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phage application in food safety
Bacteriophage and its Applications in Food
By
Rasha Mohamed Mahmoud
Supervised by
Prof. Dr. Zakaria Yehia Daw
Dr. Mona Hussein Badawi
Introduction to Bacteriophage
Classification
Phage Life Cycle
Where we can find phages?
Desirable properties of phages used as biocontrol agents in foods
Bacteriophages for biocontrol of pathogens in food
Some reviews on Bacteriophage in food safety
Large Scale Production and Purification Strategy
Bacteriophages in Detection
Advantages of Phage
Challenges in Bacteriophage and Measures to control
Introduction
 Obligate intracellular parasites that multiply inside bacteria by
making use of some or all of the host biosynthetic machinery
Felix d'Herelle
Independently discovered
again 1917
Frederick Twort
In 1915 Discovered Bacteriophage
Classification
 Taxonomy by, International Committee on Taxonomy of Viruses (ICTV)
 3 orders, 73 families, 9 sub -families, 287 genera and 1938 virus species
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
phage application in food safety
Phage Attack
 In humans and animal intestines
 In running water
 In the soil
 Effluent outlets
 Sewage from corpses
 Dr. "Grammy bar" from the University of San Diego,
California, says that Bacteriophag viruses live in the human
intestine does not harm him (Research 2013). On the contrary,
is stick to the mucous layer of the intestines and are covered,
if approached her cell bacteria hung out and penetrate the cell
wall and enter them and multiply. Then explodes bacteria cell
and out of a large number of viruses Bacteriophage . Thus,
service to the people of this virus leads, at the same time
provide them with the appropriate center for human living
Desirable properties of phages used as
biocontrol agents in foods
 Phage should be strictly lytic
 Phage should have a broad host range
 Determine the complete genome sequence of phages
 Lack of transduction of bacterial DNA
 Oral feeding studies of phages should show no adverse effects
 Phage preparation should be stable over storage and application
 Phage should be amenable to scale up for commercial production
Bacteriophages for biocontrol of pathogens in food From
Farm
To
Fork
Some reviews on Bacteriophage in food safety
Pathogen Description References
Listeria monocytogenes 8 log reduction in Melon juice, 2.10 log 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
Large Scale Production and Purification
Strategy
Cultivation of
Bacteria
Inoculating the
Bacteriophage
Separation of lysate by
filtration
Primary Purification
Secondary Purification
(Size Exclusion 
Chromatographic
focussing)
Final
(Freeze Drying)
Bacteriophages in Detection
Bacteriophage will help to attach bacteria specifically, which will be
detected by a sensor or by any means
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 antimicrobial agents
Challenges in Bacteriophage and
Measures to control
 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
Some products and companies involved
in the phage business
Any Questions?
References
 http://www.biocontrol-ltd.com/ with thanks to Dr. David R. Harper
 • http://www.phagetherapycenter.com/
 • http://en.wikipedia.org/wiki/Phage_therapy
 • http://recursive-nature.com/bacteriaphoge/
 Haq I U et al (2012) Bacteriophages and their Implications on Future Biotechnology: A Review J
Virol 9:
 Wang Y, Ye Z, Ying Y. New trends in impedimetric biosensors for the detection of foodborne
pathogenic bacteria. Sensors (Basel) 2012; 12:3449-71;PMID:22737018; http://dx.doi.org/10.3390/
s120303449
 Stewart GS, Jassim SA, Denyer SP, Newby P, Linley K, Dhir VK. The specific and sensitive detection
of bacterial pathogens within 4 h using bacteriophage amplification. J Appl Microbiol 1998; 84:777-
83; PMID:9674131; http://dx.doi.org/10.1046/j.1365-2672.1998.00408.x
 Power point presentation (2015) www.slideshare.net/
phage application in food safety

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phage application in food safety

  • 2. Bacteriophage and its Applications in Food By Rasha Mohamed Mahmoud Supervised by Prof. Dr. Zakaria Yehia Daw Dr. Mona Hussein Badawi
  • 3. Introduction to Bacteriophage Classification Phage Life Cycle Where we can find phages? Desirable properties of phages used as biocontrol agents in foods Bacteriophages for biocontrol of pathogens in food Some reviews on Bacteriophage in food safety Large Scale Production and Purification Strategy Bacteriophages in Detection Advantages of Phage Challenges in Bacteriophage and Measures to control
  • 4. Introduction  Obligate intracellular parasites that multiply inside bacteria by making use of some or all of the host biosynthetic machinery Felix d'Herelle Independently discovered again 1917 Frederick Twort In 1915 Discovered Bacteriophage
  • 5. Classification  Taxonomy by, International Committee on Taxonomy of Viruses (ICTV)  3 orders, 73 families, 9 sub -families, 287 genera and 1938 virus species 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
  • 8.  In humans and animal intestines  In running water  In the soil  Effluent outlets  Sewage from corpses  Dr. "Grammy bar" from the University of San Diego, California, says that Bacteriophag viruses live in the human intestine does not harm him (Research 2013). On the contrary, is stick to the mucous layer of the intestines and are covered, if approached her cell bacteria hung out and penetrate the cell wall and enter them and multiply. Then explodes bacteria cell and out of a large number of viruses Bacteriophage . Thus, service to the people of this virus leads, at the same time provide them with the appropriate center for human living
  • 9. Desirable properties of phages used as biocontrol agents in foods  Phage should be strictly lytic  Phage should have a broad host range  Determine the complete genome sequence of phages  Lack of transduction of bacterial DNA  Oral feeding studies of phages should show no adverse effects  Phage preparation should be stable over storage and application  Phage should be amenable to scale up for commercial production
  • 10. Bacteriophages for biocontrol of pathogens in food From Farm To Fork
  • 11. Some reviews on Bacteriophage in food safety Pathogen Description References Listeria monocytogenes 8 log reduction in Melon juice, 2.10 log 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
  • 12. Large Scale Production and Purification Strategy Cultivation of Bacteria Inoculating the Bacteriophage Separation of lysate by filtration Primary Purification Secondary Purification (Size Exclusion  Chromatographic focussing) Final (Freeze Drying)
  • 13. Bacteriophages in Detection Bacteriophage will help to attach bacteria specifically, which will be detected by a sensor or by any means
  • 14. 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 antimicrobial agents
  • 15. Challenges in Bacteriophage and Measures to control  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
  • 16. Some products and companies involved in the phage business
  • 18. References  http://www.biocontrol-ltd.com/ with thanks to Dr. David R. Harper  • http://www.phagetherapycenter.com/  • http://en.wikipedia.org/wiki/Phage_therapy  • http://recursive-nature.com/bacteriaphoge/  Haq I U et al (2012) Bacteriophages and their Implications on Future Biotechnology: A Review J Virol 9:  Wang Y, Ye Z, Ying Y. New trends in impedimetric biosensors for the detection of foodborne pathogenic bacteria. Sensors (Basel) 2012; 12:3449-71;PMID:22737018; http://dx.doi.org/10.3390/ s120303449  Stewart GS, Jassim SA, Denyer SP, Newby P, Linley K, Dhir VK. The specific and sensitive detection of bacterial pathogens within 4 h using bacteriophage amplification. J Appl Microbiol 1998; 84:777- 83; PMID:9674131; http://dx.doi.org/10.1046/j.1365-2672.1998.00408.x  Power point presentation (2015) www.slideshare.net/