phage therapy: a new paradigm for food industry€¦ · 1945 –“golden era of antibiotics” in...
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Phage therapy: a new paradigm for food
industry
Catalin Iancu, PhD
Scientist, Micreos Food Safety
April 2016, Timisoara, Romania
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Why me and phages for food safety?
Common point: Food-born pathogens
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History of bacteriophage therapy
Félix d'Herelle
1917 – “an invisible, antagonistic microbe of the dysentery bacillus"
1921 – First administration of phages at Hopital des Enfants-Malade, Paris
“Golden Era of Antibiotics” in the Western world – phage therapy left behind1945 –
2006 – First phage-based antilisterial product approved by FDA
“You can't really move forward until you look back”.Cornel West
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Facts about phages
Natural enemy of bacteria, forced to kill
Ubiquitous in the environment and in the human
body;
Most abundant micro organism on earth (>1032);
Outnumber bacteria 10 to 1;
Up to a billion phages in 1 ml seawater
2 distinct “lifestyles”: lytic and lysogenic
Moineau, S. et al., 2010. Nature Reviews
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MICREOS
Est. in 2005
Privately owned
50 employees
Technology leader
Targeted
anti-bacterial solutions
Food Safety Human Health
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Phages and food safety
RTE Meats Smeared Cheese Fish Food Contact Surfaces
Vegetables Raw Poultry
Phages are best on surface or liquid applications;
Two main phage providers the food safety arena: and
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Rules of the game
Strictly lytic phages;
Do not carry undesirable genes (e.g. pathogenicity; antibiotic resistance)
Do not transduce host DNA (transduction experiments)
Food regulations compliance;
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Phages vs common antimicrobials
Compared to other antimicrobials the numbers are in reality tiny:
→ Nisin 12.5 mg/kg 2.24 x 1015 molecules/g
→ Octanoic acid 400ppm 1.6 x 1018 molecules/g
→ 1 g of sodium lactate treats a small hotdog to prevent outgrowth of Listeria
→ 2 uL of phage particles (5 x 106 PFU/cm2) can kill 105 of Listeria cells;
→ 1 g of phages will treat between 2-200 million hotdogs
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Resistance and sustainable use
Inherent phage resistance mechanisms (R/M systems, CRISPRs) are easily circumvented by testing candidate phages against a large strain collection or are of no concern (ABIs, super-infection exclusion systems)
Mutations are generally detrimental (only phage pressure gives an advantage)
1 in 106 or 107 contaminations will not be removed but if food is treated there is no feed-back loop to the environmental niche
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LISTEX™
Extremely broad host range;
Receptors: N-acetylglucosamine and rhamnose
substituents of cell wall teichoic acids;
High affinity towards Listeria cells;
FDA approved - GRAS status;
Used mainly in RTE and cheese;
P100 phage
(TEM)
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LISTEX™ on Hot dogs
1
10
100
1000
10000
100000
0 1 2 3 4 5 6
time in days
Lis
teri
a C
FU
/g
101
102
103
104
Hotdogs, L monocytogenes WSLC1001 SV1/2c
Control
3x108 PFU/g 3x107 PFU/g
1
10
100
1000
10000
100000
0 1 2 3 4 5 6
time in days
Lis
teri
a C
FU
/g
101
102
103
104
Hotdogs, L monocytogenes WSLC1001 SV1/2c
Control
3x108 PFU/g 3x107 PFU/g
Hot dogs weight : surface ratio 1:2;1.5 x 107 and 1.5 x 108 PFU/cm2
Guenther, 2008, Appl Env Microbiol.
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Efficacy of LISTEX™on read-smear soft
cheese during ripening
Phage dosage: 3 x 108 PFU/cm2
Data generated by ETH (Swiss Federal Institute of Technology Zurich)
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LISTEX™ as an additional hurdle
LISTEX™ in combination with Potassium Lactate-Sodium Diacetate
Extend the shelf life of RTE products
Chibeu et al 2013, Int J Food Microbiol.
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SALMONELEX™
Cocktail of two phages
S16 - T4-like phage which attaches to OmpC
protein – an osmoregulatory membrane
protein present in all strains regardless of
serovar;
Felix O1 - Attaches to a sugar residue of the
outer core of the LPS molecule ;
FDA – GRAS status;
Used mainly in raw meats (poultry, pork, beef);
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SALMONELEX™ on chicken skin
4°C
Phage concentrations 1x107 pfu/cm² or 2x107 pfu/cm²;
Contamination with ~1x104 cfu/cm²
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SALMONELEX™ on chicken breast fillet
4°C
Phage concentrations 1x107 pfu/cm² or 2x107 pfu/cm²;
Contamination with ~1x104 cfu/cm²
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SALMONELEX™ on raw pork
Phage concentrations 1x107 pfu/cm² or 2x107 pfu/cm²;
Contamination with ~1x104 cfu/cm²
4°C
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Concluding Remarks
WHY?Phage therapy (food safety)
Natural solution
Effective
Friendly
Clean label
Sustainable
Available
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THANK YOU!