In this article we will discuss about the two types of low temperature storage, i.e, chilling and freezing technique for preservation of food.

Chilling Technique:

Chilled foods are those foods stored at temperatures near, but above their freezing point, typically 0-5 °C. This commodity area has shown a massive increase in recent years as traditional chilled products such as fresh meat and fish and dairy products have been joined by a huge variety of new products including complete meals, prepared and delicatessen salads, dairy desserts and many others.

Three main factors have contributed to this development:

(1) The food manufacturers’ objective of increasing added value to their products;

(2) Consumer demand for fresh foods while at the same time requiring the convenience of only occasional shopping and ease of preparation; and

(3) The availability of an efficient cold chain – the organization and infrastructure which allows low temperatures to be maintained throughout the food chain from manufacture/harvest to consumption.

Chill storage can change both the nature of spoilage and the rate at which it occurs. There may be qualitative changes in spoilage characteristics as low temperatures exert a selective effect preventing the growth of mesophiles and leading to a microflora dominated by psychrotrophs.

This can be seen in the case of raw milk which in the days of milk churns and roadside collection had a spoilage microflora comprised largely of mesophiliclactococci which would sour the milk.

Nowadays in the UK, milk is chilled almost immediately it leaves the cow so that psychrotrophic Gram-negative rods predominate and produce an entirely different type of spoilage. Low temperatures can also cause physiological changes in micro­organisms that modify or exacerbate spoilage characteristics.

Two such examples are the increased production of phenazine and carotenoid pigments in some organisms at low temperatures and the stimulation of extracellular polysaccharide production in Leuconostoc spp. and some other lactic acid bacteria.

In most cases, such changes probably represent a disturbance of metabolism due to the differing thermal coefficients and activation energies of the numerous chemical reactions that comprise microbial metabolism.

Though psychrotrophs can grow in chilled foods they do so only relatively slowly so that the onset of spoilage is delayed. In this respect temperature changes within the chill temperature range can have pronounced effects.

For example, the generation time for one pseudomonad isolated from fish was 6.7 hours at 5°C compared with ‘26.6 hours at 0°C. Where this organism is an important contributor to spoilage, small changes of temperature will have major implications for shelf-life.

The keeping time of haddock and cod fillets has been found to double if the storage temperature is decreased from 2.8 °C to — 0.3°C. Mathematical modelling techniques of the sort can be useful in predicting the effect of temperature fluctuations on shelf-life, but, as a general rule, storage temperature should be as low, and as tightly controlled, as possible.

The ability of organisms to grow at low temperatures appears to be particularly associated with the composition and architecture of the plasma membrane. As the temperature is lowered, the plasma membrane undergoes a phase transition from a liquid crystalline state- to a rigid gel in which solute transport is severely limited.

The temperature of this transition is lower in psychrotrophs and psychrophiles largely as a result of higher levels of unsaturated and short chain fatty acids in their membrane lipids. If some organisms are allowed to adapt to growth at lower temperatures they increase the proportion of these components in their membranes.

There seems to be no taxonomic restriction on psychrotrophic organisms which can be found in the yeasts, moulds, Gram-negative and Gram-positive bacteria. One feature they share is that in addition to their ability to grow at low- temperatures, they are inactivated at moderate temperatures.

A number of reasons for this marked heat sensitivity have been put forward including the possibility of excessive membrane fluidity at higher temperatures. Low thermal stability of key enzymes and other functional proteins appears to be an important factor, although thermo-stable extracellular lipases and proteases produced by psychrotrophic pseudomonads can be a problem in the dairy industry.

Though mesophiles cannot grow at chill temperatures, they are not necessarily killed. Chilling will produce a phenomenon known as cold shock which causes death and injury in a proportion of the population but its effects are not predictable in the same way as heat processing.

The extent of cold shock depends on a number of factors such as the organism (Gram-negatives appear more susceptible than Gram- positives), its phase of growth (exponential-phase cells are more susceptible than stationary phase cells), the temperature differential and the rate of cooling (in both cases the larger it is, the greater the damage), and the growth medium cells grown in complex media are more resistant).

The principal mechanism of cold shock appears to be damage to membranes caused by phase changes in the membrane lipids which create hydroppores through which cytoplasmic contents can leak out. An increase in single-strand breaks in DNA has also been noted as well as the synthesis of specific cold-shock proteins.

Since chilling is not a bactericidal process, the use of good microbiological quality raw materials and hygienic handling are key requirements for the production of safe chill foods. Mesophiles that survive cooling, albeit in an injured state, can persist in the food for extended periods and may recover and resume growth should conditions later become favourable.

Thus chilling will prevent an increase in the risk from mesophilic pathogens, but will not assure its elimination. There are however pathogens that will continue to grow at some chill temperatures and the key role of chilling in the modern food industry has focused particular attention on these.

Risks posed by these organisms, may increase with duration of storage but this process is likely to be slow and dependent on the precise storage temperature and composition of the food.

Some foods are not amenable to chill storage as they suffer from cold injury where the low temperature results in tissue breakdown which leads to visual defects and accelerated microbiological deterioration. Tropical fruits are particularly susceptible to this form of damage.

Freezing Technique:

Freezing is the most successful technique for long term preservation of food since nutrient content is largely retained and the product resembles the fresh material more closely than in appertized foods.

Foods begin to freeze somewhere in the range — 0.5 to — 3 °C, the freezing point being lower than that of pure water due to the solutes present. As water is converted to ice during freezing, the concentration of solutes in the unfrozen water increases, decreasing its freezing point still further so that even at very low temperatures, e.g. — 60 °C, some water will remain unfrozen.

The temperatures used in frozen storage are generally less than — 18 °C. At these temperatures no microbial growth is possible, although residual microbial or endogenous enzyme activity such as lipases can persist and eventually spoil a product.

This is reduced in the case of fruits and vegetables by blanching before freezing to inactivate endogenous polyphenol oxidases which would otherwise cause the product to dis-colour during storage.

Freezer burn is another non-microbiological quality defect that may arise in frozen foods, where surface discolouration occurs due to sublimation of water from the product and its transfer to colder surfaces in the freezer. This can be prevented by wrapping products in a water-impermeable material or by glazing with n layer of ice.

Low temperature is not the only inhibitory factor operating in frozen foods; they also have a low water activity produced by removal of water in the form of ice. Table 4.11 describes the effect of temperature on water activity. As far as microbiological quality is concerned, this effect is only significant when frozen foods are stored at temperatures where microbial growth is possible (above — 10 C).

Effect of freezing on the water activity of pure water/ ice

In this situation, the organisms that grow on a product are not those normally associated with its spoilage at chill temperatures but yeasts and moulds that are both psychrotrophic and tolerant of reduced water activity.

Thus meat and poultry stored at — 5 to — 10 °C may slowly develop surface defects such as black spots due to the growth of the mould Cladosporium herbarum, white spots caused by Sporotrichum carnis or the feathery growth of Thamnidium elegans.

Micro-organisms are affected by each phase of the freezing process. In cooling down to the temperature at which freezing begins, a proportion of the population will be subject to cold shock.

At the freezing temperature, further death and injury occur as the cooling curve levels out as latent heat is removed and the product begins to freeze. Initially ice forms mainly extracellularly, intracellular ice formation being favoured by more rapid cooling.

This may mechanically damage cells and the high extracellular osmotic pressures generated will dehydrate them. Changes in the ionic strength and pH of the water phase as a result of freezing will also disrupt the structure and function of numerous cell components and macromolecules which depend on these factors for their stability.

Cooling down to the storage temperature will prevent any further microbial growth once the temperature has dropped below — 10 °C. Finally, during storage there will be an initial decrease in viable numbers followed by slow decline over time. The lower the storage temperature, the slower the death rate.

As with chilling, freezing will not render an unsafe product safe – its microbial lethality is limited and preformed toxins will persist. Frozen chickens are, after all, an important source of Salmonella.

Survival rates after freezing will depend on the precise conditions of freezing, the nature of the food material and the composition of its microflora, but have been variously recorded as between 5 and 70%. Bacterial spores are virtually unaffected by freezing, most vegetative Gram-positive bacteria are relatively resistant and Gram-negatives show the greatest sensitivity.

While frozen storage does reliably inactivate higher organisms such as pathogenic protozoa and parasitic worms, food materials often act as cryoprotectants for bacteria so that bacterial pathogens may survive for long periods in the frozen state. In one extreme example Salmonella has been successfully isolated from ice cream stored at — 23 °C for 7 years.

The extent of microbial death is also determined by the rate of cooling.

Maximum lethality is seen with slow cooling where, although there is little or no cold shock experienced by the organisms, exposure to high solute concentrations is prolonged. Survival is greater with rapid freezing where exposure to these conditions is minimized. Food freezing processes are not designed however to maximize microbial lethality but to minimize loss of product quality.

Formation of large ice crystals and prolonged exposure to high osmotic pressure solutions during slow cooling also damage cells of the food material itself causing greater drip loss and textural deterioration on thawing, so fast freezing in which the product is at storage temperature within half an hour is the method of choice commercially.

The rate of freezing in domestic freezers is much slower so, although microbial lethality may be greater, so too is product quality loss.

Thawing of frozen foods is a slower process than freezing. Even with moderate size material the outside of the product will be at the thawing temperature some time before the interior. So with high thawing temperature, mesophiles may be growing on the surface of a product while the interior is still frozen. Slow thawing at lower temperature is generally preferred.

It does have some lethal effect as microbial cells experience adverse conditions in the 0 to — 10 °C range for longer, but it will also allow psychrotrophs to grow. Provided the product is not subject to contamination after thawing, the microflora that develops will differ from that on the fresh material due to the selective lethal effect of freezing.

Lactic acid bacteria are often responsible for the spoilage of defrosted vegetables whereas they generally comprise only about 1% of the microflora on fresh chilled produce which is predominantly Gram-negative.

Freezing and defrosting may make some foods more susceptible to microbiolo­gical attack due to destruction of antimicrobial barriers in the product and condensation, but defrosted foods do not spoil more rapidly than those that have not been frozen. Injunctions against refreezing defrosted products are motivated by the loss of textural and other qualities rather than any microbiological risk that is posed.