Download Power Point

Howard Thomas
Citroxx
All plants have some
form of defense
mechanism against
damage or injury
Smell
Colour
Taste
Seville Orange
What is Citroxx?

Citroxx is a patented
combination of small & large
flavonoids molecules with
known anti-microbial / viral
activity: Neohesperidin
 Narangin
 Poncirin
 Rhiofolen

Enhanced activity by addition
of natural fruit acids: Malic
 Citric
 Ascorbate
Approvals
Citroxx has been developed over a number of years
and has been approved by the British Government’s
MAFF (now DEFRA) to control the onset of various
diseases such as:Newcastle’s Disease found in chickens
Foot and Mouth Disease
Tuberculosis
Swine Fever
Approvals
Citroxx is also approved by the Department of Health
(HPA) for use as a disinfectant.

Citroxx is also approved to decontaminate fresh
produce for Britain’s leading supermarkets.

Conforms to EU Organic Farming Regulations for the
treatment of fresh produce.
Effectiveness
Citroxx has been independently tested and found to be effective
(i.e. killing pathogens with an efficiency rate of 5 logs-up to
99.999*%)
The list of organisms against which Citroxx has been tested and
found to be effective is long:
Bacteria
Alternaria spp
Aspergillus flavus
Aspergillus niger
Aspergillus oizae
Aspergillus terrus
Campylobacter jejuni
Candida albicans
Chaetomium golbosum
Dipiodia natalensis
Escherichia coli
F. sp. tuberosa
Fusarium sambucinum
Geotrichum candidium
Klebsiella pneumonia
Lactobacillus pentoaceticus
Legionella pneumophila
(NCTC11192)
Effectiveness
Listeria monocytogenes
MRSA (clinical strain)
Mycobacterium fortutium (NCTC 8573 for indication of tuberculocidal
activity)
Penicillium digitatum
Penicillium funiculosum
Penicillium italicum
Penicillium roqueforti
Penicillium sp.
Phomopsis ortl
Proteus vulgaris
Pseudomonas aeruginosa (ATCC 15442)
Pullularia pullulans
Salmonella chloreraesuis
Salmonella typhimurium (DT004)
Scerotinia laxa
Staphylococcus aureus (NCTC 6571)
Staphylococcus pyogenes
Staphylococcus sp.
Sterptococcus faecalis
Trichophyton iterdigitale
Effectiveness

Viruses
H5N1 (Avian Influenza A)
Human Rhinovirus Influenza A Human Immunodeficiency Virus (HIV) Urbani SARS African swine fever
Foot & mouth disease
Gumboro virus
Herpes virus type 1
Herpes virus type 2
Herpes zoster
Hepatitis A & B
Effectiveness

Yeast and Fungi
Aspergillus flavus
Aspergillus niger
Aspergillus terreus
Botrytis cinerea
Candida albicans
Candida glabrata
Chaetonium globosum
Cladosporium
Collectotricum sp.
Fusarium sp.
Mucor sp.
Penicillium sp.
Pullularia pullulans
Pythium sp.
Trichophyton interdigitale
Trichophyton mentagrophytes
Protozoa
Histomonas meleagradis
Giardia lamblia
Entamoeaba histolytica
Blastocystis hominis
Broth Micro Dilution
MIC values for each micro organism in the presence of Citrox BC30.
BC30
A odontolyticus
1.000
A viscosus
C albicans
0.800
C glabrata
C krusei
0.600
C parapsilosis
C tropicalis
0.400
C difficile
P gingivalis
0.200
P buccae
P intermedia
0.000
0
0.
00
7
0.
01
5
0.
03
1
0.
06
2
0.
12
5
0.
25
0
0.
50
0
1.
00
0
2.
00
0
4.
00
0
8.
00
0
Absorbance (Turbidity)
1.200
% Citrox (v/v)
S gordonii
S sanguinis
Broth Micro Dilution
Candida albicans
1.200
Absorbance
1.000
0.800
BC30
MDC30
0.600
0.400
0.200
0.000
0
0.007 0.015 0.031 0.062 0.125 0.250 0.500 1.000 2.000 4.000 8.000
% Citrox (v/v)
Broth Micro Dilution
Candida glabrata
1.200
Absorbance
1.000
0.800
BC30
0.600
MDC30
0.400
0.200
0.000
0
0.007 0.015 0.031 0.062 0.125 0.250 0.500 1.000 2.000 4.000 8.000
% Citrox
Broth Micro Dilution
Candida krusei
0.900
Absorbance
0.800
0.700
0.600
0.500
BC30
MDC30
0.400
0.300
0.200
0.100
0.000
0
0.007 0.015 0.031 0.062 0.125 0.250 0.500 1.000 2.000 4.000 8.000
% Citrox (v/v)
Broth Micro Dilution
Candida parapsilosis
1.200
Absorbance
1.000
0.800
BC30
MDC30
0.600
0.400
0.200
0.000
0
0.007 0.015 0.031 0.062 0.125 0.250 0.500 1.000 2.000 4.000 8.000
% Citrox (v/v)
Broth Micro Dilution
Candida tropicalis
1.000
0.900
Absorbance
0.800
0.700
0.600
BC30
0.500
MDC30
0.400
0.300
0.200
0.100
0.000
0
0.007 0.015 0.031 0.062 0.125 0.250 0.500 1.000 2.000 4.000 8.000
% Citrox (v/v)
Broth Micro Dilution
Porphyromonas gingivalis
0.700
Absorbance
0.600
0.500
0.400
BC30
MDC30
0.300
0.200
0.100
0.000
0
0.007 0.015 0.031 0.062 0.125 0.250 0.500 1.000 2.000 4.000 8.000
% Citrox (v/v)
Broth Micro Dilution
Clostridium difficile
0.800
0.700
Absorbance
0.600
0.500
BC30
MDC30
0.400
0.300
0.200
0.100
0.000
0
0.007 0.015 0.031 0.062 0.125 0.250 0.500 1.000 2.000 4.000 8.000
% Citrox (v/v)
Biofilm Assay
Biofilm assay - BC30
2.00
Actinomyces odontolyticus
Actinomyces viscosus
1.50
Candida albicans
Candida glabrata
1.00
Candida krusei
Candida parapsilosis
Candida tropicalis
0.50
Clostridium difficile
Porphyromonas gingivalis
0.00
Prevotella buccae
0.
00
0%
0.
00
7%
0.
01
5%
0.
03
2%
0.
06
2%
0.
12
5%
0.
25
0%
0.
50
0%
1.
00
0%
2.
00
0%
4.
00
0%
8.
00
0%
Relative growth
Candida dubliniensis
-0.50
Citrox BC30 concentration (v/v)
Prevotella intermedia
Streptococcus gordonii
Streptococcus sanguinis
Biofilm Assay
Porphyromonas gingivalis
2.50
1.50
1.00
0.50
Citrox BC30 concentration (v/v)
8.
00
0%
4.
00
0%
2.
00
0%
1.
00
0%
0.
50
0%
0.
25
0%
0.
12
5%
0.
06
2%
0.
03
2%
0.
01
5%
-0.50
0.
00
7%
0.00
0.
00
0%
Relative growth
2.00
Biofilm Assay
Clostridium difficile
1.20
Relative growth
1.00
0.80
0.60
0.40
0.20
0.00
%
-0.20 00
0
0.
-0.40
7 % 15 % 32 % 62 % 25 % 50 % 00 % 00 % 00 % 00 % 00 %
0
0
0
0
0
1
2
5
0
0
0
0
0.
0.
0.
0.
0.
0.
0.
1.
2.
4.
8.
Citrox BC30 concentration (v/v)
Biofilm Assay
Citrox BC30 concentration (v/v)
8.
00
0%
4.
00
0%
2.
00
0%
1.
00
0%
0.
50
0%
0.
25
0%
0.
12
5%
0.
06
2%
0.
03
2%
0.
01
5%
0.
00
7%
0.90
0.80
0.70
0.60
0.50
0.40
0.30
0.20
0.10
0.00
-0.10
-0.20
0.
00
0%
Relative growth
Candida glabrata
Biofilm Assay
Candida parapsilosis
1.20
0.80
0.60
0.40
0.20
-0.40
Citrox BC30 concentration (v/v)
8.
00
0%
4.
00
0%
2.
00
0%
1.
00
0%
0.
50
0%
0.
25
0%
0.
12
5%
0.
06
2%
0.
03
2%
0.
01
5%
-0.20
0.
00
7%
0.00
0.
00
0%
Relative growth
1.00
Biofilm Assay
Candida tropicalis
1.40
1.00
0.80
0.60
0.40
0.20
Citrox BC30 concentration (v/v)
8.
00
0%
4.
00
0%
2.
00
0%
1.
00
0%
0.
50
0%
0.
25
0%
0.
12
5%
0.
06
2%
0.
03
2%
-0.40
0.
01
5%
-0.20
0.
00
7%
0.00
0.
00
0%
Relative growth
1.20
Biofilm Assay
Candida krusei
1.40
1.00
0.80
0.60
0.40
0.20
Citrox BC30 concentration (v/v)
8.
00
0%
4.
00
0%
2.
00
0%
1.
00
0%
0.
50
0%
0.
25
0%
0.
12
5%
0.
06
2%
0.
03
2%
0.
01
5%
0.
00
7%
0.00
0.
00
0%
Relative growth
1.20
Use of Citrox as an antimicrobial
agent
Elaine Waters
UCD School of Biomolecular and
Biomedical Science
Staphylococcus aureus

Gram+ cocci with many virulence factors
(including biofilm formation)

Important hospital pathogen

Drug resistant
Citrox is an effective
antimicrobial agent against
S. aureus

Very low MIC 0.008% (tested 15 S. aureus
clinical isolates)
Effect of Citrox on biofilms


1.
2.
3.
4.
5.
6.
Sub-inhibitory concentrations of Citrox does not
inhibit biofilm production
0.25% Citrox solution kills biofilm cells in <10 mins
Control
Tetracycline (200µg/ml)
Citrox (0.1%)
Citrox (0.25%)
Citrox (0.5%)
Citrox (1%)
1 2 3 4 56
Biofilm susceptibility assay (Alamar blue)
• Pink-metabolically active
• Blue – metabolically inactive
Future Work:

Effect of Citrox on mature biofilms –
Examine against more clinical strains and
compare to other classes of antimicrobial
drug (b-lactams, nucleic acid synthesis
inhibitors)
Clostridium difficile

Anaerobic, spore forming bacteria

Healthcare associated infection (ethanol doesn’t inhibit
spores)

High mortality rate

Collaborating with Prof. Séamus Fanning and Dr. Katie
Solomon to examine the effect of Citrox on C. diff spores
Use of Citrox to decontaminate a
room using the nebulair system
Prevent infections spreading
Disperses citrox as a 'dry' mist
to “fog” the room
Area bacteria are found in a hospital
room
Citrox is effective at cleaning a microbial
contaminated room using the Nebulair
Use of Nebulair in Beaumont
Hospital

In collaboration with Prof. Hilary Humphreys and Dr
Anthony Dolan

Use of Citrox/nebulair remove known concentrations
of methicillin-resistant Staphylococcus aureus (MRSA)
and vancomycin resistant Enterococcus faecalis
(VRE) from mattress cover

Compare with fogging a room with Hydrogen Peroxide
Use of Nebulair in Beaumont
Hospital

105 colony forming units (CFU) per 100cm2
killed in 3 hours using a 5% Citrox solution

Impressed with the size and convenience
of Nebulair
Future work with the Nebulair

Continue experiments with the Nebulair –
test on different surfaces with a known
number of bacteria – MRSA, VRE and C.
diff spores (in collaboration with Dr. Katie
Soloman)
Using Citrox to coat surfaces
In collaboration with Dr Denis Dowling and Dr Maria
Katsikogianni (UCD Engineering surface group)
-currently looking at coating food packaging surfaces (PET
Film)
-In the future hopefully use the technology to coat medical
devices

Lab line system

Try to increase Citrox attachment to surface

Establish how long Citrox will be still active on the
attached surface

pre-treating film with He/O2
Non pre-treated 100 passes
He/O2 pre-treated 100 passes
Citrox remaining attached after
1 hour in water
Optical Microscope
Citrox coating is more active against S. aureus
after 21 days on He/02 pre-treated film than
untreated film
6000
5000
4000
Pre-treated film (plantonic cells)
3000
Untreated film (plantonic cells)
Pre-treated film (Attached cells)
2000
Untreated film (Attached cells)
1000
0
Control
50 Passes
100 Passes
150 Passes
Future Work:

Continue work with PET Film – currently
examining S. aureus, Salmonella and E.
coli over a 35 day period
Future work: EnBIO

Use Co-blast system

Potential coat Hip replacements/titanium
surfaces with citrox to prevent infection
Acknowledgments








Dr Jim O’Gara
Sarah Rowe
Brian Conlon
Sarajane McTiernan
Shunyao Yang
Patrick Houston
Dr Linda Holland
Dr Clarissa Pozzi
Collaborators:
 Dr Denis Dowling
 Dr Maria Katsikogianni
 Dr Katie Soloman
 Liam Mannion
Citrox
Dr Howard Thomas
Funding:
Irish Research Council for
Science Engineering and
Technology (IRCSET)
Citrox
Thank You
Howard Thomas