Food Research International 44 (2011) 12241230

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Food Research International

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Combined effect of active coating and MAP to prolong the shelf life of minimally processed kiwifruit (Actinidia deliciosa cv. Hayward)
Marianna Mastromatteo b, Marcella Mastromatteo a, Amalia Conte a,b, Matteo Alessandro Del Nobile a,b,
a b

Istituto per la Ricerca e le Applicazioni Biotecnologiche per la Sicurezza e la Valorizzazione, dei Prodotti Tipici e di Qualit, BIOAGROMED, Via Napoli, 52, 71100 Foggia, Italy Department of Food Science, University of Foggia, Via Napoli, 25, 71100 Foggia, Italy

a r t i c l e

i n f o

a b s t r a c t
In this work different strategies aimed to prolong the shelf life of minimally processed kiwifruits are presented. First, the effectiveness of several treatments in delaying the quality loss of the investigated produce packaged under passive MAP was addressed; afterward, the treatments that have shown the best performances were used to assess the effectiveness of active MAP in prolonging the packaged produce shelf life. Different treatments such as coating with sodium alginate in combination with dipping into an hydro-alcoholic solution (Coat-dipp-EtOH), dipping into an hydro-alcoholic solution (Dipp-EtOH) and coating with sodium alginate containing grape fruit seed extract solution (Coat-GFSE) were investigated. The untreated samples were used as control. Headspace gas concentrations, pH, mass loss, sensory quality and viable cell load of main spoilage microorganisms were monitored in both the experimental steps. Results suggested that the best performances under passive MAP were recorded with the coating treatments, justifying the choice of this treatment in the second step. In fact, the coatings were more effective in delaying dehydration and slowing down respiratory activity of minimally processed kiwifruits both in passive and active MAP. The combination of active compounds with alginate-based coating delayed the microbial growth whereas the sole dipping treatment was inefcient. In particular, a viability loss of the mesophilic and psychrotrophic bacteria of about 2 log cycle for the coated samples with respect to control and dipped samples was found. However, as the microbial load was always found below the threshold value imposed by law, the sensorial acceptability limit of the packaged fresh-cut produce coincided with its shelf life. Alginate-based coating reduced respiratory activity, as well as sensory decay, increasing the sensorial acceptability limit of the samples packaged under passive MAP up to 12 days with respect to the control (8 days). For the samples packaged under active MAP, the coating treatments reduced the excessive dehydration of the produce due to the MAP conditions. In fact, when the active MAP was used alone a very short shelf life of the uncoated samples occurred (2.7 days). Whereas, the combined use of active MAP and coating treatments prolonged the produce shelf life up to 13 days. 2010 Elsevier Ltd. All rights reserved.

Article history: Received 5 July 2010 Accepted 1 November 2010 Keywords: Kiwifruit Minimally processing Antimicrobial compounds MAP

1. Introduction Minimally processed (MP) fruits are products that maintain their attributes and quality similar to those of fresh products (Alzamora, Lopez-Malo, & Tapia, 2000). However, minimal processing alters the integrity of fruit and induces wounding stress and spoilage. Physical damage or wounding caused by slicing, peeling, and/or other mechanical injuries in minimally processed fruits results in increased respiration rates and ethylene production within minutes (Abe & Watada, 1991). Increases occur in biochemical reactions related to

Corresponding author. Department of Food Science, University of Foggia, Via, Napoli, 25, 71100, Foggia, Italy. Tel./fax: + 39 881 589 242. E-mail address: ma.delnobile@unifg.it (M.A. Del Nobile). 0963-9969/$ see front matter 2010 Elsevier Ltd. All rights reserved. doi:10.1016/j.foodres.2010.11.002

changes in color, avor, texture, nutritional quality and susceptibility to dehydration. These responses occur in disrupted tissues where cellular decompartmentation leads to intermixing of enzymes and substrates, as well as the release of acid and hydrolyzing enzymes (Watada, Abe, & Yamuchi, 1990). During storage such products have a very limited shelf life (Brackett, 1994; O'Connor-Shaw, Roberts, Ford, & Nottingham, 1994). Kiwifruit (Actinidia chinensis, Planch and Actinidia deliciosa, A. Chev.) is a native of the mountains of southern China. Commercial development of the fruit took place in New Zealand, with a number of cultivars being selected from seeds of A. deliciosa. Long storage life was regarded as essential for the development of an export industry in New Zealand, so the cultivar Hayward, which has a storage life of at least six months at 0 C, was adopted for cultivation among others (Scott, Spraggon, & McBride, 1986). The rmness of a kiwifruit strongly inuences its sensory qualities at the moment of consumption, including perceived aroma intensity, sweetness, acidity and

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ripeness (Stec, Hodgson, Macrae, & Triggs, 1989). Enzymatic activities, softening and ripening of kiwifruit are promoted by ethylene (Arpaia, Mitchell, Kader, & Mayer, 1985; Wegrzyn & MacRae, 1992). The loss of cellular compartmentation, due to peeling and cutting, causes mixing of metabolites of the ethylene generating system stimulating ethylene production (Mazliak, 1983; Watada et al., 1990). Therefore, it is difcult to maintain the quality of minimally processed kiwifruit once they have been cut (Chien & Buta, 2003). In order to extend the shelf life of minimally processed products a number of technologies are available. Temperature and headspace atmosphere are two important factors to prolong their shelf life. Modied atmosphere (MAP) can be interpreted as a dynamic system with two gas uxes, the respiration rate of the fresh product and the gas exchange through the packaging lm (Van de Velde & Kiekens, 2002). In general, gas compositions inside a MAP package are low in O2 and high in CO2, depending primarily on temperature, product weight and respiration rate, O2 and CO2 lm transmission rates and package total surface area. MAP and low temperature storage are usually not sufcient to extend the shelf life of pre-cut produce as the excessive physiological stress and increased susceptibility towards microbial spoilage caused by processing operations like cutting and slicing, reduce the shelf life signicantly. Edible coatings, containing antimicrobial agents, are gaining importance as potential treatments to reduce the deleterious effects imposed by minimal processing on fresh-cut fruit (Conte, Scrocco, Brescia, & Del Nobile, 2009; Del Nobile, Conte, Scrocco, Laverse, et al., 2009). The use of edible coatings for a wide range of food products, including fresh and minimally processed fruit, has received increased interest, because coatings can serve as carrier for a wide range of food additives, including anti-browning agents, colorants, avors, nutrients, spices and various antimicrobials that can extend product shelf life and reduce the risk of pathogen growth on food surface (Cagri, Uspunol, & Ryser, 2004; Pranoto, Salokhe, & Rakshit, 2005). Incorporating antimicrobial compounds into edible lms or coatings provides a novel way to improve the safety and shelf life of ready-toeat foods (Cagri et al., 2004). Some of the more commonly used antimicrobials include benzoic acid, sorbic acid, lysozyme, bacteriocins (nisin and pediocins) and plant-derived secondary metabolites, such as essential oils and phytoalexins. Essential oils are regarded as alternatives to chemical preservatives, and their use in foods meets the demands of consumers for minimally processed natural products, as reviewed by Burt (2004). Essential oils can extend shelf life of unprocessed or processed foods by reducing microbial growth rate or viability (Beuchat & Golden, 1989). These compounds can be added to edible lms and coatings to modify avor, aroma, and odor, as well as to introduce antimicrobial properties (Cagri et al., 2004). However, there are a few studies about the effectiveness of the incorporation of those compounds into edible coatings applied to fruit for ensuring quality and safety has been published (Eswaranandam, Hettiarachchy, & Meullenet, 2006; Raybaudi-Massilia, Mosqueda-Melgar, & Martn-Belloso, 2008; Rojas-Gra et al., 2007). Ethanol is the most common microbiological and medical disinfectant. Recently, many studies dealing with table grape preservation techniques have found that the use of ethanol, as a common food additive with antimicrobial activity, suppressed microbial growth and prevented berry decay (Del Nobile, Conte, Scrocco, Brescia, et al., 2009; Del Nobile et al., 2008; Pinto, Lichter, Danshin, & Sela, 2006). Treatment of table grapes with ethanol has the potential not only to kill spoilage microorganisms, but also to inhibit hazardous food borne pathogens. The objective of this work was to evaluate the effectiveness of the natural essential oil, such as Grape Fruit Seed Extract (GFSE) incorporated into an edible coating, and ethanol on the shelf life of packed minimally processed kiwifruit under both passive and active MAP.

2. Materials and methods 2.1. Sample preparation Fresh italian kiwifruits (A. deliciosa cv. Hayward) kindly provided by a local farm (Ermes, Noicattaro, Italy), were transported within 2 h to the laboratory under refrigerated conditions (4 C) and immediately processed. Kiwifruit were washed with tap water, treated for 5 min with chlorinated water (20,000 ppm) and peeled using a manual vegetable cutter. The minimally processed kiwifruit were subjected to the following treatments: a) Coat-dipp-EtOH: kiwifruits were dipped for 5 min into a hydroalcoholic solution (30% v/v in ethanol) and then air-dried for 10 min (Del Nobile, Conte, Scrocco, Brescia, et al., 2009; Del Nobile et al., 2008). Afterward kiwifruits were dipped into a sodium alginate (4% w/v) water solution. The sodium alginate solution was prepared by dissolving sodium alginic acid in distilled water at 50 C for 2 h. The coated samples were immersed into a 5% (w/v) calcium chloride (CaCl2) solution for 1 min to promote the alginate gel forming process and then into a hydro-alcoholic solution (30% v/v in ethanol) for 5 min (Conte et al., 2009; Del Nobile, Conte, Scrocco, Laverse, et al., 2009). Sodium alginic acid and CaCl2 were provided by Sigma-Aldrich Co. Inc. (USA), Ethanol by Baker (Holland). b) Dipp-EtOH: kiwifruit were dipped for 5 min into a hydro-alcoholic solution (30% v/v in ethanol) and then air-dried for 10 min. c) Coat-GFSE: kiwifruit were dipped into a sodium alginate (4%, w/v) solution containing grape fruit seed extract water solution (GFSE, 5000 ppm) (Biocitro, Probena s.l., Zaragoza, Spain) and then immersed into a 5% (w/v) CaCl2 solution for 1 min (Cagri et al., 2004; Rojas-Gra et al., 2007). After dipping or coating treatment, two kiwifruits per package (~200 g per pack) were packaged in an Oriented Polypropylene lm (OPP thickness 20 m) bags with a surface area of 396 cm2. Samples simply peeled were also packaged and used as control (Ctrl). The samples were sealed by means of a S100-Tecnovac equipment (Tecnovac, San Paolo D'Argon, Bergamo, Italy) with the following atmospheres: passive and active modied atmosphere packaging (MAP: 10% O2, 10% CO2). The main parameters of the Tecnovac equipment were: 99.0% gas, 99.0% vacuum, 2.3 min sealer. All samples were stored at 4 C for about 1521 days. Three samples for each treatment were randomly selected for analysis. 2.2. Headspace gas composition The changes in headspace O2 and CO2 concentration of packaged minimally processed kiwifruit were measured using a PBI Dansensor O2/CO2 analyzer (Checkmate 9900, Denmark). The volume taken from the package headspace for gas analysis was about 10 cm3. The package headspace volume was determined by the difference between the total volume of the packages and the volume of the sample. The total volume was measured by dipping the packages containing the fruit into a graduated water container and by observing the increase in the water level. Similarly, the volume of the samples was calculated by immersion of the clusters in a graduated cylinder with water, and by measuring the increase in water level. To avoid modications in the headspace gas composition due to gas sampling, each package was used only for a single measurement of the headspace gas composition. Three bags were used for each measurement. 2.3. Microbiological analysis and pH evaluation For microbiological analysis, about 25 g of sample was aseptically removed from each package, placed in a stomacher bag, diluted with 0.9% NaCl solution and homogenized with a Stomacher LAB Blender

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400 (Pbi International, Milan, Italy). Decimal dilutions were carried out using the same diluent. Mesophiles and psychrotrophs were determined on Plate Count Agar (PCA) with incubation at 30 C for 2448 h and 7 C for 10 days, respectively. For Enterobacteriaceae, Violet Red Bile Glucose Agar (VRBGA) was used and plates were incubated at 37 C for 1824 h. Lactic acid bacteria (LAB) were plated on MRS Agar and incubated anaerobically at 30 C for 24 days. Yeasts and moulds were determined on Sabouraud Dextrose Agar (SDA), supplemented with chloramphenycol (0.1 g l 1) (C. Erba, Milan, Italy) with incubation at 25 C for 48 h and 5 days, respectively. The pH was evaluated on the homogenized kiwifruit by a pH meter (Crison Instruments, Barcelona, Spain). The pH measurement was carried out twice, on two different batches. 2.4. Mass loss The mass loss percent was determined according to the following expression: %MLt = M0 M t d 100 M0 1

color, odor, rmness and overall quality by the panelists was stopped as soon as visible moulds were detected. 2.6. Permeation tests The Water Vapor Transmission Rate (WVTR) of the selected lm was determined by means of a water vapor permeability analyser (Lyssy, Model 80-5000, Dansensor, Ringsted, Denmark). Two samples of the lm with a surface area of 5 cm2 were tested at 23 C and 85% of relative humidity (RH). A ow rate of 100 ml/min of nitrogen was used. The Oxygen Transmission Rate (OTR) was determined by means of an Ox-Tran (Mocon, Model 2/20). Two samples of the lm with a surface area of 5 cm2 were tested at 10, 16 and 23 C and 0% RH at the upstream and the downstream side of the sample. The Carbon Dioxide Transmission Rate (CDTR) was determined by means of a Permatran (Mocon, Model C 4/41). Two samples of the lm with a surface area of 5 cm2 were tested at 10, 16 and 23 C and 0% RH at the upstream and the downstream side of the sample. 2.7. Statistical analysis The values of the parameters relative to microbiological analysis and sensory quality were compared by one-way analysis of variance (ANOVA). A Duncan's multiple range test with the option of homogeneous groups (P b 0.05) was used to determine signicance between treatments. To this aim, STATISTICA 7.1 for Windows (Stat-Soft, Inc, Tulsa, OK, USA) was used. 3. Results and discussion 3.1. Film barrier properties The measured values of OTR and CDTR for the tested lm are listed in Table 1. The gas permeation tests were conducted at temperatures different to 5 C, that was the storage temperature of kiwifruit, as it is not possible with the actual equipments to make permeation tests at temperatures lower than 10 C. In order to determine the gas barrier properties of the selected lm in the real working conditions, the Arrhenius equation was used:   E P = P0 d exp a : RT 3

where: %ML(t) is the mass loss percent at time t, M0 is the initial sample mass and M(t) is the sample mass at time t. The sample mass was determined by a digital precision balance (0.1 g) (Gibertini Europe, Italy). At each sampling time, the mass was measured twice, on two different batches. 2.5. Sensory evaluation A panel consisting of seven untrained evaluators using a ve point hedonic scale (5: excellent; 4: good; 3: acceptable limit of marketability; 2: poor and 1: extremely poor) (Larmond, 1977; Mastromatteo, Danza, Conte, Muratore, & Del Nobile, 2010; Mastromatteo, Lucera, Sinigaglia, & Corbo, 2009; Nowak, Von Muefing, Grotheer, Klein, & Watkinson, 2007) was used in this study to quantitatively determine the overall quality according to the procedure reported by Gimenez et al. (2003). Panelists were asked to base their decision on the sample overall quality only taking into account its color, odor, and rmness. Therefore, the samples overall quality has to be considered as an average of the above-mentioned sensorial attribute values (i.e., color, odor, and rmness) as weighted by the panelist. Moreover, panelists were also asked to score color, odor, and rmness of each sample. A score equal to 3 was used as the threshold for produce acceptability. During the test sessions, two samples for each treatment were randomly presented for sensory analysis. To determine the sensory acceptability limit (SAL) of the investigated minimally processed produce, a rst order kinetic equation (Conte et al., 2009) was tted to the experimental data: SAt = SAmin SA0 d expkd SAL 1 expkd SAL   SA SA0 d expkd SAL + SA0 min d expkd t 1 expkd SAL 2

where: SA(t) is the kiwifruit sensory attribute at time t, k is the kinetic constant, SA0 is the initial value of the kiwifruit sensory attribute, SAmin is the limit for minimally processed produce acceptability, SAL is the sensory acceptability limit (i.e., the time at which SA(t) is equal to SAmin), and t is the storage time. Panelists were also asked to search for visual moulds, thus allowing determining the day between the latest storage time at which moulds were not visible and earliest storage time at which moulds were visible, hereinafter referred to as VMT (Visual Moulds Time) (Costa, Lucera, Mastromatteo, Conte, & Del Nobile, 2010). Due to safety reasons, the evaluation of minimally processed kiwifruit

The tests were conducted at 23 C, 16 C and 10 C and by using the above equation the permeability values at 5 C were obtained by extrapolation. As expected the permeability increased as the temperature of the permeation test increased. A similar trend was also found for CO2 permeability. The extrapolated permeability values (molcm/cm2atmh) for both O2 and CO2 were then converted to the respective values of OTR and CDTR (cc/(m2day)). As far as the WVTR is concerned, the above-mentioned approach was not adopted due to the fact that the apparatus devoted to measure water vapor transport properties cannot make measures at more than two temperatures, thus not allowing any extrapolation. For this reason, the permeation tests to water vapor were carried out only

Table 1 Values of Oxygen Transmission Rate (OTR) and Carbon Dioxide Transmission (CDTR) of Oriented Polypropylene lm (OPP thickness 20 m). Temperature (C) 23 16 10 5 OTR (cm3m 2day 1) 2481.74 241.54 1674.61 125.87 1364.13 300.88 1014.72 9.5 CDTR (cm3m 2day 1) 6982.53 574.15 4926.5 341.52 3552.0 268.55 2700 176.7

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at one temperature and the values recorded should be used to sole comparative purposes. WVTR value equal to 0.6 g m 2 day 1 was obtained for Oriented Polypropylene lm (OPP thickness 20 m). 3.2. Step I: packaging under passive MAP 3.2.1. Headspace gas composition Fruit and vegetables consume oxygen and produce carbon dioxide while packed, giving rise to a modication of the headspace gas composition (Jayas & Jeyamkondan, 2002). The respiration of the product and the gas permeability of the lm inuence the change in gaseous composition of the environment surrounding the product. Fig. 1a, b shows the evolution over storage of O2 and CO2 concentrations in the OPP20 bag headspace for Coat-dipp-EtOH, Dipp-EtOH, Coat-GFSE and Ctrl samples packaged in passive MAP. As expected, a decrease in the headspace oxygen concentration, as well as an increase in the headspace carbon dioxide concentration was observed. In particular, a faster decrease in the headspace oxygen concentration for Dipp-EtOH (5.55) and Ctrl (6.64) samples compared to coated samples (about 10.011.0) was observed (Fig. 1a). These data suggest that coating treatment can reduce the respiration activity of minimally processed kiwifruit. Similar results were also reported in the literature for minimally processed lampascioni and fresh-cut apples. In fact, it was found that edible coating based on sodium alginate, whey proteins concentrate, carrageenan or polysaccharides/ lipid formulation, in combination with anti-browning agents, reduced the respiration rate of minimally processed lampascioni and sliced apples (Conte et al., 2009; Lee, Park, Lee, & Choi, 2003; Wong, Tillin, Hudson, & Pavlath, 1994). In addition, Wong et al. (1994) indicated that the diffusion of the headspace oxygen to the tissue apple pieces was slowed down by the oxygen resistance of the coating. Moreover, a higher increase of carbon dioxide concentration for coated samples

Table 2a Evolution of mesophilic and psychrotrophic bacteria (log cfu/g) for Step I kiwifruit packed under passive MAP. Samples Mesophiles log cfu/ginitial Coat-dipp-EtOH Dipp-EtOH Coat-GFSE Control 2.00 0.00 2.40 0.35a,b 2.00 0.00a 2.48 0.00a,b
a

Psychrotrophs log cfu/g8 days 2.10 0.17 4.13 0.84b 2.40 0.35a 4.20 0.25b
a

log cfu/ginitial 2.00 0.00 2.00 0.00a 2.00 0.00a 2.00 0.00a
a

log cfu/g8 days 2.33 0.58a 3.76 0.24b 2.48 0.83a 4.21 0.53b

Mean values standard deviation. ad Means in the same column followed by different superscript upper cases are signicantly different (P b 0.05).

(10.0) with respect to the uncoated samples (about 7.08.0) was observed (Fig. 1b). Olivas and Barbosa-Cnovas (2005) indicated that coatings with selective permeability to gases are capable of decreasing the interchange of O2 and CO2 between coated fruit and the environment, slowing down the metabolism by decreasing internal O2 concentration and increasing CO2 concentration. 3.2.2. Microbiological stability and pH Table 2a reports the cell load of mesophilic and psychrotrophic bacteria at 0 and 8 days of sampling for treated and untreated samples packed under passive MAP. The initial cell load for mesophiles and psychrotrophs was the same for all samples and did not exceed 2.5 log cfu/g. It is interesting to note that at 8 days of sampling a viability loss of the mesophilic bacteria of about 2 log cycle for the coated samples (Coat-dipp-EtOH and Coat-GFSE) with respect to Dipp-EtOH and Ctrl samples was found. The Dipp-EtOH samples showed the same behavior of the untreated samples. Probably, the coating treatment improved the ethanol efcacy as it avoided its evaporation. For the psychrotrophic bacteria, signicant differences between the cell load of the coated samples and uncoated samples were also observed (P b 0.05). Many studies have found that ethanol pre-treatment was the most effective as it successfully reduced the cell load of the main spoilage microorganisms of minimally packed produce (Del Nobile et al., 2008; Del Nobile, Conte, Scrocco, Brescia, et al., 2009). Shapero, Nelson, and Labuza (1978) examined the inhibition of a strain of Staphylococcus aureus by ethanol at low water activities (about 0.88) and concluded that the inhibition was due not only to the lowering of water activity by ethanol but also to some other effects on the bacteria. Ballesteros, Chirife, and Bozzini (1993) came to the same conclusion in an electron microscopic study of two different strains of S. aureus, which indicated that cell wall changes were partly responsible for the antibacterial action of ethanol. This is supported by the work of Ingram (1981) and Ingram and Buttke (1984), and these authors suggested that ethanol inhibits cross-linking during peptidoglycan biosynthesis by decreasing the strength of hydrophobic interactions. Moreover, the inclusion of essential oils into edible coatings signicantly inhibited the growth of psychrophilic aerobes, yeasts and moulds of fresh-cut apples (Rojas-Gra et al., 2007). It is worth noting that at 8 days the cell load of mesophiles was always found below the threshold value (5 107 log cfu/g) imposed by the French Regulation (Corbo et al., 2004) during the entire storage time.
Table 2b Evolution of mesophilic and psychrotrophic bacteria (log cfu/g) for Step II kiwifruit packed under active MAP. Samples Mesophiles log cfu/ginitial Coat-dipp-EtOH Coat-GFSE Control 2.00 0.00 2.00 0.00a 2.89 0.77a,b
a

Psychrotrophs log cfu/g8 days 2.20 0.35 2.88 0.79a 4.00 0.26b
a

log cfu/ginitial 2.00 0.00 2.00 0.00a 2.00 0.00a

a

log cfu/g8 days 2.00 0.00a 3.11 0.23b 4.16 0.18c

Fig. 1. Evolution of oxygen (a) and carbon dioxide (b) plotted as a function of storage time for minimally processed kiwifruit belonging to Step I.

Mean values standard deviation. ad Means in the same column followed by different superscript upper cases are signicantly different (P b 0.05).

1228 Table 3a Mass loss percentage for Step I kiwifruit. Time (day) 1 4 6 8 11 13 15 Samples Coat-dipp-EtOH 0.308 0.049 0.354 0.084a 0.514 0.085a 0.610 0.080a 0.503 0.054a 0.483 0.038a 0.555 0.166a
a

M. Mastromatteo et al. / Food Research International 44 (2011) 12241230 Table 4a Shelf life of Step I kiwifruit packaged under passive MAP assumed as the lowest value among the calculated microbial and sensorial acceptability limits (MAL, SAL) and visual moulds time (VMT). Coat-GFSE
a

Dipp-EtOH 0.310 0.063 0.535 0.074b 0.519 0.101a 0.492 0.063a 0.517 0.117a 0.528 0.029a

Control
b

Samples Coat-dipp-EtOH Dipp-EtOH Coat-GFSE Control

SALOQ (day) 12.30 0.55 10.20 1.07b 11.75 0.56a 8.22 0.52c
a

VMT (day) N 15 14 N 15 14

Shelf life (day) 12.30 0.55a 10.20 1.07b 11.75 0.56a 8.22 0.52c

0.437 0.041 0.470 0.097a,b 0.518 0.159a 0.592 0.146a 0.558 0.111a 0.575 0.063a 0.612 0.076a

0.145 0.027c 0.186 0.049c 0.175 0.099b 0.256 0.058b 0.283 0.028b 0.327 0.256a

Mean values standard deviation. ac Means in the same row followed by different superscript upper cases are signicantly different (P b 0.05). The samples were not acceptable.

Mean values standard deviation. ad Means in the same column followed by different superscript upper cases are signicantly different (P b 0.05).

Lactic acid bacteria and Enterobacteriaceae (b 1 log cfu/g), yeasts and moulds (b 2 log cfu/g) were always below the level of detection. The pH values were about 3.4 in all samples and remained constant during the entire storage period (data not shown). 3.2.3. Mass loss Concerning the effect of different treatments on mass loss, signicant differences between the coated and dipped samples with respect to the control samples were observed (P b 0.05). In particular, the higher mass loss percent of the coated and dipped samples was probably due to the high water content of the alginate coating and the dipping treatment. However, for these samples the mass loss percent did not exceed 0.6% for the entire storage period (Table 3a). 3.2.4. Sensory evaluation Fig. 3a shows the evolution during storage of the minimally processed kiwifruit overall quality for all investigated samples packed under passive MAP. As can be inferred from the gure, the uncoated samples packaged under passive MAP reached rst the overall quality threshold value with respect to the coated samples. SAL values were determined for minimally processed kiwifruit overall quality following the procedure reported in the Materials and methods section. Curves shown in the gure were obtained by tting Eq. (2) to the experimental data, results are listed in Table 3a; whereas, the solid horizontal line is the overall quality threshold value. SALOQ is the time at which the investigated minimally processed produce was no more marketable from a sensory point of view. As can be inferred from the data listed in Table 4a, for the control and DippEtOH samples, a SALOQ value of about 810 days was measured, due to a general spoiling in terms of color, odor and consistence. Conversely, alginate-based coating reduced respiratory activity, as well as sensory decay, increasing the SALOQ value up to 12 days. Also, the combination of active compounds with alginate-based coating delayed the appearance of visible moulds. It is worth noting that as the microbial

load was always found below the threshold value imposed by law (Corbo et al., 2004), in this study SALOQ coincides with minimally processed produce shelf life. In particular, the highest shelf life value was obtained with coating treatments (12.3 and 11.75 for Coat-dippEtOH and Coat-GFSE, respectively) with respect to dipping treatment (10.2). For this reason, in the second step only the combined use of coating treatments with active MAP was investigated. The active MAP conditions were chosen on the basis on the headspace gas concentration found at equilibrium for Coat-dipp-EtOH and CoatGFSE samples; i.e., 10% O2 and 10% CO2. Nitrogen percent concentration was set as complement of oxygen and carbon dioxide percent concentration to 100. 3.3. Step II: packaging under active MAP 3.3.1. Headspace gas composition Fig. 2a, b shows the evolution over storage of O2 and CO2 concentrations in the OPP20 bag headspace for Coat-dipp-EtOH, Coat-GFSE and Ctrl samples packaged in active MAP. It is worth noting that for control samples the oxygen concentration decreased rapidly during the rst 12 days of storage. Whilst for the coated samples, as expected, an appropriate equilibrium between the respiration of the product and the gas permeability of the lm was obtained. Data suggest that the coating treatment was effective to avoid a drastic reduction of oxygen concentration as it occurred in non-coated minimally processed kiwifruit. This result is a direct consequence of the coating inuence over the oxygen diffusion between the fruit and environment (Baldwin, Nisperos-Carriedo, Chen, & Hagenmaier, 1996). The carbon dioxide headspace concentrations of the samples packaged under active MAP level off to values higher than those of the kiwi stored under passive MAP. In fact, for coating and control samples a carbon dioxide nal value of about 15 and 12% respectively, was obtained. 3.3.2. Microbiological stability and pH Table 2b reports the cell load of mesophilic and psychrotrophic bacteria at 0 and 8 days of sampling for treated and untreated samples packed under active MAP. Data show that by means the combined use of active MAP and active coating, no growth of mesophiles and psychrotrophs at 8 days of storage was observed. On the contrary, the control samples showed an increase of the cell load of about 2 log
Table 4b Shelf life of Step II kiwifruit packaged under active MAP assumed as the lowest value among the calculated microbial and sensorial acceptability limits (MAL, SAL) and visual moulds time (VMT). Samples Coat-dipp-EtOH Coat-GFSE Control SAL
OQ

Table 3b Mass loss percentage for Step II kiwifruit. Time (day) 1 4 6 8 11 13 15 Samples Coat-dipp-EtOH 0.664 0.074a 0.644 0.099a 0.729 0.147a 0.796 0.037a 0.960 0.093a 1.209 0.155a 1.441 0.261a Coat-GFSE 0.775 0.204a 1.092 0.546a 1.141 0.189b 1.257 0.289b 1.49 0.053b 1.288 0.148a 1.288 0.055a Control 0.761 0.086a 0.928 0.146a 0.80 0.146a 1.094 0.204a,b 1.031 0.196a 1.119 0.171a

(day)
a

VMT (day) N 21 18.5 13

Shelf life (day) 11.45 1.13a 13.52 1.03b 2.70 0.67c

Mean values standard deviation. a b Means in the same row followed by different superscript upper cases are signicantly different (P b 0.05). The samples were not acceptable.

11.45 1.13 13.52 1.03b 2.70 0.67c

Mean values standard deviation. ad Means in the same column followed by different superscript upper cases are signicantly different (P b 0.05).

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Fig. 2. Evolution of oxygen (a) and carbon dioxide (b) plotted as a function of storage time for minimally processed kiwifruit belonging to Step II.

Fig. 3. Overall quality of Step I (a) and Step II (b) minimally processed kiwifruit.

cycle. The combined use of active coating and active MAP was most effective as no growth was observed during the storage. Literature data report that the combined use of antimicrobial compounds and active MAP affected signicantly and positively the microbiological stability of the sliced strawberries and carrots (Amanatidou, Slump, Gorris, & Smid, 2000; Campaniello, Bevilacqua, Sinigaglia, & Corbo, 2008). As reported for the samples packaged under passive MAP, also in this case the nal cell load of mesophiles was always found below the threshold value (5 107 log cfu/g) imposed by the French Regulation (Corbo et al., 2004). 3.3.3. Mass loss Concerning the effect of active MAP on mass loss, all samples showed a sudden increase in the mass loss percent at the early stage of storage. Most probably, this result is directly related to both the evaporation of moisture from produce during the severe initial vacuumization that was applied prior to injecting the gas in the package to realize the modied atmosphere, and to the absence of humidity in the gas mixture injected into the bag. However, the mass loss percentage did not exceed 1.5 for all samples (Table 3b). 3.3.4. Sensory evaluation Fig. 3b shows the evolution during storage of the minimally processed kiwifruit overall quality for all investigated samples packed under active MAP. Curves shown in the gure were obtained by tting Eq. (2) to the experimental data, whereas the solid horizontal line is the overall quality threshold value. SAL values for the minimally processed kiwifruit overall quality packaged under active MAP were reported in Table 4b. As can be inferred from the data, for samples packaged under active MAP a SALOQ value of

11.45 and 13.52 days for Coat-dipp-EtOH and Coat-GFSE, respectively, was observed. The active MAP alone worse the shelf life; in fact, a value of about 2.7 days for the control samples was obtained. This was due to a general spoiling in terms of overall quality such as color, odor and consistence. The coating allowed a good preservation of the product by reducing the respiratory activity, as well as sensory decay. The combined use of active coating and active MAP delayed the moulds growth until over 21 and 18.5 days for Coat-dipp-EtOH and Coat-GFSE, respectively. Dantigny, Guilmart, Radoi, Bensoussan, and Zwietering (2005) reported that ethanol was an effective additional barrier to inhibit fungal growth in food products. Moreover, active MAP involving reduced O2 levels and elevated levels of CO2, was used in combination with other environmental factors to control growth and mycotoxin production by several moulds (El Halouat & Debevere, 1997).

4. Conclusions Results of this study suggested that the coatings were the best treatments and could be used to control dehydration and respiration of minimally processed kiwifruits both in passive and active MAP, thus extending its shelf life. The combination of coating with hydroalcoholic solution and GFSE inhibited the microbial growth whereas the dipping samples showed the same behavior of the untreated sample. Probably, the coating treatment improved the ethanol efcacy as it avoided its evaporation. Moreover, the microbial loads did not limit the shelf life of the investigated produce. It is worth noting that, when the active MAP was used alone a very short shelf life of the uncoated samples occurred. However, the combined use of active MAP and coating treatments allowed a good preservation of the

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M. Mastromatteo et al. / Food Research International 44 (2011) 12241230 Gimenez, M., Olarte, C., Sanz, S., Lomas, C., Echvarri, J. F., & Ayala, F. (2003). Relation between spoilage microbiological quality in minimally processed artichoke packaged with different lms. Food Microbiology, 20, 231242. Ingram, L. O. (1981). Mechanism of lysis of Escherichia coli by ethanol and other chaotropic agents. Journal of Bacteriology, 146(1), 331336. Ingram, L. O., & Buttke, T. M. (1984). Effects of alcohols on microorganisms. Advances in Microbial Physiology, 23, 779789. Jayas, D. S., & Jeyamkondan, S. (2002). Modied atmosphere storage of grains meats fruits and vegetables. Biosystems Engineering, 82(3), 235251. Larmond, E. (1977). Laboratory methods for sensory evaluation of foods (pp. 1637). Ottawa Department of Agriculture Publication. Lee, J. Y., Park, H. J., Lee, C. Y., & Choi, W. Y. (2003). Extending shelf-life of minimally processed apples with edible coatings and antibrowning agents. Lebensmittel Wissenschaft und Technologie, 36, 323329. Mastromatteo, M., Lucera, A., Sinigaglia, M., & Corbo, M. R. (2009). Combined effects of thymol, carvacrol and temperature on the quality of non conventional poultry patties. Meat Science, 83, 246254. Mastromatteo, M., Danza, A., Conte, A., Muratore, G., & Del Nobile, M. A. (2010). Shelf life of ready to use peeled shrimps as affected by thymol essential oil and modied atmosphere packaging. International Journal of Food Microbiology, 144(2), 250256. Mazliak, P. (1983). Plant membrane lipids: Changes and alterations during ageing and senescense. In M. Liebermann (Ed.), Postharvest physiology and crop preservation (pp. 123140). NewYork: Plenum Press. Nowak, B., Von Muefing, T., Grotheer, J., Klein, G., & Watkinson, B. M. (2007). Energy content, sensory properties, and microbiological shelf life of German bologna-type sausages produced with citrate or phosphate and with inulin as fat replacer. Journal of Food Science, 72, S629S638. O'Connor-Shaw, R. E., Roberts, R., Ford, A. L., & Nottingham, S. M. (1994). Shelf-life of minimally processed honeydew, kiwifruit, papaya, pineapple and cantaloupe. Journal of Food Science, 59(12021206), 1215. Olivas, G. I., & Barbosa-Cnovas, G. V. (2005). Edible coating for fresh-cut fruits. Critical Review in Food Science and Nutrition, 45, 657670. Pinto, R., Lichter, A., Danshin, A., & Sela, S. (2006). The effect of an ethanol dip of table grapes on populations of Escherichia coli. Postharvest Biology and Technology, 39, 308313. Pranoto, Y., Salokhe, V., & Rakshit, K. S. (2005). Physical and antibacterial properties of alginate-based edible lm incorporated with garlic oil. Food Research International, 38, 267272. Raybaudi-Massilia, R. M., Mosqueda-Melgar, J., & Martn-Belloso, O. (2008). Edible alginate-based coating as carrier of antimicrobials to improve shelf-life and safety of fresh-cut melon. International Journal of Food Microbiology, 121, 313327. Rojas-Gra, M. A., Raybaudi-Massilia, R. M., Soliva-Fortuny, R., Avena-Bustillos, R. J., McHugh, T. H., & Martn-Belloso, O. (2007). Apple puree alginate coating as carrier of antimicrobial agents to prolong shelf life of fresh-cut apples. Postharvest Biology and Technology, 45, 254264. Scott, K. J., Spraggon, S. A., & McBride, R. L. (1986). Two new maturity tests for kiwi fruit. CSIRO Food Research, Q46, 2531. Shapero, M., Nelson, D. A., & Labuza, T. P. (1978). Ethanol inhibition of Staphylococcus aureus at limited water activity. Journal of Food Science, 43(5), 14671469. Stec, M. G., Hodgson, J. A., Macrae, E., & Triggs, C. (1989). Role of fruit rmness in the sensory evaluation of kiwi fruit (Actinidia deliciosa cv Hayward). Journal of the Science of Food and Agriculture, 47, 417433. Van de Velde, K., & Kiekens, P. (2002). Biopolymers: Overview of several properties and consequences on their applications. Polymer Testing, 21, 433442. Watada, A. E., Abe, K., & Yamuchi, N. (1990). Physiological activities of partially processed fruits and vegetables. Food Technology, 44(5), 116122. Wegrzyn, T. F., & MacRae, E. A. (1992). Pectinesterase, polygalacturonase and galactosidase during softening of ethylene-treated kiwifruit. Horticultural Science, 27, 900902. Wong, W. S., Tillin, S. J., Hudson, J. S., & Pavlath, A. E. (1994). Gas exchange in cut apples with bilayer coatings. Journal of Agricultural and Food Chemistry, 42, 22782285.

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