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bioRxiv2026

Survival of Streptococcus anginosus in blood is linked to hydrogen peroxide production

Streptococcus anginosus is an opportunistic pathogen causing bacteremia, respiratory tract infections, abscesses, odontogenic infections and has recently been associated with gastric cancer. While numerous genes encoding putative virulence factors have been detected in whole genome sequences, a detailed molecular characterization is missing for most of them. To address putative virulence determinants, we investigated hydrogen peroxide production by S. anginosus and assessed its functional significance for survival in human blood. In Streptococcus pneumoniae the pyruvate oxidase, encoded by the spxB gene, is responsible for H2O2 production and represents a major virulence factor. Interestingly, S. anginosus harbors two genetic variants of the spxB gene, one with high homology to the S. pneumoniae spxB (sequence identity: approx. 94 %) gene, while the other appears to be specific for S. anginosus. Strains carrying the S. pneumoniae variant demonstrate higher H2O2 production (10-20 M) compared to strains harboring the anginosus variant (<1 M). Incubation of S. anginosus strains in human blood showed that bacterial survival was correlated to the amount of hydrogen peroxide production. Assessing H2O2 production in an S. anginosus strain carrying a deletion of the CcpA regulator revealed the control of spxB and H2O2 production by carbon catabolite repression. In conclusion, we found evidence for the presence of a major S. pneumoniae virulence factor in S. anginosus, and we were able to demonstrate that it increases the ability of S. anginosus to survive in human blood.

microbiology↗

bioRxiv2026

Fluoroquinolone-Triggered Prophage Induction in Streptococcus anginosus Reveals Lytic Cycle, CRISPR-Prophage Interplay, and the potential for Cross-Species Horizontal Gene Transfer

Streptococcus anginosus (S. anginosus) has long been considered a commensal of the human microbiome but is increasingly associated with invasive infections and malignant processes. For understanding evolutionary dynamics, it is essential to investigate its mobile genetic elements, such as prophages, which are known to impact virulence, antibiotic resistance, and horizontal gene transfer. While many S. anginosus strains carry prophages, lysogen induction by external stimuli has not been demonstrated, and phage-mediated infection or lysis of this species has not been reported. To analyze the prevalence and diversity of prophages in S. anginosus genomes, we screened 140 clinical isolates by PCR revealing that 31.4% of strains were lysogenic. Correlating these findings with the presence of CRISPR immunity, we observed that S. anginosus strains carrying a CRISPR-Cas type II-A system were less likely to harbor prophages. Using a PCR-based approach, the spontaneous excision of several prophages of S. anginosus could be demonstrated and a fluoroquinolone-triggered prophage induction could successfully be established. Induction by ciprofloxacin and levofloxacin resulted in significant, concentration-dependent phage release and bacterial lysis. Transmission electron microscopy revealed viruses exhibiting the morphology characteristic of siphoviruses. Further analysis of the susceptibility of S. anginosus isolates and other oral and pyogenic streptococci to the isolated S. anginosus phages demonstrated a broad host range and the potential for cross-species horizontal gene transfer. In conclusion, a lytic cycle of S. anginosus phages could be induced, highlighting their functional relevance to pathogenicity and horizontal gene transfer, while demonstrating potential clinical implications of antibiotic-mediated prophage activation.

microbiology↗