The effect of formalin-killing of Pasteurella multocida on the antigenicity and extractability of its lipopolysaccharide.

The extraction of lipopolysaccharides (LPS) from formalin-killed (FK) Pasteurella multocida strain X-73 and from cells not exposed to formalin (NF) were compared by the Westphal and phenol-chloroform-petroleum ether (PCP) extraction procedures. The LPS was determined by: (1) serologic analyses with antiserum specific for LPS; (2) analyses for toxicity; and (3) chemical analyses for components expected to be in LPS (such as hexoses, heptoses, amino sugars, 3-deoxyoctulosonic acid, and fatty acids). Strain X-73, the strain most virulent for chickens, was markedly affected by formalin killing. Unlike many strains, which readily yield LPS into the aqueous phase when extracted with phenol at 68 degrees by the Westphal procedure, strain X-73 did so only with FK and not with NF cells. With the NF cells, LPS was extracted by EDTA from the precipitate obtained during the Westphal procedure. With the PCP procedure, LPS was extracted readily from NF cells, but not from FK cells. The change in extractability of LPS as a result of formalin-killing was the same for both the encapsulated form of X-73 and a nonencapsulated variant derived from it. Although formalin-killing affected the extractability of LPS, no antigenic differences could be detected by immunodiffusion. However, the chick-embryo toxicity of LPS extracted from NF cells was greater than that of LPS from FK cells.