Bacillus sphaericus strain P-1 has previously been shown to have a tetragonally arrayed (T layer) protein which forms the outer layer of the cell wall. The T layer was quantitatively extracted from whole cells by 6 M urea, and the T layer subunits were purified by electrophoresis of the extracts on acrylamide gels containing 0.1% sodium dodecyl sulfate or 6 M urea. Using ethylene diacrylate cross-linked gels, the T layer was found to make up 16% of the total cellular protein. A virulent bacteriophage which is inactivated by purified T layer was isolated from soil. Twenty-four phage-resistant mutants were isolated, of which 17 had T layer subunits of increased mobility on sodium dodecyl sulfate acrylamide gels. No mutants devoid of' T layer were found. Mutants were grouped into six classes according to the molecular weight of their T layer subunits. These ranged from that of the wild type, 150,000 down to 86,000. Two mutants from different classes were examined in detail. Cells of the mutant strains did not adsorb phage nor did cell walls isolated from these mutants inactivate phage. The amino acid composition of the T layers from mutants differed little from that of' the wild-type T layer. 1491

A medium for the single membrane enumeration of Escherichia coli and coliforms from potable water was developed by the modification of the standard UK membrane filtration medium. The medium, membrane‐Lactose Glucuronide Agar (m‐LGA), employs a chromogenic substrate for the detection of β‐glucuronidase activity and sodium pyruvate to enhance recovery of chlorine‐stressed coliforms. Escherichia coli identification was significantly improved on m‐LGA with 98.6% of presumptive isolates confirming. Recovery of coliforms from drinking water samples was also significantly improved.

Enzyme immunoassays for the detection of chlamydial antigens are commonly used to diagnose Chlamydia trachomatis infection. As is true for ail nonculture methods, the specificities of these tests are a concern. A confirmatory blocking assay (Abbott Laboratories, North Chicago, Ill.) was evaluated at four sexually transmitted disease test sites. This assay is designed to confirm true-positive Chlamydiazyme (CZ) specimens and to identify false-positive CZ reactions caused by cross-reacting bacteria. Cervical specimens were collected from 2,891 women. Chlamydia prevalence by tissue culture (TC) was 9.2% (266 of 2,891 specimens). Compared with TC, the sensitivity and specificity of CZ were 78.9% (210 of 266 specimens) and 98.2% (2,577 of 2,625 specimens), respectively. There were 48 CZ false-positive reactions. The direct fluorescent-antibody test (DFA) was positive for 31 of 48 false-positive reactions, indicating culture misses. Thus, when the standard was both TC and DFA, CZ sensitivity was 81.1% and CZ specificity was 99.3%. Of the 17 CZ-positive patients who were negative by both TC and DFA, 3 were negative on repeat CZ and Il of 14 were identified as false positive by the confirmatory assay. The confirmatory test was positive for CZ-positive women who were positive by TC or DFA. Use of the confirmatory test, which increased the specificity to 99.9%, would increase confidence in positive CZ results and make the test more useful for screening populations with a low prevalence of C. trachomatis infection.

A Streptococcus (Diplococcus) pneumoniae autolysin, partially purified from cellular autolysates, was optimally active at pH 7.0 and was stimulated by monovalent cations. Addition of autolysin to walls resulted in the appearance of only N-terminal L-alanine, whereas no glycosidase activity was observed. Walls which had been solubilized by autolysin were separated by gel filtration into a low-molecular-weight peptide containing amino acids in the same ratios found in intact walls and a high molecular fraction containing the amino acid-deficient peptidoglycan backbone. Thus, the major activity is an N-acetylmuramyl-L-alanine amidase. In addition, walls undergoing spontaneous lysis revealed no glycosidase activity but showed an increase in only N-terminal alanine. Autolysin, which was bound to walls in saline, was almost completely removed when walls were washed in distilled water, and all of the activity was recovered in the water wash fluid.

No abstract