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1Bacteriophages and its Applications in FoodMOZHIARASU S
313012
M.TECH I YEAR FSQM
NIFTEM
2OUTLINE
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
3Introdution 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.
Felix d'Herelle
Independently discovered again 1917
Frederick TwortIn 1915 Discovered
Bacteriophage
Image: Prescott’s Microbiology
Image: Wikipedia
4ClassificationTaxonomy by, International Committee on Taxonomy of Viruses (ICTV)
Image: Prescott’s Microbiology
Classification Examples
Myoviridae T-even, P1, P2, G
Siphoviridae λ, T1, T5, MM1, A2
Podoviridae P22, N4, T3, T7, C1
Inoviridae M13, fd
Microviridae ΦX174
Tectiviridae ΦNS11, PRD1
Corticoviridae PM2
Leviviridae MS2, R17, f2, Qβ
Cystoviridae Φ6
3 orders, 73 families, 9 sub -families, 287 genera and 1938 virus species
5Phage Life Cycle
Lytic and Lysogenic CycleImage: Prescott’s Microbiology
6Genomic Map of λ Phage
Image: Molecular Biology of gene by Watson’et. al
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7Bacteriophage as Biocontrol
Animal
• Phage Therapy• Reduction of Colonization
Food
Industry
• Biosanitation• Disinfection of equipments, contact
surfaces
Food
Industry
• Biocontrol• Carcasses, Raw material
Final Product
• Biopreservation• Storage and preservation
Fa rm to Fo rk
Garcia et al., 2008
8Some reviews on Bacteriophage in food safety
Pathogen Description References
Listeria monocytogenes 8 log reduction in Melon juice, 2.10log reduction in Pear juice by ListexTM p100
Oliveira et al., 2014
E coli O157:H7 Fresh cut leafy greens: EcoshieldTM
reduction upto 3.24 log unit compared with MAP results 4.24 Log reduction
Boyacioglu et al., 2013
Salmonella Reduction of 6 log units of S. typhimurium by bacteriophage Φ st1 in Chicken
Wong et al., 2014
Vibrio parahemolyticus 2.76 log reduction in oysters by phage VPp1
Rong Rong, 2014
Campylobacter 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
Kittler et al., 2013
9Pathogen Description References
Campylobacter jejuni
3 log reduction in C. jejuni in the young broilers
Wagennar et al., 2005
Shewanella putrefaciens
S.Putrefaciens in marine fish Paralichthys olivaceus by bacteriophage Spp001 yields log reduction better compared to the potassium sorbate
Meng li et al., 2013
Staphylococcus aureus
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
Uchiyama et al., 2013
Escherichia coli (APEC)
Phage therapy of custom cocktail of 6 φ phages results decrease in 45% of mortality in chicken
Oliveira A et al., 2010
Vibrio harveyi 80% survival with infected V.harveyi compared to the 25% survival in control
Vinod et al., 2006
Contd.,
10Large Scale Production and Purification Strategy
Production of Bacteria
Inoculating the Bacteriophage
Separation of lysate by filtration
Primary Purification(ATPE
Yamamoto et al., )
Secondary Purification (Size
Exclusion Chromatographic
focussing)
Final(Freeze Drying)
11Some products and companies involved in the phage business Exponential biotech (US)
Gangagen (India)
Intralytix Inc (US)
Novolytics (UK)
Phage Biotech (Isreal)
EBI Food safety (Netherland)
Biophage Inc (Canda)
12Advantages of Phage
Ubiquitous in natureHighly active and specificGenetically amenableVersatile use along the food chainTools for detecting pathogensSource of potent antimicrobials
Garcia et al., 2008
13Challenges in Bacteriophage and Measures to control over it
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) ≥103−106 (Yoichi et al. (2004))
Large-scale safer production systems
Use of non virulent bacteria in production
Enhance activity in food systems
Modelling phage behaviour
Garcia et al., 2008
14Bacteriophages in Detection
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
Concentration of pathogen (Bennett et al ., 1997) Polystyrene dipstick to separate Salmonella from other
Enterobacteriaceae
Immobilized Magnetoelastic sensor By magnetostriction principle (Lakshmanan et al., 2007)
Phage based staining method (Good Ridge et al., 1999)
Reporter phage
15
With Nature’s Antidote
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