The lipopolysaccharide outer core transferase genes pcgD and hptE contribute differently to the virulence of Pasteurella multocida in ducks
Tóm tắt
Fowl cholera caused by Pasteurella multocida exerts a massive economic burden on the poultry industry. Lipopolysaccharide (LPS) is essential for the growth of P. multocida genotype L1 strains in chickens and specific truncations to the full length LPS structure can attenuate bacterial virulence. Here we further dissected the roles of the outer core transferase genes pcgD and hptE in bacterial resistance to duck serum, outer membrane permeability and virulence in ducks. Two P. multocida mutants, ΔpcgD and ΔhptE, were constructed, and silver staining confirmed that they all produced truncated LPS profiles. Inactivation of pcgD or hptE did not affect bacterial susceptibility to duck serum and outer membrane permeability but resulted in attenuated virulence in ducks to some extent. After high-dose inoculation, ΔpcgD showed remarkably reduced colonization levels in the blood and spleen but not in the lung and liver and caused decreased injuries in the spleen and liver compared with the wild-type strain. In contrast, the ΔhptE loads declined only in the blood, and ΔhptE infection caused decreased splenic lesions but also induced severe hepatic lesions. Furthermore, compared with the wild-type strain, ΔpcgD was significantly attenuated upon oral or intramuscular challenge, whereas ΔhptE exhibited reduced virulence only upon oral infection. Therefore, the pcgD deletion caused greater virulence attenuation in ducks, indicating the critical role of pcgD in P. multocida infection establishment and survival.
Tài liệu tham khảo
Wilson BA, Ho M (2013) Pasteurella multocida: from zoonosis to cellular microbiology. Clin Microbiol Rev 26:631–655
Abreu F, Rodriguez-Lucas C, Rodicio MR, Vela AI, Fernandez-Garayzabal JF, Leiva PS, Cuesta F, Cid D, Fernandez J (2018) Human Pasteurella multocida infection with likely zoonotic transmission from a pet dog, Spain. Emerg Infect Dis 24:1145–1146
Blackall PJ, Pahoff JL, Marks D, Fegan N, Morrow CJ (1995) Characterisation of Pasteurella multocida isolated from fowl cholera outbreaks on turkey farms. Aust Vet J 72:135–138
Kardos G, Kiss I (2005) Molecular epidemiology investigation of outbreaks of fowl cholera in geographically related poultry flocks. J Clin Microbiol 43:2959–2961
Mohamed MA, Mohamed MW, Ahmed AI, Ibrahim AA, Ahmed MS (2012) Pasteurella multocida in backyard chickens in Upper Egypt: incidence with polymerase chain reaction analysis for capsule type, virulence in chicken embryos and antimicrobial resistance. Vet Ital 48:77–86
Harper M, John M, Turni C, Edmunds M, St Michael F, Adler B, Blackall PJ, Cox AD, Boyce JD (2015) Development of a rapid multiplex PCR assay to genotype Pasteurella multocida strains by use of the lipopolysaccharide outer core biosynthesis locus. J Clin Microbiol 53:477–485
Wilkie IW, Harper M, Boyce JD, Adler B (2012) Pasteurella multocida: diseases and pathogenesis. Curr Top Microbiol 361:1–22
Harper M, Boyce JD, Adler B (2012) The key surface components of Pasteurella multocida: capsule and lipopolysaccharide. Curr Top Microbiol 361:39–51
Petruzzi B, Briggs RE, Tatum FM, Swords WE, De Castro C, Molinaro A, Inzana TJ (2017) Capsular polysaccharide interferes with biofilm formation by Pasteurella multocida serogroup A. Bio 8:e01843-17
Zhao X, Liu Q, Xiao K, Hu Y, Liu X, Li Y, Kong Q (2016) Identification of the crp gene in avian Pasteurella multocida and evaluation of the effects of crp deletion on its phenotype, virulence and immunogenicity. BMC Microbiol 16:125
Steen JA, Steen JA, Harrison P, Seemann T, Wilkie I, Harper M, Adler B, Boyce JD (2010) Fis is essential for capsule production in Pasteurella multocida and regulates expression of other important virulence factors. PLoS Pathog 6:e1000750
Megroz M, Kleifeld O, Wright A, Powell D, Harrison P, Adler B, Harper M, Boyce JD (2016) The RNA-binding chaperone Hfq is an important global regulator of gene expression in Pasteurella multocida and plays a crucial role in production of a number of virulence factors, including hyaluronic acid capsule. Infect Immun 84:1361–1370
Xiao K, Liu Q, Liu X, Hu Y, Zhao X, Kong Q (2015) Identification of the Avian Pasteurella multocida phoP gene and evaluation of the effects of phoP deletion on virulence and immunogenicity. Int J Mol Sci 17:12
Cohen J (2002) The immunopathogenesis of sepsis. Nature 420:885–891
Tan Y, Kagan JC (2014) A cross-disciplinary perspective on the innate immune responses to bacterial lipopolysaccharide. Mol cell 54:212–223
Periasamy S, Praveena PE, Singh N (2018) Effects of Pasteurella multocida lipopolysaccharides on bovine leukocytes. Microb Pathog 119:225–232
Horadagoda NU, Hodgson JC, Moon GM, Wijewardana TG, Eckersall PD (2002) Development of a clinical syndrome resembling haemorrhagic septicaemia in the buffalo following intravenous inoculation of Pasteurella multocida serotype B:2 endotoxin and the role of tumour necrosis factor-alpha. Res Vet Sci 72:194–200
Harper M, Boyce JD (2017) The myriad properties of Pasteurella multocida lipopolysaccharide. Toxins 9:254
Harper M, Boyce JD, Cox AD, St Michael F, Wilkie IW, Blackall PJ, Adler B (2007) Pasteurella multocida expresses two lipopolysaccharide glycoforms simultaneously, but only a single form is required for virulence: identification of two acceptor-specific heptosyl I transferases. Infect Immun 75:3885–3893
St Michael F, Harper M, Parnas H, John M, Stupak J, Vinogradov E, Adler B, Boyce JD, Cox AD (2009) Structural and genetic basis for the serological differentiation of Pasteurella multocida Heddleston serotypes 2 and 5. J Bacteriol 191:6950–6959
Boyce JD, Harper M, St Michael F, John M, Aubry A, Parnas H, Logan SM, Wilkie IW, Ford M, Cox AD, Adler B (2009) Identification of novel glycosyltransferases required for assembly of the Pasteurella multocida A:1 lipopolysaccharide and their involvement in virulence. Infect Immun 77:1532–1542
Harper M, Cox A, St Michael F, Parnas H, Wilkie I, Blackall PJ, Adler B, Boyce JD (2007) Decoration of Pasteurella multocida lipopolysaccharide with phosphocholine is important for virulence. J Bacteriol 189:7384–7391
Harper M, Wright A, St Michael F, Li J, Deveson Lucas D, Ford M, Adler B, Cox AD, Boyce JD (2017) Characterization of two novel lipopolysaccharide phosphoethanolamine transferases in Pasteurella multocida and their role in resistance to cathelicidin-2. Infect Immun 85:e00557-e617
Harper M, Cox AD, St Michael F, Wilkie IW, Boyce JD, Adler B (2004) A heptosyltransferase mutant of Pasteurella multocida produces a truncated lipopolysaccharide structure and is attenuated in virulence. Infect Immun 72:3436–3443
Harper M, Boyce JD, Wilkie IW, Adler B (2003) Signature-tagged mutagenesis of Pasteurella multocida identifies mutants displaying differential virulence characteristics in mice and chickens. Infect Immun 71:5440–5446
Edwards RA, Keller LH, Schifferli DM (1998) Improved allelic exchange vectors and their use to analyze 987P fimbria gene expression. Gene 207:149–157
Bosse JT, Durham AL, Rycroft AN, Kroll JS, Langford PR (2009) New plasmid tools for genetic analysis of Actinobacillus pleuropneumoniae and other pasteurellaceae. Appl Environ Microbiol 75:6124–6131
Kittelberger R, Hilbink F (1993) Sensitive silver-staining detection of bacterial lipopolysaccharides in polyacrylamide gels. J Biochem Biophys Methods 26:81–86
Loh B, Grant C, Hancock RE (1984) Use of the fluorescent probe 1-N-phenylnaphthylamine to study the interactions of aminoglycoside antibiotics with the outer membrane of Pseudomonas aeruginosa. Antimicrob Agents Chemother 26:546–551
Luo HY, Liu MF, Wang MS, Zhao XX, Jia RY, Chen S, Sun KF, Yang Q, Wu Y, Chen XY, Biville F, Zou YF, Jing B, Cheng AC, Zhu DK (2018) A novel resistance gene, lnu(H), conferring resistance to lincosamides in Riemerella anatipestifer CH-2. Int J Antimicrob Agents 51:136–139
Scott PC, Markham JF, Whithear KG (1999) Safety and efficacy of two live Pasteurella multocida aro-A mutant vaccines in chickens. Avian Dis 43:83–88
Harper M, John M, Edmunds M, Wright A, Ford M, Turni C, Blackall PJ, Cox A, Adler B, Boyce JD (2016) Protective efficacy afforded by live Pasteurella multocida vaccines in chickens is independent of lipopolysaccharide outer core structure. Vaccine 34:1696–1703
Chung JY, Wilkie I, Boyce JD, Townsend KM, Frost AJ, Ghoddusi M, Adler B (2001) Role of capsule in the pathogenesis of fowl cholera caused by Pasteurella multocida serogroup A. Infect Immun 69:2487–2492
Okuda S, Sherman DJ, Silhavy TJ, Ruiz N, Kahne D (2016) Lipopolysaccharide transport and assembly at the outer membrane: the PEZ model. Nat Rev Microbiol 14:337–345
Chang PC, Wang CJ, You CK, Kao MC (2011) Effects of a HP0859 (rfaD) knockout mutation on lipopolysaccharide structure of Helicobacter pylori 26695 and the bacterial adhesion on AGS cells. Biochem Biophys Res Commun 405:497–502
Dam S, Pages JM, Masi M (2018) Stress responses, outer membrane permeability control and antimicrobial resistance in Enterobacteriaceae. Microbiology 164:260–267
van der Heijden J, Reynolds LA, Deng W, Mills A, Scholz R, Imami K, Foster LJ, Duong F, Finlay BB (2016) Salmonella rapidly regulates membrane permeability to survive oxidative stress. mBio 7:e01238-16
Bojkovic J, Richie DL, Six DA, Rath CM, Sawyer WS, Hu Q, Dean CR (2015) Characterization of an Acinetobacter baumannii lptD deletion strain: permeability defects and response to inhibition of lipopolysaccharide and fatty acid biosynthesis. J Bacteriol 198:731–741
Park BS, Song DH, Kim HM, Choi BS, Lee H, Lee JO (2009) The structural basis of lipopolysaccharide recognition by the TLR4-MD-2 complex. Nature 458:1191–1195
Conde-Alvarez R, Arce-Gorvel V, Iriarte M, Mancek-Keber M, Barquero-Calvo E, Palacios-Chaves L, Chacon-Diaz C, Chaves-Olarte E, Martirosyan A, von Bargen K, Grillo MJ, Jerala R, Brandenburg K, Llobet E, Bengoechea JA, Moreno E, Moriyon I, Gorvel JP (2012) The lipopolysaccharide core of Brucella abortus acts as a shield against innate immunity recognition. PLoS Pathog 8:e1002675
Schroeder TH, Lee MM, Yacono PW, Cannon CL, Gerceker AA, Golan DE, Pier GB (2002) CFTR is a pattern recognition molecule that extracts Pseudomonas aeruginosa LPS from the outer membrane into epithelial cells and activates NF-kappa B translocation. Proc Natl Acad Sci USA 99:6907–6912
Weiss G, Schaible UE (2015) Macrophage defense mechanisms against intracellular bacteria. Immunol Rev 264:182–203
Rubires X, Saigi F, Pique N, Climent N, Merino S, Alberti S, Tomas JM, Regue M (1997) A gene (wbbL) from Serratia marcescens N28b (O4) complements the rfb-50 mutation of Escherichia coli K-12 derivatives. J Bacteriol 179:7581–7586