Chapter 2

Principles of Food Preservation

Sudarsan Mukhopadhyay, Dike O. Ukuku, Vijay K. Juneja,

Balunkeswar Nayak, and Modesto Olanya

Abstract  Food preservation is an action or method used to maintain foods at certain

desirable properties or quality to obtain maximum benefit. A good method of food pres-
ervation is one that slows down or prevents altogether the action of the agents of spoilage
without damaging the food. To achieve this, certain basic methods are applied depending
on the food types. Food preservation has been an essential activity throughout human
history. The cycle of seasons brings periods of shortage and abundance of various foods
at different times of the year. Preservation makes it possible to consume some of these
foods during off seasons, throughout the year. Food preservation usually involves con-
trolling or preventing growth of microrganisms or minimizing the quality degradation
due to microbial spoilage or unwanted chemical changes in foods such as rancidity due
to oxidation of fats over time. Preservation of foods is no longer simple and straightfor-
ward today; it has evolved to a highly inter-disciplinary field of science. In recent years,
many new sophisticated preservation techniques have developed to extend the quality
and shelf-life, minimize risk, protect the environment, and improve functional, sensory,
and nutritional properties. Many of emerging preservation technologies have already

Mention of trade names or commercial products in this article is solely for the purpose of providing
specific information and does not imply recommendation or endorsement by the U.S. Department
of Agriculture. USDA is an equal opportunity provider and employee.
S. Mukhopadhyay (*) • V.K. Juneja
Residue Chemistry and Predictive Microbiology Research Unit, Eastern Regional Research
Center, USDA-Agricultural Research Service, Wyndmoor, PA, USA
e-mail: sudarsan.mukhopadhyay@ars.usda.gov; vijay.juneja@ars.usda.gov
D.O. Ukuku • M. Olanya
Food Safety Intervention Technologies Research Unit, Eastern Regional Research Center,
USDA-Agricultural Research Service, Wyndmoor, PA, USA
e-mail: dike.ukuku@ars.usda.gov; modesto.olanya@ars.usda.gov
B. Nayak
Food Science and Human Nutrition, School of Food & Agriculture, University of Maine,
Orono, ME, USA
e-mail: balunkeswar.nayak@maine.edu

© Springer Science+Business Media, LLC 2017 17

V.K. Juneja et al. (eds.), Microbial Control and Food Preservation, Food
Microbiology and Food Safety, https://doi.org/10.1007/978-1-4939-7556-3_2
18 S. Mukhopadhyay et al.

reached commercial adoption in specific applications while many others remain promis-
ing. Development of suitable equipment, especially for continuous processing for a vari-
ety of foods and standardization of the process parameters for easy regulatory approval
will pave the way for improved food preservation. The objective of this chapter was to
examine the science and technology involved in the manipulation of conventional as
well as sophisticated emerging preservation methods.

Keywords Thermal preservation • Advance thermal technology • Emerging

nonthermal preservation • Pulsed light • Membrane technology

1  Introduction

Preservation of foods involves controlling or preventing the growth of microorganisms

such as bacteria, fungi and yeasts, although in some instances food preservation can be
achieved by introducing benign microorganisms to the food. Preservation is also
achieved by retarding the oxidation of fats that cause rancidity. Processes that inhibit
visual deterioration of foods, for example the browning of apples due to enzymatic
reaction once apples are cut is also considered as food preservation. In some cases food
preservation involves a number of different food methods each of which independently
as a separate preservation technique. Preparation of Jam or Jelly, for example, involves
steps as heat treatment or boiling to lower the moisture content of fruit and to kill
microbes, etc., addition of sugar as preservative to control the re-growth of organisms
and lastly sealing of the food within an airtight container in order to stop recontamina-
tion of the final product. Many old preservation methods which been used traditionally
in the past have been proven to be less energy intensive and have lower carbon footprint
compared to modern intentions (Preserving food without freezing or canning 1999).
Quality maintenance, for example preservation of color, flavor, texture, micronutrients
and overall nutritional quality in an integral part of food preservation.

2  Advantage of Food Preservation

Food preservation has many advantages. It makes possible to prevent the spoilage
foods due to the action of inherent microorganisms and enzymes. Preservation
prolongs the duration of safe storage of foodstuffs and thereby creates its avail-
ability throughout the season and makes up for the deficiencies. Preservation also
makes food transportation much easier.
2  Principles of Food Preservation 19

3  Principles of Food Preservation

There are three main principles by which food preservation is achieved:

Controlling microorganisms. Microbial contamination may cause change that ren-
ders food unfit for human consumption. Primary sources of contamination are irri-
gation water, air, soil, animals, feed, and food processing equipments. Foods may be
contaminated by bacteria yeasts or molds at any stages of pre-harvest to post-har-
vest continuum. Inactivation or control of microbial population is achieved by
increasing or decreasing food temperature, adding salt, sugar or acid to food or by
removing the water or air from the food. Removing air from food inhibits oxygen
dependant enzymatic and chemical reactions and inhibits growth of aerobic micro-
organisms. Removal of moisture from produce such as leafy greens causes leaves to
dry which limits microbial growth.
Controlling enzymes. Enzymes play important role in living tissues of food. But
it catalyzes spoilage reactions following harvest or slaughter of plant or animal
based foods. In case of plant foods, certain enzymes remain active in the tissue
cells of fruits and vegetables following harvest and may continue to catalyze the
biochemical processes of ripening of these foods which may eventually lead to
rotting, as can be observed in many fruits. Oxidative enzymes in produce such
as banana or other fruits may continue to maintain cellular respiration by metab-
olizing sugar with oxygen for energy and may shorten the shelf life of fresh
fruits leading to spoilage. Respiration may be controlled by refrigerated storage
or modified-atmosphere packaging. Respiration may be controlled by refriger-
ated storage or modified-­atmosphere packaging.

3.1  C
 ontrolling Insects, Rodents, Birds and Other Physical
Causes of Food Deterioration

Insects grow in humid warm environments and the types of foods subject to pest
attack are cereal grains and products derived from cereal grains, legumes, dairy
products, dried fruits, dried and smoked meats and nuts. The presence of insects and
insect excreta causes loss of food nutritional value, promotes decay of food and cre-
ates off -flavors and this makes food unsalable, causing considerable economic loss.
Mice and rats carry disease and are frequently associated with food-borne infections
in humans. Proper sanitation in food processing and storage areas is very important
in the fight against rodents, since all packaging materials, except metal and glass
containers, can be attacked by rats and mice.
20 S. Mukhopadhyay et al.

4  Methods of Food Preservation

Food preservation is achieved by three main methods as inactivation of microorganisms

and/or enzymes, inhibition of microbial growth or deterioration of quality and avoid-
ance of recontamination of food at all steps from pre- to post-harvest continuum. Based
on the principle of action, the majority of food preservation techniques can be classified
as (a) thermal and (b) emerging nonthermal processing (Preserving food without freez-
ing or canning 1999; Ravishankar and Maks 2007). These techniques cause inactivation
of pathogenic microorganisms assuring food safety and control growth of spoilage
microorganisms extending the shelf life.

5  Thermal Processing for Food Preservation

Thermal processing refers to the application of heat energy. Thermal processing is an

effective way of preserving food because the great majority of harmful pathogens are
killed at temperatures close to the boiling point of water. Heating of food has long been
at the heart of food processing. Thermal treatment is not only an important method of
preserving foods but also a means of developing texture, flavor and color. An important
challenge for thermal processing is the design and development of effective application
of thermal technologies to achieve these objectives without altering the desirable sen-
sory, functional and nutritional qualities of food (Rahman 2007; Richardson 2001).
Depending on their strength, thermal preservation can be classified as pasteuriza-
tion or sterilization processes. Pasteurization is a thermal treatment which can destroy
vegetative cells but has almost no effect on spores (U.S. Food and Drug Admin 2006;
U.S. Dept. of Health and Human Services 2009; Walstra et al. 1999). Sterilization on
the other hand, is designed for inactivating all forms of microorganisms, including
spores (Dion and Parker 2013). In terms of kinetics of thermal inactivation of micro-
organisms, the decimal reduction time or log-linear model is generally accepted. The
model assumes the behavior of thermal destruction of microorganisms is that of first-
order kinetics. The relationship between the decimal reduction time and the tempera-
ture is also assumed to be log-linear, leading to the concept of important kinetic
parameters as D and Z values which helps in thermal calculation and defining rate of
thermal lethality of the treatment The D value is a measure of the heat resistance of a
microorganism. It is the time in minutes at a given temperature required to destroy 1
log cycle (90%) of the target microorganism. In an actual process, all other organisms
that are less heat tolerant are destroyed to a greater extent. The Z value reflects the
temperature dependence of thermal destruction and is defined as the temperature
change needed to change the D value by a factor of 10.
Both the first-order kinetics and the Arrhenius-type effect of temperature are con-
tested. The lethality of a thermal process is expressed in terms of its F0 value. The F0
value of a thermal sterilization process is the number of minutes of heating at 121 °C
required to achieve the same thermal destruction ratio of a specified target microorgan-
ism, having a z value of 10 °C. Process optimization is the selection of thermal pro-
2  Principles of Food Preservation 21

cessing conditions that results minimal damage to food quality while meeting the
required food safety criteria. These “heat and kill” sterilization systems include high-
temperature, short-time pasteurization (HTST) and ultra-high t­ emperature (UHT) pro-
cessing designed for heating liquid foods, and retorting or canning for processed solid
foods (Stabel and Lambertz 2004).

6  Advance Thermal Technologies for Food Preservation

Thermal processing technologies for treatment of foods have been around for many
years. In recent years, there has been some new development in the area of source of heat
supply which are considered emerging or novel. These emerging thermal technologies
for food preservation include microwave, radio frequency, ohmic and infrared heating.

6.1  Microwave and Radio Frequency Preservation Process

Microwave and radio frequency electric field heating refers to use of electromagnetic
(EM) waves of certain frequencies to generate heat in a material. Microwave (MW)
has EM range of 300–300,000 MHz, while electromagnetic wave for radio frequency
(RF) ranges from 0.003–300 MHz. Microwave is simply a part of the RF spectrum
that has become quite popular due to the large number of its possible uses. Microwaves
can also be used to transmit power from one point to another. Both MW and RF work
by generating heat energy in foods, dielectric media, through dipole rotation and
ionic polarization (Metaxas and Meredith 1993). Water in the food is the primary
dipolar component responsible for the dielectric heating. In an alternating current
electric field, the polarity of the field is varied at the rate of microwave frequency and
molecules attempt to align themselves with the changing field. Heat is generated
rapidly as a result of internal molecular friction. The second major mechanism of
heating with microwaves is through the polarization of ions as a result of the back
and forth movement of the ionic molecules trying to align themselves with the oscil-
lating electric field. Microwave heating is also affected by the state of the constitu-
ents, whether they are bound or free, e.g., bound ions have much lower microwave
absorbing capability (Decareau and Peterson 1986; Von Hippel 1954). Critical fac-
tors affecting the effectiveness of microwave heating are the size, shape, composition
(electrolytes, moisture, salt, etc.), multiple components (frozen meal), and liquidity/
solidity of the food being treated. Also the power level, time of heating, cycling,
presence of hot water or air around the food, dimension, shape and other electromag-
netic properties of the oven are also important. Microwave heating for pasteurization
and sterilization may be preferable to conventional heating processes because they
require less time to rise to the desired process temperature, particularly for solid and
semi-solid foods. Microwave heating has the advantage of overcoming the limitation
imposed by the slow thermal diffusion process of conventional heating (Meredith
1998). The volumetric heat generated by microwaves can significantly reduce the
22 S. Mukhopadhyay et al.

total heating time and severity at the elevated temperatures required for commercial
sterilization whereby bacterial destruction is enhanced, but thermal degradation of
the desired components is reduced (Decareau 1985).
RF heating shortens process times by approximately one-third compared to the
conventional method. RF energy inactivates heat-resistant spores in foods and the
quality of RF treated food is usually better than conventionally processed foods.
Computer models based on finite element methods have been developed to predict RF
energy in packaged foods. Commercial application of RF systems, to provide uniform
EM field patterns remains a major challenge (Wang et al. 2003; Chan et al. 2004;
Luechapattanaporn et al. 2004; Luechapattanaporn et al. 2005).
Microwave pasteurization and sterilization systems and its practical indus-
trial applications have been reported over the past few decades. However, radio
frequency heating systems for the purpose of food pasteurization or sterilization
are not yet fully commercialized. Scientists have conducted several studies into
radio frequency electric fields (RFEF) technology, subjecting liquid foods to its
high electric fields, and found it to be both efficient for pathogen inactivation
and cost-­effective. A pasteurization process that uses RFEF for inactivation of
bacteria in foods was developed (Geveke et al. 2002; Geveke and Brunkhorst
2004). In these studies, a set of RFEF operating parameters that achieved
99.999% (5 logs) reduction of E. coli in apple cider was determined, and the
kinetics of bacterial inactivation established. The effect of RFEF treatment on
bacterial membrane was investigated.

6.2  Infrared Heating

Like microwave and radiofrequency, the infrared (IR) radiation (wavelength

0.78–1000  μm) transmits thermal energy in the form of electromagnetic (EM)
waves. IR encompasses a section of the EM spectrum that borders between visi-
ble light and microwaves. By exposing food materials to infrared (IR) radiation,
the heat energy generated by IR can be absorbed by the food. Infrared uses elec-
tromagnetic radiation generated from the vibrational and rotational energy of
molecules of a hot source (quartz lamp, quartz tube or metal rod). Thermal energy
is formed following the absorption of radiating energy. The IR heating system
contains a radiator and a reflector. The radiator or the source which radiated the
heat energy to the surrounding environment while the reflector directs the heat
energy to the food surface. The flux of energy of the radiator usually ranges from
a long (50 kW.m−2) to short (4000 kW.m−2) wave range. The radiator is usually
heated by gas or electric power. A polished metal surface or quartz tube with low
absorptivity and high emission capability to reflect the infrared energy, are used
as reflector (Regier et al. 2010). The characteristics of IR heating are the high heat
transfer capacity and the direct penetration of heat into the food. This is fast pro-
cess with no heating of surrounding air. Infrared heating is safer and economical
as compared to the conventional heating process (Vicente and Castro 2007).
2  Principles of Food Preservation 23

In IR, a desired levels of heating, both at the surface and also at the core of
the food, can be achieved. This is not possible with the microwave heating
process. IR energy, both long (5  μm) and short (1  μm) waves, is a popular
method for drying foods as compared to the conventional drying process due to
its rapid drying characteristics with lower energy consumption and due to a
uniform homogeneous distribution of temperature (Tewari 2007). Infrared is a
newer technology and its application remains limited to the experimental stage.
It is an accepted and popular method of drying, heating, baking, cooking and
drying of food (Tewari 2007). IR heating has also been used for the surface
pasteurization of bread, thawing of foods and for the decontamination of pack-
aging materials. When applied to bread, IR heating has been reported to control
the bread crumb thickness, texture, and color (Regier et al. 2010).

6.3  Ohmic Heating Technology

Ohmic heating, sometimes referred to as Joule heating, electrical resistance heating

or electro conductive heating, is the process of passing an electric currents through
foods or other materials to heat them. Ohmic heating is an advanced thermal process
where food acts as an electrical resistor. The experimental design usually consists of
electrodes that contact the food, whereby electricity is passed through the substance
using a variety of voltage and current combinations. The food is heated by the dissi-
pation of electrical energy. In comparison to the conventional mode of heating, where
heat is conducted from the outside using a hot surface, ohmic heating conducts heat
throughout the entire mass of the food uniformly. The defining characteristics of
ohmic heating compared to other electrical methods, such as microwave and radio-
frequency heating, lie in the frequency and waveforms of the electric field, and that
the electrodes contact the material. The success of ohmic heating depends on the rate
of heat generation in the system, the electrical conductivity of the food, and the
method by which the food flows through the system (Leizerson and Shimoni 2005a).
The inactivation of pathogens by ohmic heating is thermal in nature. Microbial inac-
tivation curves of ohmic heating are similar to conventional heating curves except for
a difference in the slope, which can most likely be explained by the presence of the
electric field (Food and Drug Administration-Center for Food Safety and Applied
Nutrition (FDACFSAN) 2000). Some literature reported non-­thermal effects of
ohmic heating on microbial inactivation. Research indicates that a mild electropora-
tion mechanism occurs during ohmic heating (Alberts et  al. 2002). The principal
reason for the additional effect of ohmic treatment may be its low frequency (50–
60 Hz), which allows cell walls to build up charges and form pores. This is in contrast
to high-frequency methods such as radio or microwave frequency heating, where the
electric field is essentially reversed before sufficient charge buildup occurs at the cell
walls. The ohmic heating method has been applied successfully to a variety of foods
such as fruits and vegetables, juices, meats, seafood, soups, crèmes, and pasta
(Bengtsson et  al. 2010). Additionally, reports also indicate that the suitability of
24 S. Mukhopadhyay et al.

ohmic heating for sterilization to produce high-quality shelf-stable low-acid foods

such as ready-prepared meals and high-acid foods. Research indicated that ohmic
heating processed foods usually retain the quality attributes such as nutrient profile,
texture, color, and flavor, far better compared to traditional thermally processed
foods (Leizerson and Shimoni 2005b).

7  Emerging Nonthermal Food Preservation Technologies

Although thermal technologies are being used extensively for processing and pres-
ervation of foods, recent consumer awareness about good nutrition and increasing
demand for fresher tasting food have paved the way for emerging nonthermal food
processing technologies. Non-thermal food processing/preservation methods
cause minimal impact on the nutritional and sensory qualities of foods, and yet are
capable of extending shelf life of foods by inhibiting or killing spoilage microor-
ganisms. Nonthermal treatments do not require a heat source and hence are con-
sidered to be more energy efficient and capable of providing better quality
preserved foods compared to conventional thermal based treatments. The emerg-
ing non-thermal technologies also meet the need of industry as it offers value-
added products, new market opportunities and added safety margins. Recent
exploitation of these technologies in inactivating pathogens and background
microflora for preservation of food has been considered in this chapter.

7.1  Ultraviolet Light Treatment

Ultraviolet (UV) radiation is a U.S. Food and Drug Administration (FDA) approved

non-thermal microbial intervention technology that finds increased applications in
surface decontamination of food and food contact surfaces (US-FDA 2001). UV
light’s potential in destroying bacteria, viruses and parasites have been documented
by many researchers (Bintsis et al. 2000; Chang et al. 1985; Demirci 2002; Dunn
et al. 1995; Harm 1980; Huang and Toledo 1982; Rowan et al. 1999; Shechmeister
1983; Sommers et al. 2010; Mukhopadhyay et al. 2014). Like many of its counter-
part decontamination technologies, UV light has a destructive effect on bacteria. UV
light has the ability to inhibit the bacteria’s replication process by dimerization of
their thymine bases in their DNA strands (Demirci 2002; Bank et al. 1991; Sinha and
Hader 2002). Membrane destruction due to localized overheating was hypothesized
as the reason for microbial death (Fine and Gervais 2004). Krishnamurthy (2006)
suggested photo physical effects on the bacterial structure as the main reason when
UV was applied as high intensity pulses. Because the UV light systems vary in
design and depending on whether the UV light is applied continuously or through
pulsed power technology and depending on the specific wavelengths used in the
power source, it is hypothesized that a combination of these factors contributes to the
inactivation of pathogens. Electromagnetic wavelengths of the range 220–280 nm
2  Principles of Food Preservation 25

(most common: 254 nm) and several application doses (mW cm−2) were shown to
deliver the optimum effect. While the UV dose is used to represent the total absorbed
dose, UV fluence in units of μJ cm−2 describes incident UV radiation on non-flat
surfaces. Researchers have reported varied power levels, process times, UV light
source-product distances, product thicknesses to achieve varied inactivation levels in
many pathogens. Many difficulties and subtleties are involved in measuring UV dose
(Chang et al. 1985). Hence, it is difficult to compare and standardize the process
conditions. Bolton and Linden (2003) developed a protocol for bench-­scale UV light
experiments with specifications for construction, determination of fluence (UV
dose) given to microorganisms and microbiological testing.
The effect of UV light (254 nm) on inactivation of Salmonella Senftenberg on
surfaces of tryptic soy agar (TSA), pork skin and pork muscle was evaluated
using UV intensities from 20 to 1000 μW cm−2 (Wong et al. 1998). Intensities
were varied by adjusting the distance (10–30 cm) between the UV lamp and the
treatment surface. It was found that the maximum log reduction was achieved at
UV intensities of 80 μW cm−2 or higher for all surfaces. While >7 logs inactiva-
tion was possible on the agar surface, only 2–3 logs in pork skin and 1–2 logs in
pork muscle was possible at 80 μW cm−2. Greater inactivation rate at 100 μW cm−2
as indicated by lower DUV-value was reported for TSA (15 s), followed by pork
skin (595  s) and pork muscle (1163  s). Similar results were also reported for
beef steak (Stermer et al. 1987), chicken halves (Wallner-Pendleton et al. 1994),
poultry skin (Sumner et al. 1995a), and chicken meat (Kim et al. 2002). In their
study (UV: 250–500 μW cm−2; 3 min) on chicken meat, peptone water and stain-
less steel, Kim et al. (2002) reported 5-log reduction of Salmonella Typhimurium
in peptone water and stainless steel compared to only 1-log and 0.36-log reduc-
tions in chicken meat with and without skin, respectively. The source of UV
light used was a bench lamp (Model UVG-54 of Ultra-violet products, Inc.
USA). Aluminium foil was used to cover the sample tray. Report indicates 7-logs
(99.9%) of Salmonella Typhimurium could be inactivated on agar plates but
only 89.5% reduction was obtained on poultry skin (UV: 3120  μW  cm−2)
(Sumner et al. 1995a). These results indicate that UV irradiation is less effective
on irregular surfaces.
UV light was used in combination (hurdle approach) with other technologies and
antimicrobials more commonly to reap the maximum benefit. This minimizes the
negative impact of single technologies, when used alone, especially on product qual-
ity (Raso and Barbosa-Canovas 2003; Ross et al. 2003). UV light in combination with
antimicrobials (Mukhopadhyay et al. 2015; Uesagi and Moraru 2009), hydrogen per-
oxide (Hadjok et al. 2008) pulsed electric field (Gachovska et al. 2008), were a few of
the recent studies reported for the combined use of UV light with other inactivation
methods to inactivate pathogens and maintain quality for food preservation.
Hadjok et al. (2008) combined UV light with H2O2 to inactivate Salmonella and
a number of other pathogens inoculated on lettuce, spinach, cauliflower, broccoli,
onion and tomato. UV light acts as the oxidizing agent inducing the formation of
highly reactive hydroxyl radicals from H2O2 which react with the cellular
­components of the pathogens to induce their inactivation (Rosenfeldt et al. 2006).
This radical formation and subsequent reaction on cell components is positively
26 S. Mukhopadhyay et al.

influenced by temperature and the optimal temperature is 50  °C (Hadjok et  al.
2008; Yaun et  al. 2004). A combination treatment of UV light (37.8  mJ
cm−2/~0.63 mW cm−2) and H2O2 (1.5%) at 50 °C reduced surface Salmonella of
lettuce by >4 logs and internalized Salmonella by 2.84 log10CFU/mL (Hadjok et al.
2008). H2O2 at 1.5% v/v concentration performed better than 2.0% and the reason
was attributed to neutralization of excess radicals (Reidmiller et  al. 2003). UV
light—H2O2 was reported to be more effective in leafy vegetables such as lettuce
than produce such as cauliflower, broccoli, onion and tomatoes (Hadjok et  al.
2008). The reason for this was reported to be the inability of reactive radicals to
reach deeper into the interior bacterial locations on the latter vegetables.
Mukhopadhyay et al. (2015) reported >4.7 logs reduction of a bacterial composite
made of three serotypes of S. enterica (S. Newport H1275, S. Stanley H0558, and
S. Montevideo G4639) by a combined a low dose (0.6 kJ/m2) treatment of UV light
(253.7  nm) followed by 2  min immersion in a novel antimicrobial formulation
‘HEN’ composed of the antimicrobials hydrogen peroxide, nisin and a surfactant
EDTA at room temperature (22 °C).
Photo reactivation is the light-dependent DNA repairing ability of organisms. In
the presence of UV-A and visible light, the enzyme photolyase reverses the pyrimi-
dine dimers to facilitate the repair (Sinha and Hader 2002).Techniques of applying
UV light in cycles of light and dark as well as UV radiation plus photo reactivation
did not show any significant difference with UV treatment alone (Kuo et al. 1997).
Comparing the sensitivity of Salmonella against other pathogens to UV treatment
is difficult as there are many variables involved and the available data are mixed.
While Salmonella Typhimurium (Kim et  al. 2002) and Salmonella Montevideo
(Hadjok et  al. 2008) were reported to be more resistant than E. coli O157:H7,
Salmonella Senftenberg was reported more sensitive than E. coli (Wong et al. 1998)
to UV irradiation. In general, most of the vegetative bacteria exhibited similar resis-
tance to UV light and required less than 3–15 times of the UV dose requirements for
inactivation of viruses, bacterial spores and amoebic cysts (Chang et al. 1985).

7.2  Pulsed Light Technology

UV radiation could be applied either continuously or as high intensity short duration

pulses (Krishnamurthy and Demirci 2007; McDonald et al. 2000). Pulsed light (PL)
covers a broader spectrum (100–1100 nm) consisting of UV, visible and infrared
radiation. Pulsed Light (PL) is a novel, non-thermal decontamination technology for
food products which uses short time, high frequency pulses of an intense broad
spectrum of light. The spectrum of light for pulsed light treatment includes wave-
lengths in the ultraviolet (UV) to the near infrared region (Krishnamurthy and
Demirci 2007). The material to be sterilized is exposed to at least 1 pulse of light
(typically 1–20 flashes per s) with a duration range from 1 μs to 0.1 s (Dunn et al.
1991). For most applications, a few flashes provide a high level of microbial inacti-
vation. In terms of mechanism of its operation, UV-C light is effective in blocking
microorganisms’ development by altering their DNA. As a physical preservation
2  Principles of Food Preservation 27

method, UV irradiation has a positive consumer image. The US Food and Drug
Administration (FDA) and US Department of Agriculture (USDA) have concluded
that the use of UV irradiation is safe. In 2000, the FDA approved UV-light as alter-
native treatment to thermal pasteurization up to a cumulative dosage of 12 J/cm2 by
FDA Code 21CFR179.41 (FDA (Food and Drug Administration) 2015). The perfor-
mance criterion defined by FDA for fruit and vegetable juice processing is a 5-logs
reduction in the number of the target pathogen of concern (FDA (Food and Drug
Administration) 2015). In addition to the germicidal effect of UV, application of
high intensity pulses also results in rise of food surface temperature temperatures
(Dunn et al. 1991). Due to the physical and thermal effects associated with pulsed
application of UV light, microbial inactivation efficiency is 4–6 times more than
that of continuous application (Fine and Gervais 2004; Dunn 1996).
Levy et al. (2012) demonstrated that pulsed UV light is efficient in achieving more
than 5 log reduction for a range of spore-forming bacteria, for vegetative cells of non-
spore-forming bacteria, and for yeasts spread on agar media. Abshire and Dunton
(1981) found that some species (Pseudomonas aeruginosa) were more sensitive than
others (Micrococcus radiodurans and Candida albicans). The inactivation effect of
intense pulsed light (IPL) on Micrococcus roseus, an irradiation resistant bacterium
has been investigated by Kim et al. (2013). Approximately 6.6 log CFU/ml reduction
in the cell viability for irradiation resistant M. roseus was achieved by 3 min treatment
of 1000 V intensity of PL. Various factors govern the efficacy of PL. A number of
physical factors (dose, input voltage and the UV content of PL), biological factors
(microbial strains, spores, vegetative cells) and environmental factors (surface mor-
phology/quality) have been investigated for the efficacy of pulsed light in microbial
decontamination (Levy et al. 2012). Ozer and Demirci (2006) demonstrated about one
log reduction of E. coli O157:H7 or L. monocytogenes could be achieved by 60-s
treatment at a distance of 8  cm from the UV strobe without affecting the quality.
Nicorescu et al. (2013) compared the inactivation kinetics of pulsed UV light treat-
ment in the dry and liquid state and highlighted that 8 log reduction of B. subtilis
vegetative cells could be achieved in the liquid state compared to only 1 log reduction
in the dry state the same intensity of treatment. Empirical models as a function of
distance, layer thickness and treatment time have been developed for predicting the
population of Escherichia coli O157:H7 during pulsed light UV treatment (Sharma
and Demirci 2003) for eliminating pathogens from alfalfa seeds. Similar models for
deactivation of fungal spores of Aspergillus niger have been developed by Jun et al.
(2003) by varying the treatment time, voltage input and distance from the UV strobe.
Hillegas and Demirci (Hillegas and Demirci 2003) also developed models for pulsed
UV light sterilization system for the inactivation of C. sporogenes in honey. The num-
ber of pulses, the distance between the food product and lamp and depth of the honey
were varied to enhance the inactivation of C. sporogenes by 90%. Krishnamurthy
et al. (Krishnamurthy and Demirci 2007) varied the flow rate (20, 30, and 40 mL/min),
number of passes (1, 2, and 3 times) and distance of the sample from the source (5, 8,
and 11 cm) to optimize Staphylococcus aureus inactivation in milk using the pulsed
UV system. They reported complete inactivation (>7 log10CFU/mL) in only two cases.
Sample distance from the UV source window was the only variable reported by these
researchers that had a significant (P < 0.05) impact on the outcome.
28 S. Mukhopadhyay et al.

Studies on microbial decontamination of food powders (Fine and Gervais 2004)

using pulsed UV light revealed that 58 J/cm2 fluence was sufficient to decrease the
microbial population of S. cerevisiae by 7 logs on glass beads or dried powdered prod-
ucts. Pulsed UV light has been applied to strawberries, blueberries and raspberries at
varying UV doses and times (Bialka and Demirci 2007; Bialka and Demirci 2008).
Maximum reductions of E. coli O157:H7 and Salmonella were reported to be 3.9 and
3.4 log10 CFU/g at 72 and 59.2 J/cm−2 respectively on raspberries. Whereas on the
surface of strawberries, maximum reductions of 2.1 and 2.8 J/cm−2 log10 CFU/g at 25.7
and 34.2 J/cm−2 respectively have been recorded with no visible damage to the fruits.
On blueberries, 4.3 and 2.9 log10 CFU/g reduction was reported. PL treatment has also
been proved to be effective as an intervention strategy to prevent cross-contamination
between equipment and the final product and founding effective when applied along
meat processing lines for decontamination of L. monocytogenes and Escherichia coli
O157:H7 on the surface of a stainless steel meat slicing knife (Rajkovic et al. 2010).
Pulsed UV light has also been used to inactivate semi-opaque and solid systems.
Bialka et al. (2008) studied the efficacy of pulsed UV light technology in solid model
systems. Inactivation and energy penetration data obtained from the treatment of agar
(clear solid medium) and whey protein (opaque solid medium) gels after treatment
with pulsed UV light were used to construct several models to estimate the amount of
energy penetrating the sample at a given depth and the inactivation of E. coli K12. It
was found that pulsed UV light can penetrate opaque materials up to 10 mm deep with
decreasing energy levels and Weibull type model can be used to model the inactivation.
Decontamination studies of Penicillium roqueforti in agar media, bread and preformed
pizza (Mohammadbeygy 2013) revealed that 400 V, 1200 pulses at a distance of 5 cm
from the UV strobe resulted in complete inactivation of P. roqueforti. Overall, pulsed
UV light was found to have a potential use for the decontamination of spoilage micro-
organisms in a variety of products.
Pulsed light has been demonstrated to extend the shelf-life and improve the qual-
ity of several fruits and vegetables (Aguilo-Aguayo et al. 2013; Charles et al. 2013;
Gomez et al. 2012; Klein 1987; Oms-Oliu et al. 2010) and has been used in dairy
(Krishnamurthy and Demirci 2007), egg (Kuo et  al. 1997) and meat (Ozer and
Demirci 2006; Rajkovic et al. 2010; Pollock 2007; Sumner et al. 1995b) products. At
low doses (0.25–8.0  kJ/m2) UV radiation from pulsed light can induce beneficial
reactions in the plant, such as a defense response against pathogens attack via a phe-
nomenon known as hormesis (Gardner and Shama 2000). Many studies have dem-
onstrated that at low doses, the technology can control various kind of storage rots of
fruit and vegetables, delay fruit ripening during postharvest storage and simulate the
synthesis of enzymes or compounds with antimicrobial activity (Klein 1987).
From a nutritional point of view, post-harvest processing generally reduces the nutri-
tional value of fruits and vegetables. As a result, processed fruits and vegetables are
assumed to have lower health promoting capacity when compared to fresh produce (Klein
1987; Nicoli et al. 1999). Few studies have been carried out to determine the use of pulsed
light to enhance the nutritional quality of food materials. The enhancement of antioxidant
content of elderberry fruit by pulsed UV light was studied by Murugesan (2010). They
observed upto 50% increase in the total phenolic content of elderberry fruit by 1.1 J/cm2/
pulse treatment for a 10 seconds treatment time. Charles et al. (2013) studied the impact of
2  Principles of Food Preservation 29

pulsed light treatment on the physical and nutritional quality of fresh-cut “Kent” mangoes.
Pulse light treatment with a total fluence of 8 J/cm−2 maintained the firmness, the color and
the carotenoid content of the fresh-cut mangoes. It was also found to maintain the phenyl-
alanineammonialyase, (PAL) activity and increased polyphenoloxydase, (PPO) activity
after 3 days. For the nutritional aspect, pulsed light maintained phenol and total ascorbic
acid contents similar to that of the control. The effect of pulsed light on surface decontami-
nation (natural and inoculated microorganisms), physical (color, texture and weight) and
nutritional quality (ascorbic acid and major carotenoids) was investigated in red-ripe toma-
toes during 15 days of storage at 20 °C by Aguilo-­Aguayo et al. (2013). They reported that
PL treatment of fresh tomato would result in a reduction in microbiological contaminants
without compromising the nutritional value. However, some appearance defects were
noted on the treated tomatoes. Literature reports on shelf-life studies using pulsed light
treatments is scarce, although pulsed light treatment in combination with low temperature
storage (4 °C) has the potential to extend the shelf-life of various products. In an earlier
work, pulsed UV light treated cold smoked salmon products stored for 30 days exhibited
microbial free products without compromising sensory quality (Pollock 2007) at
4 °C. However, a moderate temperature of 25 °C showed growth of L. monocytogenes and
thus moderate temperatures were found to be problematic for pulsed-­light treated salmon.
The effect of pulsed light combined with an anti-browning pretreatment (ascorbic acid
plus CaCl2 solution) on the quality and shelf-life stability of fresh cut apples was studied
by Gomez et al. (2012). The anti-browning dipping solution was effective in inhibiting
browning on apple surfaces exposed to a PL dose of 71.6 J/cm−2. The results of the work
done by Oms-Oliu et al. (2010) suggested that the application of pulsed light at doses of
4.8 J/cm−2 could extend the shelf-life of fresh-cut mushrooms without appreciable changes
in the texture and antioxidant properties.

7.3  Ultrasound Processing

Ultrasound has applications in many areas of food processing and food analysis.
These include, extraction of aroma and other plant materials (Cocito et al. 1995), food
freezing (Sigfusson et al. 2004; Zheng and Sun 2006), inspection of food containers
(Garcia-Gonzalez et  al. 2007; Griffin et  al. 2001; Pascall et  al. 2002), temperature
monitoring (Haeggstrom and Luukkala 2000), food characterization and analysis
(Kulmyrzaev et al. 2000; McClements 1995; Saggin and Coupland 2001), inactivation
of microbes (Piyasena et al. 2003; Seymour et al. 2002; USDA 2000) and enzymes
(De Gennaro et al. 1999), to name a few. A number of published review papers are
available on these aspects (Hauptmann et al. 2002; Knorr et al. 2004; McClements
1995; Mason et al. 1996; Piyasena et al. 2003; Povey 1998; Samari 1994; Zheng and
Sun 2006; Withers 1996). Its application in microbial inactivation gained importance
in 1990s like any other non-thermal technologies (Sherba et al. 1991). The very sparse
available literature on its application to Salmonellae inactivation is limited to liquid
foods such as water, milk, liquid whole egg and minimally processed fruits and veg-
etables (Seymour et al. 2002; Wrigley and Llorca 1992). This may be due to ultra-
sound’s limitations in reaching deeper areas of solid foods.
30 S. Mukhopadhyay et al.

Mechanical vibrations applied at frequencies higher than 15 kHz generate

ultrasound waves (Raso et al. 1998). When applied to any liquid foods, sonic
waves create alternate compression and expansion cycles inside the liquid.
These lead to the development of bubbles. Generated bubbles grow in size till
the point when it is no longer sufficient to withstand the pressure from outside.
The bubbles collapse (implosion) causing collision between liquid molecules.
This process, called cavitation, along with the resulting shear and rise in tem-
perature (approximately 5500  °C) and pressure (approximately 50  MPa) are
responsible for disrupting the cell structure and subsequently the cell’s death
(Butz and Tauscher 2002; Neppiras 1980; Suslick 1988). Scherba et al. (1991)
suggested that transient cavitation (short violent bursts) may be the reason for
microbial inactivation at low frequency range rather than stable cavitation (less
violent with vibrating gaseous bodies). More the number of bubbles and higher
the duration/intensity of cavitation, better is the microbial inactivation. This
mechanical effect could be improved by increasing the amplitude of the sonic
waves and by applying this under pressure (manosonication) (Pagan et  al.
1999; Raso et al. 1998). Though ultrasound alone can be successful in many
food preservation applications, high treatment intensities may have adverse
effects on food quality (Lee et al. 1989; Wrigley and Llorca 1992). Application
of ultrasound combined with pressure will improve the lethality of a less-inten-
sive treatment. Raso et  al. (1998) reported exponential reduction in decimal
reduction time (DUS-values, time required to reduce the initial microbial popu-
lation by 90% by ultrasound) from 4 to 0.3 min by increased amplitude from 21
to 150 μm at 20 kHz frequency. The influence of pressure was also reported to
increase with increase of amplitude. Application of temperature (thermosoni-
cation) has a negative effect on cavitation (Pagan et al. 1999; Raso et al. 1998).
Also, heat and ultrasound are reported to act independently (Raso et al. 1998).
Thermomanosonication is the application of temperature and pressure toward
specific advantages.
Lethality of ultrasonic treatment, as expressed by its D-value is influenced by the
wave’s amplitude, pressure and temperature. Inactivation rate increased exponen-
tially with increase in amplitude of sonic waves. General equations describing their
influence were developed and reported by Pagan et al. (1999) and Raso et al. (1998).
Manas et al. (2000) investigated the resistance of three Salmonella serovars namely,
S. Enteritidis, S. Typhimurium and S. Senftenberg 775  W which are common in
liquid egg white to pressured ultrasonic waves at varied temperatures.
Salmonella Senftenberg, the most resistive of the three, exhibited greater
resistance to heat treatment than manosonication and the resistance is much
higher in liquid whole egg. Also, the resistance of all three serovars were similar
in manosonication suggesting that a uniform, milder treatment would be suffice
for making liquid white egg safe from Salmonella. Manothermosonication at
60 °C reduced Salmonella Senftenberg 775 W by 3 logs compared to 0.5 log by
heat at 60  °C.  This technology is also more effective on gram-negative than
gram-positive species (Pagan et al. 1999). In general, gram-positive microorgan-
isms are considered more tolerant of the effect of intervention technologies due
to the presence of outer peptidoglycan layer. However, Scherba et al. (1991), in
2  Principles of Food Preservation 31

their quantitative assessment of the germicidal efficacy of ultrasonic energy

(26 kHz at various exposure levels) using various bacteria of different structural
property indicated that the main target of ultrasonic damage may be the inner
cytoplasmic membrane and not the outer presence or absence of the peptidogly-
can layer. It was concluded that the percentage kill by ultrasonic energy was not
influenced by the morphological feature of the microorganisms.
The potential application of ultrasound in effectively inactivating Salmonella
spp. in high sugar chocolate was investigated by Lee et al. (1989). A 4-log reduction
was reported for Salmonella Eastbourne (DUS-value: 3 min) and Salmonella Anatum
(DUS-value: 2.1  min). These DUS values were based on first order kinetics. High
sugar (solids) and high viscosity likely inhibits microbial inactivation because of
reduced cavitation (Hulsen 1999; Wrigley and Llorca 1992).
Seymour et  al. (2002) demonstrated a reduction of 99.8% of Salmonella
Typhimurium on cut iceberg lettuce by using power ultrasound (32–40 kHz) and
chlorine (25 ppm) together. This was one log more than that of chlorine (1.7 logs)
or ultrasound (1.6 logs) used independently and two logs more than that of water dip
without agitation (0.7 logs). Similar results were reported by Kim et al. (1999) in
their experiments with chlorinated water (99% reduction), ozonated water (99.5%)
when compared to only 90 per cent of ultrasound treatment. The improved reduc-
tion by combining chlorine and ultrasound was supposed to be due to the ultra-
sound’s cavitation effect in detaching the microbial cells from the produce surface
rendering them susceptible to the sanitizing effect of chlorine. Sonication aiding the
effect of chlorine in Salmonellae inactivation on broiler breast skin was highlighted
by Lillard (1993, 1994). The irregular skin surface of the broiler skin providing
physical protection to the Salmonella spp. was indicated as the possible reason for
the decreased less effect of sonication when used alone (Sams and Feria 1991).

7.4  Bacteriophage Application

With the fresh-cut industry looking for an alternative to chlorine and chlorine based
compounds, potential use of safe, clean and novel biocontrol agents such as lytic
bacteriophages is gaining importance. As the bacteria became resistant to antibiot-
ics, medical researchers, especially in the erstwhile Soviet Union and Eastern
European countries started using bacteriophages for treating diseases due to patho-
gens including Salmonella (Alisky et al. 1998). Bacteriophages are viruses capable
of lysing specific bacteria by penetrating their cell membrane and disrupting their
metabolic processes. . Ubiquitous presence of these phages and their flexibility in
preparation to act against specific strain or serotype of any pathogenic bacteria
makes it a promising alternative to treat minimally processed food produce.
Leverentz et  al. (2001) examined the application of a phage cocktail mixture
with a specificity for Salmonella Enteritidis on fresh-cut honeydew melon (pH5.8)
and apple slice (pH4.2) surfaces. They also evaluated the effect of these bacterio-
phages at various storage temperatures (5, 10 and 20 °C) in comparison to standard
chemical sanitizers. The main limitation of this application is the development of
32 S. Mukhopadhyay et al.

resistance to phages by the target pathogens, the use of cocktail of phages to take
care of mutants is recommended. The mixture was found to be more effective than
chemical sanitizers and reduced the inoculated (1 × 106 CFU/mL) Salmonella popu-
lations by 2.5 to 3.5 logs on honeydew melon slices. High temperature storage
(20 °C) resulted in less reduction and no significant reduction was observed in apple
slices. This result was hypothesized to be the result of high acidity (pH4.2) of apple
slices inactivating the phages along with the bacteria. Higher concentrations of bac-
teriophages and development of low-pH tolerant phages may make this technology
suitable for all types of fruit and vegetables.
The ratio of host cells and phages, described as ‘multiplicity of infection
(MOI)’, ‘multiplicity of adsorption (MOA)’ and PFU/CFU ratio by various
researchers was considered to be important for the successful application of this
technology (Kasman et  al. 2002; O’Flynn et  al. 2004; Whichard et  al. 2003).
Bigwood et al. (2009) conducted experiments to determine the influence of host
and bacteriophage concentrations on Salmonella inactivation by a Salmonella
phage (P7). It was reported by the authors that the host cell concentration did not
influence the inactivation efficiency for a given phage concentration. However,
increase in phage concentration from 1.8  ×  104 to >5  ×  198 PFU/mL yielded
increased inactivation with the lowest concentration showing no inactivation to
the highest concentration showing almost complete inactivation.

7.5  Membrane Processing

Membrane processing is a unique method for liquid foods which is driven by pres-
sure difference across a microporous membrane. It is an attractive alternative to
energy intensive thermal processing. In addition, r membrane processed foods are
superior in terms of nutritive value and sensory quality. Membrane processing does
not require application of heat and provides the ‘cold pasteurization’ option by sim-
ply removing the harmful bacteria, spores and other microbial contaminants from
food. It can also be used to concentrate dilute liquid foods as milk, juice, etc.
Microfiltration is a pressure-driven membrane process which can separate fine par-
ticles, microorganisms and other contaminants from fluids such as liquid foods.
Membranes used in microfiltration process have a microporous structure that
separates particulate matters in size ranges from 0.1 to 10 μm. In cross-flow MF, the
fluid to be filtered flows parallel to the membrane surface, which reduces pore block-
age and hence facilitates a quasi-steady filtrate flow for a longer time compared to
standard filtration processes. Cross-flow microfiltration has been explored as a means
to remove natural microflora from milk. Guerra and coworkers observed a high spore
reduction in the range of 104–105 spores/mL of Bacillus cereus and lactate ferment-
ing Clostridium spores from skim milk by cross-flow MF using a 1.0 micron ceraflo
ceramic membrane (Guerra et al. 1997). In microfiltration, selection of the appropri-
ate membrane and process conditions not only provides rapid removal of pathogens
and spores from food but also minimize the loss of nutrients and quality functional
2  Principles of Food Preservation 33

properties of the food. Thus, from a safety and functionality and consumer accept-
ability point of view, cross-flow microfiltration processing is advantageous over con-
ventional thermal processing.
The performance of a membrane process is determined by the amount and the
stability of the permeate flow through a unit area of membrane in a second and is
called “permeate flux,” which is influenced by feed concentration, temperature and
feed velocity. Permeate flux increases with transmembrane pressure (TMP), which
is the pressure differential across the membrane between the feed and permeate.
Performance of a microfiltration processing usually declines over time mainly due
to membrane fouling and other factors. Microfiltration processing has been success-
fully applied for cold pasteurization of milk (Cheryan 1998; Saboya and Maubois
2000; Tomasula et al. 2011) and eggs (Mukhopadhyay et al. 2009). Reports on pro-
cessing of eggs in the form of liquid egg describes removal of natural background
microflora (Mukhopadhyay et  al. 2009), human pathogens such as Salmonella
Enteritidis (Mukhopadhyay et  al. 2010) and spores of Bacillus anthracis
(Mukhopadhyay et  al. 2011) using a ceramic GP membrane. The author in this
study reported no significant (p < 0.05) change in liquid eggs protein composition
and individual egg protein distribution due to microfiltration processing.

8  Conclusions

Many of the reviewed preservation technologies are extensively studied and await-
ing commercial adoption. Commercialization is hampered by the variation in
research results due to non-standard process equipment, procedures, test conditions,
microbial strains and their stress tolerance to the various processes. Any small
increase in the processing cost would deter the conservative food industry from suc-
cessful adoption of these technologies. Joint research initiatives among academia,
industry and regulatory authorities should clear the path for commercialization of
many of these technologies. Combination or integration of compatible preservation
technologies (hurdle approach), both conventional and novel should be identified
and research should be explored vigorously for improved efficacy of microbial inac-
tivation but minimal effect on the quality of food.

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