The presentation gives glimpse to basics of bacteriophages and its application in food

1. Bacteriophages
and its
Applications in
Food
MOZHIARASU S
313012
M.TECH I YEAR FSQM
NIFTEM
1

2. 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
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3. 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.
Felix d'Herelle
Independently
discovered
again 1917
Frederick Twort
In 1915 Discovered
Bacteriophage
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Image: Prescott’s Microbiology
Image: Wikipedia

4. Classification
Taxonomy by, International Committee on Taxonomy of Viruses (ICTV)
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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

5. Phage Life Cycle
Lytic and Lysogenic Cycle
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Image: Prescott’s Microbiology

6. Genomic Map of λ Phage 6
Image: Molecular Biology of gene by Watson’et. al
48 Kb

7. Bacteriophage as Biocontrol 7
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
F
a
r
m
to
F
o
r
kGarcia et al., 2008

8. Some 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
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9. 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.,

10. Large Scale Production and
Purification Strategy
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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)

11. Some 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)
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12. Advantages of Phage
Ubiquitous in nature
Highly active and specific
Genetically amenable
Versatile use along the food chain
Tools for detecting pathogens
Source of potent antimicrobials
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Garcia et al., 2008

13. Challenges 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
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Garcia et al., 2008

14. Bacteriophages 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
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