Skip to main content

Advertisement

SpringerLink
Log in

Isolation and characterization of vB_SenS_Ib_psk2 bacteriophage against drug-resistant Salmonella enterica serovar Kentucky

  • Original Article
  • Published:
Folia Microbiologica Aims and scope Submit manuscript

Abstract

Salmonella enterica serovar Kentucky is one of the food-borne zoonotic pathogens which is isolated in high frequency from poultry meat in the recent decades and is known for its multidrug resistance. The current study was aimed to isolate and characterize a bacteriophage against S. enterica serovar Kentucky isolate, 5925, which showed resistance to at least seven antibiotics and to study its efficiency to decontaminate S. Kentucky from chicken skin. The bacteriophage against S. enterica serovar Kentucky was isolated and was named vB_SenS_Ib_psk2 representing the place, source, and host. Electron microscopy revealed that the phage possesses isometric head and contractile tail, indicative of Siphoviridae family. Molecular detection of major capsid protein E gene yielded 511 bp, and NCBI blast analysis revealed that the phage belonged to the genus chivirus. The optimum temperature and pH for phage survival and multiplication were found to be − 20 to 42 °C and 6–10, respectively. One-step growth curve experiment of vB_SenS_Ib_psk2 revealed a latent period of 20 min and burst size of 253 phages/bacterial cell. The host susceptibility studies revealed that 83% of MDR isolates of S. enterica were susceptible to vB_SenS_Ib_psk2. Artificial spiking studies on chicken skin revealed that high multiplicity of infection (MOI) of phages of 106 pfu/mL is required for significant reduction (p ≤ 0.01) of bacterial concentration (0.14 ± 0.04) after 24-h incubation at 8 °C compared to group 1 (2.55 ± 0.89 cfu/mL).

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Data availability

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Abbreviations

NTS:

Non-typhoidal Salmonella

iNTS:

Invasive non-typhoidal Salmonella

MOI:

Multiplicity of infection

TEM:

Transmission electron microscopy

AM:

Ampicillin

AmC:

Amoxicillin-clavulanic acid

CIP:

Ciprofloxacin

LVX:

Levofloxacin

NA:

Nalidixic acid

SXT:

Trimethoprim/sulfamethoxazole

Tet:

Tetracycline

S:

Streptomycin

References

  • Adams MH (1959) Bacteriophages. Interscience Publishers Inc, New York

    Book  Google Scholar 

  • Antunes P, Mourão J, Campos J, Peixe L (2016) Salmonellosis: the role of poultry meat. ClinMicrobiol Infect 22(2):110–121

    Article  CAS  Google Scholar 

  • Augustine J, Bhat SG (2015) Biocontrol of Salmonella enteritidis in spiked chicken cuts by lytic bacteriophages ΦSP-1 and ΦSP-3. J Basic Microbiol 55(4):500–503

    Article  PubMed  Google Scholar 

  • Bao H, Zhang H, Wang R (2011) Isolation and characterization of bacteriophages of Salmonella enterica serovar Pullorum. Poult Sci 90(10):2370–2377

  • Barua H, Biswas PK, Olsen KE, Christensen JP (2012) Prevalence and characterization of motile Salmonella in commercial layer poultry farms in Bangladesh. PLoS ONE 7:e35914

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Endersen L, Coffey A (2020) The use of bacteriophages for food safety. Curr Opin FoodSci 36:1–8

    Google Scholar 

  • Fagbamila IO, Barco L, Mancin M, Kwaga J, Ngulukun SS, Zavagnin P et al (2017) Salmonella serovars and their distribution in Nigerian commercial chicken layer farms. PLoS ONE 12:e0173097

    Article  PubMed  PubMed Central  Google Scholar 

  • Fernández J, Guerra B, Rodicio MR (2018) Resistance to carbapenems in non typhoidal Salmonella enterica serovars from humans, animals and food. Vet Sci 5(2):40

    PubMed  PubMed Central  Google Scholar 

  • Foley SL, Nayak R, Hanning IB, Johnson TJ, Han J, Ricke SC (2011) Population dynamics of Salmonella enterica serotypes in commercial egg and poultry production. Appl Environ Microbiol 77(13):4273–4279

  • Haselbeck AH, Panzner U, Im J, Baker S, Meyer CG, Marks F (2017) Current perspectives on invasive nontyphoidal Salmonella disease. Curr Opin Infect Dis 30(5):498–503

    Article  Google Scholar 

  • Hawkey J, Le Hello S, Doublet B, Granier SA, Hendriksen RS, Fricke WF, Ceyssens PJ, Gomart, C, Billman-Jacobe H, Holt KE, Weill FX (2019) Global phylogenomics of multidrug-resistant Salmonella enterica serotype Kentucky ST198. Microb Genom 5(7):e000269

  • Hong SS, Jeong J, Lee J, Kim S, Min W, Myung H (2013) Therapeutic effects of bacteriophages against Salmonella gallinarum infection in chickens. J MicrobiolBiotechnol 23:1478–1483

    CAS  Google Scholar 

  • Inbaraj S, Agrawal RK, Thomas P, Mohan C, Agarwal RK, Verma MR, Chaudhuri P (2022) Antimicrobial resistance in Indian isolates of non typhoidal Salmonella of livestock, poultry and environmental origin from 1990 to 2017. Comp Immunol Microbiol Infect Dis 80:101719

    Article  CAS  PubMed  Google Scholar 

  • Jakočiūnė D, Moodley A (2018) A rapid bacteriophage DNA extraction method. Methods Protoc 1(3):27

    Article  PubMed  PubMed Central  Google Scholar 

  • Jurczak-Kurek A, Gąsior T, Nejman-Faleńczyk B, Bloch S, Dydecka A, Topka G, Necel A, Jakubowska-Deredas M, Narajczyk M, Richert M, Mieszkowska A (2016) Biodiversity of bacteriophages: morphological and biological properties of a large group of phages isolated from urban sewage. Sci Rep 6(1):1–17

    Article  Google Scholar 

  • Kang HW, Kim JW, JungTS, Woo GJ (2013) wksl3, a new biocontrol agent for Salmonella enterica serovars enteritidis and typhimurium in foods: characterization, application, sequence analysis, and oral acute toxicity study. Appl Environ Microbiol 79(6):1956–1968

  • Kim JH, Kim HJ, Jung SJ, Mizan MFR, Park SH, Ha SD (2020) Characterization of Salmonella spp.-specific bacteriophages and their biocontrol application in chicken breast meat. J Food Sci 85:526–534

    Article  CAS  PubMed  Google Scholar 

  • Kumar S, Stecher G, Li M, Knyaz C, Tamura K (2018) MEGA X: molecular evolutionary genetics analysis across computing platforms. Mol Biol Evol 35:1547–1549

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Le Hello S, Bekhit A, Granier SA, Barua H, Beutlich J, Zając M, Münch S, Sintchenko V, Bouchrif B et al (2013) The global establishment of a highly-fluoroquinolone resistant Salmonella enterica serotype Kentucky ST198 strain. Front Microbiol 4:395

    Article  PubMed  PubMed Central  Google Scholar 

  • Litt PK, Saha J, Jaroni D (2018) Characterization of bacteriophages targeting non-O157 Shiga toxigenic Escherichia coli. J Food Prot 81(5):785–794

    Article  PubMed  Google Scholar 

  • Manyi-Loh C, Mamphweli S, Meyer E, Okoh A (2018) Antibiotic use in agriculture and its consequential resistance in environmental sources: potential public health implications. Molecules (Basel, Switzerland) 23(4):795

  • Moye ZD, WoolstonJ SA (2018) Bacteriophage applications for food production and processing. Viruses 10(4):205

    Article  PubMed  PubMed Central  Google Scholar 

  • Nilsson AS (2014) Phage therapy—constraints and possibilities. Ups J Med Sci 119(2):192–198

    Article  PubMed  PubMed Central  Google Scholar 

  • Phothaworn P, Dunne M, Supokaivanich R, Ong C, Lim J, Taharnklaew R, Vesaratchavest M, Khumthong R, Pringsulaka O, Ajawatanawong P, Klumpp J, Brown N, Imam M, Clokie MRJ, Galyov EE, Korbsrisate S (2019) Characterization of flagellotropic, chi-like Salmonella phages isolated from Thai poultry farms. Viruses 11(6):520

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Salehi S, Howe K, Brooks J, Lawrence ML, Bailey RH, Karsi A (2016) Identification of Salmonella enterica serovar Kentucky genes involved in attachment to chicken skin. BMC Microbiol 16(1):168

    Article  PubMed  PubMed Central  Google Scholar 

  • Sharma J, Kumar D, Hussain S, PathakA SM, Prasanna Kumar V, Anisha PN, Rautela R, Upadhyay AK, Singh SP (2019) Prevalence, antimicrobial resistance and virulence genes characterization of nontyphoidal Salmonella isolated from retail chicken meat shops in northern India. Food Control 102:104–111

    Article  CAS  Google Scholar 

  • Sun WJ, Liu CF, Yu L, Cui FJ, Zhou Q, Yu SL, Sun L (2012) A novel bacteriophage KSL-1 of 2-Keto-gluconic acid producer Pseudomonas fluorescens K1005: isolation, characterization and its remedial action. BMC Microbiol 12:127

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • WHO: Food safety factsheet (2020) Retrieved from https://www.who.int/news-room/fact-sheets/detail/food-safety

  • Yang M, Liang Y, Huang S, Zhang J, Wang J, Chen H, Ye Y, Gao X, Wu Q, Tan Z (2020) Isolation and characterization of the novel phages vB_VpS_BA3 and vB_VpS_CA8 for lysing Vibrio parahaemolyticus. FrontMicrobiol 11:259

    Google Scholar 

  • ŻbikowskaK MM, Dolka B (2020) The use of bacteriophages in the poultry industry. Animals (basel) 10(5):872

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Division of Bacteriology and Mycology, ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly, Uttar Pradesh, 243122, India

    Sophia Inbaraj, Prasad Thomas & V. K. Chaturvedi

  2. Food Microbiology Lab, Livestock Products Section, ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly, Uttar Pradesh, 243122, India

    Ravi Kant Agrawal

  3. Division of Biological Products, ICAR-Indian Veterinary Research Institute, Uttar Pradesh, Bengaluru, Karnataka, 560024, India

    Pallab Chaudhuri

  4. ICAR-NRC Meat, Hyderabad, Telangana, 500092, India

    Sophia Inbaraj

Authors
  1. Sophia Inbaraj
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ravi Kant Agrawal
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Prasad Thomas
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Pallab Chaudhuri
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. V. K. Chaturvedi
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Sophia Inbaraj was involved in phage isolation, experimental design, and phage studies. Prasad Thomas was involved in Salmonella isolate serotyping, maintenance, and molecular studies. Ravi K Agrawal was involved in chicken skin experimental studies. Overall supervision was done by P Chaudhary and VK Chaturvedi. All authors reviewed the manuscript.

Corresponding author

Correspondence to Sophia Inbaraj.

Ethics declarations

Ethical approval

The article does not contain any studies with human participant or animals.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Inbaraj, S., Agrawal, R.K., Thomas, P. et al. Isolation and characterization of vB_SenS_Ib_psk2 bacteriophage against drug-resistant Salmonella enterica serovar Kentucky. Folia Microbiol 68, 771–779 (2023). https://doi.org/10.1007/s12223-023-01052-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12223-023-01052-0

Keywords

Search

Navigation