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

48 Kb

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

Thanks