Purpose Escherichia coli O157:H7 is one of the major foodborne pathogens of global public concern. Bacteriophages (phages) have emerged as a promising alternative to antibiotics for controlling pathogenic bacteria. Here, a lytic E. coli O157:H7-specific phage (KFS-EC) was isolated, identified, and characterized to evaluate its potential as a biocontrol agent for E. coli O157:H7. Methods KFS-EC was isolated from slaughterhouse in Korea. Morphological analysis, genomic analysis and several physiological tests were performed to identify and characterize the KFS-EC. Results A specificity test indicated KFS-EC was strictly specific to E. coli O157:H7 strains among 60 bacterial strains tested. Morphological and phylogenetic analyses confirmed that KFS-EC belongs to the Rb49virus genus, Tevenvirinae subfamily, and the Myoviridae family of phages. KFS-EC genome consists of 164,725 bp and a total of 270 coding sequence features, of which 114 open reading frames (ORFs) were identified as phage functional genes. KFS-EC does not contain genes encoding lysogenic property and pathogenicity, which ensure its safe application. KFS-EC was relatively stable (~1 log decrease) under stressed conditions such as temperatures (20 °C-50 °C), pHs (3-11), organic solvents (ethanol and chloroform), and biocides (0.1% citric acid, 1% citric acid, and 0.1% peracetic acid). KFS-EC was able to inhibit E. coli O157:H7 efficiently at a multiplicity of infection (MOI) of 0.01 for 8 h with greater inhibitory effect and durability and was stable at 4 °C and 22 °C over a 12-week storage period. Conclusions Our results suggest that KFS-EC could be used as a biocontrol agent to E. coli O157:H7.

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This study investigated the feasibility of the lytic, tailed Bacillus cereus-specific phage for use in a ferromagnetoelastic (FME) biosensor as a novel recognition element. The phage was immobilized at various concentrations through either direct adsorption or a combination of 11-mercapto-1-undecanoic acid (11-MUA) and [N-(3-dimethylaminopropyl)-N'-carbodiimide hydrochloride and N-hydroxysuccinimide (EDC/NHS)]. The effects of time and temperature on its lytic properties were investigated through the exposure of B. cereus (4 and 8 logCFU/ml) to the phage (8 logPFU/ml) for various incubation periods at 22°C and at various temperatures for 30 and 60 min. As the phage concentration increased, both immobilization methods also significantly increased the phage density (p < 0.05). SEM images confirmed that the phage density on the FME platform corresponded to the increased phage concentration. As the combination of 11-MUA and EDC/NHS enhanced the phage density and orientation by up to 4.3-fold, it was selected for use. When various incubation was conducted, no significant differences were observed in the survival rate of B. cereus within 30 min, which was in contrast to the significant decreases observed at 45 and 60 min (p < 0.05). In addition, temperature exerted no significant effects on the survival rate across the entire temperature range. This study demonstrated the feasibility of the lytic, tailed B. cereus-specific phage as a novel recognition element for use in an FME biosensor. Thus, the phage could be placed on the surface of foods for at least 30 min without any significant loss of B. cereus, as a result of the inherent lytic activity of the B. cereus-specific phage as a novel recognition element.

Yersinia enterocolitica is a gram-negative, non-spore-forming, coccobacilli, psychrotrophic, and facultative anaerobe, which is one of three Yersinia species that are pathogenic to humans, along with Y. pestis, and Y. pseudotuberculosis [1, 2]. The most predominant natural hosts for Y. enterocolitica are animals (especially pigs). Y. enterocolitica also exists ubiquitously in water, soil, plant surfaces, and foods [3]. Although animals are the major source of Y. enterocolitica, many cases have recently been reported in which outbreaks of Y. enterocolitica were associated with fresh produce such as salad, bean sprouts and leafy vegetables [4, 5]. Yersinia infection, commonly known as yersiniosis, begins with some common symptoms such as fever, diarrhea (often bloody), and abdominal pain, which is sometimes confused with appendicitis. It is also associated with some severe complications such as skin rash, meningitis, mesenteric lymphadenitis, and sepsis [2, 3]. A European Union Summary Report [6] classified yersiniosis as the third most common zoonosis in Europe [4]. Moreover, recent findings revealed that Yersinia species had developed resistance against penicillin, ampicillin, cephalosporin, and macrolides due to the production of beta-lactamases [7]. Thus, a safe, ecofriendly and effective "green" approach is required to control Y. enterocolitica to ensure food safety and public health [8]. Bacteriophages (phages) are the most abundant entities (10 3 1-10 3 2) in nature and have recently gathered more attention as green biocontrol agents owing to several advantages, including excellent target specificity, the ability to multiply in the presence of hosts, preparation and cost efficiencies, stability in wide-ranging pH levels and temperatures, and harmlessness to humans, animals, and plants [9-11]. The necessity of novel biocontrol agents has prompted us to isolate numerous phages (mainly lytic phages) from various environments and foods [12]. Unlike lysogenic phages, the lytic phage can lyse the target bacteria by integrating their DNA into the bacterial chromosome and then replicating themselves inside the host, a trait that is preferred for their use as biocontrol agents [2, 13, 14].

Previously, our research group isolated Bifidobacterium breve IDCC4401 from infant feces as a potential probiotic. For this study, we evaluated the safety of B. breve IDCC4401 using genomic and phenotypic analyses. Whole genome sequencing was performed to identify genomic characteristics and investigate the potential presence of genes encoding virulence, antibiotic resistance, and mobile genetic elements. Phenotypic analyses including antibiotic susceptibility, enzyme activity, production of biogenic amines (BAs), and proportion of D-/L-lactate were evaluated using E-test, API ZYM test, high-performance liquid chromatography (HPLC), and D-/L-lactic acid assay respectively. The genome of B. breve IDCC4401 consists of 2,426,499 bp with a GC content of 58.70% and 2,016 coding regions. Confirmation of the genome as B. breve was provided by its 98.93% similarity with B. breve DSM20213. Furthermore, B. breve IDCC4401 genes encoding virulence and antibiotic resistance were not identified. Although B. breve IDCC4401 showed antibiotic resistance against vancomycin, we confirmed that this was an intrinsic feature since the antibiotic resistance gene was not present. B. breve IDCC4401 showed leucine arylamidase, cystine arylamidase, α-galactosidase, β-galactosidase, and α-glucosidase activities, whereas it did not show production of harmful enzymes such as β-glucosidase and β-glucuronidase. In addition, B. breve IDCC4401 did not produce any tyramine, histamine, putrescine, cadaverine, or 2-phenethylamine, which are frequently detected BAs during fermentation. B. breve IDCC4401 produced 95.08% of L-lactate and 4.92% of Dlactate. Therefore, our findings demonstrate the safety of B. breve IDCC 4401 as a potential probiotic for use in the food industry.

The purpose of this study was aimed to isolate a Salmonella Typhimurium-specific phage (KFS-ST) from washing water in a poultry processing facility and to investigate the feasibility of the KFS-ST as a novel bio-receptor for the magnetoelastic (ME) biosensor method. KFS-ST against S. Typhimurium was isolated, propagated, and purified using a CsCl-gradient ultracentrifugation. Morphological characteristics of KFS-ST were analyzed using transmission electron microscopy (TEM). Its specificity and efficiency of plating analysis were conducted against 39 foodborne pathogens. The temperature and pH stabilities of KFS-ST were investigated by the exposure of the phage to various temperatures (−70℃-70℃) and pHs (1-12) for 1 h. A one-step growth curve analysis was performed to determine the eclipse time, latent time and burst size of phage. The storage stability of KFS-ST was studied by exposing KFS-ST to various storage temperatures (−70℃, −20℃, 4℃, and 22℃) for 12 weeks. KFS-ST was isolated and purified with a high concentration of (11.47 ± 0.25) Log PFU/mL. It had an icosahedral head (56.91 ± 2.90 nm) and a non-contractile tail (225.49 ± 2.67 nm), which was classified into the family of Siphoviridae in the order of Caudovirales. KFS-ST exhibited an excellent specificity against only S. Typhimurium and S. Enteritidis, which are considered two of the most problematic Salmonella strains in the meat and poultry. However, KFS-ST did not exhibit any specificity against six other Salmonella and 27 non-Salmonella strains. KFS-ST was stable at temperature of 4℃ to 50℃ and at pH of 4 to 12. The eclipse time, latent time, and burst size of KFS-ST were determined to be 10 min, 25 min and 26 PFU/ infected cell, respectively. KFS-ST was relatively stable during the 12-week storage period at all tested temperatures. Therefore, this study demonstrated the feasibility of KFS-ST as a novel bio-receptor for the detection of S. Typhimurium and S. Enteritidis in meat and poultry products using the ME biosensor method.

Bacteriophages have gained substantial attention as biocontrol and biorecognition agents, substituting antibodies. In this study, a Enteritidis-specific bacteriophage, KFS-SE1, was isolated, identified, and characterized. This phage infects S. Enteritidis with high specificity. This phage is highly stable under various pH (5-11), temperature (4-60°C), and organic solvent conditions. The KFS-SE1 genome consisted of 59,715 bp with 73 predicted open reading frames and 57.14% GC content; it had a complete set of genes required for phage reconstruction. Comparative phylogenetic analysis of KFS-SE1 revealed that it was very similar to the other phages in the family. These characteristics suggest that KFS-SE1 with its high specificity and host lysis activity toward S. Enteritidis may have various potential applications.