When phages infect bacteria cultured in the presence of sublethal doses of antibiotics, the sizes of the phage plaques are significantly increased. This phenomenon is known as phage-antibiotic synergy (PAS). In this study, the observation of PAS was extended to a wide variety of bacterium-phage pairs using different classes of antibiotics. PAS was shown in both Gram-positive and Gram-negative bacteria. Cells stressed with β-lactam antibiotics filamented or swelled extensively, resulting in an increase in phage production. PAS was also sometimes observed in the presence of other classes of antibiotics with or without bacterial filamentation. The addition of antibiotics induced expression in various bacteria, but a deletion mutant strain of also showed filamentation and PAS in the presence of quinolone antibiotics. The phage adsorption efficiency did not change in the presence of the antibiotics when the cell surfaces were enlarged as they filamented. Increases in the production of phage DNA and mRNAs encoding phage proteins were observed in these cells, with only a limited increase in protein production. The data suggest that PAS is the product of a prolonged period of particle assembly due to delayed lysis. The increase in the cell surface area far exceeded the increase in phage holin production in the filamented host cells, leading to a relatively limited availability of intracellular holins for aggregating and forming holes in the host membrane. Reactive oxygen species (ROS) stress also led to an increased production of phages, while heat stress resulted in only a limited increase in phage production. Phage-antibiotic synergy (PAS) has been reported for a decade, but the underlying mechanism has never been vigorously investigated. This study shows the presence of PAS from a variety of phage-bacterium-antibiotic pairings. We show that increased phage production resulted directly from a lysis delay caused by the relative shortage of holin in filamented bacterial hosts in the presence of sublethal concentrations of stress-inducing substances, such as antibiotics and reactive oxygen species (ROS).

No abstract

Enterococcus faecalis is a Gram-positive pathogen which colonizes human intestinal surfaces, forming biofilms, and demonstrates a high resistance to many antibiotics. Especially, antibiotics are less effective for eradicating biofilms and better alternatives are needed. In this study, we have isolated and characterized a bacteriophage, PBEF129, infecting E. faecalis. PBEF129 infected a variety of strains of E. faecalis, including those exhibiting antibiotic resistance. Its genome is a linear double-stranded DNA, 144,230 base pairs in length. Its GC content is 35.9%. The closest genomic DNA sequence was found in Enterococcus phage vB_EfaM_Ef2.3, with a sequence identity of 99.06% over 95% query coverage. Furthermore, 75 open reading frames (ORFs) were functionally annotated and five tRNA-encoding genes were found. ORF 6 was annotated as a phage endolysin having an L-acetylmuramoyl-l-alanine amidase activity. We purified the enzyme as a recombinant protein and confirmed its enzymatic activity. The endolysin’s host range was observed to be wider than its parent phage PBEF129. When applied to bacterial biofilm on the surface of in vitro cultured human intestinal cells, it demonstrated a removal efficacy of the same degree as cefotaxime, but much lower than its parent bacteriophage.

No abstract

BACKGROUND: Although antibiotics have contributed to treatment of bacterial infection, the antibiotic abuse can lead to antibiotic resistant bacteria. Impact of human activities on distribution of antibiotic resistance has been intensively issued and occurrence of antibiotic resistant bacteria in contaminated environments would not be a surprise. Nonetheless, anthropogenic contamination with the dissemination of antibiotic resistance along uncontaminated environments has been less considered. The aim of this study is to investigate antibiotic resistant bacteria across Ulleungdo, known as antibiotic resistance free and anthropogenic pollution free environment in Rep. of Korea. METHODS AND RESULTS: Antibiotic resistant bacteria in coastal seawater of Ulleungdo were investigated in July 2021. Antibiotic susceptibility test using the disk diffusion method was applied with six drugs according to the Clinical and Laboratory Standards Institute (CLSI) guideline. Total 43 bacterial isolates were tested and 20 isolates among of them showed multidrug resistance. Particularly, the number and ratio of resistant bacteria were relatively high in a densely populated area of Ulleungdo. The bacte-rial communities were investigated using 16S rRNA gene metabarcoding approach in the coastal seawater and soils of Ulleungdo. In the bacterial communities, Firmicutes were selectively distributed only in seawater, suggesting the possibility of anthropogenic contamination in coastal seawater of Ulleungdo. CONCLUSION(S): We found antibiotic resistant bacteria in a populated area of Ulleungdo. The occurrence of antibiotic resistant bacteria in Ulleungdo seems to result from the recent anthropogenic impact. Consistent monitoring of antibiotic resistant bacteria in the uncontaminated environment needs to considered for future risk assessment of antibiotics.