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18 Essential oils: natural antimicrobials forfish preservation

Barakat S.M. Mahmoud and Kazuo Miyashita

18.1 Introduction

Fish are highly susceptible to spoilage, which is caused mainly by microbial growth andmetabolism that produce amines, sulphides, alcohols, aldehydes, ketones, and organic acids.Spoiled products have unpleasant and unacceptable off-flavours, making fish that is notwell protected unsuitable for human consumption. Gram-negative bacteria are mostly foundin fresh fish and are responsible for fish spoilage. Gram-negative, fermentative bacteriasuch as Vibrionaceae, cause spoilage of unpreserved fish, whereas psychrotolerant gram-negative bacteria Pseudomonas spp. and Shewanella spp. grow in chilled fish. Meanwhile,Photobacterium phosphoreum and lactic acid bacteria spoil packed fish. Lactobacillus andCarnobacterium associated with gram-negative fermentative bacteria, such as P. phospho-reum and psychrotrophic Enterobacteriaceae, are the dominant bacteria in lightly salted,acidified, and chill-stored vacuum packed fish. Spore-forming bacteria such as Clostridiumor Bacillus may grow in unsalted heat-treated fish products [1–4].

Improving the safety and quality of seafood is important for both the consumers and theseafood industry. Synthetic or natural preservatives can control microbial growth and improvethe safety and quality during fish storage. Consumers prefer natural preservatives to theirartificial counterparts, which may potentially have undesirable effects on health. Essentialoils produced from some kinds of plants, such as herbs and spices, are representative naturalpreservatives for fish or fish products [5–8].

This chapter introduces antibacterial properties of essential oils against fish spoilagebacteria, focusing on the active components of essential oils and their effective applicationfor the preservation of fish and fish products.

18.2 Essential oils

18.2.1 Chemistry of essential oils

Essential oils contain many kinds of chemical compounds that have a variety of biologicalactivities. The main components are terpenes, alcohols, hydrocarbons, phenols, aldehydes,

Handbook of Seafood Q uality, Safety and Health Applications

Edited by Cesarettin Alasalvar, Fereidoon Shahidi, Kazuo Miyashita and Udaya Wanasundara

© 2011 Blackwell Publishing Ltd. ISBN: 978-1-405-18070-2


218 Seafood Quality, Safety and Health Applications

Cinnamaldehyde

Eugenol Thymol

Allyl iso-thiocyanate

OH

Carvacrol

CHO

N=C=S

O

OH

OH

Fig. 18.1 Structures of active compound from essential oils.

esters, and ketones, which occur naturally as major components in plants. Phenols foundin clove, oregano, and thyme oils, are highly antimicrobial against a wide range of micro-organisms. Terpenes (including monoterpenes, diterpenes, sesquiterpenes, etc.) are the mostabundant components of the essential oils from many plants. There is no doubt that herbsand spices are the most important sources for production of essential oils.

18.2.2 Active components of essential oils

Essential oils are plant extracts from herbs and spices such as allspice, anise, basil, bay,caraway, cardamom, cinnamon, clove, coriander, cumin, eucalyptus, fennel, garlic, ginger,horseradish, lemongrass, mace, mandarin, marjoram, mint, mustard, nutmeg, onion, orange,oregano, parsley, rosemary, saffron, sage, and thyme, etc. They have different ranges ofantimicrobial activities against spoilage and pathogenic bacteria [9–19]. Phenolic compoundssuch as carvacrol, eugenol, and thymol (Fig. 18.1) have the strongest antimicrobial properties[20–29]. Cinnamaldehyde and allyl isothiocyanate (Fig. 18.1) also show strong antimicrobialactivities, as reported in some studies [30].

18.2.3 Bacterial sensitivity to essential oils andtheir components

Gram-positive bacteria are more sensitive to essential oils than gram-negative bacteria[31–39]. Gram-negative bacteria possess an outer membrane surrounding the cell wall, whichrestricts diffusion of hydrophobic components through its lipopolysaccharide constituents[8,39–41].

18.2.4 Phenolic compounds

The antimicrobial activity of essential oils cannot be explained by one specific mecha-nism, because there are several molecular targets in the bacterial cell for active componentsof essential oils [39,42]. Phenolic compounds are the main group responsible for the an-timicrobial activity of essential oils. They can dissolve the hydrophobic domain (lipids)


Essential oils for fish preservation 219

of the bacterial membrane, disintegrate the outer membrane, increase the permeability ofthe cytoplasmic membrane to adenosine-5′-triphosphate (ATP), and inactivate the cytoplas-mic enzymes. Antimicrobial effects of phenolic compounds are based on these biologicalactivities [9,39,43–47].

18.3 Application of essential oils to fish preservation

Essential oils are soluble in alcohol, but poorly soluble in water. They are widely used in thefood industry as flavourings and preservative agents [48]. Fish is one of the most commonfood materials that are traditionally combined with many spices and herbs [9,48]. Fromthe industrial and marketing point of view, the shelf-life of fish is very important. If theshelf-life can be prolonged by one or two days, it allows a better marketability and moreprofit [48]. Essential oils are relatively inexpensive compared with other preservative agentsor technologies such as smoking, and irradiation, etc. However, essential oils influence thesensory characteristic of fish and should be added at low concentrations for preserving fish[44].

Most studies have evaluated the antimicrobial effect of essential oils and/or their activecompounds in vitro. However, few investigations have reported their potential role in fish[49–55]. In general, higher concentrations of essential oils or their active compounds arerequired to achieve the same antimicrobial effect in food as compared to that in culture media[26,56–58]. This is due to the interaction between active compounds of essential oils andnutrients of foods [7,10,45,55,59].

This chapter describes the effect of essential oils or their active compounds on bacterialgrowth in culture media and on fish preservation. The combination effect of essential oilswith other preservatives is also reviewed.

18.3.1 Effect of essential oils on fish spoilage bacteria

Mahmoud et al. [19] reported the antimicrobial activity of garlic essential oil, allyl isothio-cyanate, carvacrol, cinnamaldehyde, citral, cuminnaldehyde, eugenol, isoeugenol, linalool,and thymol against 14 bacterial strains (in culture media) isolated from carp (Cyprinus car-pio) using the paper disc diffusion method. The diameter of the growth inhibition zone wasused as the criterion for measuring the antimicrobial activity of garlic oil and its active com-pounds. Phenolic compounds, thymol and carvacrol, aromatic aldehyde, and cinnamaldehydeshowed the strongest antimicrobial activity against the bacterial isolates from carp. Theseresults are in agreement with those obtained by other researchers [10,21,27]. The above phe-nolic compounds that contain a hydroxe group and a molecular mass of 150 to 160, have beenreported as the most effective antimicrobial components found in essential oils [9,56]. Thecombination of these phenols was also effective in the inhibition of bacterial growth [19]. Thecombination of carvacrol and thymol has the strongest antimicrobial activity, followed bycinnamaldehyde and thymol, and then carvacrol and cinnamaldehyde. These results indicatethat a combination of two compounds has a stronger antimicrobial effect than the individualone [19,60].

Mejlholm and Dalgaard [44] evaluated the antimicrobial effect of nine essential oils (basil,bay, cinnamon, clove, lemongrass, marjoram, oregano, sage, and thyme) on the growth ofa mixture of five strains of P. phosphoreum, a specific spoilage organism responsible forspoilage of modified atmosphere-packaged cod fillets, in a liquid medium. All tested essential



oils reduce the growth rate of P. phosphoreum. The antimicrobial effect depends on the oiltype and the concentration; essential oil concentrations of 0.005 to 0.1% (v/v) are required forinhibition of the growth of P. phosphoreum. Oregano and cinnamon oils have the strongestantimicrobial activity followed by lemongrass, thyme, clove, bay, marjoram, sage, and basiloils.

Prasad and Seenayya [61] studied the effect of essential oils from 20 spices at concen-trations of 0.1 to 2% (w/v) against 18 strains (isolates from salt cured fish), namely 12Salinococcus roseus, 5 Halococcus turkmenicus, and 1 Halococcus morrhuae. Clove pow-der and its essential oil were the most effective agents, even at a low concentration (0.1%).Onion, coriander, garlic, asafetida, mustard, and spilanthes showed excellent growth control,followed by red chillies, turmeric, ginger, cumin seed, and fenugreek.

Yano et al. [62] studied the antimicrobial effects of essential oils from 18 spices andherbs at final concentrations of 2.5% against a food-borne pathogen bacterium, Vibrio para-haemolyticus (fish contaminants bacteria) incubated at different temperatures (5 and 30◦C).Basil, clove, garlic, horseradish, marjoram, oregano, rosemary, and thyme exhibited goodantibacterial activities against tested bacterium. The lowest minimum inhibitory concentra-tion (MIC) was 0.125% observed in clove and marjoram at 30◦C and 0.063% in marjoramand turmeric at 5◦C. V. parahaemolyticus was more sensitive at low temperatures [63]. Theseresults suggest that spices, herbs or their extract can be practical for protecting fish from therisk of contamination by V. parahaemolyticus.

18.3.2 Effect of essential oils on shelf-life of fish

Utilisation of essential oil and their active compounds to extend the shelf-life of fish haspreviously been reported (Table 18.1). Treatment of carp fillets with 0.5% carvacrol + 0.5%thymol solution for 15 min significantly decreased the initial total microbial count [19].Harpaz et al. [46] reported that treatments with 0.05% oregano and/or thyme lead to remark-able reduction in the bacterial growth of Pseudomonas spp. and Salmonella putrefaciens(mostly responsible for the spoilage of fish during cold storage) on Asian sea bass fish.Tassou et al. [7] studied the effect of oregano oil against inoculated Staphylococcus aureusand S. enteritidis on fish fillets under an aerobic and modified atmosphere of 40% CO2,30% O2, and 30% N2 or air at 0◦C. The treatment had a good bacteriostatic effect on bothnatural flora and inoculated bacteria on the fillets. Tassou et al. [7] also reported the effectof mint essential oil (0.5, 1.0, 1.5, and 2.0%, v/w) on inoculated S. enteritidis and Liste-ria monocytogenes on three models of fish foods, namely tzatziki (pH 4.5), taramasalata

Table 18.1 Essential oils and essential oil compounds for fish preservation

Fish Essential oil/compounds Reference

Carp (Cyprinus carpio) Carvacrol, thymol [19,47]Sea bass (Lates calcarifer) Oregano, thyme [46]Cod (Gadus morhua) Oregano [44]Shrimp (Penaeus spp.) Thyme, cinnamon [67]Sea bream (Sparus aurata) Oregano [7]Cod (Gadus morhua) Mint [7]Mackerel muscle extracts Cardamom, allspice, chilli, cinnamon,

clove, cumin, black pepper, nutmeg,sage, thyme

[64]



(pH 5.0), and pate (pH 6.8) at 10◦C for one week. The population of S. enteritidis wascompletely inhibited in tzatziki and significantly decreased in the other two fish foods. Thepopulation of L. monocytogenes decreased during the storage period.

Mejholm and Dalgaard [44] screened the effect of nine essential oils (basil, bay, cinnamon,clove, lemongrass, marjoram, oregano, sage, and thyme) against the spoilage fish bacteria, P.phosphoreum in vitro. Among all tested essential oils, oregano oil showed the best inhibitioneffect on bacterial growth. Treatment with oregano essential oil yielded a pleasant flavourto cold fillets and significantly increased the shelf-life to 26 days as compared with 12 daysfor the control at 2◦C. On the other hand, treatment with 0.05% oregano essential oil had aminor antimicrobial effect on salmon fillets, while the treatment was more effective on codfillets. The difference in the antimicrobial effect of oregano essential oil on cod and salmonfillets is due to the higher lipid content of salmon fillets compared to cod fillets.

Wendakoon and Sakaguchi [64] studied the effect of cardamom, allspice, chilli, cinna-mon, clove, cumin, black pepper, nutmeg, sage, thyme, and their extracts on the growthof Enterobacter aerogenes and Morganella morganii in mackerel muscle extracts at 30◦C.Clove and cinnamon showed the strongest antimicrobial effect against tested bacteria. All-spice, sage, and thyme powders and extracts were more effective against M. morganii thanE. aerogenes. The antimicrobial activity of clove and cinnamon was apparently due to eugenoland cinnamaldehyde, respectively.

18.3.3 Antimicrobial effect of combined treatment of essentialoils with other antimicrobial agents

The efficacy of essential oils can be enhanced by their combination with other preservatives[28,47,65]. Mahmoud et al. [47,66] studied the preservative effects of a combined treatmentwith 0.5% carvacrol + 0.5% thymol (Cv+Ty) and electrolysed water solutions composed ofcathodic solution (EW-) and anodic solution (EW+) on carp fillets at 5◦C for 20 days. Com-bined treatment of the fillets with Cv+Ty and electrolysed solution resulted in a significantreduction (P ≤ 0.05) in the total microbial counts immediately after treatment (day 0) ascompared with the control. The total microbial count of control samples exceeded an accept-able limit (6.0 log10 cfu/g) after 4 days at 5◦C. On the other hand, when the samples werepre-treated with electrolysed solution, and then treated with (Cv+Ty), the total microbialcount exceeded the acceptable limit after 16 days at 5◦C. Wendakoon and Sakaguchi [64]examined the combined effect of sodium chloride and clove on the growth of Enterobacteraerogenes in mackerel muscle. The presence of NaCl (2%) with clove (0.5%) completelyinhibited the growth of E. aerogenes in mackerel broth, though clove alone was inactiveon the growth of E. aerogenes. Later, Ouattara et al. [67] studied the combined effect oflow-dose gamma irradiation and antimicrobial coating (thyme oil, 0.5 and 0.75%) and trans-cinnamaldehyde (0.3 and 1.5%) on the shelf-life of pre-cooked shrimp. A synergistic effect(P ≤ 0.05) between gamma irradiation and coating treatments was observed. The shelf-lifeof treated shrimp was extended by 12 days compared to the control.

18.4 Conclusions

The most important compounds responsible for the antimicrobial activity of essential oils infish or fish product were determined. Phenolic compounds were the main group responsible



for the antimicrobial activity of essential oils. Further investigations are needed to understandhow phenolic compounds and other active compounds in essential oils interact with bacterialcells. In addition, interactions among different active compounds of essential oils and betweenthese compounds and other nutrients of fish or fish products will be of continued interestto researchers working on the antimicrobial effect of essential oils. Hence, the needs toincorporate such challenging areas into the research and evaluate essential oils individuallyand in combination with other preservation techniques remains to be of priority. Thesemay encourage the development of novel approaches in the preservation of fish and fishproducts.

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