Alexander Sulakvelidze

Transcription

Alexander Sulakvelidze
Intralytix, Inc
Bacteriophages: Safety by Nature –
Biological Control at Its Best
Alexander Sulakvelidze, Ph.D.
Intralytix, Inc.
.
3rd International Symposium on Biological Control of
Plant Bacterial Diseases (Biocontrol 2016)
11-14 April 2016, Belgrade, Serbia
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Bacteriophages / Phages
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Viruses that attack bacteria
Estimated 3 billion years old
Discovered by Frederick Twort (1915) and Felix d’Herelle (1917)
D'Herelle coined the name bacteriophage (bacteria-eater)
Most ubiquitous organisms in nature
Highly specific for bacterial host
Play a key role in maintaining balanced bacterial populations in all ecosystems where bacteria exist
Some of the commercial phage products
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Laboratoire du Bactériophage: Bacté-coli-phage, Bacté-rhino-phage,
Bacté-intesti-phage, Bacté-pyo-phage, and Bacté-staphy-phage
Eli Lily: Colo-lysate, Ento-lysate, Neiso-lysate, and Staphylo-lysate,
Colo-jel, Ento-jel, and Staphylo-jel
ImBio: Coli-protein, Samoneluese
CMBP: PhagoBioDerm
Eliava Institute: IntestiPhage, PyoPhage
EBI Food Safety (now Micreos): Listex
Intralytix, Inc: ListShield, EcoShield, SalmoFresh
Phages are small…
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… and they are the most ubiquitous organisms on
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Earth
10 Phage per microbe
In the U.S.A., approx. 3 x 109 coliphages shed per person per day
1 mL of non-polluted water contains 2x108 PFU of phages
Total number of phages on Earth is estimated to be 1x1030 – 1x1032
(100 000 000 000 000 000 000 000 000 000 000 phages) - outweigh the world
population of elephants by a thousand-fold or more
Phages have been found in commercial sera and human vaccines
Phages are common in human mouth, where they are harbored in dental plaque and
saliva
The second most abundant category of microorganisms in human GI tract (after
bacteria).
Phages are common in foods
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 Fresh ground beef and canned corned beef (likely 100%)
 Fresh pork sausage and chicken meat. 100% of examined fresh chicken and pork sausage
contained phages
 Delicatessen meat
 Farmed freshwater fish, common carp and marine fish
 Oil sardines
 Raw skim milk
 Cheese
 Frozen mixed vegetables
 The levels ranged from ca. 4 x 1010 PFU/100 g of fresh chicken, up to 4 x 1010 PFU/100 g of fresh
pork, and up to 3 x 1010 PFU/100 g of roast turkey breast samples.
 67 – 83% of all animal feed, feed ingredients, and poultry diets examined by the Texas A&M
contained phages (including samples stored at -20°C for 14 months)
Potential Applications
Food safety
“Green” industrial and household cleaners / bio-pesticides
Probiotic products / phage preparations for fine-tuning GI microbiome
Cosmetics and dental care
Pet food & food supplements
Hospital decontamination & biofilm reduction
Plant infections
Animal health products (bacterins & drugs)
Drugs / human therapeutics (e.g., wound infections)
Diagnostic uses
Phage enzymes
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First Phage Product Approved by the FDA
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This box contains a
solution of 100% naturally
occurring, Listeriamunching bacteriophages.
Good to animals and
plants. Good to the
environment. And good to
you. Safety by nature.TM
Simple Application
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Timing of Lysis
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Source: Karam JD et al (ed.), Molecular Biology of
Bacteriophage T4, ASM Press, Washington, DC
Effect of phage treatment on L. monocytogenes levels in
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(A) Lettuce, (B) Smoked salmon, and (C) Frozen entrees
Perera et al. Food Microbiology, Volume 52, December 2015, Pages 42–48
EcoShield reduces E. coli O157:H7 levels in
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ground beef stored refrigerated and frozen
(Based on triplicate samples; bars = SEM)
Approx. 94 - 98% reduction at all time points
Effect of phage treatment on Salmonella incidence
in chicken meat (86% reduction) Intralytix, Inc
86% reduction in positives
Plate # 1, six positives: B8, C7, D6, E2, E5, F5
Plate # 2, five positives: B3, B5, C7, D6, F7
Phages: Safety By NatureTM
Pros
L. monocytogenes phages released and
propagated in the environment as a result
Safe and effective
of ListShield’s being used in foodprocessing facilities was estimated, based
Do not affect taste, appearance, or smell of foods
on several “worst case scenarios,” to be
No impact on label (in most cases)
ca. 6 x 1011 PFU. This number
represents ca.
Do not alter Organic, Halal, or Kosher designation of foods
0.00000000000000000006%
of the estimated 1032 phages naturally
No adverse impact on normal microflora
present in the environment
 Environmentally-friendly, GREEN





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 Flexibility to address bacterial resistance
 Target specific bacterial pathogens
Cons
 Only effective against targeted pathogen
 Commonly used disinfectants may also inactivate phage
 Require refrigerated storage
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From: Bacteriophage, Volume 3,
issue 3
Panel A: SalmoFresh™ treated stainless steel and glass surfaces contaminated with serotype Kentucky S800; Panel
B: SalmoFresh™ treated glass surfaces contaminated with serotypes Brandenburg S806 or Paratyphi B S661; Panel
C: SalmoLyse™ treated glass surfaces contaminated with serotypes Brandenburg S806 or Paratyphi B S661; Panel D:
SalmoLyse™ treated stainless steel and glass surfaces contaminated with serotype Kentucky S800.
Phage cocktails can be rapidly
updated
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From: Woolston et al.,
Bacteriophage 2013; 3:e25697.
Intralytix phages are produced using state-of-the-art
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production technologies
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Plant Applications
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First plant application in 1925: Bacteriophages were first proposed to be potential agents
for controlling bacterial diseases of plants (blackleg disease of potato)
First field trial in 1935: Phages were successfully used to control Stewart's Disease in corn.
Treating seeds resulted in a reduction of disease incidence from 18% (untreated) to 1.4%
(phage).
In the late 1960s and early 1970s, phages were used to reduce (by 86 to 100%) Bacterial
Spot Disease of peach seedlings by Xanthomonas, and for managing other plant infections;
e.g., E. amylovora-associated fire blight
Additional examples of plant applications include application of bacteriophages for
managing Ralstonia solanacearum and Xanthomonascampestris pv. vesicatoria, the two
bacterial pathogens responsible for the two main diseases of tomato plants, known as
Bacterial Wilt and Bacterial Spot, respectively
Tomato plants treated with phages (center and
right) vs. phage-free control (left) Intralytix, Inc
Tomato plants treated with phages 3 and 5
days prior to inoculation with Ralstonia
solanacearum (center and right,
respectively) vs. phage-free control (left)
Source: Jones et al., Bacteriophage, Volume 2, Issue 4, 2012
Phage Biocontrol of Plant Diseases: Intriguing but
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not an easy Task
 Phage persistence / availability (e.g., Inactivation by UV light)
 Delivery optimization (translocate?)
 Bacterial resistance (field applications?)
 Cost
Additional Reading
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Elizabeth Kutter, Alexander Sulakvelidze. Bacteriophages:
Biology and Applications. CRC Press, 2005
Jones et al. Considerations for using bacteriophages for plant
disease control. Bacteriophage, Volume 2, Issue 4, 2012
For More Information
Intralytix, Inc
www.intralytix.com
https://www.youtube.com/watch?v=fFoo1OVa11E
https://www.youtube.com/watch?v=L3Ky3g4ZYQA
Alexander “Sandro” Sulakvelidze
asulakvelidze@intralytix.com
(+1) 410-625-2533
Safety By Nature™

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