Postharvest Biology and Technology 63 (2012) 5054

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Postharvest Biology and Technology

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A preliminary study on the effect of metabolic stress disinfection and disinfestation (MSDD) on ripening physiology and quality of kiwifruit and apple
F. Zulhendri a,b , L.E. Jamieson a , J. Feng a , C.O. Perera b , S.Y. Quek b , A.B. Woolf a,
a b

The New Zealand Institute for Plant & Food Research Limited, 120 Mt Albert Road, Sandringham, 1025 Auckland, New Zealand Food Science Programme, Chemistry Department, The University of Auckland, 23 Symonds St., Auckland, New Zealand

a r t i c l e

i n f o

a b s t r a c t
Metabolic stress disinfection and disinfestation (MSDD) is a relatively new quarantine treatment in which fruit are exposed to rapid decompression and compression cycles and high CO2 atmosphere, followed by exposure to ethanol vapour under decompression. This study evaluated the ripening response of Hayward kiwifruit (Actinidia deliciosa) and Pink Lady apple (Malus x domestica) to MSDD treatment, which can control longtailed mealybug (Pseudococcus longispinus). Following the treatments, fruit were held at 20 C for 7 d for shelf-life assessment, while the remainder were refrigerated at 0.5 C for 16 weeks. Respiration rate, volatile (ethylene, ethanol and acetaldehyde) production rates, rmness and disorders were measured at regular time intervals. MSDD treatments did not affect the metabolic activities and quality of Pink Lady apples. However, rmness was reduced by 4.9 N in non-refrigerated MSDD treated fruit. MSDD treatments did not control supercial scald disorder in refrigerated Pink Lady apples. For Hayward kiwifruit, treatments increased the respiration rate and ethylene production of short-term refrigerated fruit, promoted endogenous production of ethanol and acetaldehyde in both short-term and long-term refrigerated fruit. MSDD treatments also increased the incidence of rots in refrigerated Hayward kiwifruit. MSDD treatments accelerated the softening of short-term refrigerated kiwifruit, but retarded the softening of Hayward kiwifruit during the 16 weeks of refrigerated storage. MSDD could potentially be used as a quarantine treatment of apples. Further studies are required on the sensory effect of MSDD treatment. 2011 Elsevier B.V. All rights reserved.

Article history: Received 28 April 2011 Accepted 5 September 2011 Keywords: Fruit quality Methyl bromide alternatives Postharvest Quarantine treatments

1. Introduction Metabolic stress disinfection and disinfestation (MSDD) is an alternative technology that uses modication of atmospheric pressure combined with GRAS (Generally Recognized As Safe) chemicals to disinfest and sterilize fresh produce (Lagunas-Solar et al., 2006). It has two phases, physical and chemical. The physical phase employs a cycle of rapid decompression (vacuum) and compression using pure CO2 . Upon completion of the physical phase, a volatile chemical (ethanol) is added and pressure in the chamber is again reduced and held for a period of time (Lagunas-Solar and Essert, 2006; Lagunas-Solar et al., 2006). During the physical phase, there is an increase in partial pressure of CO2 and a decrease in that of O2 which may have similar effects to CA (controlled atmosphere)/MA (modied atmosphere) storage conditions. Standard CA storage treatments are used to extend the

Corresponding author at: Plant & Food Research Mt Albert, Private Bag 92169, Auckland Mail Centre, 1142 Auckland, New Zealand. Tel.: +64 9 925 7267; fax: +64 9 925 8628. E-mail address: allan.woolf@plantandfood.co.nz (A.B. Woolf). 0925-5214/$ see front matter 2011 Elsevier B.V. All rights reserved. doi:10.1016/j.postharvbio.2011.09.004

storage life of fruit (typically using O2 concentrations of 13 kPa and CO2 concentrations of 110 kPa over long storage period; weeks or even months). The chemical phase of MSDD is initiated by introduction of ethanol at negative pressures (and after there is no O2 in the chamber). Ethanol treatments have been shown to either inhibit or promote ripening, depending on the commodity and maturity (Beaulieu and Saltveit, 1992, 1997; Podd and Van Staden, 1998). Ethanol and acetaldehyde treatments of kiwifruit have been shown to induce ripening (Mencarelli et al., 1991). It was demonstrated that exposure to 7 h of 0.18% ethanol and 0.02 or 0.04% acetaldehyde treatments at 17 C increased the respiration rate and ethylene production of Hayward kiwifruit, but acetaldehyde had greater effect. In addition, ethanol has also been used to control specic disorders such as supercial scald in apples (Ghahramani et al., 1999). Although MSDD has been shown to be effective for treatment of a range of external pest insects (Lagunas-Solar et al., 2006; ArevaloGalarza and Follett, 2011), there is little information on the effect on MSDD on fruit physiology and quality to date. It is crucial to determine the commercial viability of MSDD. Lagunas-Solar et al. (2006) reported minimal/no physical or functional effects of MSDD on commodities. In this research note, we report the effect of MSDD to control longtailed mealybug on the quality and ripening

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Fig. 1. MSDD operational chart showing a physical phase of 30 min followed by a chemical phase of 60 min. The physical phase consisted of 5060 short cycles at 90 and 110 kPa. The chemical phase was initiated by lowering the pressure to 10 kPa and then treating with ethanol (371 mg L1 ) for 60 min.

physiology of Hayward kiwifruit (Actinidae deliciosa) and Pink Lady apple (Malus domestica). We also examined the efcacy of MSDD to control supercial scald in Pink Lady. 2. Materials and methods 2.1. Fruit Pink Lady apples (93.1 N, 13% SSC) were harvested from an orchard in Hawkes Bay, New Zealand in March 2010, and held overnight at 20 C prior to treatment. Hayward kiwifruit (from three growers) were obtained from a commercial fruit supplier in South Auckland, New Zealand in June 2010. Hayward kiwifruit had been refrigerated at 0.5 C for 3 weeks prior to MSDD treatments. Kiwifruit were held overnight at 20 C prior to treatment. Approximately 3033 fruit from each grower were pooled, and treated at once. The treatment was then replicated three times. 2.2. MSDD A 250-L MSDD prototype system was used to treat the fruit. The prototype was tted with a liquid ring vacuum pump (Model TRMB 32-75, Travaini Pumps, USA), a compound gauge (WIKA EN8371, 316 L) and a water bath-chemical heating system to convert liquid ethanol to vapour before it entered the treatment chamber. The MSDD treatment chamber was connected to a CO2 gas cylinder (food grade 99.9% v/v). A 110-mm diameter spark-proof fan was tted inside the chamber to ensure uniformity of treatment conditions (temperature and gas concentrations). The treatment temperature was maintained at 20 1 C. The treatment procedure started by drawing pressure down from 110 to 90 kPa in 10 s, followed by immediate rapid ow of CO2 gas which increased the pressure to 110 kPa in 20 s (Fig. 1). This was considered one physical cycle taking approximately 30 s. The physical phase of each MSDD treatment was carried out for 30 min, thus consisting of 5060 physical cycles. 2.3. Kiwifruit The chemical phase was initiated after the 30-min physical phase by injecting a known amount of liquid ethanol into the heat exchange system (at 90 C) and lowering the pressure to 10 kPa.

The amount of liquid ethanol was calculated for the internal volume of the empty treatment chamber (250 L) in order to achieve the target concentration. The target concentration is 371 mg L1 which is an estimate of LD99 for longtailed mealybug (Zulhendri et al., 2011). The chemical phase lasted for 60 min. The control was untreated fruit (held at standard atmosphere pressure). Each and control was replicated 3 times, and each replicate was applied to 100 kiwifruit. After being treated, ve fruit from each replicate were held at 20 C for 7 d shelf-life. Firmness, respiration rate, and volatiles (ethylene, ethanol and acetaldehyde) production rates were measured at 2 d intervals. Firmness was measured using Aweta acoustic rmness measurement (Acoustic Firmness Sensor, the Netherlands) with a setting for microphone gain and tick power of 80 and 16, respectively. Carbon dioxide and ethylene concentration measurements were carried out according to Besada et al. (2010). Ethanol and acetaldehyde were measured using Phillips PU4500 GC (Pye Unichem, Cambridge, UK) equipped with an FID detector and N2 as carrier gas. A packed stainless steel column (2 m 2 mm i.d., Carbograph 1 AW 20, 80/100) was set isothermally at 180 C. The remaining fruit were refrigerated at 0.5 C for 16 weeks and rmness of 30 fruit from each replicate was measured at 4 week intervals. After 16 weeks of refrigerated storage, fruit were held at 20 C for 14 d. During this period, ve fruit from each replicate were assessed for respiration rate, ethylene, ethanol and acetaldehyde production every 2 d. After the 14 d at 20 C, rots, esh breakdown (water-soaked appearance of the endocarp), soluble solid content (SSC) and pH were measured on all the remaining fruit. SSC and pH were measured according to Johnston et al. (2009). 2.4. Apple A similar experiment was carried out for apple. After being MSDD treated, 15 fruit from each replicate were held at 20 C for 7 d, while the remaining fruit were refrigerated at 0.5 C for 16 weeks. Respiration rate, ethanol and acetaldehyde production of ve fruit from each replicate were then analyzed. Internal ethylene concentration of 15 fruit from each replicate was measured by drawing 1 mL of gas from the seed cavity (Johnston et al., 2009). Firmness was then measured according to Johnston et al. (2009). After being refrigerated for 16 weeks, the above assessments were carried out with the same number of fruit. Quality assessment of background skin colour, SSC and pH level was carried out according to Johnston et al. (2009). In addition, disorder incidence was rated including supercial scald, internal browning and greasiness. Percentage incidence for a given disorder is dened as the percentage of fruit with a severity rating >0. Supercial scald severity was measured as slight (<25% surface area), moderate (2550% surface area) and severe (>50% surface area). Severity of internal browning and greasiness was not measured. Fruit attributes and disorders were analyzed using the analysis of variance procedure in Origin 7.5 (OriginLab, Northampton, MA). Percentage attribute and disorder were transformed using arcsine(sqrt(p)) prior to statistical analysis. 3. Results 3.1. Hayward kiwifruit The respiration rate of non-refrigerated fruit treated with MSDD was consistently higher than control fruit (Fig. 2A). During the 7 d shelf-life analysis, respiration rate was between 0.025 and 0.032 mg kg1 s1 for the treated fruit, compared to 0.017 and 0.018 mg kg1 s1 for the control fruit. Ethylene

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Fig. 2. Carbon dioxide and ethylene concentration (mean SEM) of Hayward kiwifruit and Pink Lady apple. (A and B) CO2 production (mg kg1 s1 ) of non-refrigerated and refrigerated fruit (after MSDD treatments), respectively. (C and D) Ethylene concentration of short-term and long-term refrigerated fruit, respectively (primary y-axis internal ethylene concentration ( g L1 ) of Pink Lady apple, secondary y-axis ethylene production (mg kg1 s1 ) of Hayward kiwifruit).

production of treated fruit increased from 1.944 104 to 2.417 103 mg kg1 s1 from day 1 to day 6, then decreased to 1.194 103 mg kg1 s1 by day 7. Conversely, ethylene production of the control fruit decreased from 6.667 104 to 0 mg kg1 s1

from day 1 to 5, and there was no increase in ethylene production afterwards (Fig. 2C). MSDD treatments caused a dramatic increase in ethanol and acetaldehyde production (especially from day 3); from 0 to 0.107 and 0.011 mg kg1 s1 after 7 d for ethanol and

Fig. 3. Ethanol and acetaldehyde concentration (mean SEM) of Hayward kiwifruit and Pink Lady apple. (A and B) Ethanol production (mg kg1 s1 ) of non-refrigerated and refrigerated fruit (after MSDD treatments), respectively. (C and D) Acetaldehyde production (mg kg1 s1 ) of short term and long term refrigerated fruit, respectively.

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Fig. 4. Effect of MSDD on softening of Hayward kiwifruit during shelf life test at 20 C (A) and during refrigerated storage at 0.5 C (B). Acoustic rmness (106 Hz2 kg2/3 ) was measured using an Aweta Acoustic Firmness Sensor.

acetaldehyde, respectively (Fig. 3A and B). The softening rate of treated fruit was also higher compared to control fruit; the Aweta index dropped from 39.5 to 25.2 (106 Hz2 kg2/3 ) and 39.0 to 28.4 (106 Hz2 kg2/3 ) for treated and control fruit, respectively (Fig. 4A). The Aweta index of 39.0, 28.0 and 25.0 (106 Hz2 kg2/3 ) equates to 58.8, 34.3 and 24.5 N, respectively (J. Feng, personal communication). After storage at 0.5 C for 16 weeks, treated fruit had lower CO2 production compared to control fruit (although it levelled out after day 9; Fig. 2B). There was no signicant difference in ethylene production, although the control fruit reached an earlier climacteric peak (Fig. 2D). No ethanol and acetaldehyde production could be detected in control fruit. Ethanol and acetaldehyde production in treated fruit were not detected until day 9 and 13, respectively. A marked increase in ethanol production occurred from 8.3 to 112.8 mg kg1 s1 only after 9 d. During 16 weeks of refrigeration, the softening of treated fruit was delayed compared to control fruit. The Aweta index of treated fruit dropped from 39.5 to 15.7 (106 Hz2 kg2/3 ) compared to 39.0 to 10.7 (106 Hz2 kg2/3 ) for control fruit. An Aweta index of 15.0 and 10.0 (106 Hz2 kg2/3 ) equates to 9.8 and 7.8 N, respectively. No signicant difference was observed in terms of SSC and expressed juice pH (pH and SSC levels were 4.3 and 15.0, respectively). MSDD treatments increased rot incidence by more than twofold compared to control fruit; 54% compared to 24% for control fruit.

4. Discussion MSDD was shown to promote the endogenous production of ethanol and acetaldehyde in non-refrigerated Hayward kiwifruit. MSDD treatments also increased the respiration rate and stimulated the ethylene climacteric of kiwifruit which consequently increased the rate of softening. The results reected a similar trend observed by Mencarelli et al. (1991) in which ethanol and acetaldehyde treatments (for 14 d) were shown to induce ripening of kiwifruit. A similar result was also shown in high CO2 /low O2 atmosphere treatments of kiwifruit. Irving (1992) showed that kiwifruit stored at 60% and 80% CO2 atmosphere for 2 weeks at 5 and 10 C had higher respiration rate, earlier ethylene climacteric peak and signicantly reduced rmness (compared to air storage), upon removal to 20 C air (shelf-life). Increased softening could possibly be due to accumulation of ethanol and/or acetaldehyde in the esh during storage which was not investigated by Irving (1992). Conversely, the long-term refrigerated fruit showed a different trend. MSDD treatment reduced respiration rate and delayed ethylene peak (by 2 d), upon removal to shelf-life. In addition, MSDD treatments reduced the softening rate during 16 weeks of refrigeration (an opposite trend from non-refrigerated fruit). Ethanol and acetaldehyde production were detected at day 9 and 11 after removal to shelf-life, respectively. This result suggests that the effect of MSDD treatment on ethanol and acetaldehyde production was not transient and that signicant physiological changes had occurred. MSDD treatments did not affect the physiology and quality attributes of Pink Lady apple. Generally, no differences were observed in terms of respiration rate, internal ethylene concentration, SSC and pH level in both non-refrigerated and refrigerated fruit during 7 of shelf-life assessment. However, the rmness of non-refrigerated MSDD-treated fruit was reduced by 5 N (a similar trend to that of Hayward kiwifruit). MSDD treatments did not affect the incidence of any postharvest disorders such as supercial scald, internal browning or greasiness. Ethanol treatments over long time periods (up to 16 weeks) have been shown to control supercial scald incidence in Granny Smith apples by reducing the level of -farnesene and its conjugated trienes (Ghahramani and Scott, 1998a,b). However, our results showed that ethanol diffused out of the MSDD treated fruit to undetectable level after 7 d. Consequently, the ethanol level required to control supercial scald was most likely not sufcient to be effective. In conclusion, MSDD treatments previously shown to control longtailed mealybug accelerated ripening of non-refrigerated Hayward kiwifruit, but reduced the respiration rate and ethylene

3.2. Pink Lady apple MSDD treatments did not affect respiration rate and internal ethylene concentration of non-refrigerated apples (Fig. 2C). The ethanol and acetaldehyde production of treated fruit decreased from 0.015 and 1.139 103 mg kg1 s1 (respectively) at day 1, to an undetectable level at day 7 (Fig. 3A and C). No ethanol and acetaldehyde production was detected in control fruit. After 7 d of shelf-life, MSDD treatments reduced the rmness of the fruit; 84.3 (1.7) N compared to 89.2 (2.0) N recorded for the control fruit. After storage, no difference in respiration rate and internal ethylene concentration was observed between control and treated fruit. There was also no difference in terms of rmness (57.8 N), SSC (13.0%) and pH (3.6). However, treated fruit had a higher hue angle value of background colour of 94.8 (1.1) compared to 91.1 (1.2) for control fruit. No signicant difference was recorded in terms of supercial scald (47%), internal browning 12%) or greasiness incidence (1721%). MSDD treatments also had no effect on severity of supercial scald.

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F. Zulhendri et al. / Postharvest Biology and Technology 63 (2012) 5054 Beaulieu, J.C., Saltveit, M.E., 1997. Inhibition or promotion of tomato fruit ripening by acetaldehyde and ethanol is concentration dependent and varies with initial fruit maturity. J. Am. Soc. Hort. Sci. 122, 392398. Besada, C., Jackman, R.C., Olsson, S., Woolf, A.B., 2010. Response of Fuyu persimmons to ethylene exposure before and during storage. Postharvest Biol. Technol. 57, 124131. Ghahramani, F., Scott, K.J., 1998a. The action of ethanol in controlling supercial scald of apples. Aust. J. Agric. Res. 49, 199205. Ghahramani, F., Scott, K.J., 1998b. Oxygen stress of Granny Smith apples in relation to supercial scald, ethanol, -farnesene, and conjugated trienes. Aust. J. Agric. Res. 49, 207210. Ghahramani, F., Scott, K.J., Buckle, K.A., Paton, J.E., 1999. A comparison of the effects of ethanol and higher alcohols for the control of supercial scald in apples. J. Hort. Sci. Biotechnol. 74, 8793. Irving, D.E., 1992. High concentrations of carbon dioxide inuence kiwifruit ripening. Postharvest Biol. Technol. 2, 109115. Johnston, J.W., Gunaseelan, K., Pidakala, P., Wang, M., Schaffer, R.J., 2009. Coordination of early and late ripening events in apples is regulated through differential sensitivities to ethylene. J. Exp. Bot. 60, 26892699. Lagunas-Solar, M.C., Essert, T.K., 2006. Patent No. US 2006/0269438 A1. Disinfestation and disinfection of food, perishables and other commodities. Lagunas-Solar, M.C., Essert, T.K., Pina, U.C., Zeng, N.X., Truong, T.D., 2006. Metabolic stress disinfection and disinfestation (MSDD): a new, non-thermal, residue-free process for fresh agricultural products. J. Sci. Food Agric. 86, 18141825. Mencarelli, F., Savarese, P., Saltveit Jr., M.E., 1991. Ripening of kiwifruit exposed to ethanol and acetaldehyde vapors. HortSci. 26, 566569. Podd, L.A., Van Staden, J., 1998. The role of ethanol and acetaldehyde in ower senescence and fruit ripening a review. J. Plant Growth Regul. 26, 183189. Zulhendri, F., Jamieson, L.E., Perera, C.O., McDonald, R.M., Connolly, P.G., Quek, S.Y., Woolf, A.B., 2011. The effect of metabolic stress disinfection and disinfestation (MSDD) on Hass avocado fruit physiology and mortality of longtailed mealybug (Pseudococcus longispinus). Postarvest Biol. Technol., doi:10.1016/j.postharvbio.2011.06.006.

production (consequently softening rate) of refrigerated kiwifruit. However, MSDD increased the incidence of rot in Hayward kiwifruit in the long-term refrigerated storage. Further study is required to assess the tolerance level of kiwifruit to MSDD, such as reducing the treatment time or ethanol concentration to an acceptable level in terms of insect mortality and fruit quality. MSDD treatments did not signicantly affect Pink Lady apples, from both physiology and quality perspectives. Further studies are required to assess the effect of MSDD across various maturity stages and harvest conditions as well as the effect on sensory properties of fruit. Acknowledgements We thank Jason Johnston for the advice on experimental design and critical review of the manuscript. We also thank Paul Brookeld and Jessica Robertson for the help in supplying fruit. The work was supported by the Foundation for Research, Science and Technology Contract (C06X0709) and a doctoral scholarship from The University of Auckland. References
Arevalo-Galarza, L., Follett, P.A., 2011. Response of Ceratis capitata, Bactrocera doralis, Bactrocera cucurbitae (Diptera: Tephritidae) to metabolic stress disinfection and disinfestation treatment. J. Econ. Entomol. 104, 7580. Beaulieu, J.C., Saltveit Jr., M.E., 1992. Acetaldehyde and ethanol inhibition of tomato fruit ripening. HortSci. 27, 623.