Pneumococcal surface protein A (PspA) has been shown previously to elicit antibodies protective against pneumococcal infection and to be necessary for full pneumococcal virulence in mice. The protein was originally defined by the two mouse monoclonal antibodies Xi64 and Xi126, which together recognized PspA on 14% of pneumococcal isolates. Some PspA molecules reacted with both antibodies, but most reacted with only one or the other. In the present study we demonstrated that PspA is produced by all pneumococci, confirming our hypothesis that there are variants of PspA which are not detected by Xi64 and Xi126. We produced a rabbit antiserum and five additional monoclonal antibodies specific for PspA for these studies. The rabbit antiserum reacted with each of 95 pneumococcal isolated tested, comprising 16 capsular serotypes. One or more of the seven monoclonal anti-PspA antibodies reacted with 95% (53 of 57) of pneumococcal isolates tested. The specificity of the monoclonal and polyclonal antibodies to PspA was confirmed in two ways: (i) by detection of molecules on wild-type pneumococci that are identical in molecular weight to those detected in Western blots (immunoblots) with Xi64 and Xi126 and (ii) by the use of mutants of Streptococcus pneumoniae that fail to produce PspA or that produce a truncated form of PspA. By using the seven monoclonal antibodies, we observed 31 PspA types among the 57 isolates. When the 53 strains reactive with the monoclonal antibodies were analyzed by capsular type as well as by serologic type and molecular weight of PspA, we observed 50 different clonotypes of pneumococci.

The lesions observed in channel catfish (Ictalurus punctatus) as a consequence of exposure to Edwardsiella ictaluri via gut and water were studied. Channel catfish fingerlings were exposed to 1010 bacterial cells via the gut and placed in tanks with unexposed fish. Both groups of fish were sampled periodically over a 35-day period. The gut-exposed fish exhibited gross signs of a systemic infection within 2 wk. Enteritis, hepatitis, interstitial nephritis, and myositis, all of which began as acute reactions and developed into chronic lesions, were observed histologically. Some of the contact fish developed systemic infections beginning in the intestine, thus demonstrating horizontal transmission. However, only contact fish developed the characteristic "hole in the head" lesion, which began in the olfactory sac and progressed to a granulomatous meningioencephalitis. These results indicate that both gut and nares are sites of primary infection in natural outbreaks of E. ictaluri.

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A selective medium, called Edwardsiella ictaluri medium (EIM), has been formulated for the isolation of Edwardsiella ictaluri. The medium inhibits the growth of most gram-negative bacteria, except Proteus sp., Serratia marcescens and some isolates of Aeromonas hydrophila and Yersinia ruckeri. The bacteria that grow on the EIM are easily differentiated from E. ictaluri based on colony morphology. The EIM inhibits gram-positive bacteria with the exception of enterococci. The addition of fungizone to EIM suppressed the growth of most fungi. The EIM allows the evaluation of environmental reservoirs, levels of contamination and carrier states of E. ictaluri.

A total of 119 isolates of Edwardsiella ictaluri collected over the last 7 years from several states were biochemically characterized. Although not very reactive biochemically, this bacterium shows a high degree of homogeneity. Differences were found only in the production of gas from formate or from glucose at 37C and in the production of hydrogen sulfide as detected by lead acetate paper. Analysis of E. ictaluri by year or geographic area indicated that some differences existed, but no clear-cut biotypic variations were found. All isolates studied were capable of degrading chondroitin sulfate, a major component of cartilage, which may be an important virulence factor in the formation of the hole-in-the-head lesion characteristic of infected fish.

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