The Attenuation Mechanism and Live Vaccine Potential of a Low-Virulence Edwardsiella ictaluri Strain Obtained by Rifampicin Passaging Culture
1
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
The rifampicin-resistant strain E9-302 of Edwardsiella ictaluri strain 669 (WT) was generated by continuous passage on BHI agar plates containing increasing concentrations of rifampicin. E9-302 was attenuated significantly by 119 times to zebrafish Danio rerio compared to WT in terms of the 50% lethal dose (LD50). Zebrafish vaccinated with E9-302 via intraperitoneal (IP) injection at a dose of 1 × 103 CFU/fish had relative percentage survival (RPS) rates of 85.7% when challenged with wild-type E. ictaluri via IP 14 days post-vaccination (dpv). After 14 days of primary vaccination with E9-302 via immersion (IM) at a dose of 4 × 107 CFU/ml, a booster IM vaccination with E9-302 at a dose of 2 × 107 CFU/ml exhibited 65.2% RPS against challenge with wild-type E. ictaluri via IP 7 days later. These results indicated that the rifampicin-resistant attenuated strain E9-302 had potential as a live vaccine against E. ictaluri infection. A previously unreported amino acid site change at position 142 of the RNA polymerase (RNAP) β subunit encoded by the gene rpoB associated with rifampicin resistance was identified. Analysis of the whole-genome sequencing results revealed multiple missense mutations in the virulence-related genes esrB and sspH2 in E9-302 compared with WT, and a 189 bp mismatch in one gene, whose coding product was highly homologous to glycosyltransferase family 39 protein. This study preliminarily explored the molecular mechanism underlying the virulence attenuation of rifampicin-resistant strain E9-302 and provided a new target for the subsequent study of the pathogenic mechanism of E. ictaluri.
Abstract
The rifampicin-resistant strain E9-302 of Edwardsiella ictaluri strain 669 (WT) was generated by continuous passage on BHI agar plates containing increasing concentrations of rifampicin. E9-302 was attenuated significantly by 119 times to zebrafish Danio rerio compared to WT in terms of the 50% lethal dose (LD50). Zebrafish vaccinated with E9-302 via intraperitoneal (IP) injection at a dose of 1 × 103 CFU/fish had relative percentage survival (RPS) rates of 85.7% when challenged with wild-type E. ictaluri via IP 14 days post-vaccination (dpv). After 14 days of primary vaccination with E9-302 via immersion (IM) at a dose of 4 × 107 CFU/ml, a booster IM vaccination with E9-302 at a dose of 2 × 107 CFU/ml exhibited 65.2% RPS against challenge with wild-type E. ictaluri via IP 7 days later. These results indicated that the rifampicin-resistant attenuated strain E9-302 had potential as a live vaccine against E. ictaluri infection. A previously unreported amino acid site change at position 142 of the RNA polymerase (RNAP) β subunit encoded by the gene rpoB associated with rifampicin resistance was identified. Analysis of the whole-genome sequencing results revealed multiple missense mutations in the virulence-related genes esrB and sspH2 in E9-302 compared with WT, and a 189 bp mismatch in one gene, whose coding product was highly homologous to glycosyltransferase family 39 protein. This study preliminarily explored the molecular mechanism underlying the virulence attenuation of rifampicin-resistant strain E9-302 and provided a new target for the subsequent study of the pathogenic mechanism of E. ictaluri.
참고문헌 (73)
- Klesius PH, "Advances in Veterinary Medicine", pp.523~537 (1999), Elsevier
- Zhang Y, "Identification and characterization of an intervening sequence within the 23S ribosomal RNA genes of Edwardsiella ictaluri", Syst. Appl. Microbiol, 30, pp.93~101 (2007)
- Wagner BA, "The epidemiology of bacterial diseases in food-size channel catfish", J. Aquat. Anim. Health, 14, pp.263~272 (2002)
- da Costa A, "Interspecies transmission of Edwardsiella ictaluri in Brazilian catfish (Pseudoplatystoma corruscans) from exotic invasive fish species", Dis. Aquat. Organ, 145, pp.197~208 (2021)
- Liu JY, "Isolation and characterization of Edwardsiella ictaluri strains as pathogens from diseased yellow catfish Pelteobagrus fulvidraco (Richardson) cultured in China: E. ictaluri isolates from diseased yellow catfish", Aquac. Res, 41, pp.1835~1844 (2010)
- Zhou X, "The dynamic immune response of yellow catfish (Pelteobagrus fulvidraco) infected with Edwardsiella ictaluri presenting the inflammation process", Front. Immunol, 12, pp.625928~ (2021)
- Soto E, "Edwardsiella ictaluri as the causative agent of mortality in cultured Nile Tilapia", J. Aquat. Anim. Health, 24, pp.81~90 (2012)
- Keskin O, "Edwardsiella ictaluri infection in rainbow trout (Oncorhynchus mykiss)", Turkish J. Vet. Anim. Sci, 28, pp.649~653 (2004)
- Takeuchi H, "Environmental factors affecting Edwardsiella ictaluri ‐induced mortality of riverine ayu, Plecoglossus altivelis (Temminck & Schlegel)", J. Fish Dis, 44, pp.1065~1074 (2021)
- Hawke JP, "Edwardsiellosis caused by Edwardsiella ictaluri in laboratory populations of Zebrafish Danio rerio", J. Aquat. Anim. Health, 25, pp.171~183 (2013)
- Heuer Ole E, "Human health consequences of use of antimicrobial agents in aquaculture", Clin. Infect. Dis, 49, pp.1248~1253 (2009)
- Li J, "Generation and evaluation of virulence attenuated mutants of Edwardsiella tarda as vaccine candidates to combat edwardsiellosis in flounder (Paralichthys olivaceus)", Fish Shellfish Immunol, 43, pp.175~180 (2015)
- Goldstein BP, "Resistance to rifampicin: a review", J. Antibiot, 67, pp.625~630 (2014)
- Pridgeon JW, "Identification of in vitro upregulated genes in a modified live vaccine strain of Edwardsiella ictaluri compared to a virulent parent strain", Comp. Immunol. Microbiol. Infect. Dis, 33, pp.e31~e40 (2010)
- Olivares-Fuster O, "Development and characterization of rifampicin-resistant mutants from high virulent strains of Flavobacterium columnare: Rifampicin-resistant mutants of Flavobacterium columnare", J. Fish Dis, 34, pp.385~394 (2011)
- Schurig GG, "Biological properties of RB51; a stable rough strain of Brucella abortus", Vet. Microbiol, 28, pp.171~188 (1991)
- Qi Q, "Linking system-wide ompacts of RNA polymerase mutations to the fitness cost of rifampin resistance in Pseudomonas aeruginosa", mBio, 5, pp.e01562-01514~ (2014)
- LaFrentz BR, "Isolation of rifampicin resistant Flavobacterium psychrophilum strains and their potential as live attenuated vaccine candidates", Vaccine, 26, pp.5582~5589 (2008)
- Sun Y, "Isolation and analysis of the vaccine potential of an attenuated Edwardsiella tarda strain", Vaccine, 28, pp.6344~6350 (2010)
- Pridgeon JW, "Development and efficacy of novobiocin and rifampicin-resistant Aeromonas hydrophila as novel vaccines in channel catfish and Nile tilapia", Vaccine, 29, pp.7896~7904 (2011)
- Andersson DI, "Antibiotic resistance and its cost: is it possible to reverse resistance?", Nat. Rev. Microbiol, 8, pp.260~271 (2010)
- Hao B, "Role of alternative sigma factor 54 (RpoN) from Vibrio anguillarum M3 in protease secretion, exopolysaccharide production, biofilm formation, and virulence", Appl. Microbiol. Biotechnol, 97, pp.2575~2585 (2013)
- Leung K, "Mini-Tn5 induced growth- and protease-deficient mutants of Aeromonas hydrophila as live vaccines for blue gourami, Trichogaster trichopterus (Pallas)", Aquaculture, 158, pp.11~22 (1997)
- Villain-Guillot P, "Progress in targeting bacterial transcription", Drug Discov. Today, 12, pp.200~208 (2007)
- Zaw MT, "Mutations inside rifampicin-resistance determining region of rpoB gene associated with rifampicin-resistance in Mycobacterium tuberculosis", J. Infect. Public Health, 11, pp.605~610 (2018)
- Kurepina N, "Mutations compensating for the fitness cost of rifampicin resistance in Escherichia coli exert pleiotropic effect on RNA polymerase catalysis", Nucleic Acids Res, 50, pp.5739~5756 (2022)
- McCallum N, "In vivo survival of teicoplaninresistant Staphylococcus aureus and fitness cost of teicoplanin resistance", Antimicrob. Agents Chemother, 50, pp.2352~2360 (2006)
- Han F, "Fitness cost of macrolide resistance in Campylobacter jejuni", Int. J. Antimicrob. Agents, 34, pp.462~466 (2009)
- Pridgeon JW, "Development and efficacy of a novobiocin-resistant Streptococcus iniae as a novel vaccine in Nile tilapia (Oreochromis niloticus)", Vaccine, 29, pp.5986~5993 (2011)
- Sander P, "Fitness cost of chromosomal drug resistance-conferring mutations", Antimicrob. Agents Chemother, 46, pp.1204~1211 (2002)
- Hamilton MA, "Trimmed spearman-karber method for estimating median lethal concentrations in toxicity bioassays", Environ. Sci. Technol, 11, pp.714~719 (1977)
- Dias MKR, "Lethal dose and clinical signs of Aeromonas hydrophila in Arapaima gigas (Arapaimidae), the giant fish from Amazon", Vet. Microbiol, 188, pp.12~15 (2016)
- Pridgeon JW, "Development of a novobiocin-resistant Edwardsiella ictaluri as a novel vaccine in channel catfish (Ictalurus punctatus)", Vaccine, 29, pp.5631~5637 (2011)
- Lin B, "Acute phase response in zebrafish upon Aeromonas salmonicida and Staphylococcus aureus infection: Striking similarities and obvious differences with mammals", Mol. Immunol, 44, pp.295~301 (2007)
- O'Toole R, "Visualisation of Zebrafish infection by GFP-labelled Vibrio anguillarum", Microb. Pathog, 37, pp.41~46 (2004)
- Pressley ME, "Pathogenesis and inflammatory response to Edwardsiella tarda infection in the zebrafish", Dev. Compar. Immunol, 29, pp.501~513 (2005)
- Hohn C, "Zebrafish kidney phagocytes utilize macropinocytosis and Ca2+-dependent endocytic mechanisms", PLoS One, 4, pp.e4314~ (2009)
- Petrie-Hanson L, "Evaluation of zebrafish Danio rerio as a model for Enteric Septicemia of Catfish (ESC)", J. Aquat. Anim. Health, 19, pp.151~158 (2007)
- Santander J, "The Aspartate-semialdehyde dehydrogenase of Edwardsiella ictaluri and its useas balanced-lethal system in fish vaccinology", PLoS One, 5, pp.e15944~ (2010)
- Maiden MCJ, "Multilocus sequence typing: a portable approach to the identification of clones within populations of pathogenic microorganisms", Proc. Natl. Acad. Sci. USA, 95, pp.3140~3145 (1998)
- Maiden MCJ, "Multilocus sequence typing of bacteria", Ann. Rev. Microbiol, 60, pp.561~588 (2006)
- Pridgeon JW, "Attenuation of a virulent Aeromonas hydrophila with novobiocin and pathogenic characterization of the novobiocin-resistant strain", J. Appl. Microbiol, 113, pp.1319~1328 (2012)
- Pridgeon JW, "Attempt to develop live attenuated bacterial vaccines by selecting resistance to gossypol, proflavine hemisulfate, novobiocin, or ciprofloxacin", Vaccine, 31, pp.2222~2230 (2013)
- Heckman TI, "Development and efficacy of Streptococcus iniae live-attenuated vaccines in Nile tilapia, Oreochromis niloticus", Fish Shellfish Immunol, 121, pp.152~162 (2022)
- Pridgeon JW, "Fitness cost, gyrB mutation, and absence of phosphotransferase system fructose specific IIABC component in novobiocin-resistant Streptococcus iniae vaccine strain ISNO", Vet. Microbiol, 165, pp.384~391 (2013)
- Klein JL, "Rifampin Resistance in Mycobacterium kansasii is associated with rpoB mutations", Antimicrob. Agents Chemother, 45, pp.3056~3058 (2001)
- Jin DJ, "Mapping and sequencing of mutations in the Escherichia coli rpoB gene that lead to rifampicin resistance", J. Mol. Biol, 202, pp.45~58 (1988)
- Choi J, "Salmonella pathogenicity island 2 expression negatively controlled by EIIANtr-SsrB interaction is required for Salmonella virulence", Proc. Natl. Acad. Sci. USA, 107, pp.20506~20511 (2010)
- Tan YP, "Role of type III secretion in Edwardsiella tarda virulence", Microbiology, 151, pp.2301~2313 (2005)
- Wang X, "Edwardsiella tarda T6SS component evpP is regulated by esrB and iron, and plays essential roles in the invasion of fish", Fish Shellfish Immunol, 27, pp.469~477 (2009)
- Liu Y, "Transcriptomic dissection of the horizontally acquired response regulator EsrB reveals its global regulatory roles in the physiological adaptation and activation of T3SS and the cognate effector repertoire in Edwardsiella piscicida during infection toward turbot", Virulence, 8, pp.1355~1377 (2017)
- Otten EG, "Ubiquitylation of lipopolysaccharide by RNF213 during bacterial infection", Nature, 594, pp.111~116 (2021)
- Xu T, "Ubiquitination-mediated inflammasome activation during bacterial infection", Int. J. Mol. Sci, 20, pp.2110~ (2019)
- Berglund J, "Modification of the host ubiquitome by bacterial enzymes", Microbiol. Res, 235, pp.126429~ (2020)
- Wan M, "A bacterial effector deubiquitinase specifically hydrolyses linear ubiquitin chains to inhibit host inflammatory signalling", Nat. Microbiol, 4, pp.1282~1293 (2019)
- Hensel M, "Salmonella pathogenicity island 2", Mol. Microbiol, 36, pp.1015~1023 (2000)
- Haraga A, "A Salmonella enterica serovar typhimurium translocated leucine-rich repeat effector protein inhibits NFkappa B-dependent gene expression", Infect. Immun, 71, pp.4052~4058 (2003)
- Kidwai AS, "Diverse secreted effectors are required for Salmonella persistence in a mouse infection model", PLoS One, 8, pp.e70753~ (2013)
- Halici S, "Functional analysis of the Salmonella pathogenicity island 2-mediated inhibition of antigen presentation in dendritic cells", Infect. Immun, 76, pp.4924~4933 (2008)
- Bhavsar AP, "The Salmonella type III effector SspH2 specifically exploits the NLR co-chaperone activity of SGT1 to subvert immunity", PLoS Pathog, 9, pp.e1003518~ (2013)
- Quezada CM, "A family of Salmonella virulence factors functions as a distinct class of autoregulated E3 ubiquitin ligases", Proc. Natl. Acad. Sci. USA, 106, pp.4864~4869 (2009)
- Hicks Stuart W, "Subcellular targeting of Salmonella virulence proteins by hostmediated S-palmitoylation", Cell Host Microbe, 10, pp.9~20 (2011)
- Wedlich-Soldner R, "Closing the loops: new insights into the role and regulation of actin during cell polarization", Exp. Cell Res, 301, pp.8~15 (2004)
- Walch P, "Global mapping of Salmonella enterica-host proteinprotein interactions during infection", Cell Host Microbe, 29, pp.1316~1332.e1312 (2021)
- Miao EA, "Salmonella effectors translocated across the vacuolar membrane interact with the actin cytoskeleton: Salmonella intracellular effectors", Mol. Microbiol, 48, pp.401~415 (2003)
- Navarro-Garcia F, "Actin cytoskeleton manipulation by effector proteins secreted by diarrheagenic Escherichia coli pathotypes", BioMed Res. Int, 2013, pp.1~22 (2013)
- Stevens JM, "Actin-dependent movement of bacterial pathogens", Nat. Rev. Microbiol, 4, pp.91~101 (2006)
- Lu Q, "Sweet talk: Protein glycosylation in bacterial interaction with the host", Trends Microbiol, 23, pp.630~641 (2015)
- Tra VN, "Glycans in pathogenic bacteria--potential for targeted covalent therapeutics and imaging agents", Chem. Commun. (Camb), 50, pp.4659~4673 (2014)
- Valguarnera E, "Sugar and spice make bacteria not nice: protein glycosylation and its influence in pathogenesis", J. Mol. Biol, 428, pp.3206~3220 (2016)
- Jia L, "Effect of protein O-mannosyltransferase (MSMEG_5447) on M. smegmatis and its survival in macrophages", Front. Microbiol, 12, pp.657726~ (2021)
- Liu C-F, "Bacterial protein-O-mannosylating enzyme is crucial for virulence of Mycobacterium tuberculosis", Proc. Natil. Acad. Sci. USA, 110, pp.6560~6565 (2013)
- Cain JA, "Proteomics reveals multiple phenotypes associated with Nlinked glycosylation in Campylobacter jejuni", Mol. Cell. Proteomics, 18, pp.715~734 (2019)
