The Red Blood Cell: An Essential Component of the Toxicity of Strangulation Intestinal Obstruction

Abstract

It has been demonstrated by several investigators that the peritoneal exudate from dogs which have a strangulated loop of intestine is lethal when given to normal animals( 1.2). Such fluid is not always fatal and we have attempted to find out why.Our data from many dogs and rats revealed that 2 elements were constantly present in any strangulation fluid that was subsequently lethal to normal recipients: an E. coli level of at least 10'' bacteria/ml and a hemogobin level of et least 3 g %. It did not appear that the presence of other bacteria regardless of concentration made any detectable difference. It did appear, however, that i l f the E . coli level fell below that stated, or if the hemoglobin level fell below 3 g c/b the fluid began to lose its lethal effect. The following experiments were designed to test this hypothesis.Methods. Ff the above hypothesis is correct it should be possible to manufacture this lethal fluid in the laboratory. The following groups of experiments were done and the results checked by intraperitoneal injection of the fluid into the rat in a dose of 5 ml/kg. This is the same quantity af fluid used in our earlier experiments. All studies were carried out in male rats of the Sprague-Dawley strain and weighing 200-250 g.I. The first group of 45 rats were given 5 ml/kg of a 24-hour broth culture of E . coli. The inoculum was adjusted to contain 1 X 108 organisms/ml. 11.The second group of 15 rats was given whole blood intraperitoneally 5 ml/kg. 111. The third group of 29 rats was given an intraperitoneal injection of E . coli plus whole blood. E. coli were grown in nutrient broth for 24 hours in presence of added whole blood so that hemoglobin concentration was at least 4 g %.TV. The fourth group of 40 rats was given an I.P. injection of los E . coli grown in nutrient broth to which had been added rat plasma. The plasma was that amount present in the blood with a hemoglobin concentration of 4 g %. The amount of whole blood necessary to achieve this level was determined, then centrifuged and the resultant supernate added to the broth culture.V. The fifth group of 174 rats received an 1.P. injection of E . coli (10s) in nutrient broth to which 4 g % of hemoglobin had been added in the form af washed red cells (red blood cells were separated by centrifugation and washed 3 times with d i n e sdution). VI. The final group of 10 rats were given an I.P. injection identical to group V except that Clostridia welchii replaced E . coli.The number of bacteria injected was estimated in a Coleman Jr. spectrolphotometer at 660 mp. The actual number of viable bacteria injected was determined by subsequent dilution and plating of an aliquot of the injected culture. Quantitative counts were performed in a Quebec colony counter at 24 hours.Hemoglobin level was determined using the cyanmethemoglobin technique read in a Klett colorimeter using a 420 filter.Results. The results are summarized in Table I. When E. coli or the hernoglobin solutions were given independently of each other there was no lethal effect...

Free hemoglobin concentration in severe sepsis: methods of measurement and prediction of outcome

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High Susceptibility of Strain A Mice to Endotoxin and Endotoxin-Red Blood Cell Mixtures

“…A further experiment showed that this lethal effect was produced only when the vaccine contained both SRBC and killed salmonellae. It appeared possible that this phenomenon was related to the earlier findings of Davis and Yull (1961) who, in attempting to reproduce experimentally the symptoms of shock following intestinal obstruction, detected a synergistic lethality between hemoglobin and living Escherichia coli in Sprague-Dawley rats (Davis and Yull, 1961). A study was undertaken to elucidate the present observation.…”

Enhancement of Experimental Anaerobic Infections by Blood, Hemoglobin, and Hemostatic Agents

“…Hemostatic agents, hemoglobin, and whole blood have been shown previously to alter the infectivity of certain other microorganisms. Hemoglobin enhanced infections with E. coli (1,3,5,9,10,12,34,35) and C. perfringens (4, 33). The infection-enhancing effect of hemoglobin was found to be due to ferric ions and could be duplicated with ferric ammonium citrate (2,5,33).…”