This article was downloaded by: Vahid Chamanara

On: 21 January 2013, At: 00:06

Publisher: Taylor & Francis
Informa Ltd Registered in England and Wales Registered Number: 1072954 Registered
office: Mortimer House, 37-41 Mortimer Street, London W1T 3JH, UK

Journal of Aquatic Food Product

Technology
Publication details, including instructions for authors and
subscription information:
http://tandfonline.com/loi/wafp20

Shelf-Life Extension of Fish Samples by

Using Enriched Chitosan Coating with
Thyme Essential Oil
a b c
Vahid Chamanara , Bahareh Shabanpour , M. Khomeiri & Saeed
b
Gorgin
a
Marine Science and Technology, The University of Hormozgan,
Hormozgan, Iran
b
Seafood Division of Fisheries Department, Gorgan University of
Agricultural Sciences and Natural Resources, Gorgan, Iran
c
Food Microbiology Division of Department of Food Science and
Technology, Gorgan University of Agricultural Sciences and Natural
Resources, Gorgan, Iran
Accepted author version posted online: 17 Jan 2012.

To cite this article: Vahid Chamanara , Bahareh Shabanpour , M. Khomeiri & Saeed Gorgin (2013):
Shelf-Life Extension of Fish Samples by Using Enriched Chitosan Coating with Thyme Essential Oil,
Journal of Aquatic Food Product Technology, 22:1, 3-10

To link to this article: http://dx.doi.org/10.1080/10498850.2011.621583

PLEASE SCROLL DOWN FOR ARTICLE

Full terms and conditions of use: http://tandfonline.com/page/terms-and-conditions

This article may be used for research, teaching, and private study purposes. Any
substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing,
systematic supply, or distribution in any form to anyone is expressly forbidden.

The publisher does not give any warranty express or implied or make any representation
that the contents will be complete or accurate or up to date. The accuracy of any
instructions, formulae, and drug doses should be independently verified with primary
sources. The publisher shall not be liable for any loss, actions, claims, proceedings,
demand, or costs or damages whatsoever or howsoever caused arising directly or
indirectly in connection with or arising out of the use of this material.
 Journal of Aquatic Food Product Technology, 22:3–10, 2013
Copyright © Taylor & Francis Group, LLC
ISSN: 1049-8850 print/1547-0636 online
DOI: 10.1080/10498850.2011.621583

Shelf-Life Extension of Fish Samples by Using Enriched

Chitosan Coating with Thyme Essential Oil

Vahid Chamanara,1 Bahareh Shabanpour,2 M. Khomeiri,3 and Saeed Gorgin2

1
Marine Science and Technology, The University of Hormozgan, Hormozgan, Iran
2
Seafood Division of Fisheries Department, Gorgan University of Agricultural
Downloaded by [Vahid Chamanara] at 00:06 21 January 2013

Sciences and Natural Resources, Gorgan, Iran

3
Food Microbiology Division of Department of Food Science and Technology, Gorgan
University of Agricultural Sciences and Natural Resources, Gorgan, Iran

The present study examined the effect of chitosan (Ch) and combination of chitosan and Thymus vulgaris
essential oil (Ch+T) on the quality and shelf life of butterfly-shaped rainbow trout stored under refrig-
erated condition (5 ± 1◦ C) for 15 days. The treatments in the present study were Ch (chitosan: 2% w/v)
and Ch+T (chitosan: 2% w/v and Thymus vulgaris essential oil: 1% w/v). The shelf life of the sam-
ples were determined using microbiological (total psychrotrophic count), chemical—pH, thiobarbituric
acid (TBA), and total volatile basic nitrogen (TVB-N)—and textural analyses. Results showed that total
psychrotrophic count decreased significantly (p < 0.05) using Ch+T coating during the entire period
of storage. The treatment Ch+T resulted in lower pH, TBA, and TVB-N values in comparison with
the other samples. Based mainly on our data, Ch+T treatment extended the product’s shelf life by
approximately 6 days in comparison with the uncoated control samples.

Keywords: chitosan, shelf life, Thymus vulgaris, rainbow trout (Oncorhynchus mykiss)

INTRODUCTION

Food quality and safety are major concerns in the food industry as consumers prefer fresher and
minimally processed products. Various food preservation techniques have been utilized to improve
the microbial safety and extend the shelf life of seafood in general—including freezing, chemical
preservation, salting, and modified atmosphere packaging. Use of preservatives in aquatic prod-
ucts is convenient and universal. In order to extend refrigerated storage time, antimicrobial and
antioxidant additives—especially of synthetic origin—are added to muscle foods. However, con-
sumers increasingly demand use of natural products as alternative preservatives in foods, as the
safety of synthetic additives has been questioned in past years (Imaida et al., 1983). Edible coat-
ings from polysaccharides, proteins, and lipids can extend the shelf life of foods by functioning
as solute, gas, and vapor barriers. Chitosan—poly-β-(1–4)-D-glucosamine—is a versatile biopoly-
mer, having a broad range of applications in the food industry (Knatt et al., 2008). Edible films

Correspondence should be addressed to Vahid Chamanara, Marine Science and Technology, The University of
Hormozgan, Hormozgan, Iran. E-mail: V.chamanara@gmail.com

3
 4 CHAMANARA ET AL.

or coatings have been investigated for their abilities to retard moisture, oxygen, aroma, and solute
transports. It is one of the most effective methods to maintain food quality. However, most natural
antimicrobials have a limited spectrum of activity and are effective only at very high concentrations.
Chitosan exhibits antimicrobial activity against a range of foodborne microorganisms and conse-
quently has attracted attention as a potential natural food preservative (Chen et al., 1998; Shahidi
et al., 1999). It has long been acknowledged that some plant essential oils exhibit antimicrobial
properties (Finnemore, 1926), and it is necessary to scientifically investigate those plants which have
been used in traditional medicine to improve the quality of health care. Essential oils are potential
sources of novel antimicrobial compounds, especially against bacterial pathogens (Prabuseenivasan
et al., 2006). Thymus vulgaris L. (thyme), locally known as the “Avishan” member of the Lamiaceae
family, is widely used in Iranian folk medicine for its expectorant, antitussive, antibroncholitic, anti-
spasmodic, anthelmintic, carminative, and diuretic properties. Iranian Thymus species have been
reported for their antibacterial activities (Rasooli and Mirmostafa, 2002).
Downloaded by [Vahid Chamanara] at 00:06 21 January 2013

The present study has tried to improve shelf life of refrigerated rainbow trout during 15 days
of chilled storage by means of chitosan biopolymer coating and thyme essential oil. Total
psychrotrophic count was calculated. Chemical—pH, thiobarbituric acid (TBA), and total volatile
basic nitrogen (TVB-N)—and texture profile analyses were also studied.

MATERIALS AND METHODS

Fish and Sample Preparation

Rainbow trout (Oncorhynchus mykiss) of commercial size—i.e., measuring 450–550 g in body
weight—were collected from a culture farm in Gorgan, Iran, and immediately transferred in ice
to the Seafood Processing Laboratory (Gorgan University of Agricultural Sciences and Natural
Resources). Fish were decapitated and gutted by hand and then butterfly shaped by removing the
backbone.

Chitosan
Medium molecular weight (190–310 kDa) chitosan (Ch) in powder form was obtained (Sigma-
Aldrich Co., Munich, Germany). Chitosan used in this study had a deacetylation degree of 75–85%.

Preparation of Coatings and Treatment of Fish

Two grams of Ch powder were dispersed in 97 g water before 1 mL acetic acid was added to the
mixture under stirring to give a Ch content of 2.0 wt%. By doing this, clotting of Ch in acetic acid
aqueous solution could be avoided. Glycerol plasticizer was added into the solution in a content of
0.75 mL/g Ch and stirred for 10 min on hotplate/magnetic stirrer. The resultant Ch coating solution
was filtrated through a Whatman No. 3 filter paper to remove any insoluble particles. To help dis-
tribute and fully incorporate it, the thyme oil mixed with Tween-20 (Merck, Darmstadt, Germany)
was added to the Ch solution. The final coat-forming solution consisted of 2% chitosan, 1% acetic
acid, 0.75% glycerol, 0.2% Tween-20, and 1% thyme oil. The final coat-forming solution was
homogenized under aseptic conditions at 21,600 rpm for 1 min (ULTRA-TURRAX, T25 digital,
IKA, Staufen, Germany). Another solution was prepared without addition of thyme oil, and control
solution was prepared only by dissolving glacial acetic acid in distilled water in concentration of 1%
(v/v). Butterfly-shaped fish samples were randomly assigned into three treatment lots consisting of
one control lot (uncoated) and two lots treated with the mentioned coating solutions. The samples
of each lot were immersed for 30 s in 600 mL of the coating solution and then allowed to stand for a
 SHELF-LIFE EXTENSION WITH CH AND CH+T 5

2 min period followed by a second immersion in the coating solution for 30 s. Then the fish samples
were removed and following air-drying for 8h at 8–10◦ C, the coated samples were placed in plastic
bags and the bags then stored at 5 ± 1◦ C to await further assessment. Chemical, microbiological,
and textural analyses were performed at 3-day intervals to determine the overall quality of fish.

Psychrotrophic Bacteria Count

The bacteriological characteristics (total psychrotrophic count) of 10 g of anterior dorsal zone of
fish muscle were obtained following homogenization in 90 mL 0.1% peptone water. Other deci-
mal dilutions were prepared from a 10−1 dilution. Psychrotrophic count was determined via the
pour plate method, with plate count agar (Acumedia Manufactures, Inc., Lansing, MI, USA) as the
medium. The plates were incubated at 10◦ C for 7 days. Two replicates in each group were analyzed.
Downloaded by [Vahid Chamanara] at 00:06 21 January 2013

All counts were presented as average values over two samples (Arashisar et al., 2004; Chien et al.,
2007).

Chemical Analysis
Determination of pH
The pH measurement was carried out using a digital pH meter (Metrohm 713 pH meter,
Filderstadt, Germany), according to the method described by Masniyom et al. (2005).

Determination of Total Volatile Basic Nitrogen

Total volatile basic nitrogen was determined according to the method described by Goulas and
Kontominas (2005).

Thiobarbituric Acid Value

The thiobarbituric acid (mg malondaldehyde/kg) was determined according to the Kirk and
Sawyer method (Kirk et al., 1997).

Texture Profile Analysis (TPA)

The measurement of the texture properties (texture profile analysis) of samples was performed using
a LFRA texture analyzer (LFRA 4500; Brookfield Engineering, Middleboro, MA, USA). The sam-
ples used for TPA measurement were cut out by 1 cm3 ; the TPA measurements were performed at
25% compression. A cylindrical probe of 20 mm diameter was used with a test speed of 0.8 mm/s.
Instrumental TPA is a measurement method which imitates the chewing process, thus giving the
possibility to observe and differentiate between single texture attributes and to characterize on this
basis the very complex impression of food texture on humans. The texture attribute of hardness is
defined as the maximum force of the first compression. Chewiness, as the quantity to simulate the
energy required to masticate a sample to a steady state of swallowing, is calculated as the product of
hardness, cohesiveness, and springiness. Springiness is a measure explaining ability of the product
to spring back after deformation on the first compression. Cohesiveness is the attribute describing
how well the product withstands a second deformation relative to how it behaved under the first
deformation (Schubring and Oehlenschlager, 1997).
 6 CHAMANARA ET AL.

Statistical Analysis
All measurements were replicated three times (except for microbial analysis) for each lot, and
mean values ± standard deviations were reported for each case. Analysis of variance (ANOVA)
and Duncan’s test were performed on SPSS 17.0 (SPSS, Inc., Chicago, IL, USA) to evaluate the
significance of differences among mean values.

RESULTS AND DISCUSSION

Psychrotrophic Bacteria Count

Variations in the values of total psychrotrophic counts (PTC) during the refrigerated storage are pre-
sented in Figure 1. The initial PTC (log10 CFU/g) in samples ranged from 2.9, 2.41, and 2.37 log10
Downloaded by [Vahid Chamanara] at 00:06 21 January 2013

CFU/g in control, Ch-coated, and chitosan and thyme- (Ch+T) coated samples, respectively. The
low initial PTC for fresh fish samples indicates very good fish quality. Ojagh et al. (2009) reported
higher initial counts (3.51 to 3.86 log10 CFU/g) for fresh filleted farmed rainbow trout (Ojagh et al.,
2009). The gram-negative psychrotrophic bacteria are the major group of microorganisms responsi-
ble for spoilage of aerobically stored fresh fish at chilled temperatures (Ibrahim Sallam, 2007). The
antimicrobial properties of Ch coating have been reported in the literature (López-Caballero et al.,
2005). Lopez-Caballero et al. (2005) reported that a coating consisting of a blend of Ch dissolved in
acetic acid and gelatin exerted an inhibitory effect on the gram-negative flora of fish patties. Various
factors affect the antimicrobial action of Ch, and its mechanism of action appears to be related to
interactions between the positively charged Ch molecules and the negatively charged microbial cell
membrane (Shahidi et al., 1999) as well as to its function as a barrier against oxygen transfer (Jeon
et al., 2002).
As time passed, psychrotrophic bacteria count increased from 2.9–7.9 log10 CFU/g in control
samples, 2.41–6.72 log10 CFU/g in Ch-coated samples, and 2.37–6.22 log10 CFU/g in Ch+T-
coated samples, within 15 days of preservation. The Ch+T treatment showed significantly lower
psychrotrophic bacteria count during preservation (p < 0.01). There was a reduction of up to 1.5 log
cycles between Ch+T-coated samples and controls after 15 days of chilled storage. Also, Tsai et al.

8
7.5 Control
7 Ch
6.5
Ch+T
6
5.5
Log CFU/g

5
4.5
4
3.5
3
2.5
2
1.5
1
0 3 6 9 12 15
Time (days)

FIGURE 1 Variations in the values of total psychrotrophic counts (PTC) (color figure available online).
 SHELF-LIFE EXTENSION WITH CH AND CH+T 7

(2002) found that pretreatment of fish fillets (Oncorhynchus nerka) for 3 h with 1% Ch solution
retarded the increase in the counts for mesophiles, psychrotrophs, coliforms, Aeromonas spp., and
Vibrio spp. (Tsai et al., 2002). In the present study, the coating with Ch+T was observed to exert a
higher inhibitory effect on the native flora in comparison with Ch-coating (p < 0.01). PTC of control
samples exceeded the value of 7 log10 CFU/g, considered as the upper acceptable microbiological
limit on Days 9–10 of storage, but other treatments remain under 7 log10 CFU/g.
Also, essential oils from some plants including the thyme oil caused a total inhibition of fun-
gal development on maize kernels (Montes-Belmont and Carvajal, 1998). The antimicrobial effects
of plant extracts on a wide range of microorganisms have been described (Hammer et al., 1999;
Dorman, 2000). Much of the antimicrobial activity of essential oils produced from spices and other
cookable plants can be related to phenolic compounds in them (Montes-Belmont and Carvajal,
1998). The means by which microorganisms are inhibited by phenolic compounds involves a sensi-
tization of the phospholipids bi-layer of the cell membrane, causing an increase in permeability and
Downloaded by [Vahid Chamanara] at 00:06 21 January 2013

leakage of vital intracellular constituents or impairment of bacterial enzyme systems (Ultee et al.,
2000; Ultee et al., 2002).

Chemical Analysis
The pH
Average pH values are shown in Figure 2. Primary pH of the samples was about 6.38. At first,
pH decreased and then increased. The same results were reported by other researchers (Alasalvar
et al., 2001; Manju et al., 2007).
The primary decrease in pH may be due to the process of CO2 dissolving in fish samples, while
it seems that increase in pH can be because of increase in produced volatile bases as well as internal
or microbial enzymes. The results showed that the difference in pH in Ch+T-coated samples had
the same process as treatment C (control), but in Ch+T-coated samples, pH gradually increased.
It can be inferred that the lower pH in Ch+T treatment can be a result of cooperation of coating in
increasing shelf life of the samples due to the inhibitory effect against internal enzymes’ activities
and also inhibitory effect against microbial growth.

6.9
Control Ch Ch+T
6.8

6.7
pH

6.6

6.5

6.4

6.3

6.2
0 3 6 9 12 15
Days of storage

FIGURE 2 Changes in pH values in 15 days of refrigerated storage (C: control, Ch: chitosan-coated samples, Ch+T:
chitosan and thyme-coated samples) (color figure available online).
 8 CHAMANARA ET AL.

Total Volatile Basic Nitrogen

As shown in Figure 3, the TVB-N content of control samples was 17.23 mg N/100 g at the
beginning; but this value increased to 26.62 mg N/100 g on the 9th day and reached to 37.16 mg
N/100 g on the 15th day. Through the 6th day the freshness was good, but it spoiled very quickly
thereafter. It is interesting to note that the TVB-N content of the samples showed almost an identical
pattern with the psychrotrophic bacteria count. TVB-N value in Ch-coated samples on Days 0, 9,
and 15 was 17.5, 25.16, and 31.68, respectively. Also, as shown in Figure 3, Ch+T-coated samples
showed the least values of TVB-N through the entire period of chilled storage (p < 0.01). The TVB-
N content of Ch+T-coated samples was 17.41, 21.80, and 26.69 on Days 0, 9, and 15, respectively.

Thiobarbituric Acid Value

Downloaded by [Vahid Chamanara] at 00:06 21 January 2013

As shown in Figure 4, the TBA increased between Days 0 and 15 in all samples, but significantly
lower TBA values were observed in Ch+T-coated samples compared to the control and the Ch-
coated samples during the entire period of chilled storage (p < 0.01). TBA content of all samples
ranged between 0.19–0.27 mg of malondaldehyde/kg at the beginning. As results show, there was
no significant difference observed between Ch-coated samples and control samples from Days 3 to
9. This result can be due to the barrier properties of Ch coating and its ability to reduce lipid oxida-
tion. Our results were in agreement with other researchers (Darmadji and Izumimoto, 1994; Shahidi
et al., 1999, 2002; Jeon et al., 2002). Furthermore, a significant difference was observed between
Ch+T and other samples. This can be due to active substances such as thymol and carvacrol in
thyme essential oil and also synergistic interaction of them on lipid oxidation.

Texture Profile Analyses

Textural properties are probably the most important attributes of fish and fish products. Texture
parameters are changed by enzymatic and chemical reactions that lead to softening, changes in
elasticity, or development of toughness under certain conditions. Texture changes are shown in

45
C
40 Ch
Ch+T
TVB-N (mg N/100g)

35

30

25

20

15
0 3 6 9 12 15
Days of storage

FIGURE 3 TVB-N values of samples during 15 days of storage (C: control, Ch: chitosan-coated samples, Ch+T:
chitosan and thyme-coated samples) (color figure available online).
 SHELF-LIFE EXTENSION WITH CH AND CH+T 9

0.8
C
0.7
Ch
0.6 Ch+T
TBA (mg MA/Kg)

0.5

0.4

0.3

0.2

0.1
Downloaded by [Vahid Chamanara] at 00:06 21 January 2013

0
0 3 6 9 12 15
Days of storage

FIGURE 4 TBA values of samples during 15 days of storage. (C: control, Ch: chitosan-coated samples, Ch+T:
chitosan and thyme-coated samples) (color figure available online).

TABLE 1
Changes in instrumental texture attributes of fish samples

Groups Hardness Adhesiveness Springiness Cohesiveness Chewiness

Control 501.42 ± 184.24a −6.20 ± 2.45a 1.1 ± 0.28a 0.44 ± 0.05a 243.7 ± 92.01a
Ch 442.76 ± 114.15b −8.04 ± 2.14a 0.99 ± 0.09b 0.41 ± 0.05a 178.2 ± 37.15b
Ch+T 557.49 ± 139.42c −6.40 ± 3.15a 1.04 ± 0.1ab 0.43 ± 0.04a 249.8 ± 58.5a

Ch: chitosan treatment, Ch+T: chitosan treatment incorporated with thyme essential oil.
Different superscript letters represent the statistical differences between groups (p < 0.01).

Table 1. Results showed that Ch treatment of samples decreased hardness and chewiness signifi-
cantly, because of its water absorbance ability. It was determined that Ch treatments at this amount
did not affect texture qualities such as adhesiveness and cohesiveness. In brief, when comparing
groups according to their shelf lives, it was determined that texture was not markedly influenced by
these treatments.

CONCLUSIONS

Considering antioxidant and antimicrobial effects of the coating and as the factors investigated were
kept within the permissible range by Day 15 through the coating, it can be concluded that Ch biofilm
enriched by thyme essential oil made a kind of edible active coating over the surface of the samples
which can be utilized for preservation of fish during refrigeration. By using this bioactive packaging,
the shelf life was consequently extended from 8–9 days to 14 or more days.
 10 CHAMANARA ET AL.

REFERENCES

Alasalvar, C., Taylor, K. D. A., Öksüz, A., Garthwaite, T., Alexis, M. N., and Grigorakis, K. 2001. Freshness assessment of
cultured sea bream (Sparus aurata) by chemical, physical and sensory methods. Food Chem. 72: 33–40.
Arashisar, S., Hisar, O., Kaya, M., and Yanik, T. 2004. Effects of modified atmosphere and vacuum packaging on
microbiological and chemical properties of rainbow trout (Oncorynchus mykiss) fillets. Int. J. Food Microbiol. 97:
209–214.
Chen, C., Liau, W., and Tsai, G. 1998. Antibacterial effects of N-sulfonated and N-sulfobenzoyl chitosan and application to
oyster preservation. J. Food Protect. 61: 1124–1128.
Chien, P.-J., Sheu, F., and Lin, H.-R. 2007. Quality assessment of low molecular weight chitosan coating on sliced red
pitayas. J. Food Eng. 79: 736–740.
Darmadji, P., and Izumimoto, M. 1994. Effect of chitosan in meat preservation. Meat Sci. 38: 243–254.
Dorman, H. J. D., and Deans, S. G. 2000. Antimicrobial agents from plants: Antibacterial activity of plant volatile oils. J.
Appl. Microbiol. 88: 308–316.
Finnemore, H. 1926. The Essential Oils. London, United Kingdom: Ernest Benn Limited.
Downloaded by [Vahid Chamanara] at 00:06 21 January 2013

Goulas, A. E., and Kontominas, M. G. 2005. Effect of salting and smoking-method on the keeping quality of chub mackerel
(Scomber japonicus): Biochemical and sensory attributes. Food Chem. 93: 511–520.
Hammer, K. A., Carson, C. F., and Riley, T. V. 1999. Antimicrobial activity of essential oils and other plant extracts. J. Appl.
Microbiol. 86: 985–990.
Ibrahim Sallam, K. 2007. Antimicrobial and antioxidant effects of sodium acetate, sodium lactate, and sodium citrate in
refrigerated sliced salmon. Food Control. 18: 566–575.
Imaida, K., Fukushima, S., Shirai, T., Ohtani, M., Nakanishi, K., and Ito, N. 1983. Promoting activities of butylated
hydroxyanisole and butylated hydroxytoluene on 2-stage urinary bladder carcinogenesis and inhibition of γ-glutamyl
transpeptidase-positive foci development in the liver of rats. Carcinogenesis 4: 895–899.
Jeon, Y.-I., Kamil, J. A. Y. V., and Shahidi, F. 2002. Chitosan as an edible invisible film for quality preservation of herring
and Atlantic cod. J. Agr. Food Chem. 20: 5167–5178.
Kirk, R. S., Sawyer, R., and Egan, H. 1997. Pearson’s Composition and Analysis of Foods. 9th ed. London, United Kingdom:
Longman Group.
Knatt, S. R., Chander, R., and Sharma, A. 2008. Chitosan and mint mixture: A new preservative for meat and meat products.
Food Chem. 107: 845–852.
López-Caballero, M. E., Gómez-Guillén, M. C., Pérez-Mateos, M., and Montero, P. 2005. A chitosan-gelatin blend as a
coating for fish patties. Food Hydrocolloid. 19: 303–311.
Manju, S., Jose, L., Srinivasa Gopal, T. K., Ravishankar, C. N., and Lalitha, K. V. 2007. Effects of sodium acetate dip treat-
ment and vacuum-packaging on chemical, microbiological, textural and sensory changes of Pearlspot (Etroplus suratensis)
during chill storage. Food Chem. 102: 27–35.
Masniyom, P., Benjakul, S., and Visessanguan, W. 2005. Combination effect of phosphate and modified atmosphere on
quality and shelf life extension of refrigerated seabass slices. LWT-Food Sci.Technol. 38: 745–756.
Montes-Belmont, R., and Carvajal, M. 1998. Control of Aspergillus flavus in maize with plant essential oils and their
components. J. Food Protect. 61: 616–619.
Ojagh, S. M., Rezaei, M., Razavi, S. H., and Hosseini, S. M. H. 2009. Effect of chitosan coatings enriched with cinnamon
oil on the quality of refrigerated rainbow trout. Food Chem. 120: 193–198.
Prabuseenivasan, S., Jayakumar, M., and Ignacimuthu, S. 2006. In vitro antibacterial activity of some plant essential oils.
BMC Complement. Altern. Med. 6: 39.
Rasooli, I., and Mirmostafa, S. A. 2002. Antibacterial properties of Thymus pubescens and Thymus serpyllum essential oils.
Fitoterapia 73: 244–250.
Schubring, R., and Oehlenschlager, J. 1997. Comparison of the ripening process in salted Baltic and North Sea herring as
measured by instrumental and sensory methods. Z. Lebensm. Unters. Forsch. A. 205: 89–92.
Shahidi, F., Arachchi, V. J. K., and Jeon, Y. J. 1999. Food applications of chitin and chitosans. Trends Food Sci. Tech. 10:
37–51.
Shahidi, F., Kamil, J., Jeon, Y.-J., and Kim, S.-K. 2002. Antioxidant role of chitosan in a cooked cod (Gadus morhua) model
system. J. Food Lipids 9: 57–64.
Tsai, G.-J., Su, W.-H., Chen, H.-C., and Pan, C.-L. 2002. Antimicrobial activity of shrimp chitin and chitosan from different
treatments and applications of fish preservation. Fisheries Sci. 68: 170–177.
Ultee, A., Bennik, M. H. J., and Moezelaar, R. 2002. The phenolic hydroxyl group of carvacrol is essential for action against
the foodborne pathogen Bacillus cereus. Appl. Environ. Microbiol. 68: 1561–1568.
Ultee, A., Kets, E. P. W., Alberda, M., Hoekstra, F. A., and Smid, E. J. 2000. Adaptation of the foodborne pathogen Bacillus
cereus to carvacrol. Arch. Microbiol. 174: 233–238.