Bacteriophages and its Applications in Food

MOZHIARASU S

313012
M.TECH I YEAR FSQM

OUTLINE

Introduction to Bacteriophage Classification Phage Life Cycle Genome of Phage Bacteriophage as Biocontrol

Some reviews on Bacteriophage in food safety

Large Scale Production and Purification Strategy Some products and companies involved in the phage business Advantages Challenges Bacteriophage in detection

Introdution to Bacteriophage
Bacteriophages or phages are (bacterial viruses) that infect prokaryotes Ernest Hanbury Hankin in 1896 found in Ganges and Yamuna rivers in India and Pass through a very fine porcelain filter.

Frederick Twort In 1915 Discovered Bacteriophage

Felix d'Herelle Independently discovered again 1917

Image: Prescotts Microbiology

Image: Wikipedia

Classification

Taxonomy by, International Committee on Taxonomy of Viruses (ICTV)

3 orders, 73 families, 9 sub -families, 287 genera and 1938 virus species

Classification Myoviridae Siphoviridae Podoviridae Inoviridae Microviridae Tectiviridae Corticoviridae

Image: Prescotts Microbiology

Examples T-even, P1, P2, G , T1, T5, MM1, A2 P22, N4, T3, T7, C1 M13, fd X174 NS11, PRD1 PM2 MS2, R17, f2, Q 6

Leviviridae Cystoviridae

Phage Life Cycle

Lytic and Lysogenic Cycle

Image: Prescotts Microbiology

Genomic Map of Phage

48 Kb

Image: Molecular Biology of gene by Watsonet. al

Bacteriophage as Biocontrol
Animal

Phage Therapy Reduction of Colonization Biosanitation Disinfection of equipments, contact surfaces

Food Industry

Food Industry

Biocontrol Carcasses, Raw material

Biopreservation Storage and preservation

Garcia et al., 2008

Final Product

F a r m to F o r k

Some reviews on Bacteriophage in food safety

Pathogen Listeria monocytogenes Description 8 log reduction in Melon juice, 2.10log reduction in Pear juice by ListexTM p100 Fresh cut leafy greens: EcoshieldTM reduction upto 3.24 log unit compared with MAP results 4.24 Log reduction References

Oliveira et al., 2014

E coli O157:H7

Boyacioglu et al., 2013

Salmonella Vibrio parahemolyticus Campylobacter

Reduction of 6 log units of S. typhimurium Wong et al., 2014 by bacteriophage st1 in Chicken 2.76 log reduction in oysters by phage VPp1 Testing by applying in the drinking water of chicken and tested for the Campylobacter in feces and also slaughtered and checked, in feces 7.5 PFU and reduction of 3.5 log unit Rong Rong, 2014 Kittler et al., 2013

Contd.,
Pathogen Campylobacter jejuni Shewanella putrefaciens Description 3 log reduction in C. jejuni in the young broilers S.Putrefaciens in marine fish Paralichthys olivaceus by bacteriophage Spp001 yields log reduction better compared to the potassium sorbate Tested with the silkworm larvae with newly isolated bacteriophageS25-3 and S13. The authors concluded are eligible as therapeutic candidates and that the silkworm larval model is valid for the evaluation of phage therapy as well as mouse models Phage therapy of custom cocktail of 6 phages results decrease in 45% of mortality in chicken 80% survival with infected V.harveyi compared to the 25% survival in control References Wagennar et al., 2005 Meng li et al., 2013

Staphylococcus aureus

Uchiyama et al., 2013

Escherichia coli (APEC) Vibrio harveyi

Oliveira A et al., 2010 Vinod et al., 2006

Large Scale Production and Purification Strategy

Production of Bacteria Inoculating the Bacteriophage Separation of lysate by filtration

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Primary Purification(ATPE Yamamoto et al., )

Secondary Purification (Size Exclusion Chromatographic focussing)

Final (Freeze Drying)

Some products and companies involved in the phage business

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Exponential biotech (US) Gangagen (India) Intralytix Inc (US)

Novolytics (UK) Phage Biotech (Isreal)

EBI Food safety (Netherland)

Biophage Inc (Canda)

Advantages of Phage
Ubiquitous

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in nature Highly active and specific Genetically amenable Versatile use along the food chain Tools for detecting pathogens Source of potent antimicrobials
Garcia et al., 2008

Challenges in Bacteriophage and Measures to control over it

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Bacterial resistance to bacteriophages

a mixture or cocktail of different phages (Abuladze et al ., 2008) broad specificity to account for differences in bacterial strains high multiplicities of infection (MOI) 103106 (Yoichi et al. (2004)) Use of non virulent bacteria in production Modelling phage behaviour

Large-scale safer production systems

Enhance activity in food systems

Garcia et al., 2008

Bacteriophages in Detection

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Bacteriophage will help to attach bacteria specifically, which will be detected by a sensor or by any means

Reason: Binding affinity is high and considered as irreversible Polystyrene dipstick to separate Salmonella from other Enterobacteriaceae By magnetostriction principle (Lakshmanan et al., 2007)

Concentration of pathogen (Bennett et al ., 1997)

Immobilized Magnetoelastic sensor

Phage based staining method (Good Ridge et al., 1999) Reporter phage

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With Natures Antidote