Salmonella enterica is one of the most important zoonotic pathogens causing foodborne gastroenteritis worldwide. Outer membrane vesicles (OMVs) are lipid-bilayer vesicles produced by Gram-negative bacteria, which contain biologically active components. We hypothesized that OMVs are an important weapon of S. enterica to initiate enteric diseases pathologies. In this study, the effects of S. enterica OMVs (SeOMVs) on intestinal microbiota and intestinal barrier function were investigated. In vitro fecal culture experiments showed that alpha diversity indexes and microbiota composition were altered by SeOMV supplementation. SeOMV supplementation showed an increase of pH, a decrease of OD630 and total short chain fatty acid (SCFA) concentrations. In vitro IPEC-J2 cells culture experiments showed that SeOMV supplementation did not affect the IPEC-J2 cell viability and the indicated genes expression. In vivo experiments in mice showed that SeOMVs had adverse effects on average daily gain (p < 0.05) and feed:gain ratio (p < 0.05), and had a tendency to decrease the final body weight (p = 0.073) in mice. SeOMV administration decreased serum interleukin-10 level (p < 0.05), decreased the relative abundance of bacteria belonging to the genera BacC-u-018 and Akkermansia (p < 0.05). Furthermore, SeOMV administration damaged the ileum mucosa (p < 0.05). These findings suggest that SeOMVs play an important role in the activation of intestinal inflammatory response induced by S. enterica, and downregulation of SCFA-producing bacteria is a possible mechanism.
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References
1.
AdambergK, AdambergS. Selection of fast and slow growing bacteria from fecal microbiota using continuous culture with changing dilution rate. Microb Ecol Health Dis, 2018; 29(1):1549922; doi: 10.1080/16512235.2018.1549922
2.
AdambergK, TomsonK, TalveT, et al.Levan enhances associated growth of Bacteroides, Escherichia, Streptococcus and Faecalibacterium in fecal microbiota. PLoS one, 2015; 10(12):e0144042; doi: 10.1371/journal.pone.0144042
3.
AlanizRC, DeatherageBL, LaraJC, et al.Membrane vesicles are immunogenic facsimiles of Salmonella Typhimurium that potently activate dendritic cells, prime B and T cell responses, and stimulate protective immunity in vivo. J Immunol, 2007; 179(11):7692–7701; doi: 10.4049/jimmunol.179.11.7692
4.
AlazzouniAS, FathallaAS, GabriMS, et al.Role of bone marrow derived-mesenchymal stem cells against gastric ulceration: Histological, immunohistochemical and ultrastructural study. Saudi J Biol Sci, 2020; 27(12):3456–3464; doi: 10.1016/j.sjbs.2020.09.044
5.
AndaloussiSEL, MägerI, BreakefieldXO, et al.Extracellular vesicles: Biology and emerging therapeutic opportunities. Nat Rev Drug Discov, 2013; 12(5):347–357; doi: 10.1038/nrd3978
6.
AshayerizadehA, DastarB, Shams SharghM, et al.Fermented rapeseed meal is effective in controlling Salmonella enterica serovar Typhimurium infection and improving growth performance in broiler chicks. Vet Microbiol, 2017; 201:93–102; doi: 10.1016/j.vetmic.2017.01.007
7.
BedkeT, MuscateF, SoukouS, et al.Title: IL-10-producing T cells and their dual functions. Semin Immunol, 2019; 44:101335; doi: 10.1016/j.smim.2019.101335
8.
BerndtA, WilhelmA, JugertC, et al.Chicken cecum immune response to Salmonella enterica serovars of different levels of invasiveness. Infect Immun, 2007; 75(12):5993–6007; doi: 10.1128/iai.00695-07
9.
BonningtonKE, KuehnMJ. Breaking the bilayer: OMV formation during environmental transitions. Microb Cell, 2017; 4(2):64–66; doi: 10.1569/8/mic2017.02.558
10.
BoxioR, WartelleJ, Nawrocki-RabyB, et al.Neutrophil elastase cleaves epithelial cadherin in acutely injured lung epithelium. Respir Res, 2016; 17(1):129; doi: 10.1186/s12931-016-0449-x
11.
BrownL, WolfJM, Prados-RosalesR, et al.Through the wall: Extracellular vesicles in Gram-positive bacteria, mycobacteria and fungi. Nat Rev Microbiol, 2015; 13(10):620–630; doi: 10.1038/nrmicro3480
12.
Cameron-VeasK, FraileL, NappS, et al.Multidrug resistant Salmonella enterica isolated from conventional pig farms using antimicrobial agents in preventative medicine programmes. Vet J, 2018; 234:36–42; doi: 10.1016/j.tvjl.2018.02.002
13.
ChangX, WangSL, ZhaoSB, et al.Extracellular vesicles with possible roles in gut intestinal tract homeostasis and IBD. Mediators Inflamm, 2020; 2020:1945832; doi: 10.1155/2020/1945832
14.
ChenY, JieK, LiB, et al.Immunization with outer membrane vesicles derived from major outer membrane protein-deficient Salmonella Typhimurium mutants for cross protection against Salmonella Enteritidis and avian pathogenic Escherichia coli O78 infection in chickens. Front Microbiol, 2020; 11:588952; doi: 10.3389/fmicb.2020.588952
15.
CrumpJA, Sjölund-KarlssonM, GordonMA, et al.Epidemiology, clinical presentation, laboratory diagnosis, antimicrobial resistance, and antimicrobial management of invasive salmonella infections. Clin Microbiol Rev, 2015; 28(4):901–937; doi: 10.1128/cmr.00002-15
16.
CuiH, SunY, LinH, et al.The outer membrane vesicles of Salmonella enterica serovar Typhimurium activate chicken immune cells through Lipopolysaccharides and membrane proteins. Pathogens, 2022; 11(3):339; doi: 10.3390/pathogens11030339
17.
CuypersWL, JacobsJ, WongV, et al.Fluoroquinolone resistance in Salmonella: Insights by whole-genome sequencing. Microb Genom, 2018; 4(7):e000195; doi: 10.1099/mgen.0.000195
18.
DeatherageBL, CooksonBT. Membrane vesicle release in bacteria, eukaryotes, and archaea: A conserved yet underappreciated aspect of microbial life. Infect Immun, 2012; 80(6):1948–1957; doi: 10.1128/iai.06014-11
19.
DehinwalR, CooleyD, RakovAV, et al.Increased production of outer membrane vesicles by Salmonella interferes with complement-mediated innate immune attack. mBio, 2021; 12(3):e0086921; doi: 10.1128/mBio.00869-21
20.
DeleuS, MachielsK, RaesJ, et al.Short chain fatty acids and its producing organisms: An overlooked therapy for IBD?. EBioMedicine, 2021; 66:103293; doi: 10.1016/j.ebiom.2021.103293
21.
DingJ, ZhouH, LuoL, et al.Heritable gut microbiome associated with Salmonella enterica serovar Pullorum infection in chickens. mSystems, 2021; 6(1):e01192-20; doi: 10.1128/mSystems.01192-20
22.
DoyleLM, WangMZ. Overview of extracellular vesicles, their origin, composition, purpose, and methods for exosome isolation and analysis. Cells, 2019; 8(7):727; doi: 10.3390/cells8070727
23.
DuW, XuH, MeiX, et al.Probiotic Bacillus enhance the intestinal epithelial cell barrier and immune function of piglets. Benef Microbes, 2018; 9(5):743–754; doi: 10.3920/bm2017.0142
24.
ElhenawyW, Bording-JorgensenM, ValguarneraE, et al.LPS remodeling triggers formation of outer membrane vesicles in Salmonella. mBio, 2016; 7(4):e00940-16; doi: 10.1128/mBio.00940-16
25.
FengW, AoH, PengC. Gut microbiota, short-chain fatty acids, and herbal medicines. Front Pharmacol, 2018; 9:1354; doi: 10.3389/fphar.2018.01354
GuptaD, ZicklerAM, El AndaloussiS. Dosing extracellular vesicles. Adv Drug Deliv Rev, 2021; 178:113961; doi: 10.1016/j.addr.2021.113961
28.
GyörgyB, SzabóTG, PásztóiM, et al.Membrane vesicles, current state-of-the-art: Emerging role of extracellular vesicles. Cell Mol Life Sci, 2011; 68(16):2667–2688; doi: 10.1007/s00018-011-0689-3
HavelaarAH, GarssenJ, TakumiK, et al.A rat model for dose-response relationships of Salmonella Enteritidis infection. J Appl Microbiol, 2001; 91(3):442–452; doi: 10.1046/j.1365-2672.2001.01399.x
31.
HishidaS, IwasaY. Spatial distribution of gut microbes along the intestinal duct. J Theor Biol, 2021; 523:110725; doi: 10.1016/j.jtbi.2021.110725
32.
KimHB, IsaacsonRE. Salmonella in swine: Microbiota interactions. Annu Rev Anim Biosci, 2017; 5:43–63; doi: 10.1146/annurev-animal-022516-022834
KuehnMJ, KestyNC. Bacterial outer membrane vesicles and the host-pathogen interaction. Genes Dev, 2005; 19(22):2645–2655; doi: 10.1101/gad.1299905
35.
LahiriA, LahiriA, IyerN, et al.Visiting the cell biology of Salmonella infection. Microbes Infect, 2010; 12(11):809–818; doi: 10.1016/j.micinf.2010.05.010
36.
MaitiS, HowladerDR, HalderP, et al.Bivalent non-typhoidal Salmonella outer membrane vesicles immunized mice sera confer passive protection against gastroenteritis in a suckling mice model. Vaccine, 2021; 39(2):380–393; doi: 10.1016/j.vaccine.2020.11.040
37.
MoeGR, Zuno-MitchellP, HammondSN, et al.Sequential immunization with vesicles prepared from heterologous Neisseria meningitidis strains elicits broadly protective serum antibodies to group B strains. Infect Immun, 2002; 70(11):6021–6031; doi: 10.1128/iai.70.11.6021-6031.2002
38.
NevermannJ, SilvaA, OteroC, et al.Identification of genes involved in biogenesis of outer membrane vesicles (OMVs) in Salmonella enterica serovar Typhi. Front Microbiol, 2019; 10:104; doi: 10.3389/fmicb.2019.00104
39.
Parada VenegasD, De la FuenteMK, LandskronG, et al.Short chain fatty acids (SCFAs)-mediated gut epithelial and immune regulation and its relevance for inflammatory bowel diseases. Front Immunol, 2019; 10:277; doi: 10.3389/fimmu.2019.00277
40.
QiQ, PengQ, TangM, et al.Microbiome analysis investigating the impacts of fermented spent mushroom substrates on the composition of microbiota in weaned piglets hindgut. Front Vet Sci, 2020; 7:584243; doi: 10.3389/fvets.2020.584243
41.
RatajczakW, RyłA, MizerskiA, et al.Immunomodulatory potential of gut microbiome-derived short-chain fatty acids (SCFAs). Acta Biochim Pol, 2019; 66(1):1–12; doi: 10.1838/8/abp.2018_2648
42.
RenW, WangP, YanJ, et al.Melatonin alleviates weanling stress in mice: Involvement of intestinal microbiota. J Pineal Res, 2018; 64(2); doi: 10.1111/jpi.12448
43.
RodenburgW, KeijerJ, KramerE, et al.Salmonella induces prominent gene expression in the rat colon. BMC Microbiol, 2007; 7:84; doi: 10.1186/1471-2180-7-84
44.
RothDR, RomanD, UlrichP, et al.Design and evaluation of immunotoxicity studies. Exp Toxicol Pathol, 2006; 57(5–6):367–371; doi: 10.1016/j.etp.2006.03.012
45.
SangpuiiL, DixitSK, KumawatM, et al.Comparative roles of clpA and clpB in the survival of S. Typhimurium under stress and virulence in poultry. Sci Rep, 2018; 8(1):4481; doi: 10.1038/s41598-018-22670-6
46.
SartorioMG, PardueEJ, FeldmanMF, et al.Bacterial outer membrane vesicles: From discovery to applications. Annu Rev Microbiol, 2021; 75:609–630; doi: 10.1146/annurev-micro-052821-031444
47.
SchoolingSR, BeveridgeTJ. Membrane vesicles: An overlooked component of the matrices of biofilms. J Bacteriol, 2006; 188(16):5945–5957; doi: 10.1128/jb.00257-06
48.
SegataN, IzardJ, WaldronL, et al.Metagenomic biomarker discovery and explanation. Genome Biol, 2011; 12(6):R60; doi: 10.1186/gb-2011-12-6-r60
49.
ShaoY, LeiZ, YuanJ, et al.Effect of zinc on growth performance, gut morphometry, and cecal microbial community in broilers challenged with Salmonella enterica serovar typhimurium. J Microbiol, 2014; 52(12):1002–1011; doi: 10.1007/s12275-014-4347-y
50.
Solano-ColladoV, Ruiz-CruzS, Lorenzo-DíazF, et al.Recognition of streptococcal promoters by the pneumococcal SigA protein. Front Mol Biosci, 2021; 8:666504; doi: 10.3389/fmolb.2021.666504
51.
SommerF, BäckhedF. The gut microbiota—masters of host development and physiology. Nat Rev Microbiol, 2013; 11(4):227–238; doi: 10.1038/nrmicro2974
52.
SunM, WuW, ChenL, et al.Microbiota-derived short-chain fatty acids promote Th1 cell IL-10 production to maintain intestinal homeostasis. Nat Commun, 2018; 9(1):3555; doi: 10.1038/s41467-018-05901-2
53.
TengY, RenY, SayedM, et al.Plant-derived exosomal MicroRNAs shape the gut microbiota. Cell Host Microbe, 2018; 24(5):637–652.e638; doi: 10.1016/j.chom.2018.10.001
54.
ThiennimitrP, WinterSE, BäumlerAJ. Salmonella, the host and its microbiota. Curr Opin Microbiol, 2012; 15(1):108–114; doi: 10.1016/j.mib.2011.10.002
55.
ThiennimitrP, WinterSE, WinterMG, et al.Intestinal inflammation allows Salmonella to use ethanolamine to compete with the microbiota. Proc Natl Acad Sci U S A, 2011; 108(42):17480–17485; doi: 10.1073/pnas.1107857108
56.
ThyagarajanS, ZhangY, ThapaS, et al.Comparative analysis of racial differences in breast tumor microbiome. Sci Rep, 2020; 10(1):14116; doi: 10.1038/s41598-020-71102-x
57.
ToyofukuM, NomuraN, EberlL. Types and origins of bacterial membrane vesicles. Nat Rev Microbiol, 2019; 17(1):13–24; doi: 10.1038/s41579-018-0112-2
58.
TrachselJM, BearsonBL, KerrBJ, et al.Short chain fatty acids and bacterial taxa associated with reduced Salmonella enterica serovar I 4,[5],12:i:- shedding in swine fed a diet supplemented with resistant potato starch. Microbiol Spectr, 2022; 10(3):e0220221; doi: 10.1128/spectrum.02202-21
59.
WangH, LiC, PengM, et al.N-Acetylcysteine improves intestinal function and attenuates intestinal autophagy in piglets challenged with β-conglycinin. Sci Rep, 2021; 11(1):1261; doi: 10.1038/s41598-021-80994-2
60.
WangZ, ZhangY, XuH, et al.Whole-genome sequencing analysis reveals pig as the main reservoir for persistent evolution of Salmonella enterica serovar Rissen causing human salmonellosis. Food Res Int, 2022; 154:111007; doi: 10.1016/j.foodres.2022.111007
61.
WuIW, LeeCC, HsuHJ, et al.Compositional and functional adaptations of intestinal microbiota and related metabolites in CKD patients receiving dietary protein restriction. Nutrients, 2020; 12(9):2799; doi: 10.3390/nu12092799
62.
XieX, WangZ, ZhangK, et al.Pig as a reservoir of CRISPR type TST4 Salmonella enterica serovar Typhimurium monophasic variant during 2009–2017 in China. Emerg Microbes Infect, 2020; 9(1):1–4; doi: 10.1080/22221751.2019.1699450
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