Biological Activities of Crude Polysaccharide Extracted from Two Different Immunotype Strains of Hemophilus gallinarum in Chickens

Abstract

Biological activities in chickens of crude polysaccharide extracted from two different immunotype strains of Hemophilus gallinarum (HG), strain 221 and S1, were studied to clarify a type-specific protecting antigen and a toxin. Crude polysaccharide materials extracted from strains 221 and S1 were not only protective but toxic to chickens. They also contained at least two heat-labile antigens. When the polysaccharide materials were subjected to gel filtration on Sepharose 6B by tracing at 280 and 490 nm, the protective and toxic activities could be fractionated as peak-1 and -2 polysaccharides, respectively. The fraction of peak-1 polysaccharides from strains 221 and S1 was eluted at void volume. Cross-protection was not found between strains 221 and S1 in the fraction of peak-1 polysaccharide. Hemagglutination-inhibition (HI) antibody to type 1 hemagglutinin of HG (trypsin-sensitive hemagglutinin) was detected in chickens immunized with this fraction from strain 221 but not with that from strain S1. Type specificity between both strains was also found in this fraction by agar-gel precipitation (AGP) test. On the other hand, the toxic fraction of peak-2 polysaccharide, which caused hydropericardium in chickens, had a lower molecular weight than did that of peak-1 polysaccharide. It did not give HI antibody in chickens. Common antigenicity between strains 221 and S1 was found in the peak-2 polysaccharide by AGP test.

Latex Agglutination Test for Measurement of Type-Specific Antibody to Haemophilus paragallinarum in Chickens

“…Crude polysaccharide including typespecific antigen was extracted from H. paragallinarum strain S1, serotype C, and strain 221, serotype A, according to the method of Iritani et al (7). Briefly, 24-hr-old cultured H. paragallinarum cells in chicken meat infusion broth were disrupted with a cell press, and absolute ethanol was added to the supernatant to remove the nucleic acids.…”

“…Recently, in this laboratory, crude polysaccharide antigens were extracted from H. paragallinarum serotypes A and C (7). The polysaccharides were found to contain type-specific antigens.…”

Vaccines against infectious coryza

“…The LPS of H. paragallinarum has been shown to be lethal for embryonated chicken eggs and to be positive ir a pyrogen test (Konno and Nakase, 1977). Iritani et al (1981) have also shown that a crudely purified polysaccharide extract contains a component that is toxic and causes hydropericardium following intravenous inoculation into chickens. Any records of adverse side reactions will be noted with the various adjuvant systems described below.…”

“…Sawata et al (1979) suggested that the capsule of H. paragallinarum contains protective antigens. A crude polysaccharide has also been reported to provide protection that is limited to the Page serovar on which the vaccine is based (Iritani et al, 1981).…”

“…It has been suggested that adverse reactions are associated with either the adjuvant used or the lipopolysaccharide (LPS) present in the H. paragallinarum cells (Iritani et al, 1981). The LPS of H. paragallinarum has been shown to be lethal for embryonated chicken eggs and to be positive ir a pyrogen test (Konno and Nakase, 1977).…”