Salmonella Enteritidis Bacteriophages Isolated from Kenyan Poultry Farms Demonstrate Time-Dependent Stability in Environments Mimicking the Chicken Gastrointestinal Tract

Mhone, Amos Lucky; Makumi, Angela; Odaba, Josiah; Guantai, Linda; Gunathilake, K. M. Damitha; Loignon, Stéphanie; Ngugi, Caroline Wangari; Akhwale, Juliah Khayeli; Moineau, Sylvain; Svitek, Nicholas

doi:10.3390/v14081788

Cited by 9 publications

(3 citation statements)

References 68 publications

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“…When there is headful packaging with pac initiation, it could be a restriction profile with all bands from a circular profile plus a submolar pac fragment ( 50 ). However, as shown previously ( 54 ), the EcoRV restriction profile of K47 resembled the circular profile (Fig. S4).…”

Section: Resultssupporting

confidence: 86%

Diversity of Salmonella enterica phages isolated from chicken farms in Kenya

Gunathilake,

Makumi,

Loignon

et al. 2024

Microbiol Spectr

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In this study, we describe the characterization of 67 Salmonella enterica phages isolated from Kenya. Specifically, the phages were isolated from chicken feces as well as from swabs and water samples obtained from chicken farms and slaughterhouses in Nairobi and Kiambu counties. Genome sequencing and analysis revealed that 59 phage isolates were distinct and could be grouped into 17 clusters belonging to four major types of phages (MT1–4). Further taxonomic analysis showed that each MT represents one phage genus. Of the 17 selected representative genomes (one per cluster), seven phages belong to the MT1 group and to the genus Jerseyvirus . The MT2 cluster contained eight representative phages that were similar to Escherichia phage T5 and belong to the Tequintavirus genus. The unique MT3 cluster with one phage representative belongs to the Zindervirus genus, while the sole MT4 phage isolate is a member of the Dhillonvirus genus. All representative phages are siphophages, except MT3 phage which is a podophage. All genomes do not contain genes associated with lysogeny and therefore, they are considered exclusively lytic. No known virulence genes or antibiotic resistance genes were detected in any of the genomes. Phage DNA termini analyses showed that MT2 phages have a direct terminal repeat-based packaging mechanism while MT1 has a headful packaging mechanism. Host range studies showed that phages belonging to MT1 and MT3 appear to only infect Salmonella strains, while MT2 and MT4 phages were also capable of infecting Escherichia and Shigella strains. IMPORTANCE Non-typhoidal Salmonella enterica infections are one of the leading causes of diarrhoeal diseases that spread to humans from animal sources such as poultry. Hence, keeping poultry farms free of Salmonella is essential for consumer safety and for a better yield of animal products. However, the emergence of antibiotic resistance due to over usage has sped up the search for alternative biocontrol methods such as the use of bacteriophages. Isolation and characterization of novel bacteriophages are key to adapt phage-based biocontrol applications. Here, we isolated and characterized Salmonella phages from samples collected at chicken farms and slaughterhouses in Kenya. The genomic characterization of these phage isolates revealed that they belong to four ICTV (International Committee on Taxonomy of Viruses) phage genera. All these phages are lytic and possibly suitable for biocontrol applications because no lysogenic genes or virulence factors were found in their genomes. Hence, we recommend further studies on these phages for their applications in Salmonella biocontrol.

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Section: Resultssupporting

confidence: 86%

Diversity of Salmonella enterica phages isolated from chicken farms in Kenya

Gunathilake,

Makumi,

Loignon

et al. 2024

Microbiol Spectr

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“…The use of antacids such as calcium carbonate or bicarbonate in order to neutralize stomach acidity has been common practice in several studies ( Tanji et al, 2005 ; Atterbury et al, 2007 ; Nale et al, 2016 ; Colom et al, 2017 ; Yen et al, 2017 ). In fact, phages included in the UPWr_S134 cocktail completely lose their ability to infect Salmonella after incubation in simulated gastric conditions, confirming the previous reports where phages were recovered after exposure to SGF ( Mhone et al, 2022 ). However, the addition of calcium carbonate resulted in a protective effect on phage activity, confirming previous studies ( Koo et al, 2000 ; Ma et al, 2008 ).…”

Section: Discussionsupporting

confidence: 87%

Effective reduction of Salmonella Enteritidis in broiler chickens using the UPWr_S134 phage cocktail

et al. 2023

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Salmonella is a poultry-associated pathogen that is considered one of the most important zoonotic bacterial agents of contaminated food of animal origin including poultry products. Many efforts are taken to eliminate it from the food chain, and phages are one of the most promising tools to control Salmonella in poultry production. We investigated the usefulness of the UPWr_S134 phage cocktail in reducing Salmonella in broiler chickens. For this purpose, we analyzed the survivability of phages in the harsh environment encountered in the chicken gastrointestinal tract, which has low pH, high temperatures, and digestive activity. Phages in the cocktail UPWr_S134 showed the ability to remain active after storage at temperatures ranging from 4 to 42°C, reflecting temperatures of storage conditions, broiler handling, and the chicken body, and exhibited robust pH stability. We found that although simulated gastric fluids (SGF) caused phage inactivation, the addition of feed to gastric juice allows maintenance of UPWr_S134 phage cocktail activity. Further, we analyzed UPWr_S134 phage cocktail anti-Salmonella activity in live animals such as mice and broilers. In an acute infection model in mice, the application of doses of 107 and 1014 PFU/ml UPWr_S134 phage cocktail resulted in delaying symptoms of intrinsic infection in all analyzed treatment schedules. In Salmonella-infected chickens orally treated with the UPWr_S134 phage cocktail the number of pathogens in internal organs in comparison to untreated birds was significantly lower. Therefore we concluded that the UPWr_S134 phage cocktail could be an effective tool against this pathogen in the poultry industry.

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“…The resistance of LAB isolates to heat was determined at 42°C, which is the average temperature of the chicken body and digestive tract [ 27 , 28 ]. It is recommended that probiotic bacterial isolates should be able to live and survive at this temperature.…”

Section: Discussionmentioning

confidence: 99%

Identification of novel probiotic lactic acid bacteria from soymilk waste using the 16s rRNA gene for potential use in poultry

Srifani,

Mirnawati,

Marlida

et al. 2024

Vet World

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Background and Aim: In-feed antibiotics have been used as antibiotic growth promoters (AGPs) to enhance the genetic potential of poultry. However, the long-term use of AGPs is known to lead to bacterial resistance and antibiotic residues in poultry meat and eggs. To address these concerns, alternatives to AGPs are needed, one of which is probiotics, which can promote the health of livestock without having any negative effects. In vitro probiotic screening was performed to determine the ability of lactic acid bacteria (LAB) isolated from soymilk waste to be used as a probiotic for livestock. Materials and Methods: Four LAB isolates (designated F4, F6, F9, and F11) isolated from soymilk waste were used in this study. In vitro testing was performed on LAB isolates to determine their resistance to temperatures of 42°C, acidic pH, bile salts, hydrophobicity to the intestine, and ability to inhibit pathogenic bacteria. A promising isolate was identified using the 16S rRNA gene. Results: All LAB isolates used in this study have the potential to be used as probiotics. On the basis of the results of in vitro testing, all isolates showed resistance to temperatures of 42°C and low pH (2.5) for 3 h (79.87%–94.44%) and 6 h (76.29%– 83.39%), respectively. The survival rate at a bile salt concentration of 0.3% ranged from 73.24% to 90.39%, whereas the survival rate at a bile salt concentration of 0.5% ranged from 56.28% to 81.96%. All isolates showed the ability to attach and colonize the digestive tract with a hydrophobicity of 87.58%–91.88%. Inhibitory zones of LAB against pathogens ranged from 4.80–15.15 mm against Staphylococcus aureus, 8.85–14.50 mm against Salmonella enteritidis, and 6.75–22.25 mm against Escherichia coli. Although all isolates showed good ability as probiotics, isolate F4 showed the best probiotic ability. This isolate was identified as Lactobacillus casei strain T22 (JQ412731.1) using the 16S rRNA gene. Conclusion: All isolates in this study have the potential to be used as probiotics. However, isolate F4 has the best probiotic properties and is considered to be the most promising novel probiotic for poultry. Keywords: lactic acid bacteria, poultry, probiotic.

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Salmonella Enteritidis Bacteriophages Isolated from Kenyan Poultry Farms Demonstrate Time-Dependent Stability in Environments Mimicking the Chicken Gastrointestinal Tract

Cited by 9 publications

References 68 publications

Diversity of Salmonella enterica phages isolated from chicken farms in Kenya

Diversity of Salmonella enterica phages isolated from chicken farms in Kenya

Effective reduction of Salmonella Enteritidis in broiler chickens using the UPWr_S134 phage cocktail

Identification of novel probiotic lactic acid bacteria from soymilk waste using the 16s rRNA gene for potential use in poultry

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