Anaerobic Infections of War Wounds in the Middle East

As has been shown in previous reports from this laboratory (10, 25, 75), toxic effects resembling traumatic shock have been observed following the intravenous injection of fluids exuding from-dog muscles after prolonged ischemia.The fact that 28 per cent of these fluids were highly toxic, whereas the remainder seemed harmless, strongly suggested that their toxic effects were not due to the presence of intracellular substances lost as the result of cell damage (70). For the same reason, it appeared doubtful that the occasional toxic effects were caused by breakdown products of muscle cell constituents resulting from ischemia, since the experimental conditions were all approximately identical. Attention has been directed, therefore, to the one constituent of these fluids which was obviously highly variable, namely, the bacterial flora.Another bit of indirect evidence pointed in this direction. On several occasions, dog muscle tissue was excised and minced with sterile precautions and incubated in sterile dog plasma at 370 C., for 5 hours. The plasma was then separated by centrifugation and injected intravenously into recipient dogs. Such material was invariably toxic, rapidly producing a shock-like state and death, and it was always infected with micro-organisms which on direct smear were seen to be large gram-positive rods. However, if a mixture of blended dog muscle and plasma were sterilized by passage through a Seitz filter before incubation, of the fluid to be tested were mixed with 4.5 ml. of beef broth, and 3 successive 1: 10 dilutions in broth were made. Two blood agar plates were then divided into quadrants and a loopful (approximately 1I4.) of each dilution was streaked onto 1 quadrant of each plate. One of these was incubated aerobically and the other in an atmosphere of 5 per cent CO2 in nitrogen. After 36 hours of incubation at 37°C., differential colony counts were made and identification of the bacterial species present was undertaken. Colony counts were multiplied by the appropriate factors to give numbers of viable bacteria per ml. of original fluid, and these were checked qualitatively with observations on direct smears. Clostridia were isolated and cultivated from various fluids.In an attempt to reproduce at will the picture of toxicity following fluid injection, 2 strains of Clostridia perfringens thus isolated were used in 3 experiments. In these experiments, at the time of muscle ligation, 2 to 5 ml. of a broth culture of these organisms were injected into each of the 2 muscle groups; after the period of occlusion of the blood supply the ligatures were released as usual and fluid collected. These have been termed "reinforced" fluids. Bacterial counts were made on the first fluid obtained from each leg, on a final sample drain-856

Section: Discussionmentioning

confidence: 90%

The Toxic Factors in Exerimental Traumatic Shock. Vi. The Toxic Influence of the Bacterial Flora, Particularly Clostridium Welchii, in Exudates of Ischemic Muscle 1

Pope¹,

Zamecnik²,

Aub³

et al. 1945

“…This correlates with the observation that edema into the injected area becomes the outstanding feature. The clinical picture which this toxin produces when injected intravenously is that of shockapparently as a consequence of 2 factors: (1) trapping of whole blood in the viscera, rendering it unavailable to the circulation as a whole; and (2) extravasation of plasma and whole blood into vital organs through damaged capillaries. Addition of fluid to the circulatory system under these conditions does not restore the animal to normal, although it has a temporary beneficial effect until the additional fluid has likewise been lost from the circulation.…”

Section: Discussionmentioning

confidence: 99%

“…oedematiens in war wounds in general and in gas gangrene in particular (1) has stimulated the study of the physiologic effects of the toxin of this organism. The availability of Cl.…”

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confidence: 99%

Physiologic Action of Clostridium Oedematiens (Novyi) Toxin in Dogs 123

Aub¹,

Zamecnik²,

Nathanson³

1947

The frequent occurrence of Cl. oedematiens in war wounds in general and in gas gangrene in particular (1) has stimulated the study of the physiologic effects of the toxin of this organism. The availability of Cl. oedematiens toxin in purified form has made it possible to investigate its action as distinct from toxins of other synergistic and symbiotic organisms which contribute to the clinical picture of anaerobic infections. Such a study is herein reported.The pathologic effects of Cl. oedematiens were described first by Novy (2); and, subsequently, on the same or closely related organisms, by Weinberg and Seguin (3), and others (4 to 6). Previous studies of the physiologic effects of this toxin have not, however, reached our attention. It has not been demonstrated that more than one toxin is produced by the organism, although analogy with Cl. welchii makes this a possibility.Recent work from this laboratory (7 to 13) has shown that the presence of clostridia may constitute an important factor in some forms of traumatic shock produced experimentally in dogs. The present communication points out that a picture similar to that of traumatic shock may be produced experimentally in dogs by the injection of Cl. oedematiens toxin. MATERIALS AND METHODSThe experiments have been carried out using several preparations of purified Cl. oedematiens (Novyi) The methods used in this study have been described previously (14), with the following exceptions: liver function was tested by the bromsulfalein excretion method; and kidney function was tested by phenolsulfonphthalein excretion. RESULTS Cl. oedematiens intravenouslyDosage: Thirty-five dogs were injected intravenously with a dosage of toxin which ranged from 3,700 down to 20 MLD per kgm. Of 9 unanesthetized dogs simply injected with varying amounts of toxin and then observed, there was only 1 survivor, the animal receiving the 20 MLD per kgm. dose. Dogs given 46 and 90 MLD per kgm. died on the third day. With larger doses of toxin (over 500 MLD per kgm.) the survival time was uniformly shortened to a matter of hours.Twenty-six dogs were anesthetized and, following carotid and tracheal cannulation, were injected intravenously with toxin. The majority of these animals were given 1,250 MLD per kgm., and only 3 out of the 26 survived longer than 8 hours.Rate of injection: There was no great differ-. ence in the effect of the toxin whether it was given rapidly in small volume (less than 10 ml.) by syringe, or diluted to 100 ml. and allowed to drip into the animal by slow infusion over a period of hours. The survival time following slow infusion was an hour or two longer, and the decline of the blood pressure more gradual, but otherwise the picture was the same by either method.

“…Moreover, our previous experiments have suggested that these and perhaps other bacteria may be responsible for a toxic factor contributing to shock in traumatizing wounds. Since clostridia are known to be frequent contaminants of war wounds (8,9) it appeared desirable to study the physiologic action of clostridial toxins in order to determine whether these toxins could produce shock 4 alone or in synergy with blood or plasma loss following trauma.…”

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confidence: 99%

“…2 The work described in this paper was done under a contract, recommended by the Committee on Medical Research, between the Office of Scientific Research and Development and the Massachusetts General Hospital. 8 Grateful acknowledgment is due the Josiah Macy, Jr. Foundation for a grant for the histopathological studies.…”

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confidence: 99%

Physiologic Action of Clostridium Welchii (Type A) Toxins in Dogs 123

Zamecnik¹,

Nathanson²,

Aub³

1947

Recent work reported from this laboratory (1 to 7) has shown that clostridia grow and produce toxins readily in anoxic muscle in zivo, and that they are readily introduced into dog muscles during the course of traumatizing procedures. Moreover, our previous experiments have suggested that these and perhaps other bacteria may be responsible for a toxic factor contributing to shock in traumatizing wounds. Since clostridia are known to be frequent contaminants of war wounds (8, 9) it appeared desirable to study the physiologic action of clostridial toxins in order to determine whether these toxins could produce shock 4 alone or in synergy with blood or plasma loss following trauma.Since the action of these toxins depends on the amount used, the route of injection, and the rate of injection, we have given attention to these variables. Wright (12) has criticized studies on the pharmacologic effects of Cl. welchii toxins in which large doses of toxin were administered rapidly, in a manner unlikely to have a parallel in the' clinical disease. Study of the effects of the Cl. welchii toxic filtrate has a particular interest in that the specific enzymatic nature of its probable principal component (the alpha toxin or lecithinase) (13)