Isolation and Characterization of Potential Salmonella Phages Targeting Multidrug-Resistant and Major Serovars of Salmonella Derived From Broiler Production Chain in Thailand

Abstract

Salmonella is a major foodborne pathogen that causes foodborne disease in humans through consumption of contaminated foods, especially those of animal origin. Multiple Salmonella strains are antibiotic-resistant due to the common use of antibiotics in farm animals, including broiler farms. In this study, an alternative strategy using phage-based treatment was evaluated against Salmonella isolated from the broiler production. The prevalence of Salmonella spp. showed up to 46.2 and 44.4% in bedding samples from the broiler farms located in eastern and southern Thailand, respectively. Overall, 21 samples (36.2%) were positive for Salmonella and eight serovars were recovered from cloacal swabs, bedding materials (rice husk), and boot swabs collected from five farms. Up to 20 Salmonella phages were isolated from seven water samples from wastewater treatment ponds, a river, and a natural reservoir in Songkhla province. Isolated phages were investigated, as well as their lysis ability on eight target Salmonella serovars derived from broiler farms, five foodborne outbreak-related serovars, and 10 multidrug-resistant (MDR) serovars. All phages showed a strong lytic ability against five serovars of Salmonella derived from broiler farms including Kentucky, Saintpaul, Schwarzengrund, Corvalis, and Typhimurium; three foodborne outbreak serovars including Enteritidis, Typhimurium, and Virchow; and eight MDR serovars including Agona, Albany, Give, Kentucky, Typhimurium, Schwarzengrund, Singapore, and Weltevreden. Three phages with the highest lysis potential including vB_SenS_WP109, vB_SenS_WP110, and vB_SenP_WP128 were selected for a phage cocktail preparation. Overall, a phage cocktail could reduce Salmonella counts by 2.2–2.8 log units at 6 h of treatment. Moreover, Salmonella did not develop a resistant pattern after being treated with a phage cocktail. Findings here suggest that a phage cocktail is an effective biocontrol to combat Salmonella derived from broiler production chain, other serovars linked to foodborne outbreaks, and MDR serovars.

Evolution of β-lactams, fluroquinolones and colistin resistance and genetic profiles in <em>Salmonella</em> isolates from pork in northern Italy

“…Antimicrobial resistance in Salmonella is increasing, likely due to the abuse/misuse of antibiotics in farmed animals, including pigs. AMR compromises the efficacy of antibiotics used to treat human infections, thus suggesting a reducing in antibiotic use in livestock (Pelyuntha et al ., 2021 ).…”

Phage Revolution Against Multidrug-Resistant Clinical Pathogens in Southeast Asia

“…In the past decade, scientists reported the isolation of diverse phages in SEA from different sample types, with potential activities against human pathogens (Table 2). Among the phages discovered in the region with notable activity against MDR organisms are: (1) phages vB_AbaM_PhT2 (Styles et al, 2020) and AB1801 (Wintachai et al, 2019), which exhibited anti-MDR A. baumannii activity, in vitro and in vivo, respectively, (2) phage UPM2146 (Assafiri et al, 2021), which exhibited activity against carbapenem-resistant K. pneumoniae, in vitro and in vivo, (3) phage C34 (Guang- Han et al, 2016), which controlled naturally resistant Burkholderia pseudomallei bacterial load, in vivo, (4) phage ΦHN10 (Phothichaisri et al, 2018), which exhibited the highest breadth of activity against various Clostridiodes difficile strains, in vitro, (5) phage ΦKAZ14 (Ahmad et al, 2015), which induced lysis of extended-spectrum betalactamase (ESBL)-producing Escherichia coli, in vitro, (6) phages UPMK_1 and 2 (Dakheel et al, 2019) and ΦNUSA-1 and 10 (Tan et al, 2020), which all exhibited lytic activity against methicillin-resistant Staphylococcus aureus (MRSA), and antibiofilm activity for the first two phages in in vitro experiments, (7) cocktail of phages vB_SenS_WP109, WP110, and WP128 (Pelyuntha et al, 2021), which induced lysis of multiple MDR Salmonella serovars, in vitro, and (8) phages KP1, 2 Poultry and farm feces, urban catchment, tissue samples Indonesia, Malaysia, Singapore Lau et al, 2012;Gan et al, 2013;Rezaeinejad et al, 2014;Ahmad et al, 2015;Vergara et al, 2015;Sellvam et al, 2018;Lukman et al, 2020;Dewanggana et al, 2021;Sjahriani et al, 2021;Waturangi et al…”

“…Dairy farm, poultry, clinical samples Malaysia, Thailand Wong et al, 2014;Wongsuntornpoj et al, 2014;Phothaworn et al, 2019Phothaworn et al, , 2020Pelyuntha et al, 2021 Shigella Staphylococcus phages (UPMK_1, UPMK_2, ΦNUSA-1, ΦNUSA-10)…”

“…Lastly, a few publications focused on the applications of phages in combating MDR organisms. For instance, SEA researchers discovered and characterized phages with activity against MRSA (Dakheel et al, 2019;Tan et al, 2020), ESBL-producing E. coli (Adamu Ahmad et al, 2016), MDR K. pneumoniae (Cornista et al, 2019;Assafiri et al, 2021), MDR A. baumannii (Wintachai et al, 2019;Styles et al, 2020), MDR Salmonella species (Pelyuntha et al, 2021), and Clostridioides difficile (Phothichaisri et al, 2018). The efficacy of these phages has yet to be confirmed clinically.…”

Isolation and Characterization of a Novel Autographiviridae Phage and Its Combined Effect with Tigecycline in Controlling Multidrug-Resistant Acinetobacter baumannii-Associated Skin and Soft Tissue Infections

“…These phenomena highlight the possibility of using phages to treat A. baumannii-associated SSTIs. In Thailand, a few studies on phage isolations have been reported, and there are no commercial phage products available nowadays [33][34][35]. Due to the emergence of MDR bacteria in Thailand similar to other countries worldwide, alternative strategies to control MDR bacterial infections, including phage and the combination of phages with other antibiotics, are attractive to investigate among researchers.…”