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Food Spoilage: P. R. Hayes (Ed.), Food Microbiology and Hygiene © Springer Science+Business Media Dordrecht 1995
Food Spoilage: P. R. Hayes (Ed.), Food Microbiology and Hygiene © Springer Science+Business Media Dordrecht 1995
FOOD SPOILAGE
3.1. INTRODUCTION
in due time become contaminated via the visceral blood supply. Total
bacterial counts for freshly cut meat surfaces are likely to vary between
103 and 10 5 organisms per cm2 . These organisms are derived mainly
from the exterior and the gut of the animal but also from knives, other
utensils, butchery tables, etc. so that variations in count often reflect the
hygienic conditions in the abattoir (Nottingham, 1982).
The storage temperatures used have a profound effect upon the mi-
crobial spoilage pattern and this aspect is now discussed in some detail.
20·C
10·C
2 34 5 7 10 14 21
Storage periods (days)
Fig. 3.t. Numbers of bacteria on fresh minced beef stored at different temp-
eratures.
112 Food Microbiology and Hygiene
7 14
Storage time (days at SOC)
7 14
Storage time (days at SOC)
Fig. 3.2. Growth of bacteria on fresh meat stored at SOC showing: (a) increas-
ing proportion of pseudomonads ( - - , total count; - - - -, Pseudomonas);
(b) changes in total psychrotrophs (- - - -) and total mesophiles (---).
The principal ingredients of any traditional cure are salt, sodium nitrate
and sodium nitrite although nitrite has largely replaced nitrate in order
to effect greater control over the processing; other possible additions to
the cure include sucrose and ascorbic acid. Curing can be combined
with other processing techniques including smoking, heating and fer-
mentation so that so-called cured meats include a variety of diverse
products prepared under different conditions (Egan & Roberts, 1987).
Salt is present as a preservative agent which acts by reducing the aw of
the meat. Pseudomonas spp., important in the spoilage of refrigerated
meats, are particularly sensitive to lowered aw levels (see Table 1.5) and
this partly accounts for the relative stability of cured meats. The role of
nitrate in spoilage control is not entirely clear although it proved useful
in the development of the red colour of meats and it is reduced by bac-
teria to nitrite. The nitrite also helps to maintain the colour and flavour
of meat but its principal role is to prevent growth of germinating spores.
On its own nitrite is not very active but its effectiveness is enhanced by
factors such as the salt concentration, pH and storage temperature
which are all important in determining the stability of cured meats.
of the body. In both these injection procedures the meats are subse-
quently immersed in a pickling brine. To increase the shelf-life of the
bacon a dry salting process has been developed whereby bacon sides are
passed through a cloud of dry salt after removal from the curing brine
(Gardner, 1983).
Bacon, ham, salted beef and salted pork are the principal non-com-
minuted meat products of the curing process and of these only the first
will be considered in any detail.
are killed on the bacon during the smoking process, the numbers de-
pending on the time and type of smoking (Handford & Gibbs, 1964).
Micrococci, yeasts and moulds are most frequently isolated on smoked
bacon although lactic acid bacteria are more likely to predominate
where liquid smoke is used. As these 'lactics' are responsible for a less
offensive souring spoilage than that associated with micrococci, and at a
later stage, the shelf-life of the product is extended.
3.3.3.3. Ham
The processes involved in the curing of hams are similar to those em-
ployed for bacon except that sugar is often added to the cures. This can
be attacked by bacteria, particularly lactobacilli, and the fermentations
produce souring of various types; however, it has been suggested that
lactobacilli may be useful in maintaining the stability of curing brines
by preventing excessive pH rise.
In general the microorganisms found on hams are similar to those on
bacon and the flora consists mainly of micrococci, streptococci and lac-
tobacilli, the proportions depending on the salt concentration and
period of storage. Higher salt concentration hams also tend to support
the growth of a greater proportion of yeasts and possibly moulds.
The packaging materials used by the food industry vary from the highly
impermeable, required for vacuum-packaging to the highly permeable
and from the opaque to the transparent. Materials may consist either of
single components such as polyethylene (polythene) or polyvinyl chlo-
ride (PVC), or of multiple components where the properties required of
the packaging material cannot be satisfied by a single film. Where multi-
ple components are used the packaging material is either made up of
laminates of two or more films, or are prepared by co-extruding poly-
mers together to form a single material. An example of a multi-compo-
nent packaging material is Cryovac BBI which is constructed from a
triple co-extrusion of EVA/PVDC/EV A (Taylor, 1985). Alternatively
coatings may be applied to one or both sides of a single component film
such as cellulose.
Most films are relatively impermeable to water vapour since weight
losses must be minimized during storage. Gas permeability may vary
enormously (see Table 3.1) and this can have a dramatic effect on the
shelf-life and spoilage characteristics of the packaged meat.
Table 3.1
Permeabilities of Meat Packaging Materials to Oxygen and Carbon Dioxidea
Material Permeabilitl to
-I
110,,1
E
-'" ~
f!
C> 7
1 08
~
ti
'"
.c 106
'0
a; J:
.c
E 104 Co
::J total count
z impermeable pack
10 2 - - - -------------------, ---0- __ - - _ - - - - -
---+5
7 14
10 10
108
E
-'...5i
I!
«I
106
ti
'"
.c
'0
Iii
.c 10 4 ~
E I
----- -----
:::I I
Z
Pseudomonas
10 2 ---,---~.--
7 14
Mature bacon normally carries 104_10 6 bacteria per cm 2 which are sub-
sequently distributed over the cut surfaces at the time of slicing before
packaging. Because of this the bacon has a rather limited shelf-life but it
can be stored at ambient temperatures without loss of colour.
As previously mentioned, micrococci are the main types of bacteria
developing on the higher salt concentration bacons when stored at am-
bient temperatures (ea 20°C). When such bacons are stored under vac-
uum-packaged conditions, micrococci still predominate and rise in
numbers to about 107 per gram after about 9 days' storage. Obvious
spoilage is apparent at about 2 weeks and is characterized by a rancid
odour. With lower salt concentration bacons the flora is initially more
mixed but counts again reach a peak at about 9 days and spoilage fol-
lows a few days later. At this time the flora consists of approximately
equal proportions of micrococci, streptococci (i.e. enterococci) and
other lactic acid bacteria (i.e. lactobacilli and leuconostocs) (Tonge et
al., 1964); with high storage temperatures (e.g. 25°C) Gram negative
forms (e.g. Vibrio spp. and Proteus spp.) may be responsible for putre-
factive spoilage.
The advantage of chill storage is that it takes between 3 and 4 weeks
for bacterial counts to reach a maximum and hence obvious spoilage may
be delayed for up to 5 weeks. Qualitatively the changes in the microbial
flora during storage reflect the influence of salt concentration and the
flora is essentially similar to that for vacuum-packed bacon stored at
higher temperatures (Dempster, 1972; 1973); lower temperatures tend to
f?vour the growth of lactic acid bacteria at the expense of micrococci, a
tendency that increases with smoked bacon (Gardner, 1983).
with time and this may create problems. Greater control of the atmo-
sphere can be achieved by periodically refilling the containers in order
to maintain the desired blend of gases; this is known as controlled at-
mospheric packaging (CAP).
Because of the inhibitory effects of carbon dioxide on the growth of
bacteria it is always included in gas blends but it can also be used in the
pure form. In the latter case, although causing discolouration of fresh
meats, carbon dioxide considerably extends the shelf-life of even retail
cuts of meat. Thus Blickstad & Molin (1983) found that pork could be
stored in carbon dioxide for up to 3 months at O°C (cf 2 weeks for aer-
obic packs) before obvious spoilage occurred, this being associated with
a total dominance of lactobacilli (10 7 per cm 2). The colour changes in-
duced in red meats in particular caused by too high a concentration of
carbon dioxide can be offset by blending the gas with either oxygen (e.g.
40% CO 2 : 60% O 2) or oxygen and nitrogen, the latter being used solely
as a 'filler' gas. It must be remembered that with MAP oxygen levels de-
crease during storage whilst carbon dioxide levels increase due to the ac-
tivities of respiratory enzymes. Bartkowski et al. (1982) found that a
minimum initial concentration of 15% carbon dioxide was critical in
controlling bacterial growth. They concluded that a gas blend of 75%
O 2 : 15% CO2 : 10% N2 was ideal since it also gave the best appearance
to the raw steaks used in the experiments; in fact the recommended mix-
ture was far more effective than conventional vacuum-packaging in con-
taining bacterial growth.
Since MAP foods are necessarily stored at refrigeration temperatures
questions have been asked about the growth of psychrotrophic
pathogens such as Clostridium botulinum type E, Listeria monocytogenes
and Yersinia enterocolitica. C. botulinum poses a special problem in
packaged fish (see Section 3.6.7) but the other two pathogens are often
isolated from meats. It would appear that both are capable of growth in
certain gas mixtures but not in others, and that various other factors
such as the pH and exact storage temperatures also playa significant
part as growth determinants. In a detailed review Farber (1991) pointed
out that MAP foods often suffer from the weakness that the growth of
aerobic spoilage bacteria is inhibited so that spoilage is not apparent
and yet growth of pathogens could have occurred; furthermore, the
extended shelf-life of MAP foods may allow dangerous levels of
pathogens to develop. There is some cause for concern, therefore, at the
rapid proliferation of MAP products and Farber correctly concludes
that additional research is needed to ensure their microbiological safety.
126 Food Microbiology and Hygiene
The term 'poultry' applies to a range of domestic fowls and whilst the
following paragraphs deal exclusively with chickens, the general princi-
ples apply to other commonly eaten fowls such as turkeys and ducks. In
addition, the discussion will be limited to commercially dressed chickens
since this is now accepted as the normal method of marketing.
When live birds are brought into the processing plant they are harbour-
ing large numbers of microorganisms of many different types on their
feathers and feet and in their intestines. The various stages in processing
Food Spoilage 127
RiZciZption of
liviZ birds 1----+1
are outlined in Fig. 3.5 and only those that have a significant effect on
the microbiological condition of the carcass will be discussed.
Scalding to loosen the feathers is performed by immersing birds for
30 s in a tank of hot water (ca 55°C). There is a reduction in the num-
bers of organisms on the carcass due to the washing effect and to the
destruction of heat-sensitive bacteria including, in particular, psy-
chrotrophic spoilage bacteria. Even where lower scalding temperatures
are needed (ca 50°C for birds to be air chilled; see below) psy-
chrotrophic bacteria are still destroyed.
The mechanical feather pluckers increase the bacterial load on the
skin of birds and also cause cross-contamination and 'aerosol' prob-
lems, particularly with Staphylococcus aureus. Evisceration also in-
creases the bacterial load on the skin by spreading faecal types onto the
surface; such bacteria can be easily transferred to other carcasses again
causing cross-contamination problems.
The microbial load reaches a maximum immediately prior to spray
washing. This process, which produces an approximately 90% reduction
of microorganisms per carcass, is followed by chilling which is per-
formed by one of two methods (Mead, 1982). In the first of these im-
mersion or 'spin' chillers are used. The system utilizes one, two or three
128 Food Microbiology and Hygiene
tissue and in the intestines from 102 to 109 per ml contents (Hobbs,
1983). However, it has been claimed that numbers of bacteria on fish in
unpolluted waters are at the lower end of the ranges quoted and that
the higher numbers result from the poor hygienic standards on board
ship during initial handling (Huss et al., 1974).
Variations in marine environments affect the types of bacteria present
on the skin and gill surfaces of newly caught fish. Thus in the cooler seas
of the northern hemisphere the bacterial flora is dominated by psy-
chrotrophic Gram negative rods. Liston (1957) found the flora of North
Sea flatfish (skate and lemon sole) to comprise PseudomonaslAlteromonas
spp. (60%), AcinetobaeterlMoraxella (14%) with the majority of the re-
mainder being other types of Gram negative rods; Georgala (1958) found
that the bacterial flora of North Sea cod consisted of pseudo monads (44%)
and acinetobacters (32%) together with a variety of miscellaneous types. In
the warmer waters off India, the east coast of South Africa, Australia and
in the Adriatic the proportion of mesophiles increases and micrococci and
coryneforms become more important. Thus Gillespie & Macrae (1975)
found micrococci (49%) to predominate on newly caught fish from
Australian waters; pseudomonads (only 18%), coryneforms (12%) and
acinetobacters (only 9%) were the other main groups isolated.
3.6.2. The Effect of Initial Processing and Storage in Ice on Board Ship
Gutting of the fish on board ship tends to spread the intestinal flora
over the surface of the fish. The principal organisms found in the in-
testines are Vibrio spp. although many other genera are represented.
Fish are then washed in sea water and either packed in crushed ice or
possibly frozen or stored in refrigerated sea water. The conventional
method is to pack in ice and the effects of this form of storage will be
considered in some detail.
The ice in which the fish are to be preserved is itself usually contami-
nated (ea 103 per ml of ice melt water) and, in addition, the holds of the
fishing vessels normally have an indigenous flora composed of
Pseudomonas and Acinetobaeter spp. (Shewan, 1961). Thus when fish,
which already contain a fairly high proportion of pseudomonads and al-
teromonads, are placed in ice they are likely to be further contaminated
with these organisms. Partly because of the relatively high proportion of
pseudomonads and alteromonads on the skin of fish initially and in the
storage environment and partly because of the relatively high pH of
many fish species, spoilage in ice is relatively rapid. Even under the best
Food Spoilage 131
8
Q)
~::>
E 7
E
!!!
-E
0>
::>
6
0
u
5
~
tlto
.Q
4 d
0>
0
...J
3
2 4 6 8 10 12 14 16
Time In days
Fig. 3.6. Numbers of bacteria on fresh fish stored at: (a) 25°C, (b) 7°C,
(c) O°C, and (d) -4 0 c.
After landing, the fish may be left on the quay for many hours un-iced in
boxes or kits. Under such conditions the temperature of the fish will rise
132 Food Microbiology and Hygiene
Two basic techniques may be used in the salting of fish, either 'dry' or
'wet' (i.e. pickling) salting. In the former, used for non-fatty fish like
cod, salt is spread over the surface of the fish and layers of fish are in-
terspersed with salt layers. In wet salting, used for fatty fish like herring,
the fish are pickled in salt in barrels; combined 'dry' and 'wet' salting
can be used and variations in the latter can be achieved by using suit-
able spices and vinegar.
Irrespective of the method used, the added salt lowers the aw of the
fish and, as has already been discussed with cured meats, this has a pro-
found effect on normal spoilage patterns. The normal predominantly
Gram negative flora of fish is relatively sensitive to high salt concentra-
tions and so bacterial numbers decline. The final flora depends on the
134 Food Microbiology and Hygiene
Before smoking, fish are gutted and given a preliminary salting treat-
ment, the NaCI concentration depending on the level desired in the fish.
The salting is often comparatively light and the preservative effect is
therefore minimal as with, for example, finnan haddock, kippers and
smoked cod. With certain fish such as 'red' herring and smoked salmon
the salting is more extreme and may playa significant part in preserving
the fish (Cutting, 1965).
Following brining the fish are either 'cold' or 'hot' smoked. In the
commonly used cold smoking process the temperature of the fish should
Food Spoilage 135
not exceed that at which protein is denatured (ea 30°C). The process,
which includes both drying of the fish and impregnation with wood
smoke, results in some reduction in the numbers of bacteria caused
mainly by phenolic substances present in the smoke; overall, however,
the effects of this process on the microbial flora are insubstantial
(Graikoski, 1973).
In hot smoking, used for specialized canned products such as brisling
(sprats) and sild (small herring) the temperature of the fish is raised to
ea 70°C for 30 min so only the more heat-resistant bacteria should sur-
vive; the microbial flora therefore consists mainly of micrococci and
Bacillus spp.
The microbiological changes occurring during the storage of smoked
fish have not been studied in any detail but the predominant microor-
ganisms at the time of spoilage depend largely on the processing condi-
tions. With lightly brined cold smoked fish pseudomonads may become
the major group but a slight increase in salt concentration could result in
micrococci predominating. One of the most common causes of spoilage
of smoked fish is due to mould contamination; both Penicillium and
Aspergillus spp., which grow readily at chill temperatures, are present in
the sawdust used for smoke production and may subsequently develop in
the stored fish. With the higher salt concentration smoked fish storage
life is prolonged for several weeks or months even at high ambient tem-
peratures and little change in the microbial flora is likely to occur.
3.6.8. Shellfish
3.6.8.1. Crustaceans
Freshly caught shrimps are highly perishable due to bacterial and enzy-
matic activity. Bacterial activity is enhanced by the high content of
amino acids but autolytic enzymes (proteases) cause rapid breakdown
of protein providing bacteria with an ideal growth substrate. Because of
their perishable nature, shrimps are preferably either frozen or boiled as
soon as possible after catching but storage in ice is also common. The ini-
tial flora is similar to that of freshly caught fish (Hobbs & Hodgkiss,
1982); storage in ice results in an increase in the proportion of
AcinetobacterlMoraxella species which account for over 80% of the flora
at the time of spoilage (Fieger & Novak, 1961). This group does not ap-
pear to be responsible for bacterial spoilage, however. The active spoilers
are Alteromonas spp. and pseudomonads play a relatively minor role in
the spoilage of this food; spoilage is accompanied by increases in ammo-
nia, trimethylamine, hypoxanthine and acetic acid (Van Spreekens, 1977).
Autolytic enzymes are particularly active in lobsters and this makes
them another highly perishable food. Use is made of these enzymes in
conditioning which involves storage in ice for 2 to 4 days. As with
shrimps, further storage results in a pronounced increase in the propor-
tion of acinetobacters and moraxellas which are predominant at the
time of spoilage (Walker et al., 1970); this is characterized by increasing
concentrations of ammonia and trimethylamine. Crab meat, too, spoils
rapidly and crabs are therefore cooked in boiling water immediately
after capture. Studies on crabs have concentrated on the bacteriology of
the cooked meat: it is likely that the flora at the time of spoilage is
again dominated by acinetobacters and related species.
Food Spoilage 137
3.6.8.2. Molluscs
Of the bivalve shellfish eaten by humans, oysters, scallops and mussels
are probably most frequently consumed. The main microbiological
problem associated with these foods is the hazard of food poisoning re-
sulting from the not infrequent pollution of their growth habitat (see
Chapter 2). It is therefore necessary to cleanse these foods by depura-
tion with chlorinated or ozonated water, with water disinfected by
iodophors or, as is practised in the United States, with water disinfected
by uv light (Richards, 1988). The natural flora of the bivalves may thus
change extensively during treatment, and spoilage characteristics may
vary depending on the efficiency of the cleansing process. An important
feature of bivalves is the significant amount of carbohydrate (3-6%) pre-
sent in their flesh and this can influence the type of spoilage obtained. If
fermentative bacteria such as Escherichia coli and other coliforms are
not removed during the cleansing process, spoilage is primarily one of
souring, acids being formed by the dissimilation of the carbohydrate
(Fieger & Novak, 1961). With properly cleansed molluscs held at chill
temperatures the spoilage is totally different and is associated with in-
creases in volatile bases and hypoxanthine, and the flora is now domi-
nated by AcinetobacterlMoraxella spp. (Thomson et al., 1974).
3.7.1. Milk
widespread, it having been shown that some 30% of British dairy herds
suffer from mastitis (Wilson & Richards, 1980); some control of this in-
fection can be achieved by applying a disinfectant to the teats after
milking and by antibiotic treatment of non-lactating cows.
As milk is an ideal growth medium for bacteria it is essential to cool
it as rapidly as possible. The introduction of refrigerated farm bulk milk
tanks over the last 25 years, coupled with the bulk collection of milk in
refrigerated tankers, has markedly influenced the bacteriological quality
of raw milk supplies. The major effect of this change has been to reduce
the quantity of milk spoiled by souring (Murray & Stewart, 1978).
Souring of milk at normal temperatures is caused by the lactic acid bac-
teria which grow mainly at temperatures above lO°C. These bacteria
produce lactic acid from the milk sugar (lactose) which induces a sour
flavour and later coagulation of the milk. Most lactic acid bacteria are
killed by pasteurization but a few (e.g. Streptococcus thermophilus) are
thermoduric and can cause post-pasteurization problems.
With rapid cooling and refrigerated storage of milk the problems are
somewhat different. Nowadays it is the psychrotrophs, mainly pseu-
domonads, which are primarily responsible for spoilage problems.
Psychrotrophic bacteria, originally derived from soil and water, are
readily isolated from farm milking equipment, pipes and bulk milk
tanks (Thomas et al., 1971). Inefficient or delayed cooling of milk
markedly increases the proportion of psychrotrophs but growth of this
group continues more slowly at recommended storage temperatures for
raw milk (3-7°C). Counts of bacteria in bulk tanks vary from <104 to
106 per ml with a mean count of 20000 per ml of milk (Panes et al.,
1979); variations reflect the extent and type of contamination and stor-
age conditions but a large majority of the bacteria will be psy-
chrotrophs. A large proportion of these produce proteases and lipases,
and many of these enzymes are unaffected by pasteurization; in fact
both sets of enzymes are resistant to a heat treatment of 140°C for 5 s
and noc for 17 s (Griffiths et al., 1981). Defects due to proteases in-
clude bitterness and the principal spoilage effect of lipases is rancidity.
3.7.1.2. Pasteurization
This process involves heating the milk to a temperature high enough to
destroy all pathogenic bacteria such as Mycobacterium tuberculosis,
Salmonella spp. and Brucella spp; in so doing the large majority of other
bacteria including spoilage bacteria are killed and the keeping quality of
the milk is thus enhanced. Most of the milk produced in the UK is pas-
Food Spoilage 139
directly injected with steam under pressure. The sterile product is then
filled aseptically in special cartons (e.g. Tetra Pak) which are then heat-
sealed. UHT milk has an appearance, flavour and nutritional quality
similar to that of conventionally pasteurized milk and it should remain
acceptable for several months without refrigeration.
Spoilage of UHT milk can be caused occasionally by the growth of
spore formers, mainly Bacillus stearothermophilus or B. subtilis, the
spores of which have either survived the UHT process or are post-treat-
ment contaminants (Murray & Stewart, 1978). More commonly,
spoilage can result from the continuing activities of heat-stable pro-
teases and lipases produced by psychrotrophs in the raw milk before
processing; such spoilage can significantly reduce the theoretical shelf-
life. Enhanced deactivation of these enzymes can be achieved by
modification of the normal heating process; typically this involves a
dual heat treatment (l40°C for 5 s followed by 60°C for 5 min) which
can extend the shelf-life of the product by several months (Bucky et al.,
1988). Gelation of stored UHT milk, which may also be caused by a
chemical process, can also be attributed to proteases.
3.7.2. Butter
pH (4·5-5) is attained. The cream is then churned but is not salted be-
cause salt and acid react together to give undesirable flavours. Large
numbers (10 7 to 108 per gram) are required to produce the acid. An al-
ternative process involves the natural souring of cream which is then
pasteurized before churning.
Thus the microbial content of fresh butters varies considerably de-
pending on the manufacturing process, 'sweet' butters containing far
fewer microorganisms than 'sour' butters.
Spoilage of butter can be of microbial, enzymatic or chemical origin;
many undesirable flavours may stem from the cream itself but this as-
pect of spoilage is not considered here.
Microbial spoilage is caused principally by psychrotrophic bacteria as
butter is usually stored under refrigeration. Pseudomonads and related
Gram negative rods which enter the product post-pasteurization are
often responsible for rancidity caused by the hydrolysis of butter fat lib-
erating fatty acids. Putrid flavours and surface taint result from the pro-
teolytic activities of Alteromonas putrefaciens growing on the surface of
the butter. Moulds also grow on the surface producing discolouration;
commonly implicated are members of the genera Alternaria,
Cladosporium, Aspergillus, Penicillium, Mucor and Rhizopus. Rancidity
may be induced by lipases present in the cream and chemical reactions
include the oxidation of unsaturated fats.
3.7.3. Cheese
There are over 400 known vane tIes of cheese grouped into about 20
general classes. Most of these can be made from the same milk by vary-
ing the microorganisms, enzymes and salt added, and by changes in the
temperature during manufacture and curing.
Cheeses are classified by their texture or the degree of hardness and
two major groups of natural cheeses are recognized. The first group, the
ripened cheeses, vary from the very hard, low moisture content grating
cheeses (e.g. Parmesan), through the hard cheeses (e.g. Cheddar) to the
higher moisture content, semi-soft cheeses (e.g. Stilton) and soft cheeses
(e.g. Camembert). The second group of cheeses are the unripened soft
cheeses with a high moisture content such as cottage cheese.
Most cheeses are manufactured using the same basic processes.
Nowadays pasteurized milk is normally used but the ripening process
occurs more slowly and, because the natural flora has been largely de-
stroyed, 'starter' cultures of bacteria must be added to the milk. In the
142 Food Microbiology and Hygiene
3.7.4. Yoghurt
Fig. 3.7. Simplified diagram of a section through an egg shell showing: (a) cuticle,
(b) cuticular plug, (c) pore, (d) matrix, (e) outer shell membrane, and (f) inner
shell membrane (after Board, 1966).
Food Spoilage 145
Table 3.2
Egg Spoilage Caused by Bacteria
Although the contents of fresh eggs are usually sterile, commercially pro-
duced egg products (liquid, frozen and dried) used to be heavily contami-
nated with bacteria. In particular, evidence accumulated that liquid whole
egg was frequently contaminated with salmonellas that were well able to
withstand the comparatively low temperatures frequently used in the bak-
ing industry. Heat treatment regulations were introduced in the UK in
1963 and since then all liquid whole egg intended for distribution in the
chilled state, for freezing or for spray drying, has been pasteurized at
64AoC for 2Y2 min and then cooled immediately. The heat-resistant
salmonellas should be destroyed by this treatment and, in particular, S.
enteritidis phage type 4, should not survive (Humphrey et at., 1990); fur-
thermore the baking properties of the egg are not impaired (Shrimpton et
at., 1962). In addition, in the chilled liquid state pasteurized whole egg
can be safely stored for at least 6 days without significant increases in
bacterial counts although post-pasteurization contamination, mainly
caused by coliforms, must be avoided.
In Britain, liquid egg albumen is now conventionally pasteurized although
heat-treatment regulations have yet to be implemented. Untreated
chilled albumen is normally spoiled by pseudomonads and related Gram
negative rods whilst faecal streptococci and lactobacilli are predominant
in pasteurized albumen (Barnes & Corry, 1969). More severe heat treat-
ment is required to destroy salmonellas in liquid egg yolk but again
heating regulations have still to be introduced.
The low pH «4·5) of most fruits means that spoilage is caused mainly
by fungi. On the other hand, the pH range of most vegetables varies be-
tween 5·0 and 7·0 and thus spoilage may be caused by either fungi or
bacteria although the former are again the more important group. In
terms of their spoilage characteristics fungi are often somewhat arbitrar-
ily divided into two groups: the plant pathogens which infect the plant
before harvesting and the saprophytic fungi which attack the commod-
ity after harvesting. An important property of most spoilage organisms,
both fungal and bacterial, is their ability to secrete pectolytic enzymes
which soften and disintegrate plant tissues. Thus the growth of fungi
on fruits and vegetables usually results in severe tissue breakdown caus-
ing mushy areas; this spoilage is termed a 'rot'. The names given to
the different rots indicate the appearance of the food when spoiled.
Many of the more important forms of fungal spoilage are listed in Table
3.3.
An important cause of spoilage is Penicillium, many species of which
are able to attack fruit; perhaps as much as 30% of all fruit decay can
be attributed to this genus. Many fruit-vegetables such as tomatoes and
cucumbers, and vegetables such as potatoes and beetroots are also sus-
ceptible. Another important disease is Rhizopus soft rot which affects a
wide range of fruits and vegetables particularly during transit under
poor refrigeration. Harvested strawberries and potatoes are often at-
tacked and spoilage is indicated by soft, mushy areas with greyish
mycelium evident in the affected areas.
Table 3.3
Fungi Important in the Spoilage of Harvested Fruits and Vegetables G
Tree fruits
Apples and pears Blue mould rot Penicillium
Grey mould rot Botrytis
Peaches and plums Rhizopus soft rot Rhizopus
Soft fruits
Raspberries and Soft rot Mucor
blackberries Rhizopus
Green mould rot Cladosporium
Strawberries Grey mould rot Botrytis
Soft rot Mucor
Rhizopus
Grapes Grey mould rot Botrytis
Sub-tropical fruits
Bananas Crown rot Fusarium
Citrus fruits Stem-end rot Alternaria
Brown rot Phytophthora
Blue and green mould rots Penicillium
Sour rot Geotrichium
Vegetable fruits
Tomatoes Grey mould rot Botrytis
Black rot Alternaria
Soft rot Rhizopus
Sour rot Geotrichium
Cucumbers Green mould rot Cladosporium
Grey mould rot Botrytis
Vegetables
Carrots Grey mould rot Botrytis
Watery soft rot Sclerotinia
Cabbage Black rot Alternaria
Brown rot Phytophthora
Potatoes Blight Phytophthora
Dry rot Fusarium
Soft rot Rhizopus
The microbial flora of harvested cereal grains such as corn, wheat and
oats contains up to many millions of bacteria and moulds per gram.
However, the low aw of grains effectively inhibits the growth of all mi-
croorganisms provided storage conditions are satisfactory but in moist
conditions mould growth is likely.
Certain steps involved in flour manufacture reduce the microbial load
and of these bleaching has the greatest effect. Mould counts remain fairly
constant, in the low thousands per gram, in properly stored flours and the
most commonly isolated species are members of the genera Penicillium,
Aspergillus and Rhizopus. Bacteria decrease in numbers during storage
and counts of <1000 per g are usual, with Bacillus spp. the predominant
group. Where the moisture content is above normal mould growth is
likely and at still higher aw levels growth of Bacillus spp. will occur.
Commercially produced bread should be of sufficiently low moisture
content to inhibit growth of most microorganisms except moulds which
Food Spoilage 151
are the principal spoilage agents; in fact, moulds are said to be responsible
for the loss of 1% of annual bread production (Seiler, 1971). Amongst the
most common are Rhizopus nigricans, the 'bread mould' which produces
characteristic black dots of sporangia, Penicillium and Aspergillus spp.,
which produce green conidia in abundance, and Neurospora sitophila, the
'red bread' mould. Mould spoilage is encouraged by slicing, wrapping the
bread when too warm and storage in a warm, moist environment.
Ropiness of bread, caused by Bacillus spp., is now rarely seen in com-
mercially produced bread. It is initially characterized by brownish spots
accompanied by an unpleasant odour and, later, disintegration of the
crumb or slices follows; the spoilage is caused by hydrolysis of flour
protein and starch which produces stickiness and stringiness in the
bread. Control is best achieved by low temperature storage, the addition
of preservative (e.g. calcium propionate or sorbic acid) and the use of
good-quality flour (Frazier & Westhoff, 1988).
Moulds are responsible for most spoilage problems in cakes although
the situation is complicated by the wide variety of ingredients that may
be incorporated some of which, such as dairy and imitation creams, cus-
tard and chocolate, have been implicated in bacterial food poisoning in-
cidents (Seiler, 1978). Generally, mould growth is again controlled by
low temperature storage and low aw levels together with the use of
preservatives as in bread.
Pastas are made from a stiff dough which is extruded in different
shapes. There is then a warm drying stage in processing which requires
careful control since both Salmonella spp. and Staphylococcus aureus
could proliferate during this period; these organisms have been impli-
cated in food poisoning outbreaks mainly involving pastas containing
egg although egg-free pastas have been shown to contain these
pathogens on occasions (Seiler, 1988). With careful drying and subse-
quent storage in dry conditions spoilage should be prevented.
3.11. BEER
The main ingredients of beer are malted barley, hops and water. The
malt is ground up with water to form a mash and the enzymes naturally
present convert the starch into an easily fermentable sugar, maltose.
This 'wort' is then boiled together with hops which are added princi-
pally for flavouring. After cooling, yeasts (strains of Saccharomyces
152 Food Microbiology and Hygiene
3.12. WINE
including undesirable yeasts and lactic acid bacteria (Go swell, 1986); a
starter culture of yeast is then added and thus greater control over the
fermentation process can be achieved. At the end of active fermentation
the wine is transferred to storage tanks for ageing after which it is
filtered and bottled or otherwise stored.
The microorganisms causing wine spoilage are principally wild yeasts
and bacteria although, as with beer, defects are frequently non-micro-
bial in origin. Important spoilage yeasts include Candida, Pichia and a
number of Saccharomyces spp. which all produce growth films on the
surface of wines. Some yeasts may be desirable in certain wines but may
cause spoilage in others where residual sugar is desired. Wine spoilage
bacteria are principally acetobacters and lactic acid bacteria. The former
produce sourness whilst the latter, represented by the genera Lacto-
bacillus, Leuconostoc and Pediococcus, produce lactic and acetic acids
from sugars; these acids are usually accompanied by turbidity, off-
flavours and possibly the evolution of carbon dioxide. Ropiness, al-
though less common than in beers, can also be caused by leuconostocs.
3.13. SAUERKRAUT
toxins and enzymes, thereby rendering the food free from possible
spoilage or harmful effects. From a biological viewpoint the process
may fail in one or both of two ways. The first is a post-process infection
involving leakage of microorganisms inwards through faulty seams and
the second is the survival of organisms as a result of inadequate heat
treatment.
An alternative canning process, used mainly for fluid foods (e.g. sauces,
soups and fruit concentrates), involves the continuous sterilization of the
product using a high-temperature, short-time (HTST) heat treatment (e.g.
130a C for 30 s). Typically the foods are passed through heat exchangers
for product heating, a holding tube where the food is maintained at the
required temperature/time, and finally into further heat exchangers where
rapid cooling is achieved; containers and lids are sterilized separately with
superheated steam and the food is added to the container which is then
lidded in the sterile environment (Frazier & Westhoff, 1988). In fact this
aseptic packaging principle has been used with various forms of flexible
packages made with plastics and aluminium foil, the pre-sterilization of
which is often done by means of hot hydrogen peroxide or uv irradiation
(Mitchell, 1988). Spoilage of these aseptically processed foods is again
caused by seam leakage or under processing.
Before making a more detailed review of the microbiological spoilage
of canned foods it should be mentioned that spoilage may also be caused
by chemical changes, the most important of which is the 'hydrogen swell'.
This results from the reaction of the can metal (iron) with acidic foods
when the hydrogen liberated causes the can to swell. The higher the acid-
ity of the food, the greater the likelihood of this problem developing al-
though suitable internal lacquers should largely eliminate this fault.
Table 3.4
The Influence of the Number of Bacteria in the Cooling
Water on the Rate of Reinfection Q
2
8
18
30
48
62
Fig. 3.8. Steps in can closure after filling: (a) positioning of lid, (b) to (e) pro-
gressive operations in the formation of the double seam.
I thicknass I
I I
I I
"'~""'i~---
~.....--body hook
(1) ensure that the construction of the double seam, the lap and the
side seam are in accordance with accepted quality standards; (2)
avoid rough handling of cans; (3) avoid excessive deformation of
can ends during sterilization and cooling due to sudden pressure
changes; (4) correctly chlorinate cooling water to a residual level of
1-2 mg/litre of free chlorine measured in the water drained from
the seam after the cooling operation. Also ensure that the cooling
water conforms to the chemical and bacteriological standards laid
down for drinking water; (5) wash and disinfect at frequent inter-
vals all surfaces of mechanical handling equipment which might
come in contact with the double seam; (6) dry cans immediately
after cooling and transport them on clean dry surfaces; (7) check
cannery hygiene by regular microbiological surveys; (8) insist on
and supervise high hygienic standards amongst employees.
Even with the advent of more modern types of cans these principles
still hold true to-day.
Where spoilage results from seam leakage the bacteria implicated usu-
ally have relatively low optimum growth temperatures (25-35°C) and
would be readily killed by the heat processing. In addition, more than
one type of bacterium is usually isolated; these types commonly include
members of the genera Pseudomonas, Alcaligenes and Flavobacterium,
together with coliforms and micrococci. The above types are common
contaminants where chlorination levels in the cooling water are minimal
or non-existent; where chlorination is only slightly sub-standard,
Bacillus and Clostridium spp. may be involved since their spores are
highly resistant to chlorination. Clostridial spores may also gain access
to cans post-cooling from dirty tracks and other items of equipment
thus again stressing the need for good cannery hygiene (Lake et al.,
1985). When it is remembered that nearly two-thirds of all forms of
spoilage in canned foods are due to reinfection after heating, the impor-
tance of control measures, especially adequate chlorination of cooling
water, cannot be overstressed.
158 Food Microbiology and Hygiene
\
\
~- - - - - - - - ~<~==:.-jD:>====:;;;>;--"
01
c:
i \
>
I-
\
~ 3
..
I-
.0
E \
~
c:
\ b
01 2
o \
..J
Fig. 3.10. Hypothetical survivor curves showing: (a) curves for heat-resistant
spore, and (b) curve for heat-sensitive spore.
Food Spoilage 159
has been suggested that this growth may result from localized regions in
the food having slightly higher pHs than those recorded.
The rate of killing bacterial spores (or vegetative cells for that matter,
although they are much less heat-resistant) is a function of temperature
and time; the higher the temperature, the greater the rate of destruction
for any given time. Bacterial death is said to be logarithmic which means
that equal percentages of surviving cells are killed in each successive unit
of time. As can be seen in Fig. 3.10, the D value, represented by the
slope of the survival curve is defined as the time at any given tempera-
ture for a 90% reduction in viability to be effected (thus for the heat-re-
sistant spore the D value is ca 10 min whilst for the heat sensitive spore
the value is less than 1 min). This means that the greater the number of
spores that are present in any given food, the longer the time necessary
for their destruction. From a practical standpoint it is therefore impor-
tant to ensure that foods being canned have a predictable number of
spores in them which can be destroyed by the normal heat treatment ap-
plied; thus the microbiological quality of raw materials and process line
sanitation standards must be carefully monitored.
also grow well in this pH range and cause spoilage, with gas production,
of canned tomatoes, pears and other fruits. The spores produced by
these clostridia are even more heat labile and since they are destroyed in
15 min at lOoDe it is unlikely that processing is inadequate.
Amongst the non-spore forming bacteria the lactic acid bacteria are
most frequently incriminated in the spoilage of these high acid foods
(Dennis, 1987). Lactobacillus brevis commonly causes fermentation of
tomato ketchup, Worcester sauce, pickles and salad dressings. Other
lactobacilli and leuconostocs occasionally cause spoilage of a range of
canned fruits and fruit juices. These lactic acid bacteria (i.e. some
Lactobacillus spp. and all Leuconostoc spp.) produce gas as well as acid
from the syrup sugar so that spoilage is accompanied by can distension;
other end products include acetic acid and ethyl alcohol so that with
these diverse products such lactic acid bacteria are termed 'heterofer-
mentative'. With 'homofermentative' lactic acid bacteria (i.e. the re-
maining lactobacilli and all Streptococcus spp.) only lactic acid is
produced by the fermentation of sugar.
Yeasts are extremely heat-sensitive and are therefore rarely involved in
spoilage of canned foods. Certain Torulopsis spp. may occasionally cause
gaseous spoilage of sweetened condensed milk, which relies upon the high
sugar content rather than upon a substantial heat treatment as the
method of preservation. Other yeasts, Saccharomyces spp., have produced
spoilage in citrus juices and pickles. Like yeasts, moulds rarely cause
spoilage but there are two notable exceptions in Byssochlamys fulva and
B. nivea the ascospores of which are unusually heat-resistant tolerating
85°e for 30min. The foods attacked are mainly strawberries and raspber-
ries which may totally disintegrate in spoilage due to the action of the
pectolytic enzymes produced by the organisms (Put & Kruiswijk, 1964).
Control of this type of spoilage is best achieved by pre-treating the in-
fected fruit with gaseous methyl bromide or peracetic acid and by careful
cleaning of both the raw material and the processing equipment.
a few specialized bacteria and even down to -1 DoC for certain moulds
(Ingram & Mackey, 1976). As the temperature falls from ODC a series of
eutectics (i.e. ice: solute mixtures) is formed which is accompanied by an
increasing concentration of dissolved solids in the unfrozen water. As
well as lowering the freezing point of the remaining unfrozen water
these increasing solute concentrations also progressively lower the aw
and this has an increasingly deleterious effect on the microbial popula-
tion; thus organisms capable of growth in foods at sub-zero tempera-
tures must also tolerate lowered aw values. A small percentage of water
remains unfrozen at temperatures well below -IOODC; however, for
practical purposes the 'freezable' water in meat and fish is totally frozen
at -50 to -70 D C whilst for fruits and vegetables the corresponding
figures are -16 to -20 D e.
80
(c)
60
~
:g
til
-; 40
E
CI)
u
liiQ.
20
,
,
o i I I I
1 10 100 1000 10000
Fig. 3.11. Effect of freezing on viability of typical Gram negative rod (after
MacLeod & Ca\cott, 1976).
Whilst the main losses in viability occur during initial freezing, further
kill-off of bacteria occurs during frozen storage. Provided the storage
temperature is low enough, death rates are minimal but at normal
frozen food storage temperatures (-20°C) some loss in viability is evi-
dent, particularly in the early days of storage. Foods show a far greater
reduction in viable counts when held at -5 to -lOoC than at -20°C but
whilst the higher storage temperatures may be an effective method of re-
ducing counts they contribute to an increased rate of deterioration of
the food resulting from other causes. Even when microbial growth is
completely inhibited the product quality can still deteriorate due to the
continued activity of released microbial enzymes or to the indigenous
enzymes present in the food; in the case of vegetables these enzymes
must be destroyed by blanching. Other harmful physico-chemical and
biochemical changes can occur during freezing and cold storage (Boegh-
Soerensen & Jul, 1985).
Food Spoilage 165
When foods are very rapidly frozen the number of microorganisms sur-
viving the freeze-thaw cycle is partly dependent on the rate of thawing,
somewhat lower recoveries being obtained with slower thawing; this is
due to the growth of the very small ice crystals within the microbial cell
causing increased cellular damage (Mackey, 1984). The survivors start
multiplying, as in the normal growth cycle, after a lag period (see pp.
17-18) but this period is extended by the inherently low temperature of
the food so that the log phase of growth may take 3-6 h to become es-
tablished. When frozen foods are allowed to defrost over a long period
at, say, 3-lO D C psychrotrophs may dominate the flora and subsequently
cause spoilage. In other cases the types of organisms growing will de-
pend on the temperature at which the thawed food is held but with
most foods the organisms predominating would be similar to those in
the equivalent unfrozen product. Particular problems arise with larger
packs, e.g. frozen turkeys, where a temperature gradient is established
between the warm surface and the cold interior. If such packs are
166 Food Microbiology and Hygiene
Many changes can occur during the storage of dried foods and most are
non-microbial in origin. The commonest and most important change is
non-enzymic (Maillard) browning which involves a complex series of
chemical reactions between reducing sugars and amino acids or pro-
teins. The aw levels that must be achieved during drying to stop this
browning are far lower than those required to inhibit microbial growth
so that microbial spoilage should not arise. In fact during storage there
is a decrease in numbers of viable organisms although the spores of bac-
teria and moulds remain unaffected. If dried foods are incorrectly pack-
aged or are stored under moist conditions, sufficient water may be
re-absorbed to enable moulds to grow but the water uptake should not
be such as to permit bacterial growth.
3.16.4. Rehydration
When dried foods are rehydrated similar responses are shown by the
contained microorganisms as with thawed frozen foods; there is a lag
phase of growth and many organisms exhibit metabolic injury (Gibbs,
1986). Clearly the temperature of the water used for rehydration may
have a marked effect on the flora and on the subsequent rate of spoilage
of the food. If boiling water is used Bacillus spp. will predominate and
cause spoilage and at progressively lower rehydration temperatures the
flora will become more varied and contain more heat-labile organisms.
When refrigerated storage is used the storage life of most rehydrated
foods is restricted to 1 or 2 days but storage at room temperature
should clearly be limited to no more than a few hours.
ture contents of 20--40%, and which do not require refrigeration for stabil-
ity. Foods in this group include dried fruits, certain bakery products and
salted meats and fish all of which have already been discussed, together
with jams, syrups and honey. Spoilage of the latter group is attributed to
osmophiles, microorganisms growing in high sugar concentrations
(65-70%) and tolerating low pH values «4·0). The most common spoilage
agents are osmophilic yeasts (Saccharomyces and Torulopsis spp.) which
ferment the sucrose with the production of alcohol. Certain moulds may
develop on the surface of jams, the most common being species of
Aspergillus and Penicillium. Since osmophiles are heat-sensitive and there-
fore readily killed during heat processing, spoilage of these products is
only possible after recontamination which may occur through faulty seal-
ing or after opening the containers. Furthermore, it is probable that mois-
ture re-absorption is necessary before growth can be initiated.
Table 3.5
Approximate Radiation Doses to Effect a Reduction of a Million III Viable
N umbers of Different Microorganisms"
Microorganisms Dose
(kGy)
many lesions on the DNA for the organism to cope with, replication
will cease. Some of the more complex enzyme repair systems are found
in all organisms including humans although the repair mechanisms are
far less efficient in higher animals; thus a human would certainly be
killed by exposure to a radiation dose of 0.005 kGy.
There are, however, certain meats such as bacon where radiation ap-
pears to be of questionable value. The indigenous flora including micro-
cocci, lactic acid bacteria and acinetobacters is inherently more resistant
to the effects of irradiation and doses of 10 kGy could prove inade-
quate; more seriously foreign odours persist in these higher lipid con-
taining foods after treatment even though the appearance is unaffected
(Dempster et ai., 1986).
In conclusion, radiation as a means of food processing produces no
unique microbiological problems-the problem of the resistant organ-
ism(s) is common to many food processing techniques. Thus the same
degree of control must be exercised with radiation processing as with
any other type of process so that the quality of raw materials and the
hygiene standards applied on the processing line, and to the finished
product, are as important here as with any other process.
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