱导活性最高，稚贝附着率达到56.42%；随着OMVs添加量的增加，细菌密度和膜厚呈增长趋势，胞外产物中α-多糖、β-多糖、脂质和蛋白质含量呈升高趋势；相关性分析显示，生物被膜中的脂质含量与贻贝附着显著相关（P<0.05）。研究表明，OMVs的添加可促进海假交替单胞菌生物被膜形成，提高生物被膜胞外产物的含量，并推测主要通过增加脂质含量提高生物被膜对贻贝附着的诱导能力。Abstract: To investigate the effects of outer membrane vesicles (OMVs) on the biofilm formation of Pseudoalteromonas marina and the settlement of mussel (Mytilus coruscus) plantigrades with shell length (0.88 mm±0.16 mm) and shell height of (0.55 mm±0.12 mm), OMVs were extracted and purified from P.marina and added into P.marina bacterial fluid to form biofilms, and the inducing capacity of biofilms on the settlement of mussel juveniles was detected at the final OMVs concentration of 0.1, 1, 10 and 20 μg/mL. The results showed that OMVs were successfully extracted from P.marina, with the more content of OMVs extracted from the bacterium than that in the supernatant. The biofilms had the maximal inducing activity to mussel plantigrades, with the settlement rate of 56.42% at the OMVs concentration of 10 μg/mL. With the increase in OMVs concentration, the bacterial density and biofilm thickness of biofilms were found to be increased, with increase in α-polysaccharides, β-polysaccharides, lipids and proteins in extracellular products. Correlation analysis showed that the lipid content in the biofilms was significantly correlated with mussel settlement (P<0.05), indicating that addition of OMVs promoted the formation of P.marina biofilms and increases the content of extracellular products of biofilms. It is speculated that the improvement in inducing ability of biofilmd to mussel settlement after adding OMVs is primarily due to the increase in lipid content.
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Key words:
- outer membrane vesicle /
- biofilm /
- Mytilus coruscus /
- plantigrade /
- settlement
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YOU Z J, XU S L, XIE Q L.Shellfish resources and its cultivation and proliferation along Zhejiang coast[J].Donghai Marine Science, 2000, 18(1):50-56.(in Chinese)
ZHOU X, GUO X P, CHEN Y R, et al.Effects of bacterial biofilms formed on low surface wettability on settlement of plantigrades of the mussel Mytilus coruscus[J].Journal of Dalian Fisheries University, 2015, 30(1):30-35.(in Chinese)
YANG J L, SHEN P J, LIANG X, et al.Larval settlement and metamorphosis of the mussel Mytilus coruscus in response to monospecific bacterial biofilms[J].Biofouling, 2013, 29(3):247-259.
CARL C, POOLE A J, VUCKO M J, et al.Optimising settlement assays of pediveligers and plantigrades of Mytilus galloprovincialis[J].Biofouling, 2011, 27(8):859-868.
COSTERTON J W, CHENG K J, GEESEY G G, et al.Bacterial biofilms in nature and disease[J].Annual Review of Microbiology, 1987, 41:435-464.
HABASH M, REID G.Microbial biofilms:their development and significance for medical device-related infections[J].The Journal of Clinical Pharmacology, 1999, 39(9):887-898.
XIAO M Y, WU R Y, TAN C M, et al.Recent advances in understanding the role of quorum sensing system and quorum sensing inhibitors in regulating bacterial biofilm formation[J].Food Science, 2020, 41(13):227-234.(in Chinese)
TU C T, WANG Y, YI L, et al.Roles of signaling molecules in biofilm formation[J].Chinese Journal of Biotechnology, 2019, 35(4):558-566.(in Chinese)
PASSOS DA SILVA D, MATWICHUK M L, TOWNSEND D O, et al.The Pseudomonas aeruginosa lectin LecB binds to the exopolysaccharide Psl and stabilizes the biofilm matrix[J].Nature Communications, 2019, 10:2183.
QI L J, CHRISTOPHER G F.Role of flagella, type IV pili, biosurfactants, and extracellular polymeric substance polysaccharides on the formation of pellicles by Pseudomonas aeruginosa[J].Langmuir, 2019, 35(15):5294-5304.
HASHIMOTO K, ELY VASQUEZ H, KITAMURA H, et al.Larval settlement of the Pacific oyster Crassostrea gigas in response to marine microbial films and monospecies bacterial films[J].Sessile Organisms, 2017, 34(2):21-27.
FRECKELTON M L, NEDVED B T, HADFIELD M G.Induction of invertebrate larval settlement, different bacteria, different mechanisms?[J].Scientific Reports, 2017, 7:42557.
YANG J L, SHEN P J, WANG C, et al.Effects of biofilms on settlement of plantigrades of the mussel Mytilus coruscus[J].Journal of Fisheries of China, 2013, 37(6):904-909.(in Chinese)
CHEN H E, HE K, HE C H, et al.Effects of alginate on biofilm formation of Pseudoalteromonas marina and larval settlement and metamorphosis of the mussel Mytilus coruscus[J].Journal of Dalian Ocean University, 2022, 37(4):620-626.(in Chinese)
WU J X, LI J Z, HU X M, et al.Effects of lipopolysaccharide on bacterial biofilm formation and metamorphosis of mussel larvae[J].Journal of Fisheries of China, 2022, 46(11):2134-2142.(in Chinese)
LIANG X, TONG H, PENG L H, et al.Regulation of formation of biofilms and larval settlement and metamorphosis of mussel Mytilus coruscus by cellulose[J].Journal of Dalian Ocean University, 2020, 35(1):75-82.(in Chinese)
LIANG X, DING W Y, ZHANG C, et al.Effects of three fatty acids on biofilm formation and settlement and metamorphosis of hard shelled mussel Mytilus coruscus[J].Journal of Dalian Ocean University, 2021, 36(4):580-586.(in Chinese)
CAI Y S, ZHANG X K, ZHU Y T, et al.Effects of Pseudoalteromonas marina flagellin on biofilm formation and settlement of Mytilus coruscus[J].Haiyang Xuebao, 2021, 43(4):75-83.(in Chinese)
BONNINGTON K, KUEHN M.Breaking the bilayer:OMV formation during environmental transitions[J].Microbial Cell, 2017, 4(2):64-66.
TURNBULL L, TOYOFUKU M, HYNEN A L, et al.Explosive cell lysis as a mechanism for the biogenesis of bacterial membrane vesicles and biofilms[J].Nature Communications, 2016, 7:11220.
RENELLI M, MATIAS V, LO R Y, et al.DNA-containing membrane vesicles of Pseudomonas aeruginosa PAO1 and their genetic transformation potential[J].Microbiology, 2004, 150(Pt 7):2161-2169.
IM H, LEE S J, SOPER S A, et al.Staphylococcus aureus extracellular vesicles (EVs):surface-binding antagonists of biofilm formation[J].Molecular BioSystems, 2017, 13(12):2704-2714.
YONEZAWA H, OSAKI T, KURATA S, et al.Outer membrane vesicles of Helicobacter pylori TK1402 are involved in biofilm formation[J].BMC Microbiology, 2009, 9(1):1-12.
FREDRIKSEN J H, ROSENQVIST E, WEDEGE E, et al.Production, characterization and control of MenB-vaccine "Folkehelsa":an outer membrane vesicle vaccine against group B meningococcal disease[J].NIPH Ann, 1991, 14(2):67-79.
GONZÁLEZ-MACHADO C, CAPITA R, RIESCO-PELÁEZ F, et al.Visualization and quantification of the cellular and extracellular components of Salmonella agona biofilms at different stages of development[J].PLoS One, 2018, 13(7):e0200011.
REMIS J P, WEI D G, GORUR A, et al.Bacterial social networks:structure and composition of Myxococcus xanthus outer membrane vesicle chains[J].Environmental Microbiology, 2014, 16(2):598-610.
KIM Y S, CHOI E J, LEE W H, et al.Extracellular vesicles, especially derived from Gram-negative bacteria, in indoor dust induce neutrophilic pulmonary inflammation associated with both Th1 and Th17 cell responses[J]. Clin Exp Allergy, 2013, 43(4):443-454.
SCHWECHHEIMER C, KUEHN M J.Outer-membrane vesicles from gram-negative bacteria:biogenesis and functions[J].Nature Reviews Microbiology, 2015, 13(10):605-619.
FRECKELTON M L, NEDVED B T, CAI Y S, et al.Bacterial lipopolysaccharide induces settlement and metamorphosis in a marine larva[J].Proceedings of the National Academy of Sciences of the United States of America, 2022, 119(18):e2200795119.
WANG J S, ZHANG W, WANG X Y, et al.Mussel settlement mediated by bacterial VgrG proteins via extracellular outer membrane vesicles[J].International Biodeterioration & Biodegradation, 2023, 180:105595.
LI C F, ZHU L F, WANG D D, et al.T6SS secretes an LPS-binding effector to recruit OMVs for exploitative competition and horizontal gene transfer[J].The ISME Journal, 2022, 16(2):500-510.
BAUMGARTEN T, SPERLING S, SEIFERT J, et al.Membrane vesicle formation as a multiple-stress response mechanism enhances Pseudomonas putida DOT-T1E cell surface hydrophobicity and biofilm formation[J].Applied and Environmental Microbiology, 2012, 78(17):6217-6224.
GUERRERO-MANDUJANO A, HERNÁNDEZ-CORTEZ C, IBARRA J A, et al. The outer membrane vesicles:secretion system type zero[J].Traffic, 2017, 18(7):425-432.
KUMAR P, LEE J H, BEYENAL H, et al.Fatty acids as antibiofilm and antivirulence agents[J].Trends in Microbiology, 2020, 28(9):753-768.
HU X M, ZHANG J B, DING W Y, et al.Reduction of mussel metamorphosis by inactivation of the bacterial thioesterase gene via alteration of the fatty acid composition[J].Biofouling, 2021, 37(8):911-921.
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