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5 ADVANCED PRESERVATION TECHNOLOGIES FOR MILK AND MILK

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 95

Recent Technologies in Dairy Science (2019): 95-117

Editors : Rekha Rani, Bhopal Singh, Chetan N Dharaiya, Hanslal and Arghya Mani
Today & Tomorrow’s Printers and Publishers, New Delhi - 110 002, India

5
ADVANCED PRESERVATION
TECHNOLOGIES FOR MILK AND
MILK PRODUCTS
Bhopal Singh*, Anindita Debnath, Rekha Rani and
Chetan N Dharaiya

ABSTRACT
Preservation of food has always been emphasized as one of the essential
aspects in food sectors since ancient times as milk and milk products
are perishable in nature. Several preservation methods are, therefore,
practiced to make the products safe and to enhance their shelf life.
The thermal treatments (such as pasteurization, sterilization etc.) are
the largely used methods for enhancing the shelf life of milk and milk
products. The other means of preservation of dairy products include
chilling, freezing, drying, fermentation, salting, smoking, addition of
chemical preservatives like BHA, BHT, sorbic acid, benzoic acid etc.
The above mentioned processes are limited by their adverse effects
either on nutritional value or on human health. The demand for fresh
products, free from any harmful additives and minimally processed is,
therefore, increased. This necessitated the use of some newer
preservation technologies. Studies revealed that emerging technologies
like membrane processing, high hydrostatic pressure, pulse electric
field, ultraviolet radiation, ultrasonication, ohmic heating, microwave
treatment can be considered as the promising technologies for
enhancing the shelf life of dairy products without causing drastic
changes in sensory and nutritional properties of the products. In
addition to these newer processing technologies, preservatives from
natural ingredients like extract or essential oils from spices and herbs
have also introduced a new way of preservation.
Keywords: Preservation, Emerging technologies, Essential oils
96

1.0 INTRODUCTION
India is the largest producer of milk in world accounting 165.4
MMT of milk annually with per capita availability of 355 g/day (NDDB,
2018). Milk is considered to be an adequate source of valuable
macronutrients (fat, protein and lactose) as well as micronutrients (vitamins
and minerals), making it a wholesome food. It can also serve as an excellent
carrier product for nutrient, and if enriched or fortified, it can satisfy the
nutritional needs of the population. It is perishable in nature hence,
converted in heat desiccated products (condensed milk, basundi, rabri,
khoa, peda, burfi, kalakand, milk cake etc.) cultured or fermented products
(dahi, yoghurt, cheese) dried products (butter powder, channa powder,
skim milk powder, whole milk powder, milk protein concentrate, whey
protein concentrate) frozen desserts (Icecream, kulfi, lollies), acid and heat-
acid coagulated products (channa, chakka, paneer, rasogolla, channa-
murki), fat rich products (cream, butter, ghee, butteroil) and desserts (kheer,
rasmalai) to increase the shelf-life. By-products of dairy industry (skim
milk, butter milk, ghee and ghee residue) can be further utilized for
preparation of longer shelf-life products. In atmospheric conditions these
products are also spoiled within a week hence that proper preservation
method is essential to increase the market value of dairy products and
economy of dairy industry and also provide the safe food to the consumer.
Food preservation is a method of maintaining desired food quality
as long as possible by minimizing the rate of quality deterioration.
Therefore, understanding the effect of each preservation method on the
quality attributes of food is an important aspect. The major food preservation
technologies to enhance the shelf life of food products either by slowing
down or inhibiting chemical deterioration and microbial growth, or by
directly inactivating bacteria, yeasts, molds, or enzymes, or by avoiding
recontamination before and after processing (Gould, 1989; Gould, 1995)
(Table 1).
Traditionally milk and milk products are preserved either by
physical means like heating such as pasteurization, sterilization, ultra-high
heat treatment (UHT), chilling, freezing and drying or by chemical means
like addition of butylated hydroxy toluene (BHT), butylated hydroxyanisol
(BHA), salts of benzoic acid, salts of sorbic acids etc., salting, smoking,
LP-system or by microbiological means like fermentation methods. Many
of these processes diminish the nutritional value of foods and/or cause
adverse effects on human health. Among the above processes thermal
processing is considered as one of the most reliable and effective methods
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Table 1 Major food preservation techniques (Gould, 1989; Gould, 1995)

Food Preservation Methods
Inhibition Inactivation Avoid recontamination
1. Low temperature storage 1. Sterilization 1. Packaging
2. Reduction of water activity 2. Pasteurization 2. Hygienic processing
3. Decrease of oxygen 3. Irradiation 3. Hygienic storage
4. Increase of carbon dioxide 4. Electrifying 4. Aseptic processing
5. Acidification 5. Pressure treatment 5. HACCP
6. Fermentation 6. Blanching 6. GMP
7. Adding preservatives 7. Cooking 7. ISO9000
8. Adding antioxidants 8. Frying 8. TQM
9. Control of pH 9. Extrusion 9. Risk analysis and
management
10. Freezing 10. Light
11. Drying 11. Sound
12. Concentration 12. Magnetic field
13. Surface coating
14. Structural modifications
15. Chemical modifications
16. Gas removal
17. Changes in phase transition

18. Hurdle technology

for extending shelf life.Nowadays the demand for fresh and natural food,
free from chemical preservatives and other harmful additives, minimally
heat processed, containing less salt or sugar and at the same time have a
reasonable shelf life with assured quality have increased. Several novel
technologies like high hydrostatic pressure, pulse electric field, UV light,
ultrasonication, microwave treatment, ohmic heating have been proposed
by various workers to meet this demand. Besides these emerging
technologies, application of different types of natural ingredients like spices
and herbs, essential oils have been practiced to enhance the shelf life of
milk and milk products.
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2.0 PRESERVATION TECHNIQUES

2.1 HIGH PRESSURE PROCESSING (HPP)
In HPP processing, the food product is placed in a pressure vessel
filled with the pressure-transmitting medium. Water is generally used as
the pressure-transmitting medium. Castor oil, silicone oil, sodium benzoate,
ethanol or glycol can also be used as medium. Application of high pressure
to food based on two major principles - Le Chatelier’s Principle and Isostatic
Rule.
As per Le Chatelier’s Principle when a system at equilibrium is
disturbed, the system responds in a way that tends to minimize the
disturbance (Pauling, 1964). According to this principle high pressure
stimulates those reactions which cause a decrease in volume and opposes
reactions that involve an increase in volume. The phenomenon like phase
transition, change in molecular configuration, chemical reaction are the
result of high pressure.
As per Isostatic Rule pressure is instantaneously and uniformly
transmitted throughout a sample under pressure, whether the sample is in
direct contact with the pressure medium or hermetically sealed in a flexible
package that transmits pressure (Olsson, 1995). Pressure is transmitted in
a uniform (isostatic) and quasi instantaneous manner throughout the
sample; the time necessary for pressure processing is therefore independent
of sample size, in contrast to thermal processing. HPP enhances shelf life
of food products by inactivating spoilage causing microorganisms. Different
mechanisms of inactivating microorganisms by high pressure have been
proposed. Mc Clements et al. (2001) reported that application of high
pressure caused modification in permeability and ion exchange properties
of cell membrane. Linton et al. (2000) proposed that denaturation of key
enzymes and disruption of ribosomes is responsible for microbial
inactivation. It has been observed that different degrees of high pressure
are needed to inactivate different types of microorganisms. The resistance
of microorganisms to high pressure varies with several factors like
processing conditions (pressure, time, temperature and cycles), food
constituents, its properties and, type and physiological state of the
microorganism (Smelt, 1998). Cheftel (1992) reported that microorganism
at their exponential stage of growth are more sensitive to high hydrostatic
pressure than at stationary growth phase. The vegetative cells are more
prone to pressure treatment than bacterial spores. The bacterial spores can
tolerate pressure of 1000 MPa (Cheftel, 1992). Cheftel (1995) reported
 99

that 200–400 MPa pressure is required to inactivate most vegetative cells.

It was reported that combination of higher pressure and temperature is
required to inactivate bacterial spores (Rovere et al., 1996; Reddy et al.,
1999 and Balasubramaniam, 2003). Lethal effect, similar to conventional
thermal treatment, against harmful vegetative bacteria in food products
was obtained by subjecting the food product to 400 - 600 MPa pressure at
a temperature of 60°C (Cheftel, 1995; Farkas and Hoover, 2000 and
Raghubeer et al., 2000). It was also reported that the combination of pressure
and temperature caused minimal nutrient degradation. The gram positive
bacteria are more resistant to high pressure processing than gram negative
bacteria. Vachon et al. (2002) reported that processing of milk by dynamic
high pressure treatment inactivated 3 major food pathogens – Listeria
monocytogenes LSD 105-1, Escherichia coli O157:H7 ATCC 35150 and
Salmonella enteritidis ATCC 1304. Krasowska et al (2005) observed that
the application of high pressure in the range of 300-500 MPa for 15 min to
milk reduced the acid producing ability of lactococci. Rademacher and
Kessler (1997) suggested application of high pressure treatment of 400
MPa for 15 min or 600 MPa for 3 min at 20°C to milk to get a shelf-life of
10 days at 10°C. It was reviewed that high hydrostatic pressure in the
range of 400– 600 MPa can give raw milk the same quality as that of
pasteurized milk.
2.2 PULSED ELECTRIC FIELD
Pulsed Electric Field is another emerging non thermal technology
can be used in place of traditional thermal process toinactivate the
microorganisms as well as enzymes inliquid type foods such as milk (Shamsi
and Sherkat, 2009). The pulsed electric field process involves application
of short pulses of high intensity electric field to liquid food flowing between
two electrodes for milliseconds (Barbosa Canovas et al., 2000 and Gaudreau
et al., 2008) to achieve inactivation of microbial load in the food products
to a desirable extent. Generally, multiple short duration pulses of 20-80
kV/cm for less than 5 µs are applied for the purpose. The electric field in
the form of exponentially decaying, square wave, bipolar, or oscillatory
pulses and at ambient, sub-ambient, or slightly above ambient temperature
were applied to different liquid food system. However, the combination of
heating (40-50ÚC) and pulsed electric field is often preferred to enhance
the shelf life of food products.Two mechanisms namely electroporation
and electrical breakdown as discussed below have been put forward by the
scientists to explain microbial inactivation by pulsed electric field
technology.
100

In electroporation theory, bacterial cell membrane regulate the flow

of nutrients into the cell and metabolic wastes out of it through the pores
present in membrane. Application of high voltage electrical field to
microorganisms causes enlargement of these pores on the cell membrane
and as a consequence the release of cell constituents into the medium. The
lipid bilayer and proteins of the cell membrane get destabilized when
exposed to the electric field. It was reported that application of pulsed
electric field forms some stable hydrophobic pores which can conduct
electric current without causing localized heating effects and converts the
lipid bilayer from a rigid gel to crystalline lipid. The cell plasma membrane
becomes more permeable. Permeation of small molecules into the cell
results in swelling and finally rupturing of the cell membrane. This
phenomenon is called electroporation in which inhibition or destruction
of microbial cell occurs mainly due to compression and poration of
membrane. Increase in temperature is moderate (less than 30°C). Increase
in the intensity of electric field and pulse duration or decrease in the ionic
strength of the medium can form larger pores in cell membrane (Tsong,
1991; Knorr et al., 1994; Ho and Mittal, 1996; Pothakamury et al., 1996
and García et al., 2007).
According to the concept of electrical breakdown theory bacterial
cell membrane is considered as a capacitor filled with dielectric material
of low electric conductance and a dielectric constant of 2. The normal
potential difference across the membrane (about 10 mV) is increased when
subjected to an external electric field. The increase in potential difference
reduces the thickness of membrane. Larger pores are formed in the
membrane when the potential difference across the cell membrane reaches
a critical level of about 1 V. Formation of large pores causes discharge of
cell material and further damage of membrane. Reversible membrane
breakdown occurs when the pores are small in relation to the total membrane
surface but the cells are destroyed when the pores are large and covers a
large membrane area (Zimmermann, 1986; Chen and Lee, 1994 and
Sepulveda et al., 2005a).
Several studies have been conducted to evaluate the effect of pulsed
electric field having different intensities in inactivation of microorganisms
in milk. Microbial reduction by 2 log cycle in raw skimmed milk was
achieved by Raso et al. (1999). Pseudomonas fluorescens, Lactococcus
lactis and Bacillus cereus showed a reduction of 0.3 to 3 log cfu/ml of
UHT skimmed milk when subjected to 35 kV/cm electrical field with 64
pulses of bipolar square wave for 188 µs and having a flowrate of 1 ml/s
(Michalac et al., 2003). Dutreux et al. (2000) observed less than 1 log
 101

cycle difference between inactivation of Escherichia coli in pulsed electric

field treated milk and in pulsed electric field treated phosphate buffer.
Smith et al. (2002) observed that microbial count of milk was reduced
from an initial count of 107 - 108 cfu / ml to a final count of < 250 cfu/ml
after treated with combination of lysozyme and/or nisin, and pulsed electric
field (80kV, 50 pulses). Bendicho et al. (2002a) accomplished pasteurization
of milk by subjecting it to pulsed electric field processing and observed
reduction in microbial load with minimal undesirable changes in milk.
Sepulveda et al. (2005b) got success in shelf life enhancement of milk by
applying pulsed electric field with intensity of 35kV/cm, 2 pulses for 2.3
µs to milk after high temperature short time (HTST) pasteurization. When
milk was subjected to pulsed electric field immediately after pasteurization,
remained acceptable for 60 days at 4°C, whereas, application of pulsed
electric field to pasteurized milk after 8 days of storage at 4°C extended
the shelf life to 78 days at 4°C. The application of high intensity pulsed
electric field (either 40 kV/cm for 60 ìs or 36 kV/cm for 84 ìs) enhanced
shelf life of skim milk to 14 days at 4ºC (Fernandez-Molina et al., 2005).
The shelf life of whole milk subjected to high intensity pulsed electric
field treatment (35.5 kV for 1000 ìs with 7 ìs bipolar pulses at 111 Hz) was
reported as 5 days at 4°C. The mesophilic aerobic count of the milk was
reduced from 3.2 log to 1 log cycle. No significant changes in acidity, pH
and free fatty acids content were observed (Odriozola-Serrano et al., 2006).
Sobrino-López and Martín-Belloso (2010) tried to evaluate the effect of
pulsed electric field on chocolate milk. The chocolate milk subjected to
high intensity pulsed electric field (30 kV/cm, 45 ìs) prior to heating at
105ºC and 112ºC for 31s, exhibited a shelf life of 119 days at 37°C. Floury
et al. (2006) compared the cumulative effect of combination of pulsed
electric field and thermal treatment with the effect of pulsed electric field
treatment alone to skim milk and observed that application of pulsed electric
field along with heat treatment reduced the Salmonella enteritidis count in
skim milk by 2.3 log cycle as compared to 1.2 log cycle reduced after
application of pulsed electric field treatment alone. Sharma et al. (2018)
compared the pulsed electric field treated milk with conventionally
pasteurized milk and reported that microbial effect of pulsed electric field
treatment was similar to conventional pasteurization; however, pulsed
electric field caused no thermal induced damages.
Pulsed electric field treatment is reported to have ability to
inactivate many enzymes. It was assumed that during pulsed electric field
treatment occurrence of unfolding, denaturation and breakdown of covalent
bonds, and oxidation reduction reactions might be responsible for enzyme
102

inactivation (Barsotti and Cheftel, 1999). The effects of pulsed electric

field treatment on activities of alkaline phosphatase (ALP), lipases,
lactoperoxidase (LPX) and proteases (e.g. plasmin) in milk and SMUF
has been reported by several researchers (Bendicho et al., 2002b; Castro
et al., 2001 and Vega-Mercado et al., 1995). A 65% reduction in alkaline
phophatase enzyme activity in simulated bovine milk ultrafiltrate and skim
milk with process conditions of 70 pulses of 740 µs at 22 kV/cm and 18.8
kV/cm, respectively (Castro et al., 2001). Van Loey et al. (2001) observed
26% reduction in ALP activity in milk after processing at 20kV/cm with
200 pulses of 2µs at 70°C. Plasmin content in simulated bovine milk ultra
filtrate decreased by 60% and 90% when process conditions of 30 or 45
kV/cm with 50 pulses of 2 µs pulse duration were used at 10°C and 15°C,
respectively (Vega-Mercado et al., 1995). Lysozyme was, however, not
inactivated by an applied electric field of 38 kV/cm strength but pepsin
activity reduced considerably (Yang et al., 2004).
2.3 OHMIC HEATING
Ohmic heating exhibits its potentiality to enhance shelf life of food
products without causing major heat damage to the quality of food. Ohmic
heating is also known as resistance heating or Joule heating or electro
heating. Food that contains water and ionic salts can conduct electricity
and at the same time exhibit electric resistance which causes heat generation
as well as temperature rise within the food product. Ohmic heating involves
dissipation of electrical energy volumetrically in the form of heat using an
electrical conductor.
Knirsch et al. (2010) repor ted that the inactivation of
microorganisms in food product subjected to ohmic heating is occurred
because of electroporation of cell membrane. Kumar et al. (2014) prepared
paneer from both ohmically heat treated milk sample and conventionally
heat treated milk sample (in both case temperature increased from 20°C to
72°C) and compared the microbial effect of both treatment. It was observed
that microbiological load was lower in paneer made from ohmically heat
treated milk than conventionally heat treated milk.
2.4 ULTRAVIOLET (UV) LIGHT
The UV light radiation is mainly used for surface sterilization.
However, the interests for use of UV light radiation for food preservation
have been growing. The UV light radiation with the wavelength of 100 to
400 nm was applied for food processing. Bintsis et al. (2000) and Sastry et
al. (2000) reported that UV light radiation with wavelength ranges from
 103

100 to 400 nm have higher energy than infrared and lower energy than X-
rays. When a food product is subjected to UV light radiation the energy is
transferred via photons to the food material. The high energy of photons,
released from UV light radiation cause ionization of molecules. As a
consequence the molecules after absorbing the energy are transformed from
their ground state into excited state.The excited molecules come back to
their ground state by releasing energy either as heat or as photons. During
releasing energy some chemical changes are taken place (Demirci and
Krishnamurthy, 2011). In addition to chemical changes, some biological
effects have also been observed.
UV radiation helped in microbial inactivation by affecting and
modifying the DNA of microbial cell resulting in either inhibition of cell
growth or cell death (Liltved and Landfald, 2000). Wright et al. (2000)
observed the bactericidal effect of UV-C (254 nm) radiation and reported
that UV-C radiation caused a cross linking between neighboring thymine
and cytosine (pyrimidine nucleoside bases) in the same DNA strand.
Kramer and Ames (1987) reported that UV-A (315-400 nm) affected
bacterial cells by causing membrane damages and/or generating active
oxygen species or H2O2 on cell structure.The UV light having wavelength
in the range of 220 to 300 nm exhibits germicidal effect against bacteria,
viruses, protozoa, molds and yeasts, and algae (Bintsis et al., 2000;
Perchonok, 2003 and Sizer and Balasubramaniam, 1999). Bachmann (1975)
reported that UV light with 250 to 270 nm wavelength provides highest
germicidal effect. In major cases, therefore, 254 nm wavelength is used
for disinfection. It was also noticed that owing to bigger cell size fungi and
yeasts are more resistant to UV light than bacteria Bachmann (1975). The
UV radiation absorption coefficient of raw milk is 290cm-1 at 253.7 nm
wave length of UV radiation. The negative impact of UV light radiation
on milk was noticed by Koutchma (2009). The photosensitive nature of
milk is the reason. Krishnamurthy et al. (2007) applied pulsed UV light at
5, 8, or 11 cm distance from a UV light strobe with 20, 30, or 40 ml min–1
flow rate up to three times by re-circulation on continuously flowed milk
and observed inactivation of pathogenic organism Staphylococcus aureus
by 0.55 to 7.26 log cfu/ml. The reduction of Mycobacterium avium subsp.
Para tuberculosi in UV light treated milk by 0.5 to 1.0 log/ml was reported
by Altic et al. (2007). Matak et al. (2007) reported about some sensorial
and nutritional changes because of hydrolytic rancidity and oxidationin
UV light treated goat milk. UV-Ctreatment caused more than 5 log cfu/ml
reduction of Listeria monocytogenes in goat milk (Matak et al., 2005). Lu
et al. (2011) reported that application of the 21.3 mJ/cm2 UV dose caused
104

3–4 log cfu/ml reduction in Salmonella spp., Shigella spp., Listeria

monocytogenes, Staphylococcus spp., Enterobacteriaceae, LAB,
pseudomonads and the total aerobic bacteria.
2.5 ULTRASONICATION
Pressure waves of more than 20 kHz frequency is used in
ultrasonication process. The process exhibits its potentiality to be used in
food preservation process by inactivation of microorganisms (Butz and
Tauscher, 2002). Ultrasonication creates cavitation in liquid. Ultrasound
passed through the medium by a series of compression and rarefaction
waves. When this rarefaction cycle exceeds the attractive force of the
molecules of liquid causes formation of cavitation bubbles. At expansion
phase the vapour or gas enters from the medium into the bubble, results in
increase in size of bubbles. Owing to the occurance of simultaneous
expansion and compression the grown bubbles collapse after completion
of few cycles. The formation, growth and collapse of bubbles generate
high localized temperatures, pressures and turbulence (Ashok kumar and
Grieser, 2007 and Brncic et al., 2010).
The microbial inactivation by ultrasonication process is occurred
due to acoustic cavitation. The generation of acoustic cavitations cause
cell membrane thinning (Sams and Feria, 1991), confined heating (Suslick,
1998), singlet electron transfer in cooling phase (Lee and Feng, 2011),
hydroxyl radical formation (Kadkhodaee and Povey, 2008) which increase
the cell membrane permeability and cause cell leakage.The resistance of
different microorganisms to ultrasonication depends on their size and
shapes, cell characteristics, growth stages etc. It was observed that smaller
size cells are more resistant than bigger size cells. The sensitivity of rod
shaped bacteria are more than cocci shaped. The Gram-positive bacteria
exhibit more resistance to ultrasonication than Gram-negative bacteria.
Aerobic bacteria are more resistant than anaerobic bacteria. The younger
cells are more sensitive than older ones and vegetative cells are more
sensitive than spores (Paci, 1953; Jacobs and Thornley, 1954; Davies, 1959
and Ahmed and Russell, 1975). The efficacies of ultrasound process together
with other processes like application of pressure, temperature have been
studied. The effects of ultrasonication (ultrasound at low temperature),
manosonication (ultrasound together with pressure), thermosonication
(ultrasound together with heat) and manothermosonication (ultrasound
together with pressure and heat) processes against microorganisms were
studied by Zheng and Sun (2006), Ercan and Soysal (2013), Mason et al.
(1996), Villamiel et al. (1999) and Chemat et al. (2011), respectively.
 105

Guerrero et al. (2001) got success in inactivation of Saccharomyces by

applying combination of heat and ultrasound. Piyasena et al. (2003)
recommended manosonication and thermosonication as the promising
means of microbial destruction. Ultrasound has also been used in
combination with other non-thermal technologies. San Martin et al. (2001)
applied pulses of 20 kHz ultrasound together with magnetic fields, high
pressure and lisozyme treatments to reduce contamination by Escherichia
coli ATCC 11775. Vercet et al. (1997) reported that manothermosonication
inactivated both extracellular lipase and protease more effectively than
heat treatment alone. Bermúdez-Aguirre and Barbosa-Cánovas (2008) and
Bermúdez-Aguirre et al. (2009) observed higher efficiency in inactivation
of Listeria innocua and mesophilic bacteria in thermosonication treated
raw whole milk than pasteurized milk. Garcia et al. (1989) reported that
the survival rate of Bacillus subtilis in milk subjected to a combination of
heat (70–95°C) and ultrasound (20 kHz, 150 W) was lesser than in milk
treated either by heat or ultrasound alone. However, thermosonication of
milk caused some off flavour generation (Riener et al., 2009). Villamiel
and de Jong (2000) reported that the functional properties of the native
enzymes present in milk were not affected by ultrasound.
2.6 MICROWAVE
Microwave heating is a dielectric heating caused by polarization
effects at a selected frequency bandin a nonconductor. Microwave heating
in foods is because of coupling of electrical energy from an electromagnetic
field in a microwave cavity with the food and its subsequent dissipation
within food product. The coupling of electrical energy and its dissipation
increases the temperature within the product. In microwave treatment, the
alternating current electric fieldcauses dipole rotation of polar solvents as
well as back and forth movement of dissolved ions. In analternating current
electric field, the polarity of thefield is changed at the rate of microwave
frequency. The polar molecules and ions change their alignment with the
changing field. As a consequence the molecular friction takes place and
generating heat (Oliveira and Franca, 2002).
Microwave treatment is a very popular means of food processing.
Microwave heating can be used for pasteurization and sterilization of food
products. The pasteurization or sterilization through microwave processing
can be achieved within shorter time as compared to the conventional
pasteurization or sterilization processes. Microwave processing, therefore,
ensures minimal quality changes in food products. Microwave
pasteurization of milk by continuous flow has been found to be an efficient
106

way to achieve a product with satisfactory microbial and sensory quality

without excessive heat damage (Villamiel et al., 1996). Microwave
pasteurization had no adverse effects on milk flavour (Valero et al., 2000).
Al-Hilphy and Ali (2013) have pasteurized cow milk by microwave flash
pasteurizer.
2.7 MICROFILTRATION (MF)
The MF also showed its efficacy in enhancing the shelf life of
liquid milk by reducing the microbial load of it without causing any changes
in its composition and sensorial qualities (Pafylias et al., 1996). Hoffmann
et al. (2006) proposed membrane filtration as a means of reducing the
microbial load of milk without affecting the milk composition. However,
Hoffmann et al. (2006) suggested to use microfiltration process for low fat
milk, though minor changes in protein, calcium and lactose were observed
after the treatment. García and Rodríguez (2014) developed extended shelf
life (ESL) milk with a shelf life of 33 days by using microfiltration together
with thermal treatment. A least compositional change was observed in ESL
milk from raw untreated milk.
2.8 NATURAL PRESERVATIVES
Studies revealed that spices and herbs can be used as an alternative
preservative in food. The use of essential oils is an age old practices. The
extracts and essential oils from spices and herbs can also be a potential
alternative to synthetic chemical preservatives. Spices acts as antimicrobial
agents by preventing the growth of spoilage microorganisms and by
inhibiting/regulating the growth of pathogens (Tajkarimi et al., 2010).
Essential oils are typically obtained by steam distillation of the plant or
plant parts. Essential oils are reported to possess antibacterial, antiviral,
anti-inflammatory, antifungal, antimutagenic, anticarcinogenic, and
antioxidant activity (São Pedro et al., 2013). The essential oils, being rich
in volatile aromatic compounds and several bioactive compounds are used
either as food flavourings or as fortifying ingredients. Nowadays the
effectivity of essential oils as food preservatives has also been studied.
Chemically, essential oils are variable mixtures of monoterpenes (C10),
sesquiterpenes (C15), diterpenes (C20), a variety of low molecular weight
aliphatic hydrocarbons, acids, alcohols, aldehydes, acyclic esters or
lactones, exceptionally N- and S-containing compounds, coumarins and
homologues of phenylpropanoids (Benchaar et al., 2008). The antimicrobial
activity of essential oil depends on the amount of these chemical
components into it. The mechanisms behind the antimicrobial activity of
essential oil involve degradation of the cell-wall as well as cytoplasmic
 107

membrane materials like membrane protein. As a result the membrane

permeability increases. The increased membrane permeability reduces the
membrane potential, causes leakage of cell contents, coagulation of
cytoplasm, reduces the proton motive force, decreases the intracellular
ATP pool via decreased ATP synthesis and enhances hydrolysis (Nazzaro
et al., 2013). Burt (2004) reported that being hydrophobic in nature,
essential oils separate lipids from the bacterial cell membrane and
mitochondria, make membrane more permeable and as a consequence cause
leakage of cell contents.The cell wall of Gram-negative bacteria is more
resistant to the activity of EOs and their components. The Gram-negative
cell wall does not allow for the entrance of hydrophobic molecules as
readily as Gram-positive bacteria; thus, EOs are less able to affect the cell
growth of the Gram-negative bacteria (Nazzaro et al., 2013). Burt (2004)
reported that the highest antimicrobial activity is exhibited by essential oil
from oregano followed by clove, coriander, cinnamon, thyme, mint,
rosemary, mustard, cilantro/sage. Ceylan and Fung (2004) evaluated the
effect of thyme, mint, bay leaves and their alcohol extracts on the growth
of Salmonella typhimurium, Vibrio parahaemolyticus and Staphylococcus
aureus in a medium. The highest inhibitory effect was exhibited by Thyme.
Among these organisms Salmonella typhimurium was reported as the least
sensitive to these spices. Souza et al. (2005) also reported about the
inhibitory action of thyme (Thymus vulgaris), mint (Mentha piperita) and
laurel (Laurus nobilis) ground leaves and their extracts against
Staphylococcus aureus, Salmonella typhimurium and Vibrio
parahaemolyticus.The inhibitory effect of garlic (Allium cepa), ginger,
mustard and ground clove was evaluated against the growth and survival
of Escherichia coli O157 and Salmonella enteric serovar enteridis in
mayonnaise. Garlic and clove exhibited bacteriostatic and bactericidal
effect, respectively, towards Salmonella enteric and Escherichia coli O157,
and Escherichia coli was more sensitive Souza et al. (2005). Sofia et al.
(2007) studied that mint have lesser antimicrobial effect than
mustard.Spices and herbs or their extracts or essential oils gain their
importance as food preservatives not only for possessing the antimicrobial
activity but also for their antioxidant property. The antioxidant property of
spices and herbs is because of the phenolic compounds present in them.
Rubió et al. (2013) reported that they act as antioxidant by scavenging
free radicals, chelating transition metals, quenching of singlet oxygen, and
enhancing the activities of antioxidant enzymes. Stoilova et al. (2007)
reported that the CO2 extract of ginger showed in vitro activity against
lipid peroxidation, comparable with that of BHT, both at 37 and 80°C.
Rubió et al. (2013), Przygodzka et al. (2014), Srinivasan (2014) have
108

evaluated the antioxidant activity of quercetine (dill), capsaicin (red chilli),

curcumin (turmeric), carvacrol (oregano, thyme, marjoram), thymol
(oregano, thyme), piperine (black pepper), gingerol (ginger).The
antimicrobial as well as antioxidative activity of spices and herbs or their
extracts or essential oils in food depends on several factors, such as type
of food product, type of spices or herbs, effects of spices and herbs on
sensorial quality of food product, composition of food product and,
concentration and biological properties of the micro-organism and
processing and storage conditions of the targeted food product (Gutierrez
et al., 2008; Naidu, 2000 and Romeo et al., 2008). The major constraints
in application of spices and herbs as food preservatives is the requirement
of high concentration of spices and herbs or their extracts for getting
desirable antimicrobial effect which may produce undesirable flavour in
food product (Proestos et al., 2008 and Silva et al., 2007). Some experiments
have been tried out to use these natural antimicrobial agents as food
preservatives in dairy products. Abdalla et al. (2007) added the extract of
mango seed kernel into the pasteurized cow milk and observed that ability
of the mango seed kernel extract to reduce total bacterial count, inhibit
coliform growth and the antimicrobial activity against Escherichia coli
strain extended the shelf life of pasteurized cow milk. Cava et al. (2007)
observed the efficacy of essential oils in milk against the targeted micro-
organisms. Saturejacilicica essential oil was added in butter as both a natural
antioxidant and aroma agent by Ozkan et al. (2007). Burt (2004) added
cinnamon, cardamom, clove and mint oils into the yoghurt and observed
that cinnamon, cardamom and clove oils had more inhibitory effect against
the growth of yoghurt starter cultures than mint oil. In another study, Burt
(2004) noticed the antimicrobial effect of mint oil against Salmonella
enteritidis in low-fat yoghurt. Kalogeropoulos et al. (2009) used ethanol
extracts of propolisto protect starter culture strains in fermented products
from pathogenic micro-organisms. Essential oils of clove, cinnamon, bay
and thyme were also used in soft cheese and evaluated for their
antimicrobial activity against Listeria monocytogenes and Salmonella
enteritidis Burt (2004). It was reported that clove oil more effective against
Salmonella enteritidis in full-fat cheese than in cheese slurry Burt (2004).
Raeisi et al. (2012) reported that Artemisia dracunculus can be used to
inhibit the growth of Staphylococcus aureus and Escherichia coli in cheese.
CONCLUSION
The emerging technologies such as high hydrostatic pressure, pulse
electric field, ultraviolet radiation, microfiltration, ohmic heating,
ultrasoniccation are basically energy efficient process to reduce microbial
 109

load in food without significantly altering its quality. The effectiveness of

these processing technologies for microbial and enzyme inactivation within
food matrixes have been studied and compared with conventional
preservation technologies by several researchers. But still these processes
are in their infantile state. There is, however, a need for synchronization
and comparison of data available from various research laboratories to use
these technologies at commercial level. The use of spices and herbs and
their extract is definitely a better means of preservation of dairy products.
But requirement of higher concentration of these ingredients for
preservation exhibited detrimental effect on sensorial quality of the product.
Viewing the potentiality of these preservatives, further studies are needed
to minimize their adverse effects on sensorial quality of dairy products.
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