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Effect of Alpinia zerumbet essential oil on the shelf life of tambaqui fillets during short-term refrigerator storage

Efeitos do óleo essencial de Alpinia zerumbet sobre a vida útil de filés resfriados de tambaqui

ABSTRACT

We investigated the effect of Alpinia zerumbet essential oil on the quality and shelf life of tambaqui (Colossoma macropomum) fillets stored under refrigeration (10.0 ± 0.5 °C) for 14 days. The treatments were A. zerumbet essential oil at 0.75% v v-1 (AEO 0.75%), A. zerumbet essential oil at 1.5% v v-1 (AEO 1.5%) and a control (no essential oil). The sample quality and shelf life were determined by the total psychrotrophic count (TPC) and chemical parameters (pH, total volatile basic nitrogen, centesimal composition and thiobarbituric acid reactive substances - TBARS) at zero, seven and 14 days of storage time. The TPC decreased significantly (p < 0.05) with an A. zerumbetessential oil level of 1.5% until seven days of storage. The concentration of A. zerumbet essential oil at 0.75% resulted in lower pH, TBARS, and TVBN values in comparison with the other treatment and the control. Thus, A. zerumbet essential oil was efficient in extending the shelf life of refrigerated tambaqui fillets up to approximately seven days.

Keywords:
antioxidant; psychrotrophic count; quality; fish storage

RESUMO

Investigamos o efeito do óleo essencial de Alpinia zerumbet sobre a qualidade e vida útil de filés de tambaqui (Colossoma macropomum) armazenados sob refrigeração (10,0 ± 0,5 °C) durante 14 dias. Os tratamentos foram óleo essencial de A. zerumbet (OEA) a 0,75% v v-1 (OEA 0,75%) e 1,5% v v 1 (OEA 1,5%), além de um controle (sem OEA). A qualidade e vida útil das amostras foram determinadas por contagem psicrotrófica total (CPT) e parâmetros químicos (pH, bases nitrogenadas voláteis totais, composição centesimal e substâncias reativas ao ácido tiobarbitúrico - SRATB), nos dias zero, sete e 14 de armazenamento. O TPC diminuiu significativamente (p < 0,05) com óleo essencial de A. zerumbet a 1,5% por até sete dias de armazenamento. O tratamento contendo OEA a 0,75% resultou em menores valores de pH, SRTAB e BNV-T em comparação ao outro tratamento e ao controle. Portanto, o óleo essencial de A. zerumbet foi eficiente para prolongar a vida útil do filé de tambaqui por aproximadamente sete dias.

Palavras-chave:
antioxidante; contagem de psicrotróficos; qualidade; estocagem de pescado.

INTRODUCTION

Fish is the main source of animal protein in the human diet due to its high nutritional value (FAO 2016FAO. 2016. The State of World Fisheries and Aquaculture (SOFIA) (The State of World Fisheries and Aquaculture (SOFIA) (http://www.fao.org/documents/card/en/c/2c8bcf47-2214-4aeb-95b0-62ddef8a982a ). Accessed on 15 Jan 2018.
http://www.fao.org/documents/card/en/c/2...
). Fish is both a subsistence food for many populations, especially in developing countries, and a generator of wealth derived from fishing and aquaculture, as it one of the most traded food commodities worldwide and is worth approximately US$ 142 billion in 2016 (FAO 2018FAO. 2018.Global Aquaculture Production. FAO, Rome. 29p.). It is estimated that fish could represent 50% of the animal protein consumption in 2030 (FAO 2018FAO. 2018.Global Aquaculture Production. FAO, Rome. 29p.).

However, fresh and minimally processed fish are susceptible to microbial spoilage and lipid oxidation due to their high water and unsaturated fat contents (Iturriaga et al.2012Iturriaga, L.; Olabarrieta, I.; Marañon, I.M. 2012. Antimicrobial assays of natural extracts and their inhibitory effect against Listeria innocua and fish spoilage bacteria, after incorporation into biopolymer edible films.International Journal of Food Microbiology, 158: 58-64. ; Secci and Parisi 2016Secci, G.; Parisi, G. 2016. From farm to fork: lipid oxidation in fish products. A review.Italian Journal of Animal Science, 15: 124-136.), which result in an undesirable off-flavor and a reduced shelf life. Traditionally, synthetic additives with antimicrobial (e.g., potassium sorbate) and antioxidant actions (e.g., BHT and EDTA) have been used to increase the shelf life of food and sustain consumer acceptance (Carocho et al.2014Carocho, M.; Calhelha, R.C.; Queiroz, M.J.R.P.; Bento, A.; Morales, P.; Sokovic, M.; Ferreira, I.C.F.R. 2014. Infusions and decoctions ofCastanea sativa flowers as effective antitumor and antimicrobial matrices. Industrial Crops and Products, 62: 42-46.; Honold et al.2016Honold, P.J.; Jacobsen, C.; Jónsdóttir, R.; Kristinsson, H.G.; Hermund, D.B. 2016. Potential seaweed-based food ingredients to inhibit lipid oxidation in fish-oil-enriched mayonnaise.European Food Research and Technology, 242: 571-584. ). However, as the excessive consumption of synthetic food additives is related to gastrointestinal, respiratory, dermatological and neurological disorders (Randhawa and Bahna 2009Randhawa, S.; Bahna, S.L. 2009. Hypersensitivity reactions to food additives.Current Opinion of Allergy Clinic and Immunology, 9: 278-83.;Wilson and Bahna 2005Wilson, B.G.; Bahna, S.L. 2005. Adverse reactions to food additives.Ann Allergy Asthma Immunology, 95: 499-507. ), their use is restricted by regulations to minimize health risks (Leuschner and Zamparini 2002Leuschner, R.G.K.; Zamparine, J. 2002. Effect of spices on growth and survival ofEscherichia coliO 157 andSalmonella entericaserovar Enteritidis in broth model systems and mayonnaise.Food Control ,13: 399-404.). Therefore, the demand for natural alternatives, such as essential oils (EOs) extracted from plants, has been increasing (Pawar et al. 2012Pawar, N.; Arora, S.; Bijoy, R.; Wadhwa, B. 2012. The effects ofAsparagus racemosus(shatavari) extract on oxidative stability of ghee, in relation to added natural and synthetic antioxidants.International Journal of Dairy Technology ,65: 293-299.). In Europe and the USA, EOs are used as food additives (CBI 2018CBI. 2018. Exporting essential oils for food to Europe. ( (https://www.cbi.eu/market-information/natural-food-additives/essentials-oils-food/ ). Accessed on 28 Aug 2018.
https://www.cbi.eu/market-information/na...
; FDA 2017FDA. 2017. Generally Recognized as Safe (GRAS). ( (https://www.fda.gov/food/ingredientspackaginglabeling/gras/ ). Accessed on 28 Aug 2018.
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). In Brazil, ANVISA regulates the use of essential oils by Resolution RDC nr. 2 (15 January 2007), and the list of approved EOs for use in food is the same as that used by the FDA and the EU (ANVISA 2007ANVISA. 2007. Regulamento Técnico sobre Aditivos em Alimentos. RDC N. 2, de 15 de Janeiro de 2007.).

In addition to flavoring properties, EOs have shown strong antioxidant and antimicrobial properties (Pesaventoet al. 2015Pesavento, G.; Calonico, C.; Bilia, A.R.; Barnabei, M.; Calesini, F.; Addona, R.; Lo Nostro, A. 2015. Antibacterial activity of oregano, Rosmarinus and Thymus essential oils against Staphylococcus aureus and Listeria monocytogenes in beef meatballs.Food Control, 54: 188-199. ; Joukiet al.2014Jouki, M.; Yazdi, F.T.; Mortazavi, S.A.; Koocheki, A.; Khazaei, N. 2014. Effect of quince seed mucilage edible films incorporated with oregano or thyme essential oil on shelf life extension of refrigerated rainbow trout fillets.International Journal of Food Microbiology, 174: 88-97.; Yeet al.2013Ye, C-L.; Dai, D-H.; Hu, W-L. 2013. Antimicrobial and antioxidant activities of the essential oil from onion (Allium cepa L.). Food Control, 30: 48-53.; Brenes and Roura 2010Brenes, A.; Roura, E. 2010. Essential oils in poultry nutrition: Main effects and modes of action.Animal Feed Science and Technology, 158: 1-14.; Burt 2004Burt, S. 2004. Essential Oils: their antibacterial properties and potential applications in foods - a review.International Journal of Food Microbiology ,94: 233-253. ), limiting the growth of food pathogens and increasing the shelf life of some foods (Gómez-Estacaet al. 2010Gómez-Estaca, J.; López de Lacey, A.; López-Caballero, M.E.; Gómez-Guillén, M.C.; Montero, P. 2010. Biodegradable gelatin-chitosan films incorporated with essential oils as antimicrobial agents for fish preservation.Food Microbiology, 27: 889-896. ; Ojaghet al. 2010Ojagh, S.M.; Rezaei, M.; Razavi, S.H.; Hosseini, S.M.H. 2010. Effect of chitosan coatings enriched with cinnamon oil on the quality of refrigerated rainbow trout.Food Chemistry, 120: 193-198. ). For example, an oregano EO showed an antimicrobial effect, emerging as an alternative to preserve sliced ham (Galindo 2017Galindo, M.V. 2017. Filmes biodegradáveis de gelatina e quitosana com adição de óleos essenciais na conservação de presunto embalado a vácuo. Master’s dissertation. Universidade Tecnológica Federal do Paraná, Brazil. 52p. ) and chicken pate (Moraes-Lovison 2017Moraes-Lovison, M. 2017. Óleo essencial de orégano nanoemulsionado: produção, caracterização físico-química, atividade antioxidante in vitro e aplicação em patê de frango. Doctoral thesis, Faculdade de Zootecnia e Engenharia de Alimentos, Pirassununga, Brazil. 89p.); a thyme EO reduced the total psychrotrophic count and extended the shelf life of fillets of trout (Oncorhynchus mykiss) (Chamanaraet al. 2013Chamanara, V.; Shabanpour, B.; Khomeiri, M.; Gorgin, S. 2013. Shelf-life extension of fish samples by using enriched chitosan coating with thyme essential oil.Journal of Aquatic Food Product Technology, 22: 3-10.); a tangerine peel EO (Citri reticulatae pericarpium) reduced the degradation process of bream (Megalobrama amblycephala) fillets during iced storage (He and Xiao 2016He, Q.; Xiao, K. 2016. The effects of tangerine peel (Citri reticulatae pericarpium) essential oils as glazing layer on freshness preservation of bream (Megalobrama amblycephala) during super chilling storage.Food Control, 69: 339-345. ); lemon and thyme EOs maintained grass carp (Ctenopharyngodon idella) fillet quality (Cai et al. 2017Cai, L.; Leng, L.; Cao, A.; Cheng, X.; Li, J. 2017. The effect of chitosan-essential oils complex coating on physicochemical, microbiological, and quality change of grass carp (Ctenopharyhgodon idella) fillets. Journal of Food Safety, 38: 1-9.); and a clove EO decreased the microbial growth and lipid oxidation of tuna (Thunnus thynnus) fillets during 17 days of storage (Echeverría et al. 2018Echeverria, I.; Lopez-Caballero, M.E.; Gomez-Guillen, M.C.; Mauri, A.N.; Montero, M.P. 2018. Active nanocomposite films based on soy proteins-montmorillonite-clove essential oil for the preservation of refrigerated Bluefin tuna (Thunnus thynnus) fillets. International Journal of Food Microbiology, 266: 142-149.).

Among the species of plants used in the extraction of EOs with potential for use in food is Alpinia zerumbet. It is an Asian plant with antimicrobial properties (Victórioet al.2009Victório, C.P.; Alviano, D.S.; Alviano, C.S.; Lage, C.L.S. 2009. Chemical composition of the fractions of leaf oil ofAlpinia zerumbet(Pers.) and antimicrobial activity.Revista Brasileira de Farmacognosia ,19: 697-701.) againstStaphylococcus aureus(Castroet al.2016Castro, K.N.C.; Lima, D.F.; Vasconcelos, L.C.; Santos, R.C.; Pereira, A.L.M.; Fogaça, F.H.S.; Calvet, R.M. 2016. Chemical composition and efficacy of essential oil and ethanol extract ofAlpinia zerumbetonStaphylococcus aureus.Arquivos do Instituto Biológico, 83: 1-7.). The extract of A. zerumbet had antioxidant and antimicrobial activities in minced meat by inhibiting lipid oxidation and increasing microbial stability (Cheah and Gan 2000Cheah, P.B.; Gan, S.P. 2000. Antioxidative/antimicrobial effects of galangal and (alpha)-tocopherol in minced beef. Journal of Food Protection, 63: 404- 407.). Alpinia zerumbet is widely cultivated and distributed in tropical and semitropical areas, including Brazil (Thenmozhiet al. 2011Thehenmozhi, S.; Sureshikumar, S.; Venugopalan, R. 2011. Evaluation of analgesic and anti-inflammatory activity ofAlpinia speciosaK. Schum rhizomes.Journal of Pharmacy Research, 4: 728-729.). It is a very popular medicinal plant in Brazil (Costaet al.2016Costa, G.M.; Suga, R.G.; Oliveira, P.L.; Magalhães, P.P.; Farias, L.M.; Duarte, L.P.; Silva, F.C. 2016. Bioactivity of extracts fromAlpinia zerumbet(Pers.) B.L. Burtt & R.M. Sm. against sinusitis causing bacterial pathogens.Etnofarmacologia, 9: 185-194. ). Alpinia zerumbetEO does not present cytotoxic or genotoxic action in vivo or in vitro (Oliveira 2008Oliveira, C.C. 2008. Estudo toxicológico pré-clínico do extrato aquoso e do óleo essencial das folhas de Alpinia zerumbet (Pers.) Burtt & Smith. Master’s dissertation, Universidade Federal do Ceara, Brazil. 93p.). It is used in traditional Japan cuisine and herbal medicine, and there is no data on the maximum amounts tolerated by humans without signs of toxicity (Teschke and Xuan 2018Teschke, R.; Xuan, T.D. 2018. Viewpoint: a contributory role of shell ginger (Alpinia zerumbet) for human longevity in Okinawa, Japan? Nutrients, 10: 1-22.).

Tambaqui (Colossoma macropomum) is an Amazonian fish that has an efficient performance in intensive farming, and it reaches high market values (Valet al.2000Val, A.L.; Rolim, P.R.; Tabelo, H. 2000. Situação atual da aquicultura na região Norte. In: Valenti, W.C. (Ed.).Aquicultura no Brasil. CNPq, Brasília, D.F., p.247-266.). However, the method of storing raw fish in ice used in the current distribution system causes quality loss and fast spoilage (Li et al.2013Li, T.; Li, J.; Hu, W.; Li, X. 2013. Quality enhancement in refrigerated red drum (Sciaenops ocellatus) fillets using chitosan coatings containing natural preservatives.Food Chemistry, 138: 821-826.). Accordingly, the shelf life of raw tambaqui is approximately 20 days (Borgeset al. 2013Borges, A.; Conte-Junior, C.A.; Franco, R.M.; Freitas, M.Q. 2013. Quality Index Method (QIM) developed for pacuPiaractus mesopotamicus and determination of its shelf life. Food Research International, 54: 311-317. ; Silva et al. 2018Silva, M.L.B.P.; Lopes, J.M.; Vieira, S.G.A.; Araujo, T.D.S.; Calvet, R.M.; Pereira, A.M.L.; Fogaca, F.H.S. 2018. Development of a quality index scheme and shelf-life study for whole tambaqui (Colossoma macropomum).Acta Amazonica, 48: 98-108.), and its fillets have a shelf life of approximately 6 days in iced storage (Bottinoet al.2017Bottino, F.O.; Rodrigues, B.L.; Ribeiro, J.D.N.; Azaro, E.A.T.; Conte-Junior, C.SA. 2017. Influence of UV-C Radiation on Shelf Life of Vacuum Package Tambacu (Colossoma macropomum × Piaractus mesopotamicus) Fillets. Journal of Food Processing and Preservation, 41: e13003.).

Therefore, based on the antimicrobial properties presented by A. zerumbetand its oriental cuisine uses, we tested A. zerumbet EO as a natural additive for fish fillets to promote a longer shelf life. The present study investigated the effect ofA. zerumbetEO on the quality and shelf life of tambaqui fillets during 14 days of refrigerated storage.

MATERIAL AND METHODS

Preparation and analysis of essential oil

Alpinia zerumbetleaves were harvested at Parnaiba, Piaui, Brazil (03º01’27.5”S, 41º44’53.5”W) in the morning (until 9 am). The essential oil (EO) was extracted by hydro-distillation in the Clevenger system for 3 h. The A. zerumbet essential oil (AEO) was analyzed using a gas chromatography-mass spectrometry (GC-MS) instrument (Varian GC-450/MS-240, Palo Alto, CA, USA) according to Castroet al.(2016Castro, K.N.C.; Lima, D.F.; Vasconcelos, L.C.; Santos, R.C.; Pereira, A.L.M.; Fogaça, F.H.S.; Calvet, R.M. 2016. Chemical composition and efficacy of essential oil and ethanol extract ofAlpinia zerumbetonStaphylococcus aureus.Arquivos do Instituto Biológico, 83: 1-7.). The GC-MS analysis identified 23 volatile compounds, of which p-cymene (32.72%), 1.8-cineole (24.05%) and 4-terpineol (20.23%) were the major components.

Fish samples

Farmed tambaqui (45 fishes; six months old; mean weight = 560 g) were purchased at an aquaculture station at Ilha Grande, Piaui, Brazil, stored immediately on ice and transported in polystyrene boxes to the laboratory of Food Analysis and Technology, Parnaiba-PI. The fish were gutted and filleted manually. The resulting fillets were divided into three different batches of 15 units each.

Sample preparation

Initially, a primary aqueous solution with 1% glacial acetic acid was prepared. The control solution was prepared by adding 1 L of distilled water in the primary solution to a maximum of 2 L. Two treatment solutions were composed of 1% acetic acid, 0.2% Tween-20 (Merck, Darmstadt, Germany), and AEO (0.75% and 1.5%, respectively), which were homogenized under magnetic stirring at 800 rpm at 25 °C.

The fish fillets were randomly placed into three batches (control, AEO 0.75% and AEO 1.5%). The batches did not differ significantly in size and weight of fillets (Table 1). Each fillet was immersed for 30 s in 600 mL of the solution assigned to its respective treatment, and, after two minutes, immersed a second time for 30 s (Chamanaraet al. 2013Chamanara, V.; Shabanpour, B.; Khomeiri, M.; Gorgin, S. 2013. Shelf-life extension of fish samples by using enriched chitosan coating with thyme essential oil.Journal of Aquatic Food Product Technology, 22: 3-10.). The fillets were then glazed with cold water (2 °C), packaged in polyethylene bags and stored in a refrigerated incubator at 10 °C during 14 days. The chemical and microbial analyses were performed at three time points: day zero (after treatment and prior to refrigeration), and at day 7 and 14 of the refrigeration period, to assess the overall quality of the fish fillets.

Table 1
Characteristics of the tambaqui (Colossoma macropomum) fillets used in the experiment [total weight (g), width (cm), height (cm) and thickness (cm)]. Values are the mean ± SD of 15 replicates, without significant differences between the treatments (p < 0.05).

Chemical analyses

The proximate composition analysis was performed according to standard analytical methods (AOAC 2005AOAC. 2005.Official Method of Analysis. 17th ed. Association of Official Analytical Chemists, Washington, 593p.). Five samples were taken from each treatment at each time point and analyzed in triplicate. The moisture content was determined as the weight difference for 2 g of muscle before and after heating in an oven (SOLAB, Piracicaba, Brazil) for 24 h at 105 °C. The total nitrogen content (CP) was determined by the Kjeldahl procedure, and the protein content was estimated using a conversion factor of 6.25. Ash content was determined as the weight difference after heating at 550 °C for 6 h in a muffle furnace (Quimis, São Paulo, Brazil), and lipid content was quantified by extraction with petroleum ether in a Soxhlet apparatus.

The pH was measured using a digital pH meter (Del Lab model DLA-pH, São Paulo, Brazil). Muscle samples (10 g) were homogenized with 40 mL of distilled water, and the homogenate was used for pH determination according to theKirschnik et al. (2013Kirschnik, P.G.; Trindade, M.A.; Gomide, C.A.; Moro, M.E.G.; Viegas, E.M.M. 2013. Estabilidade em armazenamento da carne de tilápia-do-nilo mecanicamente separada, lavada, adicionada de conservantes e congelada.Pesquisa Agropecuária Brasileira, 48: 935-942. ) method.

Lipid oxidation was evaluated through the formation of thiobarbituric acid reactive substances (TBARS), according to Vyncke (1970Vyncke, W. 1970. Direct determination of the TBA value in trichloroacetic acid extract of fish as a measure of oxidative rancidity.Fette-Scifen Anstrichmittel ,72: 1084-1087.), in samples of 10 g of muscle. A 5-ml aliquot of the distillate was used for color development and was measured at 532 nm using a spectrophotometer (HACH, model 2800, Germany). The results were expressed as mg of malondialdehyde (MDA) per kg of muscle. The oxidation potential was calculated from the standard curve y = 0.2727x - 0.2007 (r2 = 0.991).

The total volatile base nitrogen (TVB-N) determination was carried out using a methodology adapted from Savay da Silvaet al. (2008Savay da Silva, L.K.; Riggo, R.; Martins, P.E.; Galvão, J.A.; Oetterer, M. 2008. Otimização e padronização do uso da metodologia para determinação de bases nitrogenadas voláteis totais (BNVT) em camarões,Xyphopenaeus kroyeri.Brazilian Journal of Food and Technology, 20: 138-144.), in which 50 g of muscle was homogenized in 150 mL of 5% trichloroacetic acid for the precipitation of protein nitrogen. After filtration, the amount of low-molecular-weight nitrogen compounds soluble in TCA was measured using the semi-macro Kjeldahl method, and the soluble nitrogen content was estimated using a conversion factor of 6.25.

Psychrotrophic bacteria count

The total psychrotrophic count (TPC) was determined through the pour plate method using plate count agar (HIMEDIA®, LBS Marg, Mumbai, India) as a medium at days zero, 7 and 14 of storage (Evanchoet al.2001Evancho, G.M.; Sveum, H.; Moberg, L.J.; Frank, J.F. 2001. Microbiological monitoring of the food processing environment. In: Downes, F.P.; Ito, K. (Eds.). Compendium of methods for the microbiological examination of foods. American Public Health Association, Washington, D.C., p.25-36.). We homogenized 10 g of the anterior dorsal region of the fillets in 90 mL of 0.1% peptone solution. In addition, serial decimal dilutions were prepared from a 10−1dilution, as necessary. The plates were incubated at 10 °C for seven days. Two replicates in each group were analyzed. The bacterial count results were expressed as log10.

Statistical analysis

All measurements were carried out in triplicate and mean values per sample were used in the analyses. The differences among the treatments and storage times were determined by the Kruskal-Wallis test (one-way ANOVA) followed by the Dunn’s post hoc test. The Bonferroni correction was applied to adjust the significance level of multiple test results. The interactions between the treatments and storage times were determined by the Tukey-Kramer test. Significance was defined at p < 0.05. The analyses were carried out with the ASSISTAT version 7.7 software package.

Ethical and legal aspects

The Ethics Committee on the Research Animals Use (CEUA) of Embrapa Meio Norte (protocol 02/2014) approved the study. The use of Alpinia zerumbet was authorized by CGEN through the Embrapa special authorization for genetic heritage access for scientific research purposes (nr. 002/2008, IBAMA process 2001.001558/2006-21).

RESULTS

Proximal composition

The proximate composition of the fillets varied among control and treatments (Figure 1). Moisture varied significantly among all groups at all time-points, with highest values for AEO 1.5% at day 14 (Figure 1a). The ash content was significantly higher in the fillets with AEO compared to the control group (Figure 1b). There was no significant effect of storage time on moisture and ash content. CP was significantly lower in the treatments until day 7, but significantly higher in AEO 1.5% at day 14 (Figure 1c). Lipid contents tended to increased significantly in all groups throughout storage time, yet did not vary significantly among the groups at any time-point (Figure 1d).

Figure 1
Changes in the proximate composition of control and AEO-treated tambaqui fillets during refrigerated storage (0 = day zero prior to refrigeration, 7 and 14 = after seven and 14 days of refrigeration, respectively). AEO 0.75% = Alpinia zerumbet essential oil at 0.75%; AEO 1.5% = Alpinia zerumbet essential oil at 1.5%. Columns represent the mean (N = 5), bars the standard error of the mean. The same small letters on the bars indicate no significant difference among the treatments within each time point. The same capital letters on the bars indicate no significant differences among time points within each treatment (p < 0.05).

pH

There was a significant effect of AEO at day 7 and day 14, and on the interaction of AEO x storage time on the pH for AEO 0.75%. At day zero, all samples had similar pH. At day 7, a significant decrease in pH was observed among the treatments and control (control > AEO 0.75% > AEO 1.5%). On day 14, there was a significant difference only between the control and AEO 0.75%, which showed lower pH values (Figure 2).

Figure 2
Changes in the pH of control and AEO-treated tambaqui fillets during refrigerated storage (0 = day zero prior to refrigeration, 7 and 14 = after seven and 14 days of refrigeration, respectively). AEO 0.75% = Alpinia zerumbet essential oil at 0.75%; AEO 1.5% = Alpinia zerumbet essential oil at 1.5%. Columns represent the mean (N = 5), bars the standard error of the mean. The same small letters on the bars indicate no significant difference among the treatments within each time point. The same capital letters on the bars indicate no significant differences among time points within each treatment (p < 0.05).

TBARS

TBARS were similar among the groups at day zero, but decreased in the treatments relative to the control at days 7 and 14 (Figure 3). Throughout storage time, the control showed higher values of TBARS (25.9% and 23.1% higher than AEO 0.75% and AEO 1.5%, respectively). At day 14 the lowest values of TBARS (1.5 mg MDA/kg fillet) were determined for 0.75%.

Figure 3
Changes in the TBARS of control and AEO-treated tambaqui fillets during refrigerated storage (0 = day zero prior to refrigeration, 7 and 14 = after seven and 14 days of refrigeration, respectively). AEO 0.75% = Alpinia zerumbet essential oil at 0.75%; AEO 1.5% = Alpinia zerumbet essential oil at 1.5%. Columns represent the mean (N = 5), bars the standard error of the mean. The same small letters on the bars indicate no significant difference among the treatments within each time point. The same capital letters on the bars indicate no significant differences among time points within each treatment (p < 0.05). Missing letters indicate that all comparisons were non-significant.

TVB-N

There was an effect of the AEO and storage time on TVB-N. The TVB-N values showed a similar behavior as did the TBARS at day zero, with no significant difference between the treatments. However, at day 7, the control showed lower values, followed by AEO 0.75% and AEO 1.5% (Figure 4). At day 14, there was no significant difference among samples.

Figure 4
Changes in the TVB-N of control and AEO-treated tambaqui fillets during refrigerated storage (0 = day zero prior to refrigeration, 7 and 14 = after seven and 14 days of refrigeration, respectively). AEO 0.75% = Alpinia zerumbet essential oil at 0.75%; AEO 1.5% = Alpinia zerumbet essential oil at 1.5%. The same small letters on the bars indicate no significant difference among the treatments within each time point. The same capital letters on the bars indicate no significant differences among time points within each treatment (p < 0.05). The horizontal line represents the limit established by Brazilian legislation.

Total bacterial count

AEO and storage time significantly affected TPC (Figure 5). At day zero, there was no significant difference among the groups. At day 7 AEO 1.5% showed a significantly lower bacterial count compared to the control and AEO 0.75%. At day 14 TPC increased significantly in all groups, when the fillets became unfit for consumption according to Brazilian law (Brazil 2001).

Figure 5
Development of the total bacterial count of control and AEO-treated tambaqui fillets during refrigerated storage. AEO 0.75% = Alpinia zerumbet essential oil at 0.75%; AEO 1.5% = Alpinia zerumbet essential oil at 1.5%. Bars represent the standard deviation. The horizontal line represents the limit established by Brazilian legislation.

DISCUSSION

During cold storage, there are changes in the physicochemical properties of fish fillets, with a reduced water-holding capacity that promotes moisture decrease and nutrient loss (Aubourg 2001Aubourg, S. 2001. Damage detection in horse mackerel (Trachurus trachurus) during chilled storage.Journal of American Oil Chemistry Society ,78: 857-862.). Previous studies showed a positive effect of the essential oil of thyme (Thymus daenensis) and savory (Satureja bachtiarica) on moisture loss of trout (Oncorhynchus mykiss) fillets (Abediet al. 2016Abedi, E.; Naseri, M.; Ghanbarian, G.A.; Vazirzadeh, A. 2016. Coverage of polyethylene film with essential oils of thyme (Thymus daenensis celak)and savory (Satureja bachtiarica bunge) for lipid oxidation control in rainbow trout (Oncorhynchus mykiss) fillets during short-term storage in the refrigerator.Journal of Food Processing and Preservation, 40: 483-491.). The major components of thyme oil are p-cymene (8.4%), γ-terpinene (30.9%) and thymol (47.6%) (Borugãet al.2014Borugã, O.; Jianu, C.; Miscã, C.; Golet, I.; Horhat, F.G. 2014. Thymus vugaris essential oil: Chemical composition and antimicrobial activity.Journal of Med Life, 7: 56-60.), and the major components of savory oil are p-cymene (7.3%), y-terpinene (23.9%), thymol (44.5%), β-caryophyllene (5.3%) and borneol (4.2%) (Sefidkon and Ahmadi 2000Sefidkom, F.; Ahmadi, Sh. 2000. Essential oil ofSatureja khuzistanicaJamzad.Journal of Essential Oil Research, 12: 427-428. ). Thymol and p-cymene increase superoxide dismutase and glutathione peroxidase activities, which have protective effects that maintain the integrity of muscle cells (Hashemipour et al. 2013Hashemipour, H.; Kermanshabi, H.; Golian, A.; Veldkamp, T. 2013. Effect of thymol and carvacrol feed supplementation on performance, antioxidant enzyme activities, fatty acid composition, digestive enzyme activities, and immune response in broiler chickens.Poultry Science, 92: 2059-2069. ). A similar effect was observed in our tambaqui fillets, with a lower moisture percentage in the control group, which indicates that the p-cymene (32.7%, Castroet al.2016Castro, K.N.C.; Lima, D.F.; Vasconcelos, L.C.; Santos, R.C.; Pereira, A.L.M.; Fogaça, F.H.S.; Calvet, R.M. 2016. Chemical composition and efficacy of essential oil and ethanol extract ofAlpinia zerumbetonStaphylococcus aureus.Arquivos do Instituto Biológico, 83: 1-7.) in the AEO probably acted as a preservative, coating the fillets and keeping their water-holding capacity. The higher ash content in the fillets treated with AEO indicates a preservation effect on mineral content. Since AEO prevented water loss during storage, it also reduced the cellular rupture that promotes protein loss. Crude protein represents the main nutritional value of seafood and is the main factor responsible for its texture and water-holding capacity (Sikorski and Kolakowska 2016Sikorski, Z.E.; Kolakowska, A. 2016. Seafood Quality Issues. In: Daczkowska-Kozon, E.G.; Pan, B.S. (Eds.). Environmental Effects on Seafood Availability, Safety, and Quality. CRC Press, Boca Raton, p.19-36.).

Fat content is one of the most important indices for seafood quality determination (Abediet al. 2016Abedi, E.; Naseri, M.; Ghanbarian, G.A.; Vazirzadeh, A. 2016. Coverage of polyethylene film with essential oils of thyme (Thymus daenensis celak)and savory (Satureja bachtiarica bunge) for lipid oxidation control in rainbow trout (Oncorhynchus mykiss) fillets during short-term storage in the refrigerator.Journal of Food Processing and Preservation, 40: 483-491.). A decrease in lipid content could be related to lipid oxidation and enzymatic degradation of the muscle phospholipids (Hendrikset al. 2006Hendriks, W.H.; Cottam, Y.H.; Thomas, D.V. 2006. The effect of storage on the nutritional quality of meat and bone meal.Animal Feed Science and Technology, 127: 151-160. ). The increase in lipid content observed in this study may be related to the decrease observed in crude protein content, since centesimal composition involves a relation among moisture, fat, crude protein and ash contents (Sikorski and Kolakowska 2016Sikorski, Z.E.; Kolakowska, A. 2016. Seafood Quality Issues. In: Daczkowska-Kozon, E.G.; Pan, B.S. (Eds.). Environmental Effects on Seafood Availability, Safety, and Quality. CRC Press, Boca Raton, p.19-36.) where the sum of components must be approximately 100%.

The effect of AEO on the relative proportions of the components of centesimal composition may have been related to the pH changes during storage time. In general, an increase in pH may be due to the production of volatile bases or microbial enzyme action (Li et al. 2012Li, T.T.; Hu, W.Z.; Li, J.R.; Zhang, X.G.; Zhu, J.L.; Li, X.P. 2012. Coating effects of tea polyphenol and rosemary extract combined with chitosan on the storage quality of large yellow croaker (Pseudosciaena crocea).Food Control, 25: 101-106.). The decrease in pH observed in our treatments might have resulted from the antioxidant and antibacterial characteristics of the AEO, which inhibits the activity of the endogenous proteases and contributes to the extension of the fish sample preservation (Fanet al. 2008Fan, W.; Chi, Y.; Zhang, S. 2008. The use of a tea polyphenol dip to extend the shelf life of silver carp (Hypophthalmichthys molitrix) during storage in ice.Food Chemistry, 108: 148-153.). Similar results were observed for silver carp (Hypophthalmicthys molitrix) fillets coated withPimpinella affinisessential oil (PAO), although the fillets were coated with twice the concentration of essential oil (1.5% v/v) (Ariaiiet al. 2015Ariaii, P.; Tavakolipour, R.M.; Rad, A.H.E.; Bahram, S. 2015. Effect of methylcellulose coating enriched withPimpinella affinisoil on the quality of silver carp fillet during refrigerator storage condition.Journal of Food Processing and Preservation, 39: 1647-1655.).

Essential oils are lipophilic, thus they can easily enter cells and disrupt and/or permeabilize the cell membrane (Perricone et al. 2015Perricone, M.; Arace, R.; Corbo, M.R.; Sinigaglia, M.; Bevilacqua, A. 2015. Bioactivity of essential oils: a review on their interaction with food components. Frontiers in Microbiology, 6: 1-7.), which can cause depolarization of mitochondrial membranes, influence the Ca2+ channels, and reduce the pH gradient (Bakkali et al. 2008Bakkali, F.; Averbeck, S.; Averbeck, D.; Idaomar, M. 2008. Biological effects of essential oils - a review. Food Chemistry and Toxicology, 46: 446-475.). In some cases, EOs can cause the coagulation of the cytoplasm as well as damage to lipids and proteins (Burt 2004Burt, S. 2004. Essential Oils: their antibacterial properties and potential applications in foods - a review.International Journal of Food Microbiology ,94: 233-253. ). This may explain the differences observed between control and treatments in lipid and crude protein contents and pH.

TBARS are the predominant products of secondary lipid oxidation and are considered good chemical indicators of quality assurance and estimatiors of the extent of lipid oxidation during chilled storage (Jeonet al. 2002Jeon, Y.I.; Kamil, J.Y.V.A.; Shahidi, F. 2002. Chitosan as an edible invisible film for quality preservation of herring and Atlantic cod.Journal of Agricultural and Food Chemistry, 20: 5167-5178.; Goulas and Kontominas 2007Goulas, A.E.; Kontominas, M.G. 2007. Combined effect of light salting, modified atmosphere packaging and oregano essential oil on the shelf-life of sea bream (Sparus aurata): biochemical and sensory attributes. Food Chemistry, 100: 287-296. ). Therefore, the lower TBARS level in AEO 0.75% indicates that this treatment was the most efficient in lipid oxidation preservation of the tambaqui fillets. The use of an EO to protect food against lipid oxidation has been reported elsewhere (e.g., Leeet al. 2005Lee, S.J.; Umano, K.; Shibamoto, M.; Lee, K.G. 2005. Identification of volatile components in basil (Ocimum basilicumL.) and thyme leaves (Thymus vulgarisL.) and their antioxidant properties.Food Chemistry , 91: 131-137. ; Luet al. 2010Lu, F.; Ding, Y.; Ye, X.; Liu, D. 2010. Cinnamon and nisin in alginate-calcium coating maintain quality of fresh northern snakehead fish fillets.LWT - Food Science and Technology ,43: 1331-1335.; Ojaghet al. 2010Ojagh, S.M.; Rezaei, M.; Razavi, S.H.; Hosseini, S.M.H. 2010. Effect of chitosan coatings enriched with cinnamon oil on the quality of refrigerated rainbow trout.Food Chemistry, 120: 193-198. ). In silver carp, EO resulted in a reduction of 18% in TBARS (Ariaiiet al. 2015Ariaii, P.; Tavakolipour, R.M.; Rad, A.H.E.; Bahram, S. 2015. Effect of methylcellulose coating enriched withPimpinella affinisoil on the quality of silver carp fillet during refrigerator storage condition.Journal of Food Processing and Preservation, 39: 1647-1655.). In our study, the reduction in TBARS in AEO 0.75% was approximately 51%, suggesting that AEO has a significan potential to be used as an antioxidant in fish.

In addition to lipid oxidation, fish decomposition implies progressive proteolysis by the action of microorganisms and, to a lesser degree, by autolytic enzymes (Howgate 2010Howgate, P. 2010. A critical review of total volatile bases and trimethylamine as indices of freshness of fish. Part 2: formation of bases, and application in quality assurance.Electronic Journal of Environmental, Agricultural and Food Chemistry, 9: 58-88.;Ocaño-Higuera et al. 2011Ocaño-Higueraa, V.M.; Maeda-Martínezb, A.N.; Marquez-Ríosc, E.; Canizales-Rodrígueza, D.F.; Castillo-Yañeza, F.J.; Ruíz-Bustosa, E. 2011. Freshness assessment of ray fish stored in ice by biochemical, chemical and physical methods.Food Chemistry, 125: 49-54. ), such as TVB-N. Therefore TVB-N has traditionally been used as a quality indicator in refrigerated aquatic products and includes trimethylamine, dimethylamine, ammonia and other compounds, which come from the degradation of proteins and nonprotein nitrogenous compounds by endogenous enzymes (Li et al. 2013Li, T.; Li, J.; Hu, W.; Li, X. 2013. Quality enhancement in refrigerated red drum (Sciaenops ocellatus) fillets using chitosan coatings containing natural preservatives.Food Chemistry, 138: 821-826.). The Brazilian legislation sets a maximum acceptable limit for fish of 30 mg TVB-N/100 g of product (Brasil 2001Brasil. 2001. Regulamento da Inspeção 69 Industrial e Sanitária de Produtos de Origem Animal (RIISPOA) . MAPA, Brasília, 59p.). According to the quality index, a TVB-N value of less than 25 mg is very good, 30 mg is good, 35 mg is marketable, and more than 35 mg is considered inconsumable (Varlıket al. 2000Varlik, C.; Erkan, N.; Metin, S.; Baygar, T.; Ozden, O. 2000. Determination of the shelf life of marinated fish balls.Turkistan Journal of Veterinary Animal Science, 24: 593-597.; Cakli 2007Cakli, S.; Kilinc, B.; Cadun, A.; Dincer, T.; Tolasa, S. 2007. Quality differences of whole ungutted sea bream (S. aurata) and sea bass (D. labrax) while stored in ice.Food Chemistry, 18: 391 -397.; Dogan and Izci 2016Dogan, G.; Izci, L. 2016. Effects on quality properties of smoked rainbow trout (Oncorhynchus mykiss) fillets of chitosan films enriched with essential oils.Journal of Food Processing and Preservation, 41: 1-12.).

In rainbow trout treated with EO levels of approximately 0.2% to 1%, the treatments using rosemary or thyme EO at 1% resulted in lower TVB-N values relative to the control (Erkan 2012Erkan, N. 2012. The effect of thyme and garlic oil on the preservation of vacuum-packaged hot smoked rainbow trout (Oncorhynchus mykiss).Food Bioprocess Technology, 5: 1246-1254.; Yildiz 2015Yildiz, P.O. 2015. Effect of essential oils and packaging on hot smoked rainbow trout during storage.Journal of Food Processing and Preservation ,39: 806-815.). The inhibition of TVB-N production in fish may be attributed to the effects of the phenolic and organosulfur constituents of EO (Burt 2004Burt, S. 2004. Essential Oils: their antibacterial properties and potential applications in foods - a review.International Journal of Food Microbiology ,94: 233-253. ), that react with the sulfhydryl and amino groups of proteins. In our study, the high TVB-N for AEO 1.5% at day 7 was likely related to the significant antibacterial activity detected in this treatment at the same time-point, and points to a capacity of AEO to reduce bacterial growth, which causes the oxidative deamination of nonprotein nitrogen compounds (Bankset al. 1980Banks, H.; Nickelson, R.; Finne, G. 1980. Shelf life studies on carbon dioxide packaged finfish from Gulf of Mexico.Journal of Food Science ,45: 157-162.).

Fish and seafood in general are highly perishable food products, mainly due to microbiological growth and lipid oxidation, which are known to be the principal causes of the quality deterioration in these products (Hassoun and Çoban 2017Hassoun, A.; Çoban, O.E. 2017. Essential oils for antimicrobial and antioxidant applications in fish and other seafood products.Trends in Food Science & Technology, 68: 26-36. ). The antimicrobial activity of some common EOs in fish, either alone or in combination with other preservative systems, depends on the oil type and its concentration of antimicrobial compounds, in addition to the amount used in bioactive packaging. For example,Lippia multifloraEO was only effective in inhibiting microbial development in smoked mackerel (Scomber scombrus) at concentrations greater than 0.5% (Cyrilleet al. 2017Cyrille, G.K.R.; Adjehi, D.; Yaya, S.; Désiré, K.N.; Benjamin, K.A.B.; Marcellin, D. 2017. Antimicrobial and preservative activities of Lippia multiflora essential oil on smoked mackerel (Scomber scombrus) fish.Archives of Clinical Microbiology, 8: 1-5.). Rosemary (Rosmarinus officinalis), cinnamon (Cinnamomum verum), fennel (Foeniculum vulgare), and cardamom (Elettaria cardamomum) EOs also reduced the initial total bacterial count, psychrophilic bacteria, and total mold and yeast as well as prolonged the shelf life of fillets of carp (Cyprinus carpio) during cold storage at 4 °C (Hasan et al. 2017Hasan, M.; Abdeldaiem, M.; Gamal, H.; Ali, M.; Ramadan, M.F. 2017. Impact of different essential oils on the characteristics of refrigerated carp (Cyprinus carpio) fish fingers.Journal of Food Measurement and Characterization, 11: 1412-1420. ). These oils have common components (cineole, α-pinene, α-terpineol, sabinene and limonene) with proven antimicrobial effects (Abdelwahabet al. 2017Abdelwahab, S.I.; Mariod, A.A.; Taha, M.M.E.; Zaman, F.Q.; Abdelmageed, A.H.A.; Khamis, S.; Awang, K. 2017. Chemical composition and antioxidant properties of the essential oil ofCinnamomum altissimumKosterm. (Lauraceae).Arabian Journal of Chemistry, 10: 131-135.;Chegini and Abbasipour 2017Chegini, S.G.; Abbasipour, H. 2017. Chemical composition and insecticidal effects of the essential oil of cardamom,Elettaria cardamomumon the tomato leaf miner,Tuta absoluta.Toxin Reviews, 36: 12-17.;Takayama et al. 2016Takayama, C.; Meirade-Faria, F.; Almeida, A.C.A.; Dunder, R.J.; Manzo, L.P.; Rabelo Socca, E.A.; Souza-Brito, A.R.M.; Ferreira, A.L. 2016. Chemical composition ofRosmarinus officinalis essential oil and antioxidant action against gastric damage induced by adsolute ethanol in the rat. Asian Pacific Journal of Tropical Biomedicine, 6: 677-681. ; Owolabi et al. 2009Owolabi, M.S.; Ogundajo, A.; Lajide, L.; Oladimeji, M.O.; Setzer, W.N.; Palazzo, M.C. 2009. Chemical composition and antibacterial activity of the essential oil ofLippia multifloraMoldenke from Nigeria.Records of Natural Products ,3: 170-177.;García-Jiménezet al. 2000Garcia-Jiménez, N.; Péres-Alonso, M.J.; Velasco-Negueruela, A. 2000. Chemical composition of fennel oil, Foeniculum vulgare Miller, from Spain.Journal of Essential Oil Research, 12: 159-162.). Our results for tambaqui are probably related to the presence of 1.8-cineole (24.1%) and 4-terpineol (20.2%) as major components in AEO, which likely mediated the lower bacterial count at day 7 in the treatment with highest EO level. At day 14, all treatments had CFU/g values above the maximum count for fish of 107CFU g-1 recommended by FAO (Huss 1997Huss, H.H. 1997.Garantia da qualidade dos productos da pesca. Food and Agricultural Organization of the United Nations, Rome, 30p.).

Considering the different groups of chemical compounds present in EOs, it is very likely that the antibacterial and antioxidative activity of EOs is attributable to mechanisms targetting different cell structures and processes, such as the cytoplasmic membrane, perturbations on the proton-proton force, electron flow, and transport and coagulation of cell contents (Burt 2004Burt, S. 2004. Essential Oils: their antibacterial properties and potential applications in foods - a review.International Journal of Food Microbiology ,94: 233-253. ). This diffuse action mode could explain why the lower AEO level (0.75%) was more efficient in preventing lipid oxidation and the higher AEO level (1.5%) was more efficient in preventing bacterial growth, while both levels preserved the nutritional value of the tambaqui fillets in comparison to the control.

CONCLUSIONS

Treatment with 1.5% Alpinia zerumbet essential oil (AEO) reduced the bacterial growth in tambaqui fillets refrigerated at 10 oC and prolonged the shelf life of the fillets for up to seven days. AEO 1.5% is recommended as an antimicrobial agent in tambaqui fillets during short-term refrigerator storage. Lower levels of AEO (0.75%) presented antioxidant effects.

ACKNOWLEDGMENTS

The authors are thankful to CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico) and CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior) (grant 421/2010) for financial support; CODEVASF (Companhia de Desenvolvimento dos Vales dos Rios São Francisco e Parnaíba) for the use of their equipment; fish farmers for donating the fish samples and Vegeflora for donation of the Alpinia essential oil.

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  • CITE AS:

    Sousa, A.K.A. de; Fogaça, F.H. dos S.; Araujo, T.D.S.; Castro, K.N. de C.; Pereira, A.M.L.; Canuto, K.M.; Brito, E.S. de; Mai, M.G.; Magalhães, J.A. de. 2019. Effect of Alpinia zerumbet essential oil on the shelf life of tambaqui fillets during short-term refrigerator storage. Acta Amazonica 49: 152-161.
  • ERRATA

    On page 152: Where it reads: “Thais Danielly Santos ARAUJO”: Ana Karolinne Anastacio de SOUSA1, Fabíola Helena dos Santos FOGAÇA2*, Thais Danielly Santos ARAUJO1, Karina Neoob de Carvalho CASTRO3, Alitiene Moura Lemos PEREIRA4, Kirley Marques CANUTO5, Edy Sousa de BRITO5, Mônica Giacometti MAI4, João Avelar de MAGALHÃES4 Please read as: “Thais Danyelle Santos ARAUJO”: Ana Karolinne Anastacio de SOUSA1, Fabíola Helena dos Santos FOGAÇA2*, Thais Danyelle Santos ARAUJO1, Karina Neoob de Carvalho CASTRO3, Alitiene Moura Lemos PEREIRA4, Kirley Marques CANUTO5, Edy Sousa de BRITO5, Mônica Giacometti MAI4, João Avelar de MAGALHÃES4

Edited by

ASSOCIATE EDITOR:

João Vicente Souza

Publication Dates

  • Publication in this collection
    06 May 2019
  • Date of issue
    Apr-Jun 2019

History

  • Received
    02 Apr 2018
  • Accepted
    05 Dec 2018
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