Characterization of a Salmonella choleraesuis isolate after repeated neutrophil exposure

Michael B. Roof From NOBL Laboratories, Inc, Sioux Center, IA 51250 (Roof) and the Department of Microbiology, Immunology, and Preventive Medicine, Iowa State University, Ames, IA 50011 (Kramer, Roth, Minion).

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Theodore T. Kramer From NOBL Laboratories, Inc, Sioux Center, IA 51250 (Roof) and the Department of Microbiology, Immunology, and Preventive Medicine, Iowa State University, Ames, IA 50011 (Kramer, Roth, Minion).

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James A. Roth From NOBL Laboratories, Inc, Sioux Center, IA 51250 (Roof) and the Department of Microbiology, Immunology, and Preventive Medicine, Iowa State University, Ames, IA 50011 (Kramer, Roth, Minion).

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F. Chris Minion From NOBL Laboratories, Inc, Sioux Center, IA 51250 (Roof) and the Department of Microbiology, Immunology, and Preventive Medicine, Iowa State University, Ames, IA 50011 (Kramer, Roth, Minion).

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Summary

Salmonella choleraesuis strain 38 (glycerol-positive fermentation) was repeatedly exposed to porcine neutrophils in an attempt to mimic in vivo conditions of the host immune system. After phagocytosis, viable intracellular S choleraesuis were isolated and the process was repeated at least 5 times. A fifth-passage strain-38 neutrophil-adapted clone, 38PMNa-5X, was isolated, and was compared with the parent wild-type strain 38 for changes. Strain 38PMNa-5X had increased resistance to killing by hydrogen peroxide and phagocyte killing by porcine neutrophils, as measured by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide reduction. Strain 38PMNa-5X was less invasive than the parent strain on Vero cell monolayers, and had been cured of a 50-kb plasmid. The 50-kb plasmid was marked with bacteriophage mini-Mu (kanamycin resistant) and was reinserted into strain 38PMNa-5X. Strain 38PMNa-5X was avirulent in mice, but the isolates with reinserted plasmids had intermediate resistance to neutrophil and hydrogen peroxide killing and had restored invasiveness and mouse virulence. Differences in complement sensitivity and enzymatic activity were not observed between the strains.

Summary

Salmonella choleraesuis strain 38 (glycerol-positive fermentation) was repeatedly exposed to porcine neutrophils in an attempt to mimic in vivo conditions of the host immune system. After phagocytosis, viable intracellular S choleraesuis were isolated and the process was repeated at least 5 times. A fifth-passage strain-38 neutrophil-adapted clone, 38PMNa-5X, was isolated, and was compared with the parent wild-type strain 38 for changes. Strain 38PMNa-5X had increased resistance to killing by hydrogen peroxide and phagocyte killing by porcine neutrophils, as measured by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide reduction. Strain 38PMNa-5X was less invasive than the parent strain on Vero cell monolayers, and had been cured of a 50-kb plasmid. The 50-kb plasmid was marked with bacteriophage mini-Mu (kanamycin resistant) and was reinserted into strain 38PMNa-5X. Strain 38PMNa-5X was avirulent in mice, but the isolates with reinserted plasmids had intermediate resistance to neutrophil and hydrogen peroxide killing and had restored invasiveness and mouse virulence. Differences in complement sensitivity and enzymatic activity were not observed between the strains.

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