Abstract
Temperature-sensitive (ts) mutants ofStaphylococcus aureus were isolated after mutagenesis with nitrosoguanidine and two cycles of enrichment with Penicillin G and D-Cycloserine. The mutants expressed tight, coasting, and leaky phenotypes on solid media. In broth, however, most exhibited coasting for a limited number of generations. The reversion frequency of selected ts mutants was less than 10−6. Intraperitoneal (i.p.) immunization with ts mutant G/1/2 conferred significant protection (0 dead/6 total vs. 7/7, immunized vs. control; p=0.0006) from lethal i.p. challenge with the parental wild-type (wt)S. aureus suspended in 5% porcine mucin, performed 28 days after i.p. administration of 108 colony-forming units. Protection induced by mutants of coasting phenotype was higher and lasted longer than that induced by mutants of the tight phenotype. The results of this study demonstrate that ts mutants ofS. aureus can be obtained and that ts mutants are able to induce protective immunity from subsequent challenge with the parental wt strain.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
Literature Cited
Albus A, Arbeit RD, Lee JC (1991) Virulence ofStaphylococcus aureus mutants altered in type 5 capsule production. Infect Immun 59:1008–1014
Cerquetti MC, Gherardi MM, Sordelli DO (1990) Evaluation of different temperature-sensitive mutant phenotypes ofSalmonella enteritidis as vaccine potentials. Curr Microbiol 21:225–228
Eberhart RJ, Harmon RJ, Hasper DE, Natzke RP, Nickerson Sc, Reneau JK, Row EH, Smith KL, Spencer SB (1987) Current concepts of bovine mastitis, p 6–8. The National Mastitis Council, 3rd ed., Arlington, Virginia.
Fattom A, Schneerson R, Szu SC, Vann W, Shiloach J, Karakawa WW, Robbins JB (1990) Synthesis and immunologic properties in mice of vaccines composed ofStaphylococcus aureus type 5 and type 8 capsular polysaccharides conjugated toPseudomonas aeruginosa exotoxin A Infect Immun 58:2367–2374
Foster TJ (1991) Potential for vaccination against, infections caused byStaphylococcus aureus. Vaccine 9:221–227
Fournier JM (1991)Staphylococcus aureus. In: Cryz S (ed) Vaccines and immunotherapy. New York: Pergamon Press, pp 166–177
Groisman EA, Fields PI, Heffron F (1990) Molecular biology ofSalmonella pathogenesis. In: Iglewski BH, Clark VL (ed) Molecular basis of bacterial pathogenesis San Diego, California: Academic Press, Inc., pp 251–272
Haertl R, Bandlow G (1990) Application of small fragment restriction endonuclease analysis (SF-REA) to the epidemiological fingerprinting ofStaphylococcus aureus. J Med Microbiol 33:91–96
Karnell A, Stocker BAD, Katakura S, Reinholt FP, Lindberg AA (1992) Live oral auxotrophicShigella flexneri SFL124 vaccine with a deletedaroD gene: characterization and monkey protection studies. Vaccine 10:389–394
Laemmli, UK (1970) Cleavage of structural proteins during the assembly of the head of, bacteriophage T4. Nature 227:680–685.
Lee JC, Perez NE, Hopkins CA, Pier GB (1988) Purified capsular polysaccharide-induced immunity toStaphylococcus aureus infection. J Infect Dis 157:723–730
Majumdar D, Avissar YJ, Wyche JH (1991) Simultaneous and rapid isolation of bacterial and eukaryotic DNA and RNA: a new approach for isolating DNA. Bio Techniques 11:94–101
Mellenberger RW (1977) Vaccination against mastitis. J Dairy Sci 60:1016–1021
Morris Hooke A (1987). Attenuated live bacteria: vaccine efficacy and shortcomings. In: Bell R, Torrigiani G (eds) Towards better carbohydrate vaccines. New York: John Wiley & Sons, pp 167–184
Morris Hooke A, Bellanti JA, Oeschger MP (1985) Genetically-attenuated bacterial vaccines: new approaches for safety and efficacy. Lancet 1:1472–1474
Poutrel B, Boutonnier A, Sutra L, Fournier JM (1988) Prevalence of capsular polysaccharide types 5 and 8 amongStaphylococcus aureus isolates from cow, goat, and ewe milk. J Clin Microbiol 26:38–40
Rather PN, Davis AP, Wilkinson BJ (1985) Slime production by bovine milkStraphylococcus aureus and identification of coagulase-negative staphylococcal isolates. J Clin Microbiol 23:858–862
Robbins JB, Schneerson R (1990) Polysaccharide-protein conjugates: a new generation of vaccines. J Infect Dis 161:821–832
Sambrook J, Fritsch EF, Maniatis T (1989) Digesting DNA with restriction enzymes. In: Molecular cloning, a laboratory manual. Cold Spring Harbor, New York: Cold Spring Harbor Laboratory Press, pp. 5.28–5.32
Watson DL (1992) Letter. Staphylococcal mastitis vaccine. Vaccine 10:359
Watson DL, Schwarzkoff CL (1990) A field trial to test the efficacy of staphylococcal mastitis vaccine in commrical dairies in Australia. Proceedings of the International Symposium on Bovine Mastitis, Indianapolis, Indiana, 1990:73
Yoshida K, Ichiman Y, Narikawa S, Evans WB (1984) Staphylococcal capsular vaccine for preventing mastitis in two herds in Georgia. J Dairy Sci 67:620–627
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Sordelli, D.O., Iglesias, M.F., Cerquetti, M.C. et al. Temperature-sensitive mutants ofStaphylococcus aureus: Isolation and preliminary characterization. Current Microbiology 27, 125–129 (1993). https://doi.org/10.1007/BF01576008
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF01576008