With widespread abuse of antibiotics, bacterial resistance has increasingly become a serious threat. Acinetobacter baumannii has emerged as one of the most important hospital-acquired pathogens worldwide. Bacteriophages (also called “phages”) could be used as a potential alternative therapy to meet the challenges posed by such pathogens. Endolysins from phages have also been attracting increasing interest as potential antimicrobial agents. Here, we isolated 14 phages against A. baumannii, determined the lytic spectrum of each phage, and selected one with a relatively broad host range, named vB_AbaP_PD-6A3 (PD-6A3 for short), for its biological characteristics. We over-expressed and purified the endolysin (Ply6A3) from this phage and tested its biological characteristics. The PD-6A3 is a novel phage, which can kill 32.4% (179/552) of clinical multidrug resistant A. baumannii (MDRAB) isolates. Interestingly, in vitro, this endolysin could not only inhibit A. baumannii, but also that of other strains, such as Escherichia coli and methicillin-resistant Staphylococcus aureus (MRSA). We found that lethal A. baumannii sepsis mice could be effectively rescued in vivo by phage PD-6A3 and endolysin Ply6A3 intraperitoneal injection. These characteristics reveal the promising potential of phage PD-6A3 and endolysin Ply6A3 as attractive candidates for the control of A. baumannii-associated nosocomial infections.

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In our previous research, 3-methyl-butanal and 3-methyl-butanoic acid were identified as representative and specific volatile organic compounds released by Staphylococcus aureus in broth. In this study, we explored the production of the 2 volatiles and their correlation to Staph. aureus growth in milk under different conditions. We found significant correlations between the production of 3-methyl-butanoic acid and cell counts of 5 Staph. aureus strains in sterile milk, and there were no obvious differences for its production among 5 tested strains. The intensities of the 2 volatiles were similar and positively correlated with bacterial counts in cultures at 25°C and 37°C despite delayed production of volatiles at 25°C; however, neither compound could be detected at 4°C. The production of 3-methyl-butanoic acid was strongly correlated with growth of Staph. aureus mixed with Streptococcus agalactiae, Escherichia coli O157:H7, and Shigella flexneri, whereas correlations for 3-methyl-butanal were not statistically significant. Compared with the monoculture of Staph. aureus, in mixed cultures, production of 3-methyl-butanal was decreased and that of 3-methyl-butanoic acid was comparatively higher. In pasteurized and raw milks, production of 3-methyl-butanoic acid was correlated with growth of Staph. aureus, and 3-methyl-butanoic acid could be detected when Staph. aureus populations reached 10 to 10 cfu/mL in pasteurized milk and 10 to 10 cfu/mL in raw milk; the correlations for 3-methyl-butanal were not statistically significant. Our results suggest that 3-methyl-butanoic acid is a more suitable marker for high counts of Staph. aureus in milk, whereas 3-methyl-butanal is a transient metabolite and easily depressed by environmental factors.

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