Diagnosis by polymerase chain reaction of Erysipelas septicemia in a flock of ring-necked pheasants.

A flock of 810 pheasants experienced 6.2% mortality over 6 days. Affected birds were weak and lethargic for up to 24 hr before death. Examined birds were thin, and gross lesions consisted of thick opaque crops and cecal cores. Histologically, there was capillariasis of the crop and multifocal ulcerative typhlitis with Heterakis spp. infection, and numerous systemic intravascular monocytes were filled with clusters of blue rod-shaped organisms. The organisms were gram-positive bacilli by Brown and Brenn staining and ultrastructural analysis. Liver bacterial cultures were negative for pathogenic bacteria. Erysipelas septicemia was diagnosed by an Erysipelothrix species-specific polymerase chain reaction method with the substrate DNA isolated from formalin-fixed, paraffin-embedded liver.

Potential errors in recognition of Erysipelothrix rhusiopathiae.

Here we describe four isolations of Erysipelothrix rhusiopathiae associated with polyarthralgia and renal failure, septic arthritis, classic erysipeloid, and peritonitis. Although the biochemical identification was straightforward in each case, recognition presented a challenge to the clinical microbiologist, since in three cases E. rhusiopathiae was not initially considered due to unusual clinical presentations, in two cases the significance might not have been appreciated because growth was in broth only, and in one case the infection was thought to be polymicrobic. Because the Gram stain can be confusing, abbreviated identification schemes that do not include testing for H(2)S production could allow E. rhusiopathiae isolates to be misidentified as Lactobacillus spp. or Enterococcus spp. in atypical infections.

Presence of a capsule in Erysipelothrix rhusiopathiae and its relationship to virulence for mice.

Three avirulent insertional mutants of Erysipelothrix rhusiopathiae were obtained by the technique of transposon mutagenesis with the self-conjugative transposon Tn916. The interactions between murine polymorphonuclear leukocytes and parent and mutant strains were studied in vitro. In the presence of normal serum, the virulent parent strain was resistant to phagocytosis, whereas the avirulent mutant strains were efficiently phagocytosed. In the presence of immune serum, the parent and the mutant strains were both efficiently phagocytosed. Electron microscopic examination of the parent strain demonstrated the presence of a structure resembling a capsule which was absent on the mutant strains, suggesting that a capsule may be involved in virulence. This was confirmed in studies in which an avirulent mutant strain reverted to virulence following acquisition of a capsule when the transposon was lost by spontaneous excision. These results strongly suggest that virulence of E. rhusiopathiae is associated, at least in part, with resistance to phagocytosis by polymorphonuclear leukocytes and that this antiphagocytic ability of the bacterium results from its possession of a capsule.

Correlation between adherence of Erysipelothrix rhusiopathiae strains of serovar 1a to tissue culture cells originated from porcine kidney and their pathogenicity in mice and swine.

Adherence of four virulent and four avirulent strains of Erysipelothrix rhusiopathiae, serovar 1a, to porcine kidney cell lines, PK-15 and ESK cells, was examined in an in vitro system. The virulent strains adhered well to the cells (range of means, 9.95 +/- 0.87-36.01 +/- 1.10 per cell). In contrast, the avirulent strains showed negligible adherence to the cells (range of means, 0.11 +/- 0.04-1.41 +/- 0.13 per cell). Pretreatment of bacteria with heat, trypsin, or antiserum resulted in a marked decrease in adherence. Scanning electron microscopic examination revealed that the bacteria attached directly to the microvilli of cells.

Ultrastructural characterization of stable L-form cells from Erysipelothrix rhusiopathiae and of accompanying artifacts.

The stable L-form of Erysipelothrix rhusiopathiae is a typical protoplast type L-form. Cells are surrounded by a trilamellar cytoplasmic membrane only. They grow in form of aggregations in liquid media and their diameters vary between 0.1 and 2 micrometer. Always a large portion of cells undergoes lysis. It seems to be characteristic for L-form cultures of E. rhusiopathiae that always many artifact structures are formed. The artifacts are spherical particles with diameters of 0.1 micrometer to more than 3 micrometer. They can be differentiated from L-form cells only by electron microscopy. The artifacts consist of electron dense amorphous material and their surface is irregular without a clear boundary line. Obviously, these artifacts are produced from protein components of the medium and from cytoplasmatic components of the lysing L-form cells.

Electron microscopic studies on altered forms of Erysipelothrix rhusiopathiae obtained under the effect of novobiocin.

Altered and L-forms of Erysipelothrix rhusiopathiae were obtained through in vitro and in vivo treatments with novobiocin. Electron microscopic studies were carried out on the changes taking place in the morphology and ultrastructure of strains showing a varying degree of resistance. When lower concentrations of the antibiotic were employed (15, 25, 200, 400 mcg) strongly elongated and threadlike forms were established manifesting characteristic changes in the cell wall and the inner structure. The use of high concentrations of novobiocin (1000 and 2000 mcg) incited the production of L-forms the morphology and ultrastructure of which are dealt with in detail.