Innovative Food Science and Emerging Technologies 45 (2018) 412–417

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Innovative Food Science and Emerging Technologies

journal homepage: www.elsevier.com/locate/ifset

The effect of non-thermal plasma on the lipid oxidation and microbiological T

quality of sushi
⁎
Piotr Kulawika, , Carlos Alvarezb, Patrick J. Cullenc, Ramon Aznar-Rocad, Anne Maria Mullenb,
Brijesh Tiwarid
a
Department of Animal Product Technology, Faculty of Food Technology, University of Agriculture, Krakow, Poland
b
Department of Food Quality and Sensory Science, Teagasc Food Research Centre, Ashtown, Dublin, Ireland
c
Dublin Institute of Technology, Dublin, Ireland
d
Department of Food Biosciences, Teagasc Food Research Centre, Ashtown, Dublin, Ireland

A R T I C L E I N F O A B S T R A C T

Keywords: The study examined the effect of non-thermal plasma (NTP) on the total viable count and lipid oxidation of two
Sushi common sushi products: nigiri and hosomaki. Sushi samples were treated with NTP using a dielectric barrier
Non-thermal plasma discharge system with 70 and 80 kV of potential differences for 5 min. The samples were stored at 4 °C for
Lipid oxidation 11 days and analysed for total aerobic count, moisture and protein content, TBA index and fatty acids compo-
Microbiological quality
sition. Although the effect of NTP on the total aerobic counts was not statistically significant, a tendency in log
Nigiri
Maki
reduction could be observed, with 1–1.5 log cfu/g reduction. Moisture and protein content, as well as fatty acids
composition of sushi was not affected by the treatment. The TBA index of treated samples increased significantly
by 0.4–1.5 mg/kg, with hosomaki reaching higher TBA index than nigiri. Although NTP in the studied conditions
can, to a limited degree, increase the microbiological quality of the sushi samples, it also increases the oxidation
rate.

1. Introduction important sushi ingredient, but cannot be used for many unprocessed
fish species, including salmon, since it can cause changes in physico-
Globally there is a growing trend for the consumption of ready-to- chemical properties such as color and texture (S. Zhu, Ramaswamy, &
eat (RTE) sushi meals. According to Altintzoglou, Heide, Wien, and Simpson, 2004), and increase lipid oxidation and free fatty acids for-
Honkanen (2016) consumers from Norway and Japan purchase RTE mation (Sequeira-Munoz, Chevalier, LeBail, Ramaswamy, & Simpson,
sushi meals 0.47 and 1.75 times per week, respectively, and the Nor- 2006). Processing techniques which are used in sushi processing, in-
wegian Seafood Council reports that 18% of Germans who frequently clude modified atmosphere packaging (MAP), and treatment with
consume sushi, purchase it in the form of a RTE meal from supermarket ozone, electrolyzed water, ultrasound and UV radiation. Those methods
(NSC, 2015). can be used separately or in combination, however, the highest noted
RTE sushi meal producers are challenged by the short shelf-life of shelf-life improvement was from 3 to 7 days (Steffen, Duerst, & Rice,
their product, with microbiological spoilage and rice retrogradation 2010); nevertheless, using the combination of all those methods re-
being one of the main factors affecting the product's shelf-life. Sushi quires high investment and maintenance costs.
contains raw fish and seafood, therefore the bacterial concentration Starch retrogradation is a process in which amylose and amylo-
cannot be reduced using thermal treatments and the use of pre- pectin chains, disrupted during thermal treatment in the presence of
servatives is typically not employed due to the desire from consumers water, reassociate creating different crystalline structures. This results
for a product with a ‘clean label’ classification (Don, 1997). Moreover in texture changes and might affect the sensory parameters and con-
not all minimal processing techniques, however, can be implemented sumer acceptance of the final product (Wang, Li, Copeland, Niu, &
into sushi processing. For example high-pressure processing (HPP), can Wang, 2015). Starch retrogradation can be divided into short-term and
be used to effectively reduce microbiological counts and browning for long-term retrogradation. The first appears usually within hours up to
avocado (Jacobo-Velázquez & Hernández-Brenes, 2010; López-Malo, 2 days of storage, and is caused by recrystallization of amylose, while
Palou, Barbosa-Cánovas, Welti-Chanes, & Swanson, 1998), an the latter might last several weeks and is caused by formation of

⁎
Corresponding author at: Department of Animal Products Technology, Faculty of Food Technology, University of Agriculture, ul. Balicka 122, 30-149 Cracow, Poland.
E-mail address: [email protected] (P. Kulawik).

https://doi.org/10.1016/j.ifset.2017.12.011
Received 5 September 2017; Received in revised form 15 December 2017; Accepted 20 December 2017
Available online 22 December 2017
1466-8564/ © 2017 Elsevier Ltd. All rights reserved.
P. Kulawik et al. Innovative Food Science and Emerging Technologies 45 (2018) 412–417

amylopectin crystals around the crystal nuclei formed by amylose (Fu, round polyethylene terephthalate (PET) containers and sealed in air
Chen, Luo, Liu, & Liu, 2015). Rice starch retrogradation decreases using traysealer machine (Olympus, Italian Pack, Como, Italy).
sensory properties and consumer acceptance (Kwak, Kim, & Jeong,
2015) which in turn affects shelf-life of stored RTE sushi meals.
Non-thermal plasma (NTP) is a novel preservation method in which 2.3. Plasma treatment
the treated product is exposed to a direct plasma discharge or its
afterglow (gas reactive species). Decontamination of the product's Plasma was generated using a dielectric barrier discharge system, as
surface can result from action of reactive species (RS) and UV radiation described by Misra et al. (2014). PET containers containing sushi
(Boudam et al., 2006; Ehlbeck et al., 2011; Moreau, Orange, & samples were placed between two 158 mm diameter aluminium disc
Feuilloley, 2008). The exact type of formed RS depends on the atmo- electrodes with two polypropylene dielectrics – 10 mm thick acrylic
sphere used to form the plasma which for atmospheric air includes glass used at the high voltage electrode and 2 mm thick polypropylene
reactive oxygen species, reactive nitrogen species, excited oxygen and at the ground electrode. The distance between the electrodes was
nitrogen, hydrogen peroxide, OH radicals, H2O+ and OH– ions (Scholtz, 52 mm (height of the container plus the dielectrics thickness). Previous
Pazlarova, Khun, & Julak, 2015). Non-thermal plasmas have demon- tests were performed to establish the most adequate treatment time
strated efficacy for various food pathogens, including Escherichia coli, (data not shown). Sushi samples were stored in ice and treated for 1, 3,
Staphylococcus aureus or Bacillus cereus achieving reduction rates of up 5 and 10 min with 70 and 80 kV discreet voltages generated by high
to 7 log cfu/g (Ehlbeck et al., 2011). Any associated increase in the voltage step-up transformer (Phoenix Technologies Inc., Accident, MD,
temperature of the treated surface is limited (Fridman et al., 2007) USA) at 50 Hz frequency. Afterwards, the samples were stored for 1 day
which suggests that non-thermal plasma could be effectively used to and analysed for total viable count (as per method described in Section
increase the shelf-life of sushi products. Conversely, NTP can increase 2.4). Based on the results of the preliminary analysis the shortest, most
lipid oxidation in complex food matrices (Van Durme, Nikiforov, effective treatment time was chosen to be 5 min of plasma treatment.
Vandamme, Leys, & De Winne, 2014), and might not be suitable for The sushi samples used for all subsequent analyses were treated in the
food products containing high levels of unsaturated fatty acids, such as same manner as in preliminary tests but with only 5 min as a treatment
fish (Misra, Tiwari, Raghavarao, & Cullen, 2011) or sushi. To date there time. After the plasma treatment the samples were stored at 4 °C until
is no data which would allow establishing whether NTP can be a sui- subsequent analysis.
table method for prolonging the shelf-life of sushi. Therefore the aim of
this study was to establish the effect of NTP on the bacterial con- 2.4. Microbiological analysis
centration and lipid oxidation of two common sushi products: nigiri and
hosomaki, using a dielectric barrier discharge (DBD) system with two The analysis of total viable count (TVC) was performed according to
different electric potential differences: 70 and 80 kV. ISO standard no. 4833–1:2013 with a slight modification related to the
amount of sample used. In case of nigiri exactly 6.25 g of rice and 3.75 g
2. Materials and methods of salmon from each nigiri was homogenized with 90 g of maximum
recovery diluent (Sigma-Aldrich, Saint Louis, MO, USA). In case of
2.1. Materials hosomaki, a whole hosomaki piece (13.5–14.0 g) was homogenized
with maximum recovery diluent in ratio of 1:9. The homogenization
Sushi rice (Oryza sativa japonica), nori (dried Porphyra tenera was carried out using Stomacher device for 2 min followed by pre-
sheets), rice vinegar, sugar and salt were acquired from the local market paration of appropriate decimal dilutions. The analysis was performed
located in Dublin, Ireland. Salmon (Salmo salar) was cultivated, by the pour plate method, using plate count agar (Sigma-Aldrich). The
slaughtered, filleted and skinned by an Irish fish processor and skinless plates were incubated at 30 °C for 48 h.
fillets were purchased from local fishmonger located in Dublin, Ireland. The microbiological analysis was performed on day 1, 3, 5, 8 and 11
The fillets were transported for 15 min in ice to food production la- of storage at 4 °C and the results were expressed as log cfu/g of sushi
boratories located in Teagasc Food Research Centre (Ashtown, Dublin, sample.
Ireland) and immediately used for sushi production. The total time from
fish slaughter through to sushi production was 4 days.
2.5. TBA analysis
2.2. Sushi production
The 2-thiobarbituric acid (TBA) index analysis was performed ac-
Sushi rice was washed in running water for 5 min and left to drain cording to the aqueous extraction method described by Pikul,
for an additional 5 min. Next the rice was transferred to Gastronorm Leszczynski, and Kummerow (1989). The sample was homogenized
sizes (GN) containers with covers, mixed with water using 0.8:1 w/w until reaching uniformity and then 10 g was measured and homo-
ratio and placed in convection oven (Rational SCC 61, Rational AG, genized with 34.25 ml of 4% perchloric acid and 0.75 ml of butylated
Landsberg am Lech, Germany) for 55 min at 110 °C with 100% hu- hydroxytoluene (BHT), filtered through Whatman no. 1 filter paper
midity. Afterwards the rice was mixed with vinegar mixture (rice vi- (Whatman PLC, Maidstone, UK) and adjusted to 50 ml using 4% per-
negar, sucrose, salt in ratio of 0.52:0.42:0.06 w/w/w) using ratio of chloric acid. Five milliter of filtrate was transferred into glass tubes
1:0.125 v/v, gently stirred with taking special care not to damage the containing 5 ml of 0.02 M TBA and heated at 90 °C for 1 h in water
grains and left to cool down to temperature of 40 °C. bath. After heating the tubes were cooled with tap water until reaching
Nigiri-sushi was produced by forming 20 g ( ± 0.5 g) rice balls and room temperature and measured for absorbance at 532 nm wavelength
placing a rhomboid shaped salmon piece weighing 12 g ( ± 0.1 g) on using Helios Gamma spectrophotometer (Thermo Fisher Scientific,
top of it. All nigiri samples contained approximately 37.5% of raw Essex, UK). The blank sample was prepared in the same manner as the
salmon. normal samples except 5 ml of 4% perchloric acid was used instead of
Hosomaki was produced by placing 90 g of sushi rice and 20 g of filtrate. The constant coefficient K, used for calculating the thiobarbi-
salmon strip on a half of a nori sheet (1.5 g) and wrapping it using a turic acid reactive substances (TBARS) content, was calculated from the
bamboo mat into a thin circular roll. The rolls were then cut into eight standard curve.
equal pieces (approx. 13.5–14.0 g per piece). All hosomaki samples The TBA analysis was performed on sushi samples on day 1, 3, 5 and
contained approximately 17.9% of raw salmon. 8 of storage. The analysis of nigiri samples on day 11 was not performed
Each piece of nigiri and maki were placed separately into 80 mm due to high microbiological spoilage of the samples.

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Table 1
Changes in total viable count of treated and untreated nigiri and hosomaki samples during storage [log cfu/g].

Day 1 Day 3 Day 5 Day 8 Day 11

Nigiri Control 5.96aA ± 0.62 6.80aA ± 0.92 8.34bA ± 0.15 9.06bA ± 0.02 9.01bA ± 0.17
Nigiri 70 kV 4.98aA ± 0.50 6.50bA ± 0.14 7.77cA ± 0.65 8.63cA ± 0.51 8.99cA ± 0.32
Nigiri 80 kV 4.83aA ± 0.37 6.28bA ± 0.23 7.57cA ± 0.25 9.14dA ± 0.90 9.16dA ± 0.38
Hosomaki Control 4.38aA ± 0.66 4.94aA ± 0.57 4.83aA ± 0.79 6.65aA ± 2.05 6.22aA ± 1.90
Hosomaki 70 kV 4.90aA ± 0.45 4.21aA ± 0.78 3.85aA ± 1.00 5.79aA ± 1.46 4.71aA ± 0.51
Hosomaki 80 kV 4.39aA ± 0.73 4.50aA ± 1.06 4.14aA ± 0.99 5.14aA ± 0.32 5.26aA ± 0.43

a,b,c,d – different lowercase in a row indicate differences between days of storage within the same group (P < 0.05).
A,B,C,D – different uppercase in a column indicate differences between studied group on the same day (P < 0.05). The differences are measured separately for nigiri and hosomaki
groups.

Table 2
Changes in proximate composition [g/100 g] over 8 day storage period at 4 °C.

Day 1 Day 3 Day 5 Day 8

Moisture
Nigiri control 67.49aA ± 1.60 66.66aA ± 1.38 67.51aA ± 3.04 66.58aA ± 1.32
Nigiri 70 kV 65.68aA ± 1.52 66.17aA ± 1.16 67.65aA ± 0.95 66.80aA ± 0.93
Nigiri 80 kV 67.17aA ± 1.15 66.73aA ± 3.28 67.09aA ± 0.07 67.20aA ± 1.65
Hosomaki control 63.61aA ± 1.53 63.90aA ± 0.27 64.58aA ± 1.86 63.32aA ± 1.30
Hosomaki 70 kV 65.25aA ± 0.24 63.83aA ± 1.63 63.71aA ± 1.30 62.63aA ± 0.46
Hosomaki 80 kV 64.49aA ± 1.24 64.18aA ± 1.86 64.52aA ± 0.64 64.54aA ± 1.40

Protein
Nigiri control 8.93aA ± 0.59 9.36aA ± 0.19 8.50aA ± 1.17 9.04aA ± 0.67
Nigiri 70 kV 10.10aA ± 0.20 8.84aA ± 0.18 8.19aA ± 1.13 8.84aA ± 0.91
Nigiri 80 kV 9.58aA ± 0.93 9.44aA ± 0.65 8.73aA ± 1.21 9.47aA ± 1.35
Hosomaki control 5.54aA ± 0.73 5.51aA ± 0.52 5.38aA ± 0.79 6.14aA ± 0.23
Hosomaki 70 kV 6.15aA ± 0.68 6.07aA ± 0.65 5.33aA ± 0.47 6.18aA ± 0.27
Hosomaki 80 kV 6.06aA ± 0.60 6.73aA ± 0.37 5.24aA ± 0.56 6.31aA ± 0.35

Results expressed as mean ± standard deviation.

a,b – different lowercase in a row indicate differences between days of storage within the same group (P < 0.05).
A,B – different uppercase in a column indicate differences between studied group on the same day (P < 0.05). The differences are measured separately for nigiri and hosomaki groups.

2.6. Protein, moisture and fatty acids analysis min). Fatty acids were injected in full SCAN and SIM at the same time
after derivatization. The SIM mode was used to determine and quantify
Each sample was homogenized until reaching uniform distribution peak abundance the FAME. Thus, the target and the qualifier ion
of all the ingredients and then used for subsequent analysis. abundances were determined by injection of fatty acid standards after
Protein content was determined based on the measurement of ni- derivatization supplied by Sigma-Aldrich (Sigma-Aldrich, USA) under
trogen by combustion using a LECO FP628 Protein analyzer (LECO the same chromatographic conditions described above using full-scan
Corp., MI, USA) based on the Dumas method according to the AOAC with the m/z ratio ranging from 100 to 550. Then, compounds were
method 992.15 (2007). A factor of 6.25 was used to convert nitrogen to confirmed by their retention times, the identification of target and two
crude protein per cent. Moisture content were measured using a Smart qualifier ions, and the determination of qualifier to target ion ratios.
System 5 microwave drying oven (Smart Trac 5 Model 907,875, CEM Retention times must be within ± 0.2 min of the expected time and
Corporation, NC, USA) using AOAC Official Methods no. 985.16 (2007). qualifier to target ion ratios within a 20% range for positive con-
The fat for fatty acids analysis was extracted using the method de- firmation. Peak abundance was obtained by integration with Chem-
scribed by Bligh and Dyer (1959) and the methylation procedure was Station software (Agilent Technologies, UK) and expressed as the per-
performed as described by Ichihara, Shibahara, Yamamoto, and centage of each fatty acid in comparison to the total area of all fatty
Nakayama (1996). Fatty acids were extracted by mixing 1 g of the acids present in the sample. Full scan mode was run at the same time to
sample with 10 ml of hexane. The mixture was stirred for 1 h and after a have further confirmation of each fatty with the inbuilt NIST database.
centrifugation step (2000 g for 10 min) a clear supernatant was col-
lected. This step was repeated three times. The hexane from the three
extractions was pooled and evaporated under nitrogen stream and the 2.7. Statistical analysis
weight of remaining fat was measured. After this, fatty acid profile
analysis was carried out. Samples were analysed using an Agilent All analyses were performed using three independent repetitions,
7890A gas chromatography system coupled to an Agilent 5975C mass with each repetition produced from a different fish fillet and different
spectrometer (Agilent Technologies, Cheshire, UK). Samples (0.5 μl) production batch. The normality of the results and homogeneity of
were injected onto the GC/MS system via an Agilent 7963 autosampler variances was established using Saphiro-Wilk and Levene's test re-
(Agilent Technologies, Cheshire, UK), where separation was achieved spectively (P < 0.05). The significance of differences was established
using a Phenomenex BPX70 column (120 m × 0.25 mm, 0.25 μm). The by one-way ANOVA with Tukey post-hoc test (P < 0.05). The sig-
initial temperature of each run was 50 °C, which was ramped to 160 °C nificance of differences was established separately for nigiri and hoso-
(20 °C/min), followed by a further increase in temperature to 220 °C maki samples. The statistical analysis was performed using STATISTICA
(4 °C/min) and held for 5 min. Afterwards a final increase to 240 °C v.12.0 software (StatSoft, Tulsa, USA).
(4 °C/min) was carried out, and held for 13 min. The inlet temperature
was set at 250 °C, while hydrogen was used as the carrier gas (2 ml/

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Table 3
Changes in TBA index in treated and untreated nigiri and hosomaki samples during storage [mg/kg].

Day 1 Day 3 Day 5 Day 8 Day 11

Nigiri control 1.29aA ± 0.44 0.94aA ± 0.10 1.23aA ± 0.32 0.75aA ± 0.35 n/a
Nigiri 70 kV 1.69aA ± 0.17 1.71aB ± 0.02 1.94aB ± 0.12 1.52aA ± 0.59 n/a
Nigiri 80 kV 2.03aA ± 0.33 2.00aC ± 0.10 2.16aB ± 0.26 1.83aA ± 0.20 n/a
Hosomaki control 2.42abA ± 0.03 1.78abA ± 0.37 1.58aA ± 0.24 2.60abA ± 0.30 3.12bA ± 0.70
Hosomaki 70 kV 3.12aB ± 0.10 2.64aB ± 0.25 2.40aA ± 0.72 3.12aA ± 0.41 2.85aA ± 0.36
Hosomaki 80 kV 2.92aB ± 0.20 2.40aAB ± 0.07 3.11aB ± 0.69 3.20aA ± 0.14 3.62aA ± 0.50

a,b,c,d – different lowercase in a row indicate differences between days of storage within the same group (P < 0.05).
A,B,C,D – different uppercase in a column indicate differences between studied group on the same day (P < 0.05). The differences are measured separately for nigiri and hosomaki
groups.
n/a – analysis was not performed due to high microbiological spoilage of the samples (Table 1).

Table 4
The effect of non-thermal plasma treatment on the most abundant fatty acids of sushi [% of fat] measured on day 1 of storage at 4 °C.

Nigiri control Nigiri 70 kV Nigiri 80 kV Hosomaki control Hosomaki 70 kV Hosomaki 80 kV

Fatty acid Fatty acid name Mean SD Mean Mean SD Mean SD Mean SD

C14:0 Myristic acid 1.37a 0.73 1.80a 1.52a 0.22 1.47a 0.18 1.47a 1.97
C16:0 Palmitic acid 18.33a 4.80 15.76a 20.54a 3.15 21.00a 2.98 22.52a 20.14
C18:0 Stearic acid 3.45a 2.22 2.63a 2.94a 0.75 2.94a 0.48 3.06a 3.36
C18:1 Oleic acid 29.85a 4.26 31.65a 28.70a 5.16 30.78a 4.39 29.06a 6.21
C18:2 Linoleic acid 19.57a 7.74 16.48a 18.66a 1.93 16.35a 2.28 17.76a 2.11
C18:3 α-linolenic acid 4.02a 0.90 4.47a 4.01a 0.52 3.96a 0.46 4.21a 1.25
C20:5 Eicosapentaenoic acid 3.24a 0.79 6.27a 11.30a 3.10 12.29a 3.99 10.69a 3.86
C22:6 Docosahexaenoic acid 12.18a 4.57 14.22a 15.56a 2.88 17.36a 3.95 17.40a 5.02

a,b – different uppercase in a column indicate differences between studied group on the same day (P < 0.05). The differences are measured separately for nigiri and hosomaki groups.

3. Results and discussion day 1 to day 3, meanwhile the increase in untreated nigiri samples at
the same time period was not significant (approx 0.8 log cfu/g).
3.1. The effect of non-thermal plasma treatment on microbiological quality Therefore even though the initial bacterial concentration of the treated
of sushi samples was lower, the resulting increase during the first days of sto-
rage resulted in similar contamination of above 6 log cfu/g for all
The effect of NTP treatment on the microbiological quality of stored treated samples, and resulted in no shelf-life extension.
sushi samples is shown in Table 1. Throughout the storage period the On the other hand the treated hosomaki samples were still fit for
hosomaki samples (both treated and untreated) had significantly lower consumption at the last day of storage, while the untreated samples
TVC than the nigiri (data not shown). The initial contamination of nigiri exceeded the guidelines on day 8.
samples ranged from 4.83–5.96 log cfu/g and reached Kim, Puligundla, and Mok (2015), who used corona discharge
8.99–9.16 log cfu/g at day 11. Meanwhile the initial contamination of plasma jet system to treat squid shreds using NTP and reported a
hosomaki ranged from 4.38–4.9 log cfu/g and reached 2 log cfu/g reduction in aerobic bacteria counts after 3 min of treat-
4.71–6.22 log cfu/g at the last day of storage. These differences are to ment, with a D-value of 1.47 min. The treatment was also effective in
be expected, since nigiri contained higher proportions of raw fish than reducing the counts of marine bacteria, yeasts and moulds and Sta-
hosomaki. Although the initial contamination of nigiri samples might phylococcus aureus. Albertos et al. (2017) treated fresh mackerel fillets
seem high, higher results have been reported for the bacterial con- with NTP using a DBD system operating at 70 and 80 kV for 1, 3 and
centration of fresh nigiri and hosomaki produced in restaurants 5 min. Although none of the treatments significantly affected the total
(Atanassova, Reich, & Klein, 2008; Leisner et al., 2014). aerobic mesophilic counts, the treatment was effective in reducing
There was no statistically significant difference in the TVC between bacterial concentration of psychrotrophic bacteria, lactic acid bacteria
the treated and untreated nigiri and hosomaki samples throughout the and Pseudomonas, with 80 kV for 3 and 5 min treatments being the most
whole storage period. Moreover there were not statistically significant effective. According to Rød, Hansen, Leipold, and Knøchel (2012)
differences between samples treated with 70 and 80 kV of discreet ozone concentrations during NTP treatment in atmospheric pressure
voltage throughout the whole storage period. While not significant the increases for first 40 s after which it remains on a constant level. Sur-
highest inhibition of bacterial concentration in the nigiri samples was prisingly however, the higher power was used, the lower maximum
found on day 1 of storage, with the treated samples found to have a 1 ozone concentrations were acquired and consequently lower reduction
log lower cfu/g than the control. From day 3 however, the TVC for all in Listeria innocua was observed. This was attributed to surface changes
nigiri samples was similar, with differences in the bacterial con- in the plastic bag, used as a container. Due to the fact, that ozone is
centration < 1 log cfu/g. The differences in TVC between hosomaki present inside the sealed container for additional minutes after the end
samples treated with NTP were < 1 log cfu/g during the first 3 days of of the plasma treatment, they found that conducting a few short
storage but reached ~1.5 log cfu/g at the last day of storage. treatment times with 10 min intervals between the treatments was
The recommended limit for TVC in ready-to-eat food products, such more effective than a single dose treatment for a longer period (Rød
as sushi, is 106 cfu/g (ANZFA, 2001; Gilbert et al., 2000). According to et al., 2012). Therefore, it is possible that using few shorter treatments
these guidelines the studied nigiri samples were not fit for consumption with time intervals between each treatment could have been more ef-
on day 3 and the treatment with NTP failed to improve the shelf-life of fective in inhibition of bacterial growth in studied nigiri and hosomaki
nigiri samples. The treated nigiri samples, increased significantly by samples. It should be noted however, that such prolonged exposure to
approximately 1.5 log cfu/g at the beginning of storage period, from ozone and other RS might increase the oxidation thus causing the

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deterioration in the quality of the final product. lower TBA index on day 3 and 5 of storage, while untreated hosomaki
It is surprising, that NTP treatment was more effective in increasing had a significantly lower levels of TBA index on day 1, 3 and 5 of
the shelf-life of hosomaki and not the nigiri samples. Since NTP treat- storage. After day 5 the differences in TBA index for both nigiri and
ment mostly affects the microorganisms located on the surface of the hosomaki were statistically insignificant.
product, and given that raw fish is the likely primary cause of the high Hosomaki generally had higher TBA index than nigiri samples. This
bacterial concentration, it was expected that nigiri samples would be might be due to the fact that the surface area to weight of sample ratio
more affected, due to higher raw fish surface which is exposed to is greater than in nigiri. Moreover hosomaki are wrapped in nori sheet,
contact with formed RS than hosomaki. On the other hand, NTP which contains unsaturated fatty acids and might be more prone to
treatment is known to affect the composition of starch, with various RS oxidation induced by NTP treatment (Dawczynski, Schubert, & Jahreis,
causing the degradation of amylose and formation of new compounds 2007).
which in turn reduces the pH (F. Zhu, 2017). The pH of sushi rice is As established by Connell (1995), a TBA index of 2 mg/kg and
usually close to 4.3 due to the addition of rice vinegar (Adams et al., above for fish muscle is associated with spoilage and decreases in
1994) and the NTP treatment might cause a further drop in pH sensory attributes. Research performed on rainbow trout by Raeisi,
(Schlüter et al., 2013). Since the acidified rice lowers the pH of the fish Quek, Ojagh, and Alishahi (2016) showed similar pattern, where the
piece through direct contact (Lorentzen, Wesmajervi Breiland, Cooper, fish samples were rejected by the sensory panel when the TBA index
& Herland, 2012), it might affect the TVC of the sushi samples, espe- increased beyond 2 mg/kg.
cially for hosomaki, in which a greater surface area is in contact with Nigiri treated with 80 kV exceeded the proposed limit at the be-
the rice and the whole roll is more tightly squeezed. ginning of the storage period, while untreated and treated with 70 kV
remained below the 2 mg/kg threshold during the whole storage
3.2. Changes in protein and moisture content period. The initial TBA index of hosomaki ranged from 2.42–3.12 mg/
kg, therefore the hosomaki samples, both treated and untreated, ex-
Changes in the quality and sensory attributes of stored rice are one ceeded the 2 mg/kg limit on the first day of storage.
of the main problems faced by sushi industry, which affects the pro- The TBA index of all samples remained at a constant level with no
duct's shelf-life. Such changes are mostly related to the process of rice statistically significant differences between days of storage observed
retrogradation and moisture loss, resulting in texture changes, mostly with the exception of untreated hosomaki, which increased sig-
hardness, which can affect the sensory scores of the product deeming it nificantly at the last days of storage.
not suitable for consumption (Meullenet, Marks, Hankins, Griffin, & The analysis of fatty acids composition showed that the most
Daniels, 2000). Since plasma treatment can reduce the moisture content abundant fatty acids in sushi were palmitic acid (C16:0); oleic acid
of starch (Zhu, 2017) and thus affect the sensory quality of sushi, the (C18:1, n-9); linoleic acid (C18:2, n-6) eicosapentanoic acid (EPA,
changes in moisture content of both nigiri and hosomaki was measured C20:5, n-3) and docosahexanoic acid (DHA, C22:6 n-3) (Table 4).
(Table 2). There were no significant differences in the moisture content However, no significant differences in the content of each fatty acid
between the treated and untreated samples throughout the storage were found between the treated and untreated samples for both sushi
period, which suggests that in case of sushi, NTP treatment did not products. Therefore the NTP treatment and resulting increased oxida-
cause moisture loss. A second factor for rice sensory quality is starch tion, observed through TBA index analysis, did not affect the fatty acids
retrogradation. It has been reported that plasma treatment can reduce composition as measured for these sushi samples.
the degree of crystallinity of starch molecules along with the molecular As in case of protein, the surface treatment with NTP might result in
degradation of starch (Mir, Shah, & Mir, 2016); besides, NTP can reduce increased oxidation of surface fatty acids, which was not shown
starch retrogradation rate and improve texture parameters of stored through the analysis of the overall fatty acids composition found in the
rice (Thirumdas, Kadam, & Annapure, 2017; Zhang, Chen, Li, Li, & product. The significant increase in TBA index with no significant dif-
Zhang, 2015). This fact, together the absence of moisture loss, suggests ferences in fatty acids composition suggests that TBARS could be
that NTP could be a possible method to improve sensory scores of formed from other non-lipid molecules such as proteins, nucleic acids or
stored sushi rice, however more research on this subject should be carbohydrates (Janero, 1990).
performed in order to confirm this. The fatty acids composition of the studied sushi samples differed
Neither NTP treatment conditions nor the storage time affected the from the fatty acids composition usually found in salmon (Bell, Tocher,
overall protein content of the sushi samples. This was not surprising Henderson, Dick, & Crampton, 2003; Torstensen, Lie, & Frøyland,
since usually the changes in proteins are associated with moisture loss. 2000). Nori might affect the content of EPA, since it usually contains
On the other hand Matan, Puangjinda, Phothisuwan, and Nisoa (2015) high levels of this fatty acids (Dawczynski et al., 2007). This would also
found that NTP significantly increased the content of crude protein, fat, explain the difference in the EPA content between nigiri and hosomaki
carbohydrates and fiber, while not affecting the moisture content in samples. Moreover rice contains high levels of palmitic, oleic and li-
fresh-cut dragon fruit. Moreover since NTP is a surface treatment, it can noleic acid (Zhou, Blanchard, Helliwell, & Robards, 2003), which could
cause changes in the proteins on the surface of the product, which will also explain the higher levels of these fatty acids in sushi over that of
not be shown by the crude protein content analysis. The RS created by salmon.
plasma treatment can react with proteins in both rice and fish. For
example as shown by Albertos et al. (2017) NTP treatment affected the 4. Conclusions
myofibrillar network of fish meat, significantly reducing the amount of
water immobilised in such matrix and increasing the free water content, Non-thermal plasma treatment seemed to have some influence on
while Segat, Misra, Cullen, and Innocente (2015) reported the changes microbiological quality of sushi, however the effect was limited.
in structural composition of whey protein isolate, which improved Nevertheless, both 70 and 80 kV treatments for 5 min extended the
foaming and emulsifying properties of the protein isolate. shelf-life of the stored hosomaki product by an additional 4 days in
terms of microbiological spoilage. Moreover the plasma treatment did
3.3. The effect of NTP treatment on lipid oxidation and fatty acids not affected moisture content of the sushi samples, and did not alter the
composition of sushi fatty acids composition even though the TBA analysis showed an in-
crease in oxidation processes.
NTP treatment increased the TBARS content of both nigiri and Future research should focus on the effect of non-thermal plasma on
hosomaki samples, with hosomaki containing higher levels of TBA sushi stored in modified atmosphere and the effect on rice retro-
index than nigiri (Table 3). Untreated nigiri contained significantly gradation and sensory evaluation in order to fully assess the

416
P. Kulawik et al. Innovative Food Science and Emerging Technologies 45 (2018) 412–417

effectiveness and possibility of implementing this processing method 76–80.

for the sushi industry. López-Malo, A., Palou, E., Barbosa-Cánovas, G. V., Welti-Chanes, J., & Swanson, B. G.
(1998). Polyphenoloxidase activity and color changes during storage of high hy-
drostatic pressure treated avocado puree. Food Research International, 31(8),
Conflict of interest 549–556.
Lorentzen, G., Wesmajervi Breiland, M. S., Cooper, M., & Herland, H. (2012). Viability of
listeria monocytogenes in an experimental model of nigiri sushi of halibut
The authors declare that there is no interest or relationship, fi- (Hippoglossus hippoglossus) and salmon (Salmo salar). Food Control, 25(1), 245–248.
nancial or otherwise that might be perceived as influencing an author's Matan, N., Puangjinda, K., Phothisuwan, S., & Nisoa, M. (2015). Combined antibacterial
objectivity. activity of green tea extract with atmospheric radio-frequency plasma against pa-
thogens on fresh-cut dragon fruit. Food Control, 50, 291–296.
Meullenet, J. F., Marks, B. P., Hankins, J. A., Griffin, V. K., & Daniels, M. J. (2000).
References Sensory quality of cooked long-grain rice as affected by rough rice moisture content,
storage temperature, and storage duration. Cereal Chemistry, 77(2), 259–263.
Mir, S. A., Shah, M. A., & Mir, M. M. (2016). Understanding the role of plasma technology
Adams, A. M., Leja, L. L., Jinneman, K., Beeh, J., Yuen, G. A., & Wekell, M. M. (1994).
in food industry. Food and Bioprocess Technology, 9(5), 734–750.
Anisakid parasites, Staphylococcus aureus and Bacillus cereus in sushi and sashimi from
Misra, N. N., Pankaj, S. K., Walsh, T., O'Regan, F., Bourke, P., & Cullen, P. J. (2014). In-
Seattle area restaurants. Journal of Food Protection, 57(4), 311–317.
package nonthermal plasma degradation of pesticides on fresh produce. Journal of
Albertos, I., Martin-Diana, A. B., Cullen, P. J., Tiwari, B. K., Ojha, S. K., Bourke, P., ...
Hazardous Materials, 271, 33–40.
Rico, D. (2017). Effects of dielectric barrier discharge (DBD) generated plasma on
Misra, N. N., Tiwari, B. K., Raghavarao, K. S. M. S., & Cullen, P. J. (2011). Nonthermal
microbial reduction and quality parameters of fresh mackerel (Scomber scombrus)
plasma inactivation of food-borne pathogens. Food Engineering Reviews, 3(3),
fillets. Innovative Food Science & Emerging Technologies, 44, 117–122.
159–170.
Altintzoglou, T., Heide, M., Wien, A. H., & Honkanen, P. (2016). Traditional sushi for
Moreau, M., Orange, N., & Feuilloley, M. G. J. (2008). Non-thermal plasma technologies:
modern consumers: A comparison between sushi consumption behavior in Japan and
New tools for bio-decontamination. Biotechnology Advances, 26(6), 610–617.
Norway. Journal of Food Products Marketing, 22(6), 717–732.
NSC (2015). Seafood study 2014: Insight and outlook: How the Germans eat fish. Norwegian
ANZFA (2001). Guidelines for the microbiological examination of ready - to - eat foods. The
Seafood Council.
Australia New Zealand Food Authority.
Pikul, J., Leszczynski, D. E., & Kummerow, F. A. (1989). Evaluation of three modified TBA
AOAC (2007). Official methods of analysis of AOAC international (18th ed.). .
methods for measuring lipid oxidation in chicken meat. Journal of Agricultural and
Atanassova, V., Reich, F., & Klein, G. (2008). Microbiological quality of sushi from sushi
Food Chemistry, 37(5), 1309–1313.
bars and retailers. Journal of Food Protection, 71(4), 860–864.
Raeisi, S., Quek, S. Y., Ojagh, S. M., & Alishahi, A. R. (2016). Effects of cumin (Cuminum
Bell, J. G., Tocher, D. R., Henderson, R. J., Dick, J. R., & Crampton, V. O. (2003). Altered
cyminum L.) seed and wild mint (Mentha longifolia L.) leaf extracts on the shelf life and
fatty acid compositions in Atlantic Salmon (Salmo salar) fed diets containing linseed
quality of rainbow trout (Oncorhynchus mykiss) fillets stored at 4 °C ± 1. Journal of
and rapeseed oils can be partially restored by a subsequent fish oil finishing diet. The
Food Safety, 36(2), 271–281.
Journal of Nutrition, 133(9), 2793–2801.
Rød, S. K., Hansen, F., Leipold, F., & Knøchel, S. (2012). Cold atmospheric pressure
Bligh, E. G., & Dyer, W. J. (1959). A rapid method of total lipid extraction and pur-
plasma treatment of ready-to-eat meat: Inactivation of Listeria innocua and changes in
ification. Canadian Journal of Biochemistry and Physiology, 37(8), 911–917.
product quality. Food Microbiology, 30(1), 233–238.
Boudam, M. K., Moisan, M., Saoudi, B., Popovici, C., Gherardi, N., & Massines, F. (2006).
Schlüter, O., Ehlbeck, J., Hertel, C., Habermeyer, M., Roth, A., Engel, K.-H., ...
Bacterial spore inactivation by atmospheric-pressure plasmas in the presence or ab-
Eisenbrand, G. (2013). Opinion on the use of plasma processes for treatment of foods.
sence of UV photons as obtained with the same gas mixture. Journal of Physics D:
Molecular Nutrition & Food Research, 57(5), 920–927.
Applied Physics, 39(16), 3494.
Scholtz, V., Pazlarova, J., Souskova, H., Khun, J., & Julak, J. (2015). Nonthermal plasma
Connell, J. J. (1995). Control of fish quality (4th ed.). London: Wiley-Blackwell.
— A tool for decontamination and disinfection. Biotechnology Advances, 33(6, Part 2),
Dawczynski, C., Schubert, R., & Jahreis, G. (2007). Amino acids, fatty acids, and dietary
1108–1119.
fibre in edible seaweed products. Food Chemistry, 103(3), 891–899.
Segat, A., Misra, N. N., Cullen, P. J., & Innocente, N. (2015). Atmospheric pressure cold
Don, L. Z. (1997). The impact of consumer demands and trends on food processing.
plasma (ACP) treatment of whey protein isolate model solution. Innovative Food
Emerging Infectious Diseases, 3(4), 467.
Science & Emerging Technologies, 29, 247–254.
Ehlbeck, J., Schnabel, U., Polak, M., Winter, J., von TH, W., Brandenburg, R., ...
Sequeira-Munoz, A., Chevalier, D., LeBail, A., Ramaswamy, H. S., & Simpson, B. K.
Weltmann, K. D. (2011). Low temperature atmospheric pressure plasma sources for
(2006). Physicochemical changes induced in carp (Cyprinus carpio) fillets by high
microbial decontamination. Journal of Physics D: Applied Physics, 44(1), 013002.
pressure processing at low temperature. Innovative Food Science & Emerging
Fridman, G., Brooks, A. D., Balasubramanian, M., Fridman, A., Gutsol, A., Vasilets, V. N.,
Technologies, 7(1–2), 13–18.
... Friedman, G. (2007). Comparison of direct and indirect effects of non-thermal
Steffen, H., Duerst, M., & Rice, R. G. (2010). User experiences with ozone, electrolytic
atmospheric-pressure plasma on bacteria. Plasma Processes and Polymers, 4(4),
water (active water) and UV-C light (Ventafresh technology) in production processes
370–375.
and for hygiene maintenance in a Swiss sushi factory. Ozone: Science & Engineering,
Fu, Z., Chen, J., Luo, S.-J., Liu, C.-M., & Liu, W. (2015). Effect of food additives on starch
32(1), 71–78.
retrogradation: A review. Starch-Starke, 67(1–2), 69–78.
Thirumdas, R., Kadam, D., & Annapure, U. S. (2017). Cold plasma: An alternative tech-
Gilbert, R. J., de Louvois, J., Donovan, T., Little, C., Nye, K., Ribeiro, C. D., ... Bolton, F. J.
nology for the starch modification. Food Biophysics, 12(1), 129–139.
(2000). Guidelines for the microbiological quality of some ready-to-eat foods sampled
Torstensen, B. E., Lie, Ø., & Frøyland, L. (2000). Lipid metabolism and tissue composition
at the point of sale. PHLS Advisory Committee for Food and Dairy Products.
in Atlantic salmon (Salmo salar L.)—Effects of capelin oil, palm oil, and oleic acid-
Communicable Disease and Public Health, 3(3), 163–167.
enriched sunflower oil as dietary lipid sources. Lipids, 35(6), 653–664.
Ichihara, K., Shibahara, A., Yamamoto, K., & Nakayama, T. (1996). An improved method
Van Durme, J., Nikiforov, A., Vandamme, J., Leys, C., & De Winne, A. (2014). Accelerated
for rapid analysis of the fatty acids of glycerolipids. Lipids, 31, 535–539.
lipid oxidation using non-thermal plasma technology: Evaluation of volatile com-
Jacobo-Velázquez, D. A., & Hernández-Brenes, C. (2010). Biochemical changes during the
pounds. Food Research International, 62, 868–876.
storage of high hydrostatic pressure processed avocado paste. Journal of Food Science,
Wang, S., Li, C., Copeland, L., Niu, Q., & Wang, S. (2015). Starch retrogradation: A
75(6), S264–S270.
comprehensive review. Comprehensive Reviews in Food Science and Food Safety, 14(5),
Janero, D. R. (1990). Malondialdehyde and thiobarbituric acid-reactivity as diagnostic
568–585.
indices of lipid peroxidation and peroxidative tissue injury. Free Radical Biology and
Zhang, B., Chen, L., Li, X., Li, L., & Zhang, H. (2015). Understanding the multi-scale
Medicine, 9(6), 515–540.
structure and functional properties of starch modulated by glow-plasma: A structure-
Kim, J.-W., Puligundla, P., & Mok, C. (2015). Microbial decontamination of dried laver
functionality relationship. Food Hydrocolloids, 50(Supplement C), 228–236.
using corona discharge plasma jet (CDPJ). Journal of Food Engineering, 161, 24–32.
Zhou, Z., Blanchard, C., Helliwell, S., & Robards, K. (2003). Fatty acid composition of
Kwak, H. S., Kim, M., & Jeong, Y. (2015). Physicochemical properties and determination
three rice varieties following storage. Journal of Cereal Science, 37(3), 327–335.
of key instrumental quality measurement parameters of frozen-cooked Rice by cor-
Zhu, F. (2017). Plasma modification of starch. Food Chemistry, 232, 476–486.
relating consumer acceptance. Journal of Food Quality, 38(3), 192–200.
Zhu, S., Ramaswamy, H. S., & Simpson, B. K. (2004). Effect of high-pressure versus
Leisner, J. J., Lund, T. B., Frandsen, E. A., Andersen, N. B. E., Fredslund, L., Nguyen, V. P.
conventional thawing on color, drip loss and texture of Atlantic salmon frozen by
T., & Kristiansen, T. (2014). What consumers expect from food control and what they
different methods. LWT - Food Science and Technology, 37(3), 291–299.
get – A case study of the microbial quality of sushi bars in Denmark. Food Control, 45,

417