Host receptor identification of a polyvalent lytic phage GSP044, and preliminary assessment of its efficacy in the clearance of Salmonella.

Salmonella and pathogenic Escherichia coli are important foodborne pathogens. Phages are being recognized as potential antibacterial agents to control foodborne pathogens. In the current study, a polyvalent broad-spectrum phage, GSP044, was isolated from pig farm sewage. It can simultaneously lyse many different serotypes of Salmonella and E. coli, exhibiting a broad host range. Using S. Enteritidis SE006 as the host bacterium, phage GSP044 was further characterized. GSP044 has a short latent period (10 min), high stability at different temperatures and pH, and good tolerance to chloroform. Genome sequencing analysis revealed that GSP044 has a double-stranded DNA (dsDNA) genome consisting of 110,563 bp with G + C content of 39%, and phylogenetic analysis of the terminase large subunit confirmed that GSP044 belonged to the Demerecviridae family, Epseptimavirus genus. In addition, the genomic sequence did not contain any lysogenicity-related, virulence-related, or antibiotic resistance-related genes. Analysis of phage-targeted host receptors revealed that the outer membrane protein (OMP) BtuB was identified as a required receptor for phage infection of host bacteria. The initial application capability of phage GSP044 was assessed using S. Enteritidis SE006. Phage GSP044 could effectively reduce biofilm formation and degrade the mature biofilm in vitro. Moreover, GSP044 significantly decreased the viable counts of artificially contaminated S. Enteritidis in chicken feed and drinking water. In vivo tests, a mouse model of intestinal infection demonstrated that phage GSP044 was able to reduce the number of colonized S. Enteritidis in the intestine. These results suggest that phage GSP044 may be a promising candidate biologic agent for controlling Salmonella infections.

Fitness Trade-Offs in Phage Cocktail-Resistant Salmonella enterica Serovar Enteritidis Results in Increased Antibiotic Susceptibility and Reduced Virulence.

The rapid emergence of phage-resistant bacterial mutants is a major challenge for phage therapy. Phage cocktails have been considered one approach to mitigate this issue. However, the synergistic effect of randomly selected phages in the cocktails is ambiguous. Here, we rationally designed a phage cocktail consisting of four phages that utilize the lipopolysaccharide (LPS) O antigen, the LPS outer core, the LPS inner core, and the outer membrane proteins BtuB and TolC on the Salmonella enterica serovar Enteritidis cell surface as receptors. We demonstrated that the four-phage cocktail could significantly delay the emergence of phage-resistant bacterial mutants compared to the single phage. To investigate the fitness costs associated with phage resistance, we characterized a total of 80 bacterial mutants resistant to a single phage or the four-phage cocktail. We observed that mutants resistant to the four-phage cocktail were more sensitive to several antibiotics than the single-phage-resistant mutants. In addition, all mutants resistant to the four-phage cocktail had significantly reduced virulence compared to wild-type strains. Our mouse model of Salmonella Enteritidis infection also indicated that the four-phage cocktail exhibited an enhanced therapeutic effect. Together, our work demonstrates an efficient strategy to design phage cocktails by combining phages with different bacterial receptors, which can steer the evolution of phage-resistant strains toward clinically exploitable phenotypes. IMPORTANCE The selection pressure of phage promotes bacterial mutation, which results in a fitness cost. Such fitness trade-offs are related to the host receptor of the phage; therefore, we can utilize knowledge of bacterial receptors used by phages as a criterion for designing phage cocktails. Here, we evaluated the efficacy of a phage cocktail made up of phages that target four different receptors on Salmonella Enteritidis through in vivo and in vitro experiments. Importantly, we found that pressure from phage cocktails with different receptors can drive phage-resistant bacterial mutants to evolve in a direction that entails more severe fitness costs, resulting in reduced virulence and increased susceptibility to antibiotics. These findings suggest that phage cocktail therapy using combinations of phages targeting different important receptors (e.g., LPS or the efflux pump AcrAB-TolC) on the host surface can steer the host bacteria toward more detrimental surface mutations than single-phage therapy, resulting in more favorable therapeutic outcomes.

[The turbidity and pH impact analysis of low concentration water chemical oxygen demand ultraviolet absorption detection].

Configuration standard solution in the concentration range of 1 - 25 mg x L(-1) of potassium hydrogen phthalate was used as experimental subject, Ultraviolet absorption spectra was collected, the COD quantitative analysis model was established by partial least squares with different pretreatment methods and the turbidity of the compensation effect analysis was given. The results show the model uses smoothing first derivative pretreatment method, internal cross validation RMSECV root mean square value of 0.122 27, principal component number 4, the square of the prediction model correlation coefficient is 0.999 8, and the relative prediction error is in the range of 0.03%-1.7%; for 0-100 NTU's turbidity solution, the relative standard deviation RSD is 2.3% after compensation; with pH in the range of 3-10, influence can be ignored.