Penicillin-Binding Protein Gene Alterations in Streptococcus uberis Isolates Presenting Decreased Susceptibility to Penicillin

Abstract

Streptococcus uberis is an environmental pathogen commonly causing bovine mastitis, an infection that is generally treated with penicillin G. No field case of true penicillin-resistant S. uberis (MIC > 16 mg/liter) has been described yet, but isolates presenting decreased susceptibility (MIC of 0.25 to 0.5 mg/liter) to this drug are regularly reported to our laboratory. In this study, we demonstrated that S. uberis can readily develop penicillin resistance in laboratory-evolved mutants. The molecular mechanism of resistance (acquisition of mutations in penicillin-binding protein 1A [PBP1A], PBP2B, and PBP2X) was generally similar to that of all other penicillin-resistant streptococci described so far. In addition, it was also specific to S. uberis in that independent resistant mutants carried a unique set of seven consensus mutations, of which only one (Q 554 E in PBP2X) was commonly found in other streptococci. In parallel, independent isolates from bovine mastitis with different geographical origins (France, Holland, and Switzerland) and presenting a decreased susceptibility to penicillin were characterized. No mosaic PBPs were detected, but they all presented mutations identical to the one found in the laboratory-evolved mutants. This indicates that penicillin resistance development in S. uberis might follow a stringent pathway that would explain, in addition to the ecological niche of this pathogen, why naturally occurring resistances are still rare. In addition, this study shows that there is a reservoir of mutated PBPs in animals, which might be exchanged with other streptococci, such as Streptococcus agalactiae, that could potentially be transmitted to humans.Penicillin resistance has been particularly well studied for the human pathogen Streptococcus pneumoniae (5, 15). In this organism, resistance occurs through modifications of penicillin-binding proteins (PBPs), leading to a decreased affinity for the drug. These modifications include mutations and/or mosaics in PBP2X and PBP2B, as well as in PBP1A for the highly resistant isolates (20). Several studies also indicated that full expression of resistance necessitates mutations both in the pbp genes and in other genes, of which only a few have been determined, namely, in the ciaRH, cpoA, and murMN loci (8,9,14). The same mechanism has also been found in viridians streptococci, which are occasionally responsible for human infections (18,26). Very recent reports also demonstrated the implication of PBP mutations in S. agalactiae presenting a decreased susceptibility to penicillin (4, 17, 21). In contrast, the highly pathogenic Streptococcus pyogenes, which has been widely exposed to penicillin for decades, is apparently incapable of acquiring clinical resistance in vivo (22), even though laboratory mutants presenting decreased susceptibilities were reported (11). Thus, the capacity to develop penicillin resistance varies among different bacteria belonging to the same genus.S. pneumoniae is primarily a human pathogen, while S. agalactiae infects humans and ...

Novel Mutations in a Patient Isolate of Streptococcus agalactiae with Reduced Penicillin Susceptibility Emerging after Long-Term Oral Suppressive Therapy

“…2). The equilibrium inhibitor dissociation constant (K i ) is in the micromolar range and comparable to that reported previously (8,12,18). These data suggest that both PBP1a/1b and PBP2a/2b/2x mutations are contributing to reduced susceptibility.…”

Demonstration of a shift towards penicillin resistance in the Streptococcus uberis population

“…In France, the CA-SFM recommends that penicillin resistance is suspected for all non-pneumococcal streptococci isolated from animals when a decreased susceptibility to 5 mg oxacillin is detected by disc diffusion (resistant if ,21 mm). Since it was recently shown that S. uberis can develop resistance to penicillin in vitro (Haenni et al, 2010), and since isolates presenting decreased susceptibility have already been reported (Guerin-Faublee et al, 2002;Mevius et al, 2007;Rossitto et al, 2002), this work intended to further characterize field isolates classified as either oxacillin resistant or oxacillin sensitive, in order to get a better insight of the potential population shift towards penicillin resistance in S. uberis.…”

Misidentification of Streptococcus uberis as a Human Pathogen: A Case Report and Literature Review

“…10 Invitro experiments demonstrated that S. uberis can readily develop penicillin resistance, and microbial analysis of bacterial population in treated milk showed that S. uberis can grow even in cold storage. 11,12 It has been hypothesized that the stability of this pathogen under various environmental conditions and the expression of virulence factors 11-13 may expand the pathogenicity of this bacterium from cattle to humans as described for Streptococcus agalactiae, a pathogen that is associated with both animal and human diseases. 14 However, few data provide support for this hypothesis, and in many cases the tests used for the identification of S. uberis in humans have been debated.…”