Insect pathogenic properties of Serratia marcescens: phage-resistant mutants with a decreased resistance to Cecropia immunity and a decreased virulence to Drosophila.

A non-pigmented strain of Serratia marcescens (Db10) was isolated from moribund Drosophila flies. From this strain were isolated spontaneous mutants resistant to streptomycin (Db11) and nalidixic acid (Db12). Mutant Db11 was used for the isolation of two phages, phi J and phi K, which grew on Db10, Db11 and Db12, but not on three reference strains of S. marcescens. Mutant Db11 was demonstrated to fulfil koch's postulates. Strain Db10 and its antibiotic-resistant derivatives were lethal to Drosophila whether given in the food or by injection. Evidence for toxin(s) was found only in sterile supernatants from 7 d cultures. Such extracts contained proteolytic activity and inactivated the antibacterial activity in immune haemolymph from Cecropia. Phages phi J and phi K were used to isolate phage-resistant mutants of Db11. Three such mutants and their parental strain were investigated for their susceptibility to immune haemolymph from Cecropia. The parental strain was resistant to incubation with 90% haemolymph for 2 h at 37 degrees C; all phage-resistant mutants were susceptible to the immune haemolymph with "killing times" (i.e. the time required to kill 90% of the viable cells) ranging from 15 to 55 min. When the same strains were compared for their virulence to Drosophila, the phage-resistant mutants had significantly reduced virulence. It is concluded that resistance to insect immunity plays an important role in the overall pathogenicity of S. marcescens.

[1]  D. Hultmark,et al.  Insect immunity. Purification and properties of three inducible bactericidal proteins from hemolymph of immunized pupae of Hyalophora cecropia. , 2005, European journal of biochemistry.

[2]  I. A. Boman,et al.  Insect immunity. 11. Simultaneous induction of antibacterial activity and selection synthesis of some hemolymph proteins in diapausing pupae of Hyalophora cecropia and Samia cynthia , 1975, Infection and immunity.

[3]  G. Bertani,et al.  STUDIES ON LYSOGENESIS I , 1951, Journal of bacteriology.

[4]  H. G. Boman,et al.  Insect Immunity I. Characteristics of an Inducible Cell-Free Antibacterial Reaction in Hemolymph of Samia cynthia Pupae , 1974, Infection and immunity.

[5]  G. Dalhammar,et al.  Virulence factors in Bacillus thuringiensis: purification and properties of a protein inhibitor of immunity in insects. , 1979, Journal of general microbiology.

[6]  G. Bertani,et al.  Multiplication in Serratia of a bacteriophage originating from Escherichia coli: lysogenization and host-controlled variation. , 1967, Virology.

[7]  H. G. Boman,et al.  Evidence for two immune inhibitors from Bacillus thuringiensis interfering with the humoral defense system of saturniid pupae , 1976, Infection and immunity.

[8]  A. Kreger,et al.  Purification and Characterization of a Serratia marcescens Metalloprotease , 1979, Infection and immunity.

[9]  M. Kaška The toxicity of extracellular proteases of the bacterium Serratia marcescens for larvae of greater wax moth, Galleria mellonella , 1976 .

[10]  L. Reed,et al.  A SIMPLE METHOD OF ESTIMATING FIFTY PER CENT ENDPOINTS , 1938 .

[11]  J. Foulds,et al.  Properties and Characteristics of a Bacteriocin from Serratia marcescens , 1969, Journal of bacteriology.

[12]  H. Matsumoto,et al.  A generalized transducing phage of Serratia marcescens. , 1973, Japanese journal of microbiology.

[13]  A. A. Yousten,et al.  Metalloprotease from Bacillus thuringiensis , 1975, Applied microbiology.

[14]  H. G. Boman,et al.  Cell-wall lipopolysaccharides of ampicillin-resistant mutants of Escherichia coli K-12. , 1976, European journal of biochemistry.

[15]  H. G. Boman,et al.  Characterization of lipopolysaccharides from Escherichia coli K-12 mutants , 1975, Journal of bacteriology.

[16]  F. Grimont,et al.  The genus Serratia. , 1978, Annual review of microbiology.

[17]  O. Lysenko Chitinase of Serratia marcescens and its toxicity to insects , 1976 .

[18]  G. Poinar,et al.  Serratia marcescens as a pathogen of tsetse flies. , 1979, Acta tropica.