Improved method of isolating brucellosis organisms from blood clots utilizing the membrane filter

The Millipore type of membrane filter was used to culture a Brucella abortus species from artificailly infected rabbit blood clots. Attempts were made to isolate naturally occurring species of these organisms from both human and animal bloods on this filter. The blood clots were treated with both physical and enzymatic methods in an attempt to prepare a liquid that would pass through the membrane filter. The results of blood degradation with distilled water, dilute acids and sand-glass beads-distilled water were found to be poor when attempts were made to filter the supernatants prepared by these methods. There was no reisolation when the clot was artifically inoculated with Brucella species when treated by physical and chemical methods. The use of fibrinolysin (streptokinase) to dissolve the rabbit blood clots produced excellent results. Both the digestion of the clot and the production of a filterable supernatant that would pass through the membrane were obtained. Recovery of the Brucella species artificially inoculated into these enzymatically treated clots was made. No positive isolations of Brucella species were made when this streptokinase Brucella isolation technique was applied to unknown animal and human bloods suspected of harboring this organism. A variety of organisms were islated, most of which were probably contaminants introduced inot the blood before being received at this laboratory.