Related Experiment Videos
Structures suggesting cell-wall-deficient forms detected in circulating erythrocytes by fluorochrome staining
Abstract:
Cell-wall-deficient (CWD) forms of bacteria are associated with certain cases of idiopathic septicemia. In this preliminary study of blood examined immediately after venipuncture, structures with a morphology characteristic of CWD forms were seen parasitizing the erythrocytes. These inclusions were usually circumferential, but in some cases they protruded from the red cells. The CWD forms were detected by staining with Gould's rhodamine-labeled muramidase, which reacted similarly to acridine orange but with greater specificity. A blocking test, employing unlabeled muramidase, indicated the specificity of the reaction between muramidase and the microbial substrate. Reaction of the forms with muramidase indicates their bacterial, rather than mycoplasmal, nature. Thus in vivo CWD forms have a detectable component of muramic acid, at least in certain cases. Sixty-eight individuals with a diagnosis of fever of unknown origin were tested, with 51 nondebilitated individuals serving as controls. More intraerythrocytic forms reacting with muramidase were found in the patients than in the controls. Nearly 40% of the cases had a relatively high incidence of erythrocyte parasitism. In some instances when freshly drawn blood was examined, the structures, which appear to be microbial, extended in rhizoid filaments from the erythrocytes.
Insights
Cell-wall-deficient (CWD) bacteria were found within red blood cells in patients with fever of unknown origin. This preliminary study suggests CWD bacterial forms may cause idiopathic septicemia, detectable via muramidase staining.
Area of Science:
- Microbiology
- Hematology
- Infectious Diseases
Background:
- Cell-wall-deficient (CWD) bacterial forms are implicated in idiopathic septicemia.
- Previous studies suggest CWD bacteria may infect erythrocytes.
Purpose of the Study:
- To investigate the presence and morphology of CWD bacterial forms in erythrocytes of patients with fever of unknown origin.
- To differentiate CWD bacterial forms from other intracellular inclusions using specific staining.
Main Methods:
- Blood samples were examined immediately after venipuncture.
- Gould's rhodamine-labeled muramidase staining was used to detect CWD forms.
- Blocking tests with unlabeled muramidase confirmed staining specificity.
- Comparison of CWD form incidence between patients (n=68) and controls (n=51).
Main Results:
- Morphologically characteristic CWD forms were observed within erythrocytes, often circumferential or protruding.
- Muramidase staining, confirmed by blocking tests, indicated a bacterial origin (presence of muramic acid).
- Higher incidence of intraerythrocytic CWD forms was found in patients compared to controls.
- Nearly 40% of patients showed significant erythrocyte parasitism, with some forms extending into rhizoid filaments.
Conclusions:
- In vivo CWD bacterial forms possess detectable muramic acid, confirming their bacterial nature.
- The presence of these CWD forms in erythrocytes is significantly higher in patients with fever of unknown origin.
- This study provides preliminary evidence for CWD bacteria as a potential cause of idiopathic septicemia.