Food Chemistry 124 (2011) 721726

Contents lists available at ScienceDirect

Food Chemistry
journalhomepage: www.elsevier.com/locate/foodchem

Effect of ozone processing on the colour, rheological properties and phenolic

content of apple juice
B. Torres

, B.K. Tiwari

b,*

, A. Patras , Hilde H. Wijngaard , N. Brunton , P.J. Cullen

, C.P. ODonnell

Biosystems Engineering, UCD School of Agriculture, Food Science and Veterinary Medicine, University College Dublin, Belfield, Dublin 4,
b
Ireland Department of Food and Tourism, Manchester Metropolitan University, Manchester M14 6HR, UK
c
Teagasc, Ashtown Food Research Centre, Ashtown, Dublin 15, Ireland
d
School of Food Science and Environmental Health, Dublin Institute of Technology, Dublin 1, Ireland

article info
Article history:
Received 7 February 2010
Received in revised form 8 April 2010 Accepted 14 June 2010

Keywords:
Apple juice
Viscosity
Colour
Ozone
Polyphenol

of
plant
foods
1. Introduction
are
com
Apples are of interest for their nutritional monl
composition. Fruit juice obtained fromy
apples is increasingly promoted andlinke
consumed due to its reported beneficiald to
effects
on
degenerative
diseases,their
protective effects against cardiovascularpolyp
diseases and cancer ( Hollman, 2001;henol
Kahle, Kraus, & Richling, 2005; Rice-conte
2001; Robards, Prenzler, Tucker,nt (
Kahl
Swatsitang, & Glover, 1999; Sadik, Sies, &
e et
Schewe, 2003). Apples are an excellental.,
source of several phenolic compounds and2005
the presence of polyphenols in apples).
contributes towards their health promotingApart
antioxidant properties ( Khanizadeh, Tsao,from
Rekika, Yang, & DeEll, 2008; Robards et their
repor
al., 1999; Sanoner, Guyot, Marnet, Molle,
ted
& Drilleau, 1999; Van der Sluis, Dekker,healt
Skrede, & Jongen, 2002). Polyphenolsh
have attracted much attention due to their bene
fits,
antioxidant properties ( Proteggente et al.,
polyp
2002). In fact, the potential health benefitshenol
Evans,

abstra
ct
Apple
juice
samples
were
ozonated
with
processing
variables of
ozone
concentration
(14.8% w/w)
and

processing time (010 min). Effects of processing

variables on colour values (L, a and b), rheological
prop-erties and phenolic content were studied.
Significant reductions in these parameters were
observed dur-ing ozonation. Second order
polynomial regression modelling was used to
investigate the main effects of ozone concentration
and processing time on the changes in the selected
quality parameters of ozonated apple juice.
Predicted models were found to be significant (p <
0.05) with low standard error and high coefficients

s con-tribute towards theElsevier Ltd.

formation of hazes andAll rights
sediments,
the
devel-reserved. doi:
opment of characteristic10.1016/j.foo
flavours and prevent colourdchem.2010.
changes during processing06.050
( Oszmianski & Wojdylo,
2007).
Several incidents of
food borne disease have
been associated with apple
juice. In 1991, an outbreak
of
Escherichia
coli
O157:H7

1*

Corres

ponding author.
Tel.: +44 161
247 2178; fax:
+441616135.
E-mail
address:
b.tiwari@mmu.
ac.uk
(B.K.
Tiwari).
0308-8146/$ - see
front matter 2010

of determination (R ).
2010 Elsevier Ltd.
All rights reserved.

and
haemolytic
uremic
syndrome
was
linked
to
traditionally pressed
apple cider. E. coli
O157:H7
is
an
enteric
pathogen
with a low infectious
dose, which usually
causes
haemorrhagic colitis,
but has also the
potential to cause
haemolytic uremic
syndrome in young
children,
which
leads to them being
immunocompromised.
These
outbreaks led the
United States Food
and
Drug
Administration
(FDA)
to
issue
hazard analysis and
critical control point
(HACCP)
regulations for safe
and
sanitary

processing of juice ( United States Food,FDA,

2004
2001). Their primary performance standard
)
is a minimum 5-log reduction of the
highli
pathogens of concern in the juice being
ghte
processed ( USFDA, 2001). The FDAsd
approval of ozone as a direct food additive gaps
in 2001 triggered interest in ozone appli-in the
cations. A number of commercial fruit juice rese
processors in the US and Europe began arch
employing
ozone
for
pasteurisationliterat
resulting in the issuing of industryure
guidelines. However, these guidelines (with

respect to the criticaldemand

for
control
parameters
ofnutritious
foods,
ozone during microbialwhich are minimally
inacti-vation
in
liquidand
naturally
systems.
processed, has led
to interest in non
Conventional
thermalthermal technologies
processing of fruit juices
( Schilling et al.,
remains the most widely
2008). Ozone has
adopted technology for
been
investigated
shelf-life extension and
for
fruit
juice
pres-ervation
of apple
processing
juice. However, consumer
applications includ-

ing apple cider (

Choi & Nielsen,
2005; Steenstrup &
Floros,
2004).
Willams,
Sumner,
and Golden (2005)
studied the effect of
ozone
in
combination
with
dimethyl dicarbonate
and
hydrogen
peroxide

722

B. Torres et al. / Food Chemistry 124 (2011) 721726

obtainable (4.8% w/w) at

for orange juice preservation. Theythis flow rate ( Patil et al.,
Ozone
reported that a 5-log reduction of E.2009).
coli O157:H7 could be achievedconcentration in the gas
using ozone in combination withsupply was varied (1
dimethyl
dicarbonate.
Similarly,4.8% w/w of oxygen) and
recorded using an ozone
Patil, Bourke, Frias, Tiwari,
&
gas analyser (Model OLACullen, 2009 reported a 5-logDLS,
Ozoneservices).
reduction of E. coli NCTC 12900, in Ozone treatments were
<7 min in orange juice. Steenstrupperformed at 20 0.5 LC.
Dock and Floros (2004) re-ported
that the overall inactivation of E. coli 2.4. Colour determination
O157:H7 by ozone is fast enough for
colour
was
practical application in apple juice Juice
production. However, to date themeasured using a Minolta
effect of ozonation on the quality of colorimeter (Model CRKonica
Minolta
apple juice has not been reported.400,
The objective of this work was to Sensing, Inc., Osaka,
investigate the effect of ozonation onJapan) based on three
co-ordinates,
the colour, rheological prop-ertiescolour
and phenolic content of ozonated namely L, a, b. The colour
apple juice as a function of ozonevalues were ex-pressed
concentration and processing time. as L (whiteness or
brightness/darkness),
a
(redness/green-ness) and
b (yellowness/ blueness).
2. Materials and methods
The instrument (65L/0L
geometry, D25 optical
2.1. Chemicals for polyphenol
sensor, 10L observer)
analysis
was
calibrated
using
Caffeic acid, chlorogenic acid,white (L = 92.8; a = 0.8, b
0.1)
and
black
FolinCiocalteu Reagent (FCR),=
tiles.
Total
gallic acid and trans-4-hydroxy-3-reference
methoxy cinnamic acid were ob-colour difference (TCD)
which
indicates
the
tained from SigmaAldrich Chemical
magnitude
of
colour
Co. (St. Louis, MO, USA).
change after treatment
was calculated using Eq.
2.2. Preparation of apple juice
(1).
samples

(length
=
40
mm,
diameter = 26.66 mm,
gap width = 4000 lm) at
25 0.1 LC. Flow curves
were obtained for a shear
rate sweep be-tween 0.1
1
and 99 s . Shear stress
shear rate data was fitted
to a power law model (Eq.
(2)).

ga kcn1
where

ga is apparent

viscosity (Pa s), k is

consistency index, c is the
1

shear rate (s ) and n is

the flow behaviour index.
2.6. Polyphenol analysis

2.6.1. FCR assay

Total phenolic content
of
apple
juice
was
assessed using a modified version of the Folin
Ciocalteu
assay
(
Singleton, Orthofer, &
Lamuela, 1999). Gallic
acid was used as a
standard and the aqueous
gallic acid solution (200
1
mg l ) was diluted with
distilled water to give
appropriate
concentrations
for
a
standard curve. For the
anal-ysis, 100 ll of
methanolic fruit extract or
gallic acid standard, 100 ll
q

TCD
L L 2 a a of methanol, 100 ll of
Unpasteurised fresh apple juice
reagent
FolinCiocalteu

was squeezed using a Nutritas-ter
and 700 ll of Na2CO3
Colour values L, a and
juice extractor (Model N450, Rawb values were recorded
were added into 1.5 ml
Juice & Smoothies Ltd., Dub-lin,as the mean of triplicate
micro-centrifuge
tubes.
Ireland). Samples were placed inreadings.
The
samples
were
500 ml plastic bottles, and stored at
vortexed immediately and
25 LC prior to processing. Frozen2.5. Rheological analysis
the tubes were incubated
juice samples were processed within
in the dark for 20 min at
one month of juice preparation.
A controlled stress
room temperature. After
rheometer
(Carrimed
incubation all samples
2
2.3. Ozone processing
CSL

100,
TA
were
centrifuged
at
Instruments, UK) was
13,000 rpm for 3 min. The
Experiments were carried out in aused
for
rheological
absorbance
of
the
250 ml bubble column with a built-inanalysis. Samples of 3.5
supernatant
was
then
ml
were
measured
using
diffuser. Ozone was generated using
measured at 735 nm in 1
concentric
cylinder
an ozone generator (Model OL80,a
ml plastic cuvettes using
Ozoneservices, Canada). Oxygengeometry
a spectrophotometer (UVflow rate was con-trolled using a gas
1700 Pharma Spec, Shiflow regulator. The experimental
madzu,
Japan).
The
design for this work was based upon
results were expressed in
a parallel inactivation study for E. coli
mg gallic acid equivO157:H7, using the same control
alent/100 ml (mg GAE
conditions. A 5-log reduction was
1
100 ml juice).
achieved in under 5 min at an
1
optimum flow rate of 0.125 l min
2.6.2. Polyphenolic profile
and a maximum ozone concentration
(HPLCDAD)

HPLC-analysis was performed onfiltered using a PTFE

a Varian Pro Star (Varian Inc.,syringe with 0.22 lm
Walnut Creek, USA) chromatographyfilters.
The
injection
system, equipped with a module 210volume
was
10
ll.
solvent delivery system, a module Hydroxybenzoic
acids,
510 column ther-mostat, a moduledihydrochalcones,
410 autosampler and a module 335flavanones and flavanols
diode array detector (DAD) with anwere monitored at a
absorbance detection range betweenwavelength of 280 nm,
190 and 950 nm. Separations werehydroxycinnamic
acid
conducted on a Zorbax SB C18, 5derivatives at 320 nm and
lm, 150 4.6 mm column (Agilentflavonols at 360 nm. For
Technologies, Dublin, Ire-land). Thequantification
and
gradient profile was based on the identifica-tion
purposes
method of Tsao and Yang (2003).standard
curves
of
Acetic acid in 2 mM sodium acetate analytes of interest were
(final pH 2.55, v/v) was used aspre-pared
using
eluent A and 100% acetonitrile wasmethanolic solutions of
used as eluent B. The columnthe
standards
listed
temperature was set at 37 LC andabove.
1
the flow rate was 1 ml min . The
solvent gradient programme was set
as follows: initial conditions 100% A, 2.7. Experimental design
0% B; 045 min, 015% B; 4560
min, 1530% B; 6065 min, 3050% Degradation of colour
B; 6570 min, 50100% B. Prior tovalues (L, a and b), total
injection sample extracts werecolour difference (TCD)

and phenolic content was

analysed using secondorder poly-nomial models.
Polynomial
regression
equations
were
developed to describe the
effects
of
ozone
concentration (% w/w)
and process-ing time
(min) on apple juice. The
general form of the
quadratic
polynomial
model
regression
equation employed in this
study is shown in Eq. (3).

Y b0

bijXiXj;

i1

XX

2
X biiXi

iXi

i1

where Y is the predicted

response, b0 the constant
coefficient, bi the linear
coefficient,
bii
the
quadratic coefficient, bij is
the cross-product

i ji1

B. Torres et al. / Food Chemistry 124 (2011) 721726

723

phenolic
compounds
coefficients. This equation containsnamely chlorogenic acid
linear terms X1, X2 quadratic or(234 lg/ml), caffeic acid
square terms X21; X22 and(18.5 lg/ml) and cinnamic
interaction (X1X2) terms are also in-acid (130 lg/ml) were
cluded in the equation. By using this detected in fresh apple
equation linear, quadratic andjuice. Kahle et al. (2005)
interactive effects of independent
reported that polyphenol
variables Xi (04.8% w/w) and X j (0content ranges from 154
10 min) on dependent variable (Y)to 970 mg/l in dessert and
were determined. Three dimensionalcider apple juices. The
curves of the response surface weredistribution of many of the
developed using Minitab (V.15.0)phenolics in apple juice is
software while holding the variablesinfluenced by various
con-stant in the second-order
factors including genetic
polynomial model. Analysis of
or cultivar, extrac-tion
variance (ANOVA) was carried out
method, processing, and
using the PROC NLIN procedure
(SAS V.9.1, SAS Institute, NC, USA).enzyme
treatment
(
All trials were conducted in triplicate. Khanizadeh,
Tsao,
Rekika, Yang, & DeEll,
2007; Khanizadeh et al.,
3. Results and discussion
2008).
3.1. General
3.2. Colour degradation
Table 1 lists the characteristics of
Fig. 1(iiv) shows the
the apple juice prior to ozone
treatment. Total phenol content3D response surface plots
expressed as gallic acid equivalentfor L, a, b and TCD
(GAE, mg/100 ml) was 638 123 values. Colour values for
untreated apple juice
mg/100 ml of juice. Three major
were 32.6 0.79, 22.2
0.18, 35.9 0.71 for L, a
and b value respectively.
Table 1
Characteristics of fresh and ozonated apple During ozonation juice
juice.
samples were observed
Parameters
Fresh apple to be lighter in col-our i.e.
increased L and b value,
juice
bwhereas a values of apple
L value
32.6 0.79
ajuice samples were found
a value
22.2 0.18
bto decrease with increase
b value
35.9 0.71
Chlorogenic acid
226 53.98 in processing time and
a
Caffeic acid
18.5 2.40 ozone
concentration.
a
Cinnamic acid
130 22.8 Mean L and b values
a
Total phenol (GAE, mg/
638 123 significantly in-creased (p
100 ml)
a
pH
3.81 0.23 < 0.05) to 56.0 2.81 and
a
45.8

1.10
Consistency index (k)
9.11 0.03 to
while the
brespectively,
Flow behaviour index (n)
0.37 0.01
mean a value decreased
ab
Values followed by the same letter are
not significantly different (p < 0.05).

difference (TCD) value

was 30.84 at the high-est
processing
conditions
employed i.e. at ozone
concentration of 4.8%
w/w and processing time
of 10 min.
L, b and TCD values
were
significantly
influenced by both ozone
concentration
and
processing time (p <
0.0001).
Whereas,
a
value
significantly
increased with ozone
concentration
(p
=
0.0056) and processing
time (p = 0.0016) as
shown in Table 2. Table
3 shows the regression
coefficient
for
the
predicted
models
for
colour val-ues. It is
indicated
that
the
predicted
response
models
for
colour
parameters were found to
fit
well
with
the
experimental data. The
models
presented
showed high correlation
2

coefficients (R ) of 0.97,
0.89 and 0.92 for L, b and
2

TCD, however the R

value for a was 0.62.
Predicted models were
highly significant for all
colour values (p < 0.0001)
except a (p = 0.005).
Canonical
analysis
and ANOVA of second
order quadratic mod-els
revealed
that
linear
models were significant
with p < 0.0001 for

to 8.89 1.59 ( Table 1)

and the mean total colour
w/
w)

(i)

(
ii
)
2
5

50

L40
30

1.5

20

0.0Ozone

1
0

3.0

oncentration

4
.5

(%

2
0
1
5
1

0
0.
0
O
zo
ne

(
%

10

4.5
w/w)

e
im
T

(m

in

(iii)

(iv)
45

50

30

40

1
0

30
20
0.0
Ozone

1.5

3.0

concentration .5
(
%
w/
w
)

1
5

1.5

1
0

3.

0 0
. .
0c

o
On
zc
e
ont
ra
n
ti
eo
n

(
%
w
/
w
)

in

e (m
im
T

Fig. 1.

Effect of

o z o

co

at i m
n
e

B. Torres et al. / Food Chemistry 124 (2011) 721726

L, b, TCD and p < 0.001

for a value. Quadratic
models were insignif-icant
for a and b values and
significant for L (p < 0.01)
Ozone concentration
and TCD (p < 0.001)
(% w/w)values. Statistical analysis
that
the
F valueshowed
interaction (cross-product)
142
5.76among parameters was
37.0 significant for all colour
45.4
parameters in all cases.
5.11
5.02Overall colour changes
during
11.6 observed
0.45ozonation were distinct,
indicating major changes

Table 2
ANOVA of the factors obtained from RIDGE
analysis of the regression model for various
parameters.
Parameters

L value
a value
b value
TCD
Chlorogenic acid
Caffeic acid
Cinnamic acid
Total phenol (GAE, mg/100 ml)

1*

Significant
at
p
< 0.01.

1**

Significant
p
< 0.001. at

(0.25)
Quadratic
b
Table 3
Regression coefficient and
ANOVA of regression
parameters of the predicted
quadratic models.

11

33.3
(0.84)

Linear
b1
b2

12

Intercept
c

1***
cant

Signifi
at p <
0.0001.

of

3.3. Rheological
properties
Apparent
viscosities
as a function of shear rate
for ozonated ap-ple juice
are shown in Fig. 2. Flow
curves for all samples
exhibit shear thinning
behaviour
during
ozonation.
Significant
changes

0.01

Cross product
b

Factors

22

0.24
(0.10)
0.07
(0.02)

in the appear-ance
apple juice colour.

Viscosity(Pa.s)

724

R
CV (%)
Model

0.79
(0.04)
0.97
4.02
<0.0001

Total colour difference.

b Gallic acid equivalent (
Values
in parenthesis
() indicate standard error.
c1* Significant
at
p
< 0.05.

0.001

****

3.69
(0.55)
****
1.51

concentration
Fig. 2. Effect of ozone (% w/w) on

the apparent cloudy apple juice at a fixed

viscosity of processing time of 10 min.

Shearrate(s1)

10

15

B. Torres et al. / Food Chemistry 124 (2011) 721726

Tiwari, ODonnell, Patras,

were observed in consistency (k) and
flow behaviour (n) indices of apple
juice after ozonation. Cloudy apple
juice is a dilute colloidal dispersion of
solid particles in a solution of pectins,
proteins, sug-ars, organic acids, and
salts and hence has a high
consistency index (k) ( Bentez,
Genovese, & Lozano, 2007, 2009).
The
consistency
in-dex
(k)
decreased as a function of ozone
concentration from 9.11 (control) to
1.3 (ozone concentration 4.8% w/w
and processing time of 10 min) (
Table 1). A trend towards increased
Newtonian
flow
behaviour
at
increasing ozone concentration and
processing time was observed, with
ozonated
apple
juice
(ozone
concentra-tion
4.8% w/w
and
processing time of 10 min) showing
the highest n value of 0.65 compared
to 0.37 for control ( Table 1). A
decrease in apparent viscosity (Pa
s), k value and an increase in n value
during ozonation arise from the
breakdown of a colloidal suspension
which may be caused by the
depolymerisation of macromolecules
present in the juice suspension (
Tiwari,
Muthukumarappan,
ODonnell, Chenchaiah, & Cullen,
2008).

3.4. Phenolic content

Fig. 3(iiv) shows the 3D
response
surface
plots
for
chlorogenic acid, caffeic acid,
cinnamic acid and total phenol
content of apple juice during ozone
processing. During ozonation at
higher processing conditions (4.8%
w/w ozone concentration for 10 min
processing time) a decrease of
99.1%, 96.6%, 99.8% and 49.7%
was observed for chlorogenic acid,
caffeic acid, cinnamic acid and total
phenol
content
respectively.
However, a processing time required
to achieve 5-log reductions for E. coli
under similar experimental conditions
( Patil, Valdramidis, Cullen, Frias, &
Bourke, 2010) showed a 66.5%,
73.5% and 65.0% reduction for
chlorogenic acid, caffeic acid and
cinnamic acid respectively. It was
found that polyphenols are prone to
oxidation as observed in the case of
strawberry
and
blackberry
anthocyanins ( Tiwari, ODonnell,
Muthukumarappan, & Cullen, 2009;

725

Brunton, & Cullen, 2009).

This reduction is most
likely due to the strong
oxidation potential (+2.07
eV)
of
ozone.
The
degradation
of
polyphenols
during
ozonation may result from
a variety of pos-sible
chemical reactions. These
reactions may be direct
reactions of ozone with
the target compound or its
intermediates and radical
reactions
between
hydroxyl
radicals
(produced through ozone
decomposition catalysed
mainly by the hydroxide
ion (OH) ( Cullen, Tiwari,
ODonnell,
&
Muthukumarappan,
2009).
Chlorogenic
acid,
caffeic acid, cinnamic acid
and total phenol content
of ozonated apple juice
were
all
significantly
influenced by processing
time (p < 0.0001) while
ozone concentration was
not significant for total
phenol ( Table 2). Table
3 shows the regres-sion
coefficient
for
the
predicted colour value
models. The pre-dicted
response
models
for
phenolic content were
found to fit well with the
experimental data. The
models
presented
showed high correlation
2

coefficients (R ) of 0.90,
0.93, 0.92 and 0.97 for total phenol, chlorogenic
acid, caffeic acid and
cinnamic acid respectively. Predicted models
were highly significant for
phenolic content (p <
0.0001).
Canonical
analysis
and ANOVA of second
order quadratic mod-els
revealed that both linear
and quadratic models
were significant with p <
0.0001 for all parameters.
Whereas, the interaction
(cross-product)
among
parameters
was
insignificant in every case
except for cinnamic acid
(p < 0.01) indicating that
only linear and qua-dratic
effects of the independent

variables of ozone concentration andspecies, such as OH,

2
processing time caused significantHO , O2, and O 3 which
effects on the response sur-face.facilitate degradation. A
According to Xue, Chen, and Wangsimilar
oxi-dative
(2008) ozone plays an important roledegradation of ascorbic
not only in the degradation processacid in presence of
of organic dye but also in the oxygen has been reported
formation of other high-reactiveby
Kennedy, Rivera,

Lloyd, Warner, and Jumel

(1992). Zimeri and Tong
(1999)
reported
a
degradation mechanism
of
epi-gallocatechin
gallate in the presence of
dissolved oxygen in a
mod-el liquid solution.

(
i
i
)

(i)
240acid180

120

Chlorogenic

60

1
0

0
0.0
Ozone

1.5

3.0

concentration

(%

4.5 0
w/
w)

cCaffei acid

1
1
0
5
1.5

concentration 4
(
%

.
5
w
/
w
)

5
im
T

(m

in

(iii)

Cinnami
c
acid

120
80
40
0

in

0.0
Ozone

1.5

3.0

concentration

(%

4. 0
5
w/
w)

1.5

(m
T
i
m
e

Ozone
3.0
con
cent
rati
on

4
.

10

in

e (m
im
T

Fig. 3.

Effect of

o z on e

c n ce

ti o n (%

726

B. Torres et al. / Food Chemistry 124 (2011) 721726

Khanizadeh, S., Tsao, R.,

Rekika, D., Yang, R.,
4. Conclusion
Charles,
M.
T.,
&
Rupasinghe, H. P. V.
(2008).
Polyphenol
The results presented in this
composition
and
total
study demonstrate the effect of
antioxidant capacity of
selected apple genotypes
ozone concentration and processing
for processing. Journal of
time
on
colour
degradation,
Food Composition and
rheological properties and retention
Analysis, 21(5), 396401.
of polyphenols. Apple juice colour,Khanizadeh, S., Tsao, R.,
Rekika, D., Yang, R., &
rheological properties and phenolic
DeEll, J. (2007). Phenolic
content were observed to be
composition
and
significantly influenced by ozonation.
antioxidant
activity
of
selected apple genotypes.
It is concluded that while ozonation
Journal
of
Food
can be employed as a preservation
Agriculture
and
technique for processing of apple
Environment, 5(1), 6166.
juice, its impact on the nutritional and Oszmianski, J., & Wojdylo, A.
(2007). Effects of various
quality parameters of apple should
clarification treatments on
be considered.
phenolic compounds and
color of apple juice.
European Food Research
Acknowledgement
and Technology, 224(6),
755762.
Funding for this research wasPatil, S., Bourke, P., Frias, J.
provided under the Irish NationalM., Tiwari, B. K., & Cullen, P. J.
(2009). Inactivation of
Development Plan, through the Food
Escherichia coli in orange
Institutional Research Mea-sure
juice using ozone.
Innovative Food Science &
(FIRM), administered by the Irish
Emerging Technologies,
Department of Agriculture, Fisheries
10(4), 551557.

& Food (DAFF).

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