Food Microbiology: Fundamentals and Frontiers, 3rd Ed.

Edited by M. P. Doyle and L. R. Beuchat

© 2007 ASM Press, Washington, D.C.

Frédéric Carlin

Fruits and Vegetables 8

Fresh fruits and vegetables are an extraordinary dietary examples of some important spoilage molds and bac-
source of nutrients, micronutrients, vitamins, and fiber teria, their host vegetables or fruits, and symptoms of
for humans, and an essential basic raw material for the infection. More extensive descriptions of fruit and veg-
food industry. They are living organs detached from etable spoilage can be found elsewhere (The Commercial
their parent plants and have a high water content, which Storage of Fruits, Vegetables, and Florist and Nursery
contributes to their natural fragility. In addition, fruits Stocks, available at http://www.ba.ars.usda.gov/hb66)
and vegetables are widely exposed to microbial contami- (3, 26, 89, 105, 106).
nation through contact with soil, dust, and water, and
by handling at harvest or during postharvest processing,
thereby establishing conditions that may lead to spoilage MAIN CHARACTERISTICS OF FRUITS
and loss of quality. AND VEGETABLES AND THEIR ROLES
A single produce such as tomato is cultivated under IN MICROBIAL SPOILAGE
many different climates and at many latitudes from its Fruits and vegetables are the edible parts of plant organs
tropical area of origin to the colder Nordic countries. of very diverse nature: leaves (lettuce and cabbage, for
Giving an exhaustive account of microbial spoilage of instance), stems (leek and asparagus), flowers (arti-
tomato, as well as many other types of produce, presents choke, cauliflower, and broccoli), roots (beet, carrot,
some difficulties. Moreover, fruits and vegetables cover and turnip), bulbs (garlic and onion), tubers (potato),
many different species and include many different plant and fruits in their botanical meaning, e.g., simple fruit
organs at varied stages of physiological maturity in their such as tomato, cucumber, and pepper, stone fruit such
consumed forms. Reviewing microbial spoilage of fruits as peach, seed fruit such as apple, multiple fruit such as
and vegetables is a real challenge, and the literature on pineapple, aggregate fruit such as raspberry, and fruits
this topic is particularly rich. This chapter focuses on consumed in their immature form such as green beans.
the origin, description, and control of bacterial and fun- Plant tissues in fruits and vegetables consist of an assem-
gal spoilage of fruits and vegetables. Table 8.1 presents blage of cells surrounded by a pectic and cellulosic cell

Frédéric Carlin, INRA, UMR408, Sécurité et Qualité des Produits d’Origine Végétale, Avignon, F-84914, France, and University of Avignon,
Avignon, F-84029, France.

157
158 Microbial Spoilage and Public Health Concerns

Table 8.1 Important microbial agents of postharvest spoilage of fruits and vegetablesa
Type of postharvest
Spoilage agent disease or spoilage Produce affected Biology
Alternaria alternata and Black rot, black spots, Cucurbit, solanaceous vegetables, Stem-end pathogen. Penetration by flower
other Alternaria spp. dark lesions green bean, brassica, potato, or stem scars. Infection may remain
citrus, persimmon, mango, pome quiescent.
fruits
Botrytis cinerea Soft rot covered with Cucurbit, solanaceous vegeta- Wide spectrum. Infection before or after
gray mold bles, green bean, pea, brassica, harvest through damaged or senescent
artichoke, celery, lettuce, chicory, tissue. Favored by wet conditions. May
onion, garlic, carrot, citrus, apple, spread into neighboring fruits, causing
strawberry, raspberry “nesting.” Possible growth even at low
temperatures.
Colletotrichum musae Anthracnose; dark circu- Banana Quiescent infection until fruit ripening.
lar spots on ripening
fruits
Colletotrichum gloeo- Anthracnose; lesion on Cucurbit, solanaceous vegetables, Quiescent infections. May form
sporioides and other the skin. Dark spots, green bean, avocado, apple, appressoria on the plant cuticle. Devel-
Colletotrichum spp. sunken lesions. mango opment of decay during fruit matura-
tion.
Geotrichum candidum Sour rot Cucurbit, carrot, citrus, tomato Soil pathogen. Transmission by insects.
Wound pathogen.
Monilinia spp. Brown rot. Brown spots, Apple, stone fruits Survival in winter on mummified fruits.
white molds in concen- Infection may remain quiescent on im-
tric circles. mature fruits.
Penicillium spp. Blue mold, green mold Cucurbit, onion, garlic, grape, ap- Wound pathogen. Colored spores at the
covering lesions or rot ple, citrus center of the lesions. Slow development
at low temperature. May spread from
fruits to fruits.
Rhizopus spp. Soft, very wet rot. De- Cucurbit, solanaceous vegetables, Ubiquitous. Infection at, or after harvest
velopment of profuse green bean, pea, sweet potato, by wound, or by contact with soil or
mycelium with spore stone fruits, papaya infected produce. Rapid decay above
heads turning black. 20C.
Erwinia carotovora Soft rot Cucurbit, solanaceous vegetables, Infection by wounds, scars, and lenticels
brassica, asparagus, celery, let- before or after harvest, and by contact
tuce, chicory, carrot with decaying vegetable. Favored by
wet conditions and temperatures of
24–30C.
a
Adapted from references 3, 89, and 105.

wall organized in a network, and a middle lamella rich injury, and other stress assaults, and scars resulting from
in pectin cementing together cell walls (53, 117). Both detachment from plants.
the water content of the cell vacuole and the cell wall Once fruits and vegetables have detached from the
organization and composition contribute to edible plant plant, their physiological activity continues and is neg-
tissue firmness. The outer parts of fruits and vegetables atively correlated with shelf life. Commodities, such as
are characterized by layers varying in thickness accord- potato, onion, garlic, carrot, cantaloupe, and water-
ing to the type of produce, and include a hydrophobic melon, with a low respiration rate (10 ml of CO2/kg/h
cuticle consisting of cutin and wax covering an epider- at 5C) can be stored for longer periods than those such as
mis made of a layer of cells and eventually a layer of cork Brussels sprouts, spinach, broccoli, asparagus, or mush-
cells (45). Fruit and vegetable surfaces can be interrupted room, with a high respiration rate (20 ml of CO2/kg/h
by natural openings, e.g., stomata or lenticels involved at 5C) (59). For “climacteric” fruits (e.g., apple, tomato,
in respiration and transpiration of the plant organs, tri- avocado, and banana), several biochemical changes asso-
chomes, cracks, wounds caused by insects, mechanical ciated with natural respiration occur and, triggered by
8. Fruits and Vegetables 159

autocatalytic production of the plant hormone ethylene, fruits and vegetables, tomatine (a saponin in tomato),
will lead to horticultural maturity (102). Nonclimacteric sulfur-derived compounds in the Alliaceae (onion and
commodities (e.g., strawberry, citrus, grape, and cherry) garlic), or terpenoids in carrot (3, 16, 19).
are picked at their horticultural maturity, a stage of devel-
opment corresponding to the prerequisites for utilization
by consumers. The senescence process will lead, after ORIGIN OF CONTAMINATION
horticultural maturity, to irreversible changes in structure Fruits and vegetables harbor a wide range of microbial
and metabolism of the organs, and finally to deteriora- contaminants. Populations of the aerobic mesophilic
tion (92, 104). Postharvest moisture loss due to respira- bacteria ranging from 102 to 108 CFU/g of produce and
tion causes structural damage in fruits such as apples and bacterial contamination at levels of 105 to 106 CFU/g
wilting in leafy vegetables. A decrease in acidity and soft- have been reported for fruits (76, 89). Numbers of yeasts
ening is followed by cell wall degradation. Degradation and molds (although CFU quantification is probably not
of phenolic compounds during ripening of climacteric appropriate because of the possible presence of extensive
fruits is among the physiological changes that may favor mycelia or fruiting structures) ranging between 101 and
the invasion and growth of spoilage microorganisms. 106 CFU/g on vegetables and between 103 and 107 CFU/g
Fruits and vegetables have a high water content and on fruits have been reported (23, 76, 89). These varia-
contain significant amounts of nutrients essential for tions reflect the diversity of conditions prevailing during
microbial growth (Table 8.2). The main limitation pre- cultivation and postharvest storage and, to some extent,
venting growth of most bacteria is the low pH of fruits methods used for enumeration (89). With few excep-
(as low as 2.0 in some Citrus species). Spoilage of low- tions, gram-negative species of bacteria are dominant
pH fruits is restricted to molds and yeasts, which are on vegetables and basidiomycetous yeasts are among
more tolerant than bacteria to high acidity. The favor- the major species most frequently found on fruits (23,
able effects of basic nutrients in produce tissues may be 89). Postharvest spoilage microorganisms do not seem
balanced by compounds known for their antimicrobial to include a dominating species in sound fresh fruits and
activity, such as phenolic compounds and tannins in many vegetables (89). The dominating species of bacteria and

Table 8.2 Approximate pH values and water, protein, and sugar contents of some fresh
fruits and vegetablesa
Protein (g/100 g, Sugars (g/100 g,
Fruit or vegetable pH Water (g/100 g) fresh wt) fresh wt)
Asparagus 5.0–6.1 93.2 2.2 1.9
Beans (lima) 5.4–6.5 70.2 6.8 1.5
Broccoli 6.5 89.3 2.8 1.7
Carrot 4.9–6.3 88.3 0.93 4.5
Cauliflower 6.0–6.7 91.9 2.0 2.4
Corn (sweet) 5.9–7.3 76.0 3.2 3.2
Lettuce 6.0–6.4 95.6 0.9 1.7
Onion 5.0–5.8 88.5 0.9 4.3
Pepper (red) 5.3–5.8 92.0 1.0 4.2
Potato tuber 5.6–6.2 81.6 1.7 1.2
Spinach 5.1–6.8 91.4 2.9 0.4
Squash 5.0–5.4 94.6 1.2 2.2
Tomato (ripe) 3.4–4.7 94.5 0.88 2.6
Apple 2.9–3.3 85.6 0.26 10.4
Banana 4.5–5.2 74.9 1.1 2.4
Grape 3.4–4.5 80.5 0.72 15.0
Lime 1.8–2.0 88.3 0.70 1.7
Melon (cantaloupe) 6.2–6.5 90.2 0.84 7.8
Orange 3.6–4.3 86.8 0.94 9.4
a
From reference 75 and U.S. Department of Agriculture, Agricultural Research Service, 2005, National Nutrient
Database for Standard Reference, release 18. Nutrient Data Laboratory home page, available at http://www.nal.usda.
gov/fnic/foodcomp.
160 Microbial Spoilage and Public Health Concerns

yeasts are generally not known for their ability to cause to form biofilms (11, 86). Adhesion of molds to plant
decay on fresh produce, and when detected, spoilage surfaces involves very specific interactions which can
microorganisms represent only a low proportion of the involve lectins, hydrophobic contact with the plant cuti-
normal microflora. cle, or secreted adhesives (113). Fungal agents of stor-
Postharvest spoilage microorganisms may take many age diseases may then colonize a few cells in a limited
different routes to contaminate fruits and vegetables. area of plant tissue, followed by a delay before becoming
Seeds, including vegetable seeds, have been shown to be active under specific circumstances. This period without
the primary source of postharvest diseases such as Collet- growth is known as quiescence. Infection during the qui-
otrichum infections on pepper, onion bulb neck rot caused escent period can be symptomless, as that of germinated
by Botrytis spp., potato tuber soft rot caused by Erwinia spores of Colletotrichum spp. on various types of pro-
spp., and potato tuber gangrene caused by Phoma spp. duce, or can result in visible but nonexpanding symp-
(89). Rain water is a significant vehicle of microorganisms toms, as in ghost spot of tomato caused by B. cinerea
from plant to plant, plant to soil, and soil to plant through (95). The quiescent period can be observed while fruits
splashing (72). Irrigation water from different sources are still attached to the parent plant. For instance, initial
may contain pectolytic bacteria (100). Soil and its compo- infection of gray mold on strawberry or grape occurs at
nents (rhizosphere and plant debris) are the natural reser- blooming and remains dormant until fruit formation or
voirs of spoilage bacteria such as Bacillus and Clostridium during postharvest storage (3).
(74) and molds such as Sclerotinia spp. (63) and Rhizoc- Spoilage involves to some extent the penetration of
tonia solani (106), and facilitate survival, in particular in microbial cells into plant tissue. The cuticle barrier can
plant debris or on fruits and vegetables in contact with the be compromised by many postharvest spoilage microor-
rhizosphere, as shown for Erwinia carotovora (93). Air ganisms, e.g., B. cinerea on cucumber and tomato fruits,
and wind disperse spores or fruiting bodies of molds (38), Colletotrichum spp. on tomato and bell pepper (89), or
leaves and microorganisms adhering to leaves, and aerosol S. sclerotiorum on carrot (63). Despite evidence of cutin-
particles that may contain bacteria, presuming the acqui- ase activity of some fungal pathogens, the actual contri-
sition of some resistance to desiccation (72). Postharvest bution of these enzymes to tissue penetration remains
handling of produce has been shown to be a cause of con- controversial, some mutants unable to produce cutinases
tamination with spoilage microorganisms (89). Immer- still being pathogenic (3, 45). The formation at the tip
sion tank solutions have been suspected to be a cause of of the germ tube is an organ called the appressorium, a
redistribution of Phialophora malorum on pears (109). structure used by fungal pathogens to press against and
Wooden boxes previously used to store carrots may con- attach to plant surfaces in preparation for infection (24,
tain many species of molds pathogenic to carrot, includ- 82) and which contributes, for example, to the devel-
ing Rhizoctonia carotae, Sclerotinia sclerotiorum, and opment of ghost spots caused by B. cinerea on tomato
Botrytis cinerea, which can cause lesions on sound carrots fruits, to anthracnose caused by Colletotrichum in pep-
(64). The common postharvest spoilage molds B. cinerea per and by Colletotrichum musae in banana (3, 89), or to
and Penicillium spp. can be found at multiple sites in pro- spoilage of carrot caused by S. sclerotiorum (63). In the
duction areas, e.g., on fruits, in orchard litter and soil, case of bananas, appressoria formed on preharvest fruits
in orchard air, on packing lines, and in cold storage air; remain quiescent until harvest and fruit maturation to
however, in these cases, contamination on the fruit surface eventually penetrate into tissue and cause spoilage. Ger-
is critical to further decay (68). The inoculum can consist mination and appressorium formation by C. musae and
of spores or conidia, mycelium, or sclerotia. Mycelium is Colletotrichum gloeosporioides may be induced by eth-
generally infectious. Germination of spores and sclerotia ylene produced by the infected commodity (41).
generally depends on water, the presence of nutrients at Fruits and vegetables also offer a large diversity of natu-
least in small quantities that may have leached from fruit ral or accidental openings which can serve as ports of entry
and vegetable tissues, or juice released from damaged tis- for penetration of postharvest spoilage microorganisms.
sues in wounds (3, 89). Adverse conditions (e.g., wind, frost, and contact between
The time elapsed between contact of microorgan- fruits and limbs) in orchards, vineyards, and fields, as well
isms with the surface of the plant organ and initiation as harvesting and postharvest handling involving mechan-
of spoilage is extremely variable. Anyway, this process ical or human interventions, can result in wounding of
presumes the establishment of some sort of coloniza- fruits caused by stems of other fruits or abrasion during
tion on the organ surface. Fluorescent pseudomonads, transport (83). Up to 14% of hand-picked pears and up to
including pectolytic strains, form bacterial communities 30% of apples may present wounds after harvest (1, 108).
aggregated in a matrix of exopolymers and assimilated Wounds are a common site of penetration of postharvest
8. Fruits and Vegetables 161

spoilage microorganisms and are critical, for instance, for lamella and cell wall, resulting in tissue maceration, loss
infection of tomatoes by Rhizopus stolonifer, carrot by a of rigidity of plant tissue, and irreversible cell damage.
range of postharvest pathogens, and plums by Monilinia Polygalacturonase, pectin esterase, and pectin lyase are
fructicola (52, 73, 89). Natural openings, such as dead the main types of pectinases that act at different sites on
tissues at the blossom end, stem scars on apple and citrus, the d-galacturonic acid chain constituting the basic struc-
and the calyx of tomato, eggplant, and bell pepper, are ture of pectic compounds (47). Pectolytic bacteria and
also potential sites for microorganisms to penetrate tis- molds generally produce several types of pectinases, not
sues (83, 89). Soft-rot Erwinia spp. have been shown to always playing the same role (4). Eight isozymes of poly-
penetrate into potato tissue through lenticels under cer- galacturonase are produced by Phomopsis cucurbitae, a
tain conditions (hydrostatic pressure) (6). A higher density quiescent infection mold, during postharvest infection
of lenticels on various apple cultivars is correlated with and decay of cantaloupe, and their activity changes dur-
higher susceptibility to infection with Penicillium spp. (1). ing maturation of the fruit (121). An endopolygalactu-
The presence of free water in which bacteria may be sus- ronase mutant of Alternaria citri loses its ability to cause
pended can favor infiltration into plant tissues and their black rot on citrus, while a mutant of Alternaria alternata
internalization. This process can be enhanced by hydro- for the same function is still able to cause brown spot on
static pressure created by immersing warm fruits in cold citrus (55). Pectic enzymes are necessary for A. citri to
wash water. Internalization and infiltration have been well progress from the pedicel in the central axis of the citrus
described for human pathogens, but also occur for spoil- fruit to the sac juice containing nutrients (56).
age microorganisms (83). Some postharvest spoilage bacteria and molds also
produce toxins. Some molds, in particular Alternaria
spp., have very specific host-parasite interactions (30).
CAUSING THE DISEASE OR THE SPOILAGE Mycotoxins are also known for their toxicity to humans
Specific factors associated with virulence of microorgan- or animals. Implications of the nonspecific toxins pro-
isms are the primary mechanisms at the origin of post- duced by molds causing postharvest spoilage are highly
harvest spoilage of fruits and vegetables. Their role is variable (3). For instance, production of patulin by Peni-
usually evident when comparing, within species, certain cillium expansum does not appear to be involved in post-
strains able to cause spoilage to those unable to cause harvest spoilage of apples during storage. In contrast,
spoilage. A comparison of different strains of Pseudomo- the production of oxalic acid has been shown to occur in
nas fluorescens has shown, for instance, that the produc- a number of postharvest diseases caused by Sclerotium
tion of a biosurfactant (a peptidolipid named viscosin) is rolfsii or S. sclerotiorum (63, 97).
a key factor in the decay of broccoli florets caused by the In some instances, spoilage can be fully opportunistic.
bacterium, while pectolytic activity is necessary but not The physiological activity of minimally processed veg-
sufficient to cause spoilage (50, 51). The aggressiveness of etables is markedly changed by processing and storage
Mycocentrospora acerina is related to production of pec- conditions, e.g., modified-atmosphere packaging (88).
tinase and glucanase (67). Molecular biology techniques The lactic acid bacterium Leuconostoc mesenteroides is
enable the production of mutants with specific character- not known as a plant pathogen, but it has been shown
istics and tests to determine if these characteristics (and to be strongly associated with the spoilage of shredded
their genes) are necessary for microorganisms to cause carrots. In this situation spoilage is thought to be due to
spoilage. This approach has enabled researchers to dem- a shift toward anaerobic metabolism in modified atmo-
onstrate the role of pectinases in pathogenesis of soft-rot spheres with concentrations of CO2 in excess and/or to
bacteria. Mutants of the soft-rot bacterium Erwinia chry- low concentrations of O2 that induce toxicity to carrot
santhemi that do not produce pectin methylesterase or a cells and leakage of electrolytes and nutrients used by the
specific isoenzyme of pectate lyase have a reduced abil- saprophyte (15). On minimally processed green leafy sal-
ity to macerate potato tubers, whereas the production of ads, in contrast, spoilage can be explained by the devel-
another pectate lysase does not seem to be necessary for opment of pectolytic fluorescent pseudomonads (90).
tissue maceration (4). These enzymes also have different
implications in pathogenesis. Pectic enzymes are clearly
involved in postharvest spoilage of fruits and vegetables DEFENSE REACTIONS
by bacteria as well as by molds. Purified pectin-degrading Fruits and vegetables offer a range of barriers to infec-
enzymes are able to macerate plant tissue and cause cell tion by postharvest spoilage microorganisms. Some are
death without involving other enzymes or toxic factors. preformed or constitutive in the plant organ. The cuticle
Pectinases break down pectic components in the middle barrier is the most external barrier to penetration.
162 Microbial Spoilage and Public Health Concerns

Removal of waxes from the cuticle has been shown to Seedling apples produce benzoic acid in response to
increase the vulnerability of pepper fruits to infection by infection by Nectria galligena (110), and the resistance
Colletotrichum capsici and C. gloeosporioides, and of of carrot to B. cinerea has been attributed to a couma-
cabbage by B. cinerea (89). The cuticle thickness is corre- rin, 6-methoxymellein (46). Structural changes such as
lated with the resistance of tomato fruit or grape berries to accumulation of lignin or suberin or development of cal-
B. cinerea and of peaches to Monilinia fructigena (3, 43). lus have also been observed as defensive reactions (99),
In many types of produce the barrier effect of the cuticle in particular in carrot, potato tuber, and pear (5, 70,
is reinforced by epidermis or periderm tissues, which can 108). Polygalacturonase-inhibiting proteins produced
be relatively thick structures such as the rind of citrus and by plants and acting against endopolygalacturonases of
melons. Enhanced resistance to infection linked to these plant-pathogenic molds that cause wall degradation and
structures can be explained by their higher resistance to tissue maceration have been detected in a range of fruits
crack formation and therefore to penetration by spoilage and vegetables, including apple, pear, grape, raspberry,
microorganisms, larger amounts of protective material to onion, and pepper (25). An esterase produced during
be degraded, and lower diffusion or access to water and the interaction between mature peppers and C. gloeo-
nutrients required for the infection process (3). sporioides has been shown to inhibit the formation of
Preformed antimicrobial compounds may also be the fungal appressoria, and therefore decay (61). Pro-
involved in plant resistance, although demonstration duction of reactive oxygen species (including H2O2) is
of their actual effects on resistance is relatively difficult induced after infection of pepper by C. gloeosporioides.
to achieve because of difficulties in assessing inhibi- The direct inhibitory effect of these species on the patho-
tory activity and in correlating changes in concentra- gen still remains unclear, but they likely are at the ori-
tions with decay development, as critically underpinned gin of activation of the phenylpropanoid pathway and
by Prusky (95). These compounds can be extremely accumulation of the antifungal compound diene, both
diverse, as fruits and vegetables cover a wide range of implicated in resistance (7). Other mechanisms impli-
plant families. The presence of the phenolic compounds cating enzymatic activities (chitinases and lipoxygen-
catechol and protocatehuic acid has a role in the resis- ases) or accumulation of hydroxyproline-rich proteins
tance of onion to Colletotrichum circinans, and the have been proposed (3, 89). Interactions between plants
presence of the alkaloidal saponin tomatine has a role in and pathogens are a very fertile research area, and novel
the resistance of tomato to B. cinerea (3, 78). Decreases defense mechanisms are regularly discovered.
in concentrations of 5,12-cis-heptadecenyl resorcinol
and 5-pentadecenyl resorcinol in the skin of mango fruit
during ripening are related to an increase in susceptibil- CONTROLLING SPOILAGE
ity to A. alternata, lower decreases occurring in the most
resistant cultivars (95). Quiescent infections of C. gloeo- Effects of Temperature, Relative Humidity,
sporioides may be regulated by preformed epicatechin and Modified Atmosphere
acting as an inhibitor of lipoxygenase activity and con- Postharvest control of temperature, relative humidity,
sequently delaying degradation of an antifungal diene and composition of the gaseous atmosphere aims at
present in unripe avocado fruit (95). A monoterpene reducing the physiological activity of fruits and vegeta-
aldehyde, citral, present in particular in the oil cavities bles by delaying ripening and senescence, consequently
of citrus albedo, is thought to be involved in the resis- prolonging the shelf life. These environmental factors
tance of young mature green lemons to Penicillium (95). may act by giving less opportunity to the pathogen to
The resistance of carrot roots to infection by Mycocen- develop by retaining the integrity of the plant organ
trospora acerina could be attributed to falcarindiol, a and by directly inhibiting microbial growth. The most
polyacetylene compound which accumulates in the peri- suitable temperature, relative humidity, and modified
dermis and the pericyclic parenchyma of the root tissue atmosphere for preserving the quality of most fruits and
at concentrations 50-fold higher than in the core xylem vegetables are now relatively well established and have
parenchyma, and which has a pronounced inhibitory been extensively reviewed (3, 87, 101). A decrease in
effect on molds (44). temperature by 10C reduces the respiratory activity by
A wide range of defensive barriers may also be two- to fourfold (59), and a temperature close to 0C
formed in reaction to an infection. Pathogens are known is recommended for most commodities, with the excep-
to produce elicitors which stimulate these reactions of tion of those of tropical origin and a few temperate pro-
defense (48). Plant tissues may produce small molecules duce which suffer from physiological disorders (chilling
of varied nature, known as phytoalexins (78). Bramley’s injuries) when stored at refrigeration temperatures. For
8. Fruits and Vegetables 163

these produce, the optimal storage temperature is close high humidity is generally in favor of the produce. Stor-
to 10C. While growth of many postharvest spoilage age of cabbage, celery, leek, and carrot at 98 to 100%
microorganisms is still possible at very low refrigera- relative humidity instead of 90 to 95% relative humidity
tion temperature, their rate of development is reduced. results in lower losses caused by decay (114). Thorne
In addition, some major spoilage microorganisms are (111) observed that spoilage due to R. stolonifer only
inhibited at refrigeration temperatures at which produce occurs on carrots that have lost more than 3 to 8% of
is often stored. The bacterium Erwinia carotovora subsp. their fresh weight.
carotovora cannot grow at temperatures below 6C, and
the molds Rhizopus stolonifer, Phytophtora infestans, Physical Treatments
and Aspergillus niger cannot grow below 2 to 5, 4 and The possibility of using ionizing radiation to extend the
11C, respectively (3, 66, 74). shelf life of fruits and vegetables has been studied since
Modified atmospheres, used in combination to chill the 1950s (3). Postharvest spoilage bacteria and fungi
storage, also reduce the physiological activity of fresh are sensitive to ionizing radiation. Doses lower than 4
produce. For instance, reducing the O2 concentration kGy reduce 1,000-fold the germination of major post-
from 21 to 2.5% during storage of broccoli florets results harvest pathogens such as Penicillium spp., Monilinia
in approximately a 50% decrease in respiration rate (59). fructicola, R. stolonifer, and Alternaria spp. Decimal
Recommended CO2 concentrations for produce storage reduction doses for Pseudomonas and other gram-
rarely exceed 10%, and 1 to 5% O2 is tolerated (59, 60, negative bacteria are approximately 0.2 kGy (3, 91).
101). Exposure to higher CO2 (lower O2) may result However, it appears that some fruits and vegetables are
in physiological disorders, leading to a loss in quality. adversely affected by doses necessary to inactivate some
As reviewed by El Goorani and Sommer (37), either a postharvest pathogens (81). Treatment with ionizing
reduction in O2 or an increase in CO2 delays in vitro radiation is consequently useful to lower initial contam-
growth of many postharvest pathogens, without com- ination or to inhibit growth of postharvest pathogens
plete inhibition. Modified atmospheres reduce micro- and to prolong the development of disease, but not for
bial spoilage of fruits and vegetables in many instances, complete elimination of microbial contaminants. In the
although some spoilage microorganisms are not directly case of strawberry, a 2-kGy dose prolongs the shelf life
inhibited. For example, controlled-atmosphere storage by several days (3).
of apples at 1C in an atmosphere containing 5% CO2 Prestorage heat treatment has been used with some
and 3% O2 prevents the development of lesions due to success to reduce postharvest spoilage of a wide range
Pezicula alba, while in vitro, this gaseous atmosphere of fresh produce, including green pepper, apple, and cit-
has no effect (10). Generally, high concentrations of rus (39, 103). Two main types of applications can be
CO2 or low concentrations of O2, often less than con- distinguished: short-term exposure to heat, from a few
centrations tolerated by the plant organs, are needed for seconds at 60 to 62C to 60 to 120 min at about 45C,
a significant reduction of in vitro growth. The lower sus- by immersion in or rinsing and spraying with hot water;
ceptibility to postharvest pathogens of fruits and vegeta- or long-term exposure, also known as “curing” (for a
bles stored under controlled atmospheres is mainly due few hours to several days), mainly in hot air (103). Two
to delayed senescence (37). However, modified atmo- kinds of effects can be observed, either direct effects on
spheres can increase the extent of diseases in potato microbial contaminants or indirect effects on the treated
tubers, carrot, and other root crops. Reduced physi- fruit or vegetable. Treatment of tomatoes for 3 days at
ological activity slows wound healing, giving pathogens 38C before storage at 20C for 7 days results in inhi-
additional time to establish infections. Under extreme bition of B. cinerea without alteration of quality (40).
conditions (anoxia), strict anaerobes such as pectolytic Short-time exposure at 56C delays germination of
soft-rotting clostridia may cause spoilage, as shown on Penicillium digitatum, and treatment at 59C and 62C
potato, for instance (74). inhibits germination (94). A similar level of sensitivity
Water loss is a consequence of respiratory activity to heat has been observed for M. fructicola, B. cinerea,
and is highly detrimental to quality. A 10% weight loss Cladosporium herbarum, R. stolonifer, A. alternata, and
(much less for leafy vegetables) makes produce unac- Penicillium expansum (3). These treatments also reduce
ceptable to consumers (8). Storage at relative humidity 1,000-fold the natural epiphytic microflora on citrus.
higher than 90% is recommended for most commodi- On whole citrus, hot water treatment of fruits artificially
ties, with a few exceptions, e.g., garlic and onion (87). inoculated with P. digitatum has been shown to result in
High humidity also increases the availability of nutrients less than 20% decay after 4 days at 24C, while all con-
to spoilage microorganisms. Despite this, storage under trol fruits are spoiled (94).
164 Microbial Spoilage and Public Health Concerns

Chemical Treatments result in better retention of quality during storage (89).

For example, prestorage chlorine application at suitable
Fungicides concentrations and for appropriate exposure times results
Synthetic antimicrobial chemicals are still widely applied in significant reductions of black spot disease caused by
to fruits and vegetables after harvest, with substantial dif- A. alternata on persimmon (96) and spoilage of nectar-
ferences among countries, according to approval by legal ines and plums caused by Monilinia laxa (79). Sanitizers
authorities, as complements to modifications of storage are also useful to prevent buildup of microorganisms on
environments or when modifications are not possible. equipment and in wash water, and to avoid dissemination
Their spectrum of activity and possibilities of applica- from contaminated material.
tions have been reviewed elsewhere (31, 32, 80) and in
chapter 33. These chemical compounds have different Triggering Defense Reactions
modes of action. For instance, benzimidazoles have a As stated above, fruits and vegetables, as any plant
systemic action beneath the surface of the host as well as organ, have intrinsic defense mechanisms that respond
antisporulant properties, and they inhibit tubulin assem- to postharvest microbial infection. The induction of
bly and therefore mitosis (22). Imalazil belongs to a fam- these mechanisms may be systemic, as shown for carrot
ily of systemic fungicides that inhibits the biosynthesis roots, in which inoculation with B. cinerea and S. sclero-
of ergosterol, an essential compound in the membrane tiorum near the root tip reduces infection after subse-
of fungal cells (3). Dicarboximides alter the osmotic quent inoculation with these molds near the crown (84).
adaptation capabilities of molds (118). Concerns about A range of biotic and abiotic factors that induce defense
toxicity limit official authorization for use of fungicides reactions have been shown to improve quality retention
on defined commodities and create some negative atti- during storage. Appropriate fertilization of plants with
tudes among consumers. In addition, the occurrence of calcium or direct application of a calcium solution on
fungal resistance restricts possible applications of fungi- the fruit and vegetable to be stored increases the calcium
cides. Alternative chemicals such as compounds generally content in tissue. These treatments have been successful
recognized as safe or natural compounds, e.g., acetalde- in controlling the growth of P. expansum on apples and
hyde, hexanal, and essential oils, have also shown anti- soft rot caused by E. carotovora subsp. atroseptica on
microbial properties suitable for postharvest control of potatoes. The effect of calcium is attributed to a rein-
phytopathogens (112). The presence of the essential oil forcement of pectin bonds, making the cell wall more
carvone in the storage atmosphere has been shown to resistant to pectic enzymes, or a direct inhibitory effect
reduce, for instance, decay of potato tubers inoculated of calcium on the enzymes themselves (21). Accelerating
with Phoma exigua and Fusarium sulphureum but not healing of wounds by suberization of parenchyma cells
with Fusarium solani (49). as a result of exposure at moderately high, ambient, or
low temperatures is a common procedure to reduce post-
Decontamination harvest spoilage of carrot, potato, sweet potato, onion,
Decontamination aims at reducing the number of micro- yam, and pear (17, 40, 73, 89). Heat treatment may also
bial contaminants on the surface of fruits and vegetables, induce structural changes in the epicuticular wax (result-
thereby prolonging the time required to develop spoilage. ing in fewer cracks and therefore offering a mechanical
Various chemicals have been tested for possible applica- barrier to spoilage microorganisms), delay ripening, and
tion to fruits and vegetables. Hypochlorous acid (HClO), inhibit antifungal activity, phytoalexin production, or
chlorine dioxide (ClO2), ozone, and hydrogen peroxide chitinase and glucanase activities which play a role in
are among the most extensively studied chemicals (9). degradation (103).
Limitations in their action, in particular that of chlorine, Chitosan, a compound derived from chitin, has anti-
may be due to various factors: the natural hydropho- microbial properties when used, for instance, to coat
bicity of plant surfaces, inaccessibility of microorgan- fresh fruits and vegetables for the purpose of regulat-
isms within plant tissue, and possible neutralization of ing gas and moisture exchange (27, 115). In addition
the decontaminating agent upon contact with fruit and to damaging fungal hyphae, chitosan also reduces the
vegetable tissue components. Chlorine at concentrations ability of fungal pectic enzymes to macerate plant tis-
that will cause several decimal reductions of pathogens sue by a direct effect on enzymes and indirect effects by
in pure water is often slightly more efficient than wash- inhibiting the production of oxalic and fumaric acids by
ing produce with pure water (9, 88). Reduction in micro- pathogens. Chitosan may also inhibit host-specific toxin
bial populations is generally 1 to 2 log CFU/g. However, production, phytoalexin production, and structural
despite these limitations, application of disinfectants may changes in the cell wall, as shown in tomatoes infected
8. Fruits and Vegetables 165

by the black mold rot mold, A. alternata, or on bell pep- susceptible to one of the other pathogens. It has been sug-
per fruit infected by B. cinerea (33, 34, 98). Stimulation gested that Golden Delicious cultivar apples, despite their
of antifungal hydrolyses by chitinases and glucanases fragile epidermis, have a lower probability of wounding
has also been reported (115). than Granny Smith cultivar apples because of the par-
Plant regulators are involved in the development ticularity of their pedicels, stiffer and shorter in the latter
and response of produce to environmental stresses. Jas- case (107). Increased firmness in transgenic tomatoes with
monates in particular may play a role in plant defense reduced levels of polygalactoronase results in increased
responses to microbial attacks, and their postharvest resistance to Geotrichum candidum and R. stolonifer,
application on fruits is effective in controlling spoilage while tomatoes with suppressed expression of ripening-
molds such as M. fructicola and P. expansum on peaches related expansin, with a similar increased firmness, are
and P. digitatum on grapefruit (29, 119). This positive not more resistant to B. cinerea and A. alternata (12, 65).
effect has been attributed to induced resistance of the
fruit, in particular to higher activities of chitinase, glu- Biological Control
canase, phenylalanine ammonia lyase, and peroxidase Biological control refers to the application of microbial
(119). Similar activities or accumulation of antimicro- antagonists of postharvest spoilage microorganisms, in
bial compounds related to disease resistance are induced particular on wounds, which are a natural site of entry for
in peach, grapefruit, and carrot after exposure to UV-C pathogens. Biological control is based on the selection of
(28, 35, 85). However, UV-C has little activity against B. naturally occurring microorganisms particularly adapted
cinerea inoculated on wounded bell pepper, indicating to surviving or growing on fruit and vegetable surfaces.
that wounded tissues are protected against exposure to Many mechanisms are probably involved in interactions
irradiation, while complete inhibition has been observed between antagonists and pathogens (57). These may
in vitro (85). In this case and in grapefruit inoculated include antibiosis, such as the production of pyrrolnitrin
with P. digitatum, the induction of defense reactions was by Pseudomonas cepacia, shown to control blue mold rot
the likely cause of a lower rate of decay (28, 85). caused by P. expansum and gray mold rot caused by B.
cinerea on pome fruits (57), or Fusarium sambucinum
Resistance of Cultivars on potato (13). Competition for space and nutrients is
Cultivars within a plant species may exhibit differences in a seductive hypothesis to account for the antagonistic
susceptibility to infection by postharvest pathogens. Labo- effect of some biological agents, in particular on wound
ratory experiments and practical experience have enabled sites, which are often rich in nutrients. For instance,
a classification of potato cultivars according to their sus- the yeast-like Aureobasidium pullulans depletes amino
ceptibility to Phoma exigua, which causes gangrene (73). acids in vitro and inhibits germination of P. expansum
Tests done using artificial inoculation of produce show in apple juice (58). In addition to this possible competi-
some differences in susceptibility among cultivars, e.g., tion for space and nutrients, the antagonist A. pullulans
susceptibility of broccoli to Pseudomonas marginalis induces apple -1,3-glucanase, chitinase, and peroxidase
(14), onion to Aspergillus niger and Botrytis allii (62, 71), activity, which controls decay caused by B. cinerea and
pepper to C. gloeosporioides (77), and sweet potato to P. expansum (54). A similar induction process has been
Rhizopus spp. (18). Resistance of cultivars to spoilage is shown with the antagonist yeast Candida saitoana (36).
usually only delayed, not prevented (89). Evaluation of Induction of these defense reactions could explain the
cultivars may depend on testing procedures: the ranking effect of antagonists against postharvest pathogens.
of potato cultivars for resistance against E. carotovora Some yeasts also show a strong attachment to the hyphae
subsp. atroseptica, for example, depends on the inocula- of postharvest spoilage molds and production of cell
tion method (2). The resistance of cultivars to postharvest wall-degrading enzymes (116). Some of these biocontrol
spoilage microorganisms may be the result of several com- agents, in particular strains of the yeast Candida oleoph-
plex factors and interactions. A low number of pores and ila and of the bacterium Pseudomonas syringae, are reg-
thickness of external grape berry skin, for example, are istered as biopesticides in the United States (42).
positively correlated with resistance to B. cinerea, both The diversity of biocontrol methods illustrates the
factors likely giving a general protection against fungal diversity of possibilities based on a direct control of
attack (43). In contrast, among 12 apple cultivars tested pathogen contamination or of its development, indirect
for resistance against B. cinerea, P. expansum, Mucor piri- control through delayed senescence and preserved integ-
formis, and Pezicula malicorticis, no single cultivar was rity of the organ host, or induced defense mechanisms.
the most resistant to all pathogens and each cultivar that Under practical conditions, delay of spoilage depends on
was the most resistant to one pathogen was also the most combinations of several factors: prevention of wounding
166 Microbial Spoilage and Public Health Concerns

during harvest and postharvest handling, cold storage References

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which can be successfully stored for several months and postharvest environments: consequences for decay devel-
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