The WGS-based assignment of iGBS resistance features and serotypes is an accurate substitute for phenotypic testing.

Our whole genome sequence (WGS) pipeline was assessed for accurate prediction of antimicrobial phenotypes. For 2316 invasive pneumococcal isolates recovered during 2015 we compared WGS pipeline data to broth dilution testing (BDT) for 18 antimicrobials. For 11 antimicrobials categorical discrepancies were assigned when WGS-predicted MICs and BDT MICs predicted different categorizations for susceptibility, intermediate resistance or resistance, ranging from 0.9% (tetracycline) to 2.9% (amoxicillin). For β-lactam antibiotics, the occurrence of at least four-fold differences in MIC ranged from 0.2% (meropenem) to 1.0% (penicillin), although phenotypic retesting resolved 25%-78% of these discrepancies. Non-susceptibility to penicillin, predicted by penicillin-binding protein types, was 2.7% (non-meningitis criteria) and 23.8% (meningitis criteria). Other common resistance determinants included mef (475 isolates), ermB (191 isolates), ermB + mef (48 isolates), tetM (261 isolates) and cat (51 isolates). Additional accessory resistance genes (tetS, tet32, aphA-3, sat4) were rarely detected (one to three isolates). Rare core genome mutations conferring erythromycin-resistance included a two-codon rplD insertion (rplD69-KG-70) and the 23S rRNA A2061G substitution (six isolates). Intermediate cotrimoxazole-resistance was associated with one or two codon insertions within folP (238 isolates) or the folA I100L substitution (38 isolates), whereas full cotrimoxazole-resistance was attributed to alterations in both genes (172 isolates). The two levofloxacin-resistant isolates contained parC and/or gyrA mutations. Of 11 remaining isolates with moderately elevated MICs to both ciprofloxacin and levofloxacin, seven contained parC or gyrA mutations. The two rifampin-resistant isolates contained rpoB mutations. WGS-based antimicrobial phenotype prediction was an informative alternative to BDT for invasive pneumococci.

Summary.A diverse range of heterotrophic bacteria was screened for the presence of carbonic anhydrase (CA) activity, sensitivity to inhibition of growth by acetazolamide (CA inhibitor), and the presence of protein binding monospecific antibody prepared against purified Neisseria sicca CA. CA activity was demonstrated only in strains of N. sicca and N . gonorrhoeae. However, all Neisseria strains, including various isolates of N . meningitidis and N . lactamica, were sensitive to acetazolamide, when grown in air, and showed serological cross-reaction with N . sicca CA. Strains of other genera were resistant to acetazolamide. A number of strains including members of the genera Pseudomonas, Staphylococcus, Streptococcus, Serratia and Proteus also strongly expressed a gene product(s) immunologically related to CA. The presence of CA cross-reacting proteins, which lack hydrase activity, is discussed in relation to the function of the various mammalian CA isoenzymes.

SUlMMARYA defined medium is described for the growth of H-hilus i r J~a e and I?. puraifi$mae which can be used either as a liquid medium or solidified with agar.The purine and pyrimidine, vitamin, amino acid and mineral salt requirements for three strains have been analysed.

In the process of constructing a map of the pneumococcus chromosome by transformation, a search for linkage groups involving the markers ery-ra, str-rqr, amiA-rr, opt-ra and tet-A has been carried out. No linkage was found amongst the first four markers, but linkage was found between ery-ra and tet-A. The discrepancy between these results and the previously reported linkage between the loci conferring resistance to erythromycin and streptomycin is discussed. The anomalous position of the ery-ra-tet-A linkage in relation to the chromosomemap obtained by the density-shift method, and the implications to the mode of replication of the chromosome, are discussed. Two models are proposed for the replication of the pneumococcal chromosome.