J Appl Phycol

DOI 10.1007/s10811-013-0215-0

Antioxidant potential and antimicrobial efficacy of seaweed

(Himanthalia elongata ) extract in model food systems
Sabrina Cox & Grace Hamilton Turley &
Gaurav Rajauria & Nissreen Abu-Ghannam &
Amit Kumar Jaiswal

Received: 26 August 2013 / Revised and accepted: 20 November 2013

# Springer Science+Business Media Dordrecht 2013

Abstract Extracts from marine sources exhibit antimicrobial

and antioxidant properties in vitro and there has been great
interest within the food industry to move towards natural
methods of food preservation. Natural extracts from seaweeds
could potentially have a multiple functionality within the food
industry to increase safety and enhance the quality of food
products. The present study is aimed to assess the antimicrobial
activity of a hydrophilic extract from the fucoid brown alga
Himanthalia elongata in model food systems. Carbohydrate
and protein model food systems (CMFS and PMFS, respectively) were studied at varying concentrations (1 %, 5 % and
10 %) and bacterial inhibition of the extract was investigated
against Salmonella abony and Listeria monocytogenes. The
extract provided up to 100 % inhibition of the bacteria with a
bactericidal effect in CMFS, while a bacteriostatic effect was
seen in PMFS. In general, there was a significant difference (P
<0.05) between the efficacies of the extract against S. abony as
compared to L. monocytogenes with higher inhibition for S.
abony. In terms of antioxidants; the extract had a total phenolic
content of 34.0 mg GAE g1 of extract and a DPPH (2,2diphenyl-1-picrylhydrazyl) radical scavenging activity of
139.8 mg AAE g of extract. The results of the present study
S. Cox : G. Hamilton Turley : G. Rajauria : N. Abu-Ghannam (*) :
A. K. Jaiswal
School of Food Science and Environmental Health, College of
Sciences and Health, Dublin Institute of Technology, Cathal Brugha
Street, Dublin 1, Dublin, Ireland
e-mail: nissreen.abughannam@dit.ie
S. Cox
e-mail: sabrina.cox@dit.ie
G. Hamilton Turley
e-mail: gaurav.rajauria@dit.ie
G. Rajauria
e-mail: amit.jaiswal@dit.ie

are promising as it provides an insight for the inclusion of

seaweed extracts into real food systems.
Keywords Seaweeds . Antimicrobials . Food model
systems . Antioxidants . Himanthalia elongata . Phaeophyta

Introduction
The problem of oxidation and microbial contamination are
most common aspects of food preservation, especially when
the products develop undesirable flavours, unpleasant taste,
rancid odours, discoloration and other forms of spoilage responsible for the loss of quality, safety and shortening of shelf
life. Synthetic antimicrobial agents such as sodium benzoate,
sodium nitrite and sorbic acid; synthetic antioxidants such as
butylated hydroxyanisole (BHA), butylated hydroxytoluene
(BHT) and tert-butyl-hydroxyquinone (TBHQ) are widely
used in the food industry for preserving the safety and quality
of foods. However, these synthetic antioxidants have been
reported to be toxic and exerting a carcinogenic effect (Chen
et al. 1992). In recent times, there has been an increasing
tendency towards the use of natural substances instead of the
synthetic ones. With the increase in the price of raw materials,
the problem of cost benefits for chemical production is becoming more considerable.
Many secondary metabolites found in plants have a role in
defence against herbivores, pests and pathogens. The defensive forces of these secondary metabolites, are in the form of
antimicrobial agents such as alkaloids, flavanols and phenolic
compounds (Lopez-Malo et al. 2002). Many marine plants,
including seaweeds, often carry significantly less macro and
microepibionts on their thalli compared to co-occurring
biofilms on inanimate substrata (Hellio et al. 2001; Lam and
Harder 2007). Therefore, it has been assumed that seaweeds

J Appl Phycol

defend themselves against bacterial fouling by production of

secondary metabolites which prevent attachment and growth
of bacterial colonizers (Maximilien et al. 1998). Similarly,
antioxidant activity of marine algae may arise from pigments
such as chlorophylls and carotenoids, vitamins and vitamin
precursors, phenolics and other antioxidative substances,
which directly or indirectly contribute to the inhibition or
suppression of oxidation processes (Shahidi 2009). The environment in which seaweeds grow is harsh as they are exposed
to a combination of light and high oxygen concentrations.
These factors can lead to the formation of free radicals and
other strong oxidizing agents, but seaweeds seldom suffer
from any serious photodynamic damage during metabolism.
This fact implies that their cells have some protective antioxidative mechanisms and compounds (Matsukawa et al. 1997).
Research findings suggest that seaweeds are also rich in
antioxidants metabolites, which may be used to improve food
quality by increasing shelf life of food products (Bergman
et al. 2003; Sabeena Farvin and Jacobsen 2013; OSullivan
et al. 2013).
Over the last two decades, great deal of efforts have been
directed toward identifying low-cost natural products from
plant, animal and microbial origins. In particular, 'natural' products such as polyphenols have been reported to have a variety of
biological effects, including antioxidant, anticarcinogenic, antiinflammatory and antimicrobial activities (Xia et al. 2010).
Phenolic extracts from different plant origins, such as rosemary,
cocoa, olive oil (Bubonja-Sonje et al. 2011), onion, garlic
(Benkeblia 2004), essential oils (Gutierrez et al. 2009), mango
(Kaur et al. 2010), almond skins (Mandalari et al. 2010) and
seaweeds (Bansemir et al. 2006; Dubber and Harder 2008; Cox
et al. 2010) have demonstrated their antimicrobial activity
against numerous spoilage and pathogenic bacteria. Other studies carried out within food products such as fresh and cooked
pork mince (Moroney et al. 2013), chicken products (Kanatt
et al. 2010), and cod protein (Wang et al. 2010) have demonstrated the potential application of plant extracts as antimicrobial and antioxidant agents in order to prolong the shelf life of
the final products.
However, to our knowledge, reports on the antimicrobial activity of seaweed extracts within model food systems are limited. Therefore, the aim of the present study
was to investigate the antimicrobial efficacy of an Irish
seaweed extract in protein and carbohydrate model food
systems (CMFS) and also to evaluate its antioxidant
potential.

Sequential extraction and purification

Materials and methods

Preparation of carbohydrate food systems

Himanthalia elongata was purchased from Quality Sea Veg.

(Co Donegal, Ireland). The seaweeds were collected in October 2012 and stored at 18 C until further use.

Carbohydrate solutions were prepared at 1 %, 5 % and 10 %

(w/v) concentrations by dissolving glucose in sterile PBS
stock solution according to Simpson and Gilmour (1997).

H. elongata samples (5 g) were crushed with liquid nitrogen

using a mortar and pestle, and extracted sequentially with
solvents having different polarity. In the first phase of extraction, the powdered samples were mixed with equal-volume
medium to low polarity solvents mixture (50 mL) of chloroform, diethyl ether and n-hexane. The samples were flushed
with liquid nitrogen and were kept at 100 rpm, 40 C in a shaker
incubator. After 1.5 h, the supernatant was collected and the
residue of the seaweed samples was extracted again with high
polarity solvents mixture of 60 % (v/v) aqueous methanol under
the same extraction conditions. After the extraction, both the
liquid extracts obtained from medium to low polarity and high
polarity solvents were pooled and was filtered together using
Whatman #1 filter paper.
The crude extract obtained from sequential extraction was
partially purified with liquidliquid partition approach. A
biphasic solvent system of water and ethyl acetate (1:1, v/v)
was then prepared and left for 30 min before being utilized in
the purification of the supernatants. The equal amount of
biphasic solvent and the supernatant were mixed and stirred
for 1.5 h. The mixture was transferred in separatory funnels
and left overnight in order to get a clear separation of compounds between the layers. The separatory funnel were covered with tin foil and kept in the dark to avoid the degradation
of photosensitive compounds in the extract. The separated
aqueous and ethyl acetate layers were then collected after
repeated washing with water and filtered separately. The solvents were evaporated to dryness using a multi-evaporator
(Bchi Syncore Polyvap, Mason Technology, Ireland) at
37 C with a pressure ranging from 470 to 150 mbar and the
dried extracts obtained from aqueous and ethyl acetate fractions were dissolved in water and methanol, respectively. Both
the extracts were submitted for the estimation of total phenolic
content (TPC), antioxidant potential and extraction yield and
based on the results, only dried aqueous fraction was tested
against different food model systems.
Preparation of protein food systems
Bovine serum albumin (BSA; Sigma-Aldrich, Ireland) solutions were prepared at 1 %, 5 % and 10 % w/v concentrations
by dissolving suitable quantities of BSA in sterile phosphate
buffered saline (PBS) according to Simpson and Gilmour
(1997). The solutions were then filter-sterilised using 0.2-m
Nalgene syringe filters.

J Appl Phycol

The solutions were then filter-sterilised using 0.2-m Nalgene

syringe filters.
Antimicrobial activity
Microbial culture
Two species of common food pathogenic bacteria were used in
this study; Listeria monocytogenes ATCC 19115 and Salmonella abony NCTC 6017 (Medical Supply Company, Dublin,
Ireland). All cultures were maintained at 70 C in 20 %
glycerol stocks and grown in Tryptic Soy Broth (TSB) at
37 C overnight to obtain sub-cultures. Working cultures were
prepared from sub-cultures and grown at optimal conditions for
each bacterium for 18 h before analysis. Bacterial suspensions
were then prepared in saline solution (NaCl 0.85 %;
BioMrieux, France) equivalent to a McFarland standard of
0.5, using the Densimat photometer (BioMrieux Inc, France)
to obtain a concentration of 1108 colony forming units (CFU)
mL1. This suspension was then diluted in TSB to obtain a
working concentration of 1106 CFU mL1.
Antimicrobial activity assay
This assay was carried out according to the procedure of Cox
et al. (2010), with some modifications. The influence of varying
concentrations (8, 4, 2, 1, 0.5, 0.25 and 0.125 mg mL1) of
hydrophilic extract on efficacy in food model systems were
assessed against L. monocytogenes and S. abony using 96-well
microtitre plates. The dried seaweed extract was dissolved in
peptone water and an aliquot of 200 L of extract solutions were
added to the first horizontal row of each plate in quadruplicate.
All other wells were filled with 100 L of model food system.
Then, 100 L from the first horizontal row was serial diluted 2fold along each section. Bacterial suspension (100 L) was
added to the first, second and third vertical rows of each section.
The fourth vertical row was used for sample blanks, to which
peptone water was added (100 L), while the negative controls
and control blanks resided in the eighth horizontal row of each
section. Absorbance readings were taken at 0 and 24 h at 600 nm
using a microplate spectrophotometer (Powerwave, Biotek),
with 20 s of agitation before each optical density (OD) reading.
Sodium benzoate was used as a positive control. Percentage
inhibition was calculated according to the following equation:

Enumeration of bacteria
Aliquots of sample (100 L) from wells of the microtitre
plates were selectively spread plated in duplicate. Plate count
agar (PCA) was used as a growth medium and spread plating
was carried out under aseptic conditions. After incubation for
24 h at 37 C, the plates were observed for the presence or
absence of bacterial growth.
Total phenolic content
The total phenolic concentration was measured using the
FolinCiocalteau method as outlined by Cox et al. (2012).
The TPCs were expressed as mg gallic acid equivalent per
gram of extract (mg GAE g1).
DPPH radical scavenging activity
Free radical scavenging activity was measured by 2,2diphenyl-1-picrylhydrazyl (DPPH) according to the method
described by Jaiswal et al. (2012). The antioxidant potential
was expressed as mg ascorbic acid equivalent per gram of
extract (mg AAE g1).
Statistical analysis
All experiments were performed in triplicate and replicated
twice. All statistical analyses were carried out using STAT
GRAPHICS Centurion XV software (StatPoint Technologies,
Inc., USA). Statistical differences were determined using
ANOVA followed by least significant difference (LSD) testing. Differences were considered statistically significant when
P< 0.05.

Results
Initial studies were carried out to extract both hydrophilic and lipophilic compounds from the H. elongata.
However, preliminary results showed a very low yield
of lipophilic fraction (0.03 %) so based on these findings further studies were carried out with hydrophilic
fraction.
Extract concentration and efficacy


Bacterial inhibition %

OE
O


 100;

where O is (OD of the organism at 24 hOD of the organism

at 0 h) and E is (OD of the extract at 24 hblank at 24 h)
(OD of the eExtract at 0 hblank at 0 h).

Efficacy of seaweed extracts in carbohydrate model food

systems
The results of efficacy of hydrophilic seaweed extracts at
concentrations of 8, 4, 2, 1, 0.5, 0.25 and 0.125 mg mL1,
which were tested against S. abony and L. monocytogenes in
CMFS (1 %, 5 % and 10 %) are shown in Tables 1 and 2. The

J Appl Phycol
Table 1 Percentage inhibition (%) of seaweed extract against S. abony in carbohydrate model food system (CMFS) at varying concentrations
Extract concentration (mg mL1)
CMFS (%)

0.5

0.25

0.125

1
5
10

94.91.4ay
96.54.4ax
1000.0bx

90.20.5ay
78.33.3by
92.21.1ay

74.60.9az
11.55.1bz
48.41.1cz

Each value is presented as meanSD (n =6)

Means within each column with different letters (ac) differ significantly (P <0.05)
Means within each row with different letters (xz) differ significantly (P <0.05)

results highlight that higher hydrophilic extract concentrations

were positively correlated with higher percentage inhibitions
of the food-borne pathogens tested. Between 8 and 2 mg mL1
extract concentration, bacterial inhibition against S. abony
was detected in the range of 100 to 48.4 % (10 % CMFS),
however at concentrations less than 1 mg mL1, no bacterial
inhibition was observed. Percentage inhibition results against
L. monocytogenes were similar with no bacterial inhibition
detected below 1 mg mL1. There was no significant difference between 1 and 5 % CMFS inoculated with S. abony at
8 mg mL1 hydrophilic seaweed extract, or between 5 and
10 % CMFS (P >0.05). However, there was a significant
difference (P< 0.05) between 1 and 10 % CMFS for a
given concentration of extract. Concentration dependent
antibacterial activity was detected as the extract concentrations decreased and bacterial inhibition also declined.
When the efficacy of the extract against S. abony and L.
monocytogenes were compared; a significant difference
was found between the two food-borne pathogens. In
order to determine if the extract exerted bacteriostatic or
bactericidal effect spread plating was carried out. The
extract had a potential bactericidal effect in CMFS with
bacterial cell death being most probable as there were few
colonies or no growth observed on the plates.

Efficacy of seaweed extracts in protein model food Systems

(PMFS)
Similarly, as in the case of the extract incorporated into
CMFS, the greater the concentration of extract, the
higher the bacterial inhibition that was observed for
the PMFS. However, while no inhibition was detected
below 1 mg mL1 concentration in CMFS, bacterial
inhibition was exhibited at all extract concentrations in
PMFS (8, 4, 2, 1, 0.5, 0.25 and 0.125 mg mL 1).
Bacterial inhibition was high up to 100 % in 1 and
5 % PMFS against both pathogens (Tables 3 and 4). As
with CMFS, spread plating was carried out to assess the
potential mode of action of the extract in PMFS. The
qualitative spread plating method confirmed that the
extract worked in a bacteriostatic manner in PMFS, as
it exhibited an inhibitory effect preventing further
growth of the bacteria. With regard to both S. abony
and L. monocytogenes , all of the plates exhibited a
lawn of growth. It was evident, however, that
8 mg mL1 extract had a stronger bacteriostatic effect
against L. monocytogenes compared to 2 mg mL 1 as
the lawn of bacterial growth appeared thinner, in particular when compared to control plates.

Table 2 Percentage inhibition (%) of seaweed extract against L. monocytogenes in carbohydrate model food system (CMFS) at varying concentrations
Extract concentration (mg mL1)
CMFS (%)

0.5

0.25

0.125

1
5
10

78.63.4ax
81.92.6ax
90.81.6bx

68.91.2ay
58.02.8by
71.61.9 cy

67.30.7ay
23.98.9bz
50.97.7cz

14.20.1az

Each value is presented as meanSD (n =6)

Means within each column with different letters (ac) differ significantly (P <0.05)
Means within each row with different letters (xz) differ significantly (P <0.05)

J Appl Phycol
Table 3 Percentage inhibition (%) of seaweed extract against S. abony in protein model food system (PMFS) at varying concentrations
Extract concentration (mg mL1)
PMFS (%)

0.5

0.25

0.125

1
5
10

100.00.0av
100.00.0aw
77.60.2bv

100.00.0ay
100.00.0by
76.13.5cy

100.00.0av
100.00.0aw
67.36.8bw

85.55.7aw
70.90.7bx
28.63.5cx

67.01.8ax
62.33.8by
25.53.5cx

59.91.5ay
44.63.6bz
6.74.0cy

44.21.4az
44.10.9az
0.00.0bz

Each value is presented as meanSD (n =6)

Means within each column with different letters (ac) differ significantly (P <0.05)
Means within each row with different letters (vz) differ significantly (P <0.05)

Comparison of efficacy of extract against sodium benzoate

Sodium benzoate, a bacteriostatic preservative used in the
food industry, was used as a positive control in 5 % CMFS
and 5 % PMFS (Table 5). Spread plating confirmed that
sodium benzoate was bacteriostatic due to lawns of bacterial
growth after incubation, which is due to the bacteria growth
being inhibited rather than killing. The results were similar to
those produced by the extract incorporated into PMFS. Sodium benzoate also had a bacteriostatic effect in CMFS while
the extract had a bactericidal effect against the two pathogens.
At 8 and 4 mg mL1, the extract exhibited greater bacterial
inhibition and no L. monocytogenes inhibition was noted by
sodium benzoate at 4 mg mL1 or less. Sodium benzoate,
however, exhibited greater inhibition at 20.5 mg mL1 compared to the extract (Table 5). In contrast, the extract and
sodium benzoate had a bacteriostatic effect against the pathogens in PMFS; however, the extract had a much stronger
bacteriostatic effect compared to the artificial preservative.
At the conditions used in this study, the hydrophilic extract
exhibited a greater effect in CMFS and PMFS against both of
the studied bacteria as compared to sodium benzoate and there
was a significant difference (P< 0.05) between them. It was
also found that sodium benzoate had a stronger efficacy
against S. abony as compared to L. monocytogenes. At

8 mg mL 1 extract concentration (5 % PMFS), bacterial

inhibition of 100 % was displayed against S. abony and L.
monocytogenes (Tables 3 and 4), while sodium benzoate only
inhibited the pathogens by 67 and 30 %, respectively
(Table 5).
Total phenolic content and antioxidant potential of extract
The results from Table 6 indicate that the values of TPC,
antioxidant potential and extraction yield (%) was significantly higher (P <0.05) in the hydrophilic fraction as compared to
the lipophilic fraction of seaweed extract. The extraction yield
(%) of the hydrophilic fraction was 1.92 % while the TPC of
the same fraction was 34.0 mg GAE g1 of extract. The DPPH
radical scavenging activity of the same fraction was 139.8 mg
AAE g1 of extract.

Discussion
Recent studies have shown that the crude seaweed extract
from H. elongata possessed excellent antimicrobial and antioxidant properties (Rajauria et al. 2013; Cox et al. 2010). The
present study used a sequential extraction and purification
method in order to obtain hydrophilic and lipophilic extracts.

Table 4 Percentage inhibition (%) of seaweed extract against L. monocytogenes in protein model food system (PMFS) at varying concentrations
Extract concentration (mg mL1)
PMFS (%)

0.5

0.25

0.125

1
5
10

100.00.0av
100.00.0av
98.51.3av

83.77.8aw
100.00.0bv
62.69.9cw

81.83.2aw
100.00.0bv
61.99.4cw

67.83.2ax
80.15.1bw
58.61.9cx

44.43.5ay
44.60.8ax
56.35.3bx

40.21.8az
30.94.7by
39.93.3cy

39.11.5az
25.92.1bz
32.04.0cz

Each value is presented as meanSD (n =6)

Means within each column with different letters (ac) differ significantly (P <0.05)
Means within each row with different letters (vz) differ significantly (P <0.05)

J Appl Phycol
Table 5 Percentage inhibition (%) of sodium benzoate against L. monocytogenes and S. abony in 5 % protein model food system (PMFS) and
carbohydrate model food system (CMFS)
Sodium benzoate concentration (mg mL1)
8

0.5

16.00.5a
62.22.8bv

52.12.3aw

47.51.3ax

34.93.1ay

26.94.5az

30.24.1c
67.32.0dv

17.61.6bw

15.44.1bx

5 % CMFS
L. monocytogenes
S. abony
5 % PMFS
L. monocytogenes
S. abony

Each value is presented as meanSD (n =6)

Means within each column with different letters (ac) differ significantly (P <0.05)
Means within each row with different letters (vz) differ significantly (P <0.05)

The results show that the values of TPC and antioxidant

potential of hydrophilic fractions were 2.3 and 5.8 times
higher than the lipophilic fraction, respectively. Also, the same
extract showed a higher yield; therefore, based on these findings, the hydrophilic extract was focussed on as an antimicrobial for the present study.
As expected, higher extract concentrations resulted in
higher percentage inhibitions of bacteria. There was a greater
antimicrobial efficacy of the extract at 8 and 4 mg mL1
concentrations when applied to a 10 % CMFS compared to
1 %, meaning the extract can perform better with increasing
concentrations of glucose which may be related to a synergistic effect. Glucose causes a reduction in water activity (a w)
which in turn restricts microbial growth; therefore the CMFS
in combination with extract would have a potential synergistic
effect.
With regard to extract concentrations of 8, 4 and
2 mg mL1, the microtitre assay showed no significant difference (P >0.05) between the inhibition of S. abony and L.
monocytogenes in 5 % PMFS, as 100 % inhibition was
exhibited. Therefore, if the extract was applied it would be
more economical to use the lower concentration of
2 mg mL1, as it produces the same effect as the higher extract
concentrations. In general, it was found that the extract had a

greater antibacterial effect against S. abony compared to L.

monocytogenes.
Proteins are more structurally diverse than carbohydrates.
The PMFS, made up of BSA at varying concentrations, i.e.,
1 %, 5 % and 10 %, may have had a protective effect over the
pathogens. It is probable, based on the evidence obtained from
the microtitre assay and subsequent spread plating, that the
extract was able to penetrate the bacterial cells enough to
cause a bacteriostatic effect, i.e., inhibition of any further
growth. BSA is made up of three homologous domains which
are split into nine groups by 17 disulfide bonds and it also
contains tryptophan residues (Bourassa et al. 2011). Despite
its complexity compared to carbohydrates, Bourassa et al.
(2011) indicated that BSA can act as a transporter for compounds such as drugs.
In reported studies investigating the antimicrobial potency
of plant extracts, model food systems tend to decrease extract
efficacy (Cerrutti and Alzamora 1996; Del Campo et al.
2000). However, comparing results to the control (TSB in
place of model system) in the present study, results showed
that CMFS enhanced the overall antimicrobial potential.
Comparing the two bacteria tested, there was a greater percentage bacterial inhibition against S. abony (Gram-negative)
as compared with L. monocytogenes (Gram-positive). This

Table 6 Total phenolic content, antioxidant potential and extraction yield of hydrophilic and lipophilic fraction of H. elongata seaweed extract
Extract

Extraction yield (%)

TPC (mg GAE g1)

Antioxidant potential (mg AAE g1)

Aqueous fraction
Ethyl acetate fraction

1.920.07a
0.030.01b

34.01.1a
14.70.2b

139.82.7a
24.10.7b

Each value is presented as meanSD (n =6)

Means within each column with different letters (ab) differ significantly (P <0.05)
Extraction yield (%) is calculated in terms of g of dry extracts/100 g of fresh weight. TPC and antioxidant potential are expressed as mg gallic acid
equivalents/g (dw) and mg ascorbic acid equivalents/g (dw), respectively

J Appl Phycol

finding is of interest because the literature generally indicates

that Gram-positive bacterial species are more susceptible to
antimicrobial agents than Gram-negative (Rang et al. 2012).
According to Rang et al. (2012) and Russell and Chopra
(1996), the outer membrane of Gram-negative microbes possesses porins, which are essentially pores in the membrane
composed of proteins through which hydrophilic substances
can move freely. The brown seaweeds are known to contain
phlorotannins such as eckol, dieckol, and phloroglucinol
which are hydrophilic in nature. These compounds have been
reported to show a strong antibacterial activity against number
of microorganisms (Eoma et al. 2012). It is anticipated that the
partial purified aqueous extract could have enriched with
phlorotannins and showed strong antimicrobial activity
against tested bacteria. Furthermore, the peptidoglycan layer
in Gram-negative species is much thinner compared to Grampositive species (Golan et al. 2008). Therefore, within a 24 h
incubation period at 37 C, it is possible that the extract
travelled faster across the Gram-negative outer membrane
and cell wall of S. abony more easily than the thick peptidoglycan layer of L. monocytogenes.
It is known that food preservatives can either act in a
bacteriostatic or a bactericidal manner. In order to determine
which effect the extract adopted in carbohydrate and protein
conditions, a qualitative spread plating study was carried out.
With regard to extract incorporated into CMFS, a potential
bactericidal effect was observed with bacterial cell death being
most probable as there were few colonies or no growth observed on the plates. The current findings are also in line with
Ahn et al. (2004), who reported that phlorotannins from
brown seaweed showed a strong bactericidal effect against
food-borne bacteria.
Proteins are more structurally diverse than carbohydrates.
The PMFS, made up of BSA at varying concentrations, i.e.,
1 %, 5 % and 10 %, may have had a protective effect over the
pathogens. It is probable, based on the evidence obtained from
the microtitre assay and subsequent spread plating, that the
extract was able to penetrate the bacterial cells enough to
cause a bacteriostatic effect, i.e., inhibition of any further
growth. BSA is made up of three homologous domains which
are split into nine groups by 17 disulfide bonds and it also
contains tryptophan residues (Bourassa et al. 2011). Despite
its complexity compared to carbohydrates, Bourassa et al.
(2011) indicated that BSA can act as a transporter for compounds such as drugs.
There have been many studies carried out on the efficacy of
antimicrobials of plants in model food systems. However,
there have not been previous studies published in literature
in relation to marine extracts incorporated into model food
systems to challenge their antimicrobial efficacy. Gutierrez
et al. (2009) reported antimicrobial activity of essential oils
(EO) of oregano, thyme and lemon balm against L.
monocytogenes in lettuce leaf and beef model media. They

have reported that higher antimicrobial activity in the lettuce

leaf model compared to the beef model is due to the fewer
nutrients present in the lettuce media.
In a previous study, Moroney et al. (2013) examined the
effects which a spray-dried extract from brown seaweed
(Laminaria digitata) would have on the safety and quality
aspects of fresh and cooked minced pork. The results showed
no antimicrobial effect by the extract; however, the highest
concentration of extract (0.5 % w/w) exhibited antioxidant
potential against lipid oxidation to a slightly greater extent in
cooked patties compared to fresh and were significantly different (P< 0.05) to the control. However, lack of antimicrobial
activity could be due to the lower level of polyphenolic
compound present in L. digitata. Furthermore, the main components responsible for the antibacterial effects may be different in L. digitata as compared to H. elongata. Such extracts
may have a potential use as antioxidant agents in products
such as salad dressings which potentially enhance the shelf
life of the product.
Food spoilage due to the presence of bacteria causes economic losses on a global scale. The results of the present study
show that the seaweed extract could have potential as a source
for new antimicrobial agents equal to that of commercially
applied synthetic antibacterial agents. This would have potential commercial interest as Salmonella and Listeria infections
have a large impact on human health. The ubiquitous presence
of these microorganisms in the environment can lead to their
occurrence in food-processing environments and thus in the
food chain. As antioxidants are known to enhance the safety
of foods by preventing lipid oxidation, the total phenolic
content (TPC) and antioxidant potential of the extract was
evaluated. The seaweed extract had a TPC of 34.0 mg GAE
g1 of extract. These results are in line with Chandini et al.
(2008), who reported that brown seaweed extracts had a
phenolic content of 24.61 and 49.16 mg GAE g1 seaweed
extract. Wang et al. (2009) reported the TPC in different
Icelandic seaweeds to be ranging from 4 to 242 mg
phloroglucinol equivalants (PGE) g1 extract. The DPPH radical scavenging activity of the extract was 139.8 mg AAE g1
extract. In terms of percentage inhibition, at a concentration of
250 g mL1 extract, the DPPH free radical was scavenged by
47.7 %. At 500 g mL1 extract, the DPPH free radical was
scavenged by 64.2 % which is significantly different (P< 0.05).
The results of the present study are promising as algal polyphenolic compounds are effective antioxidants in delaying oil
rancidity, therefore the seaweed extracts could have potential
in food applications (Yan et al. 1996). A recent study in Iceland
by Wang et al. (2010) incorporated extract from a brown
seaweed (Fucus vesiculosus) into washed cod muscle systems
and cod protein isolates in order to evaluate its efficacy against
lipid oxidation of cod during ice storage. The results concluded
that the seaweed extract could potentially inhibit the lipid
peroxidation when used at higher amount compared to the

J Appl Phycol

synthetic preservatives. In the current study, the hydrophilic

extract of tested seaweed showed strong antioxidant and antimicrobial efficacy against different model food systems. Thus,
the study provides promising findings with potential to utilise
the extract in food or drink products, as antioxidants to enhance
food quality, and also as antimicrobial agents, increasing the
safety and shelf life of the products.
In conclusion, higher extract concentrations were positively correlated with the percentage inhibition of the food-borne
pathogens with up to 100 % inhibition of bacteria detected. A
significant difference was noted between the inhibitions of the
pathogens, as it was found overall that the antimicrobial
properties of the extract was more effective on S. abony. In
certain circumstances, there was no significant difference in
the efficacy of the extract at concentrations between 8 and
2 mg mL1. It can be concluded that hydrophilic seaweed
extract could potentially have a multi-purpose functionality
within foodstuffs to increase safety and enhance quality at low
concentrations.

References
Ahn MJ, Yoon KD, Min SY, Lee JS, Kim JH, Kim TG, Kim SH, Kim
NG, Huh H, Kim J (2004) Inhibition of HIV-1 reverse transcriptase
and protease by phlorotannins from the brown alga Ecklonia cava.
Biol Pharm Bull 27:544547
Bansemir A, Blume M, Schroder S, Lindequist U (2006) Screening of
cultivated seaweeds for antibacterial activity against fish pathogenic
bacteria. Aquaculture 252:7984
Benkeblia N (2004) Antimicrobial activity of essential oil extracts of
various onions (Allium cepa) and garlic (Allium sativum). Food
Sci Technol 37:263268
Bergman M, Perelman A, Dubinsky Z, Grossman S (2003) Scavenging
of reactive oxygen species by a novel glucurinated flavonoid antioxidant isolated and purified from spinach. Phytochemistry 62:753
762
Bourassa P, Dubeau S, Maharvi GM, Fauq AH, Thomas TJ, Tajmir-Riahi
HA (2011) Locating the binding sites of anticancer tamoxifen and its
metabolites 4-hydroxytamoxifen and endoxifen on bovine serum
albumin. Eur J Med Chem 46:43444353
Bubonja-Sonje M, Giacometti J, Abram M (2011) Antioxidant and
antilisterial activity of olive oil, cocoa and rosemary extract polyphenols. Food Chem 127:18211827
Cerrutti P, Alzamora SM (1996) Inhibitory effects of vanillin on some
food spoilage yeasts in laboratory media and fruit pures. Int J Food
Microbiol 29:379386
Chandini SK, Ganesan P, Bhaskar N (2008) In vitro antioxidant activities
of three selected brown seaweeds of India. Food Chem 107:707713
Chen C, Pearson AM, Gray JI (1992) Effects of synthetic antioxidants
(BHA, BHT and PG) on the mutagenicity of IQ-like compounds.
Food Chem 43:177183
Cox S, Abu-Ghannam N, Gupta S (2010) An assessment of the antioxidant and antimicrobial activity of six species of edible Irish seaweeds. Int Food Res J 17:205220
Cox S, Gupta S, Abu-Ghannam N (2012) Effect of different rehydration
temperatures on the moisture and phytochemical constituents of
dried edible Irish brown seaweed. Food Sci Technol 47:300307

Del Campo J, Amiot MJ, Nguyen-The C (2000) Antimicrobial effect of

rosemary extracts. J Food Prot 63:13591368
Dubber D, Harder T (2008) Extracts of Ceramium rubrum, Mastocarpus
stellatus and Laminaria digitata inhibit growth of marine and fish
pathogenic bacteria at ecologically realistic concentrations.
Aquaculture 274:196200
Eoma SH, Kimb YM, Kim SK (2012) Antimicrobial effect of
phlorotannins from marine brown algae. Food Chem Toxicol 50:
32513255
Golan DE, Tashjian AH, Armstrong EJ, Armstrong AW (2008) Principles
of pharmacology: the pathophysiological basis of drug therapy, 2nd
edn. Lippincott Williams and Wilkins, USA
Gutierrez J, Barry-Ryan C, Bourke P (2009) Antimicrobial activity of
plant essential oils using food model media: efficacy, synergistic
potential and interactions with food components. Food Microbiol
26:142150
Hellio C, De La Broise D, Dufosse L, Le Gal Y, Bourgougnon N (2001)
Inhibition of marine bacteria by extracts of macroalgae: potential use
for environmentally friendly antifouling paints. Mar Environ Res
52:231247
Jaiswal AK, Gupta S, Abu-Ghannam N (2012) Kinetic evaluation of
colour, texture, polyphenols and antioxidant capacity of Irish York
cabbage after blanching treatment. Food Chem 131:6372
Kanatt SR, Chander R, Sharma A (2010) Antioxidant and antimicrobial
activity of pomegranate peel extract improves the shelf life of
chicken products. Int J Food Sci Technol 45:216222
Kaur J, Rathinam X, Kasi M, Leng KM, Ayyalu R, Kathiresan S et al
(2010) Preliminary investigation on the antibacterial activity of
mango (Mangifera indica L: Anacardiaceae) seed kernel. Asian
Pac J Trop Med 3:707710
Lam C, Harder T (2007) Marine macroalgae effect abundance and
community richness of bacterioplankton in close proximity. J
Phycol 43:874881
Lopez-Malo A, Alzamora SM, Palou E (2002) Aspergillus flavus dose
response curves to selected natural and synthetic antimicrobials. Int
J Food Microbiol 73:213218
Mandalari G, Bisignano C, DArrigo M, Ginestra G, Arena A, Tomaino
A et al (2010) Antimicrobial potential of polyphenols extracted from
almond skins. Lett Appl Microbiol 51:8389
Matsukawa R, Dubinsky Z, Kishimoto E, Masaki K, Masuda Y, Takeuchi
T, Karube I (1997) A comparison of screening methods for antioxidant activity in seaweeds. J Appl Phycol 9:2935
Maximilien R, De Nys R, Holmstrom C, Gram L, Givskov M, Crass K,
Kjelleberg S, Steinberg PD (1998) Chemical mediation of bacterial
surface colonisation by secondary metabolites from the red alga
Delisea pulchra. Aquat Microb Ecol 15:233246
Moroney NC, O'Grady MN, O'Doherty JV, Kerry JP (2013) Effect of a
brown seaweed (Laminaria digitata) extract containing laminarin
and fucoidan on the quality and shelf-life of fresh and cooked
minced pork patties. Meat Sci 94:304311
OSullivan AM, OCallaghan YC, OGrady MN, Hayes M, Kerry JP,
OBrien NM (2013) The effect of solvents on the antioxidant
activity in Caco-2 cells of Irish brown seaweed extracts prepared
using accelerated solvent extraction (ASE). J Funct Food 5:940
948
Rajauria G, Jaiswal AK, Abu-Ghannam N, Gupta S (2013)
Antimicrobial, antioxidant and free radical-scavenging capacity of
brown seaweed Himanthalia elongata from Western coast of
Ireland. J Food Biochem 37:322335
Rang HP, Dale MM, Ritter JM, Flower RJ, Henderson G (2012)
Pharmacol, 7th edn. Elsevier, UK
Russell AD, Chopra I (1996) Understanding antibacterial action and
resistance, 2nd edn. Ellis Horwood, Hertfordshire
Sabeena Farvin KH, Jacobsen C (2013) Phenolic compounds and antioxidant activities of selected species of seaweeds from Danish coast.
Food Chem 138:16701681

J Appl Phycol
Shahidi F (2009) Nutraceuticals and functional foods: whole versus
processed foods. Trend Food Sci Technol 20:376387
Simpson RK, Gilmour A (1997) The effect of high hydrostatic
pressure on Listeria monocytogenes in phosphate-buffered
saline and model food systems. J Appl Microbiol 83:181
188
Wang T, Jonsdottir R, Olafsdottir G (2009) Total phenolic compounds,
radical scavenging and metal chelation of extracts from Icelandic
seaweeds. Food Chem 116:240248

Wang T, Jonsdottir R, Kristinsson H, Thorkelsson G, Jacobsen C,

Hamaguchi PY, Olafsdottir G (2010) Inhibition of haemoglobinmediated lipid oxidation in washed cod muscle and cod protein isolates
by Fucus vesiculosus extract and fraction. Food Chem 123:321330
Xia EQ, Deng GF, Guo YJ, Li HB (2010) Biological activities of
polyphenols from grapes. Int J Mol Sci 11:622646
Yan XJ, Li XC, Zhou CX, Fan X (1996) Prevention of fish oil rancidity
by phlorotannins from Sargassum kjellmanianum. J Appl Phycol 8:
201203