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Middle East Journal of Applied Science & Technology (MEJAST)
(Peer Reviewed International Journal) Volume 2, Issue 4, Pages 72-79, October-December 2019
72 | P a g e ISSN (Online): 2582- 0974 Website: www.mejast.com
Preservation of Milk and Dairy Products by Using Biopreservatives
Saba Ameer1
*, Soha Aslam1
& Muhammad Saeed1
1
*Institute of Molecular Biology & Biotechnology, Bahauddin Zakariya University, Multan-60800, Pakistan.
Article Received: 29 June 2019 Article Accepted: 30 October 2019 Article Published: 25 December 2019
INTRODUCTION
Some compounds having medical properties like spices, for example, black pepper, coriander, cassia bark,
cinnamon, cumin, and cardamom. In the earliest time, the human has used spices for a valuable purpose. A hundred
varieties of spices were used to create the preserved food. In different industries, some spices are used for the wide
applications which are beneficial for living organisms such as the type of spices are good for digestion and
metabolism similar to cinnamon and ginger. Hence, material are using for production of perfumes, a color of food,
and its flavor. Turmeric, ginger, red chili, garlic and small cardamom are antimicrobial effects, not all types of
spices & herbs are safe for us. [22]
Figure 1
Bacteria are unsurprisingly occurring in the sealed food items and fermented dairy food. Bacteria held with dairy
fermentation can grow over a wide temp interval from 4 to 50c. Mesophilic bacteria approximately have a growth
assortment from 25-35c another side, thermophilic variety contain range 37-45c. (Johnson Steele, 2013). Such
types of parameters like temperature, pH, and as well as water activity growth of bacterial cells start within dairy
items including the above parameters. In the fermentation process, lactic-acid bacteria are the most important
ABSTRACT
Dairy products help to meet the need of essential nutrients that are difficult to obtain in daily life without milk and its products like yogurt, cheese,
butter. Milk is almost sterile when secreted from a healthy udder. Lower temperature and boiling the milk help in retarding the growth of spoilage
organisms. Biopreservation is a method in which natural microbiota or antimicrobials are used in food to enhance the shelf-life. For this purpose
affordable microorganisms are selected to control the pathogenic activity. The main organism which is used is lactic acid bacteria and their
metabolites. They have the ability to show the antimicrobial features and maintain the sole flavor of food. Lactic acid bacteria are considered as great
biopreservatives. Other LAB such as nisin has broad range of application in food industry and approved by food and drug administration (FDA).
Other metabolites such as enterococci, Bacteriophages and endolysins used as bio-preservation and have promising role in milk storage. As these are
economically important due to their drastic advantages like non-toxic, availability, non-immunogenic and broad activity so, they are considered to be
good agent for biopreservation. This review will focus on application of biopreservatives to reduce the spoilage and increase the safety of milk
products.
Keywords: Bacteriocins, Lactic Acid Bacteria (LAB), Enterococci, Milk, Cheese, Cream.
ButterCream
Yogurt
Milk
Cheese
Ice cream
Main dairy
foods
Middle East Journal of Applied Science & Technology (MEJAST)
(Peer Reviewed International Journal) Volume 2, Issue 4, Pages 72-79, October-December 2019
73 | P a g e ISSN (Online): 2582- 0974 Website: www.mejast.com
starter culture. Lactic acid bacteria cooperate an important role and used to control the pH of dairy objects &
foodstuffs & glycolysis process in milk. Another important role of lactic Bacillus is augmentation flavor in dairy
products. [21]
Such types of dairy foods (include cheese & sour cream & yogurt) are formed through lactic acid bacteria
fermentation process. Such type of starter culture was used in the dairy industry which has the following of this
lactococcus lactic sp, L. lactic sp, and Lactobacillus Helveticas. Probiotics help to increase & addition the
nutritional value of milk and its dairy material. Some lactic acid bacteria have same nature as Probiotics. In the food
material, using are Probiotics strains & gram positive Lactobacillus sp & Bifidobacterium. In review paper, we
discuss live bacteria that occur in the gut & are vital for Probiotics that’s why due to its quantifying and envision the
colonies of bacteria in food items.[22]
In the field of food processing, different species have been utilized for the purpose of preservation milk and dairy
products in different countries through various methodologies for preserved food & milk. There are use Probiotics
for humanizing the dietary value of food and enhance the flavor and health. Prebiotics are also used for the superior
performance of Probiotics. [1]
Basically, Probiotics are living microorganisms include yeast & bacteria which is preferred to use in food due to
increasing the shelf life of food and better quality of food. A few important Probiotics are used for the management
of health issues such as food allergic reaction & colon cancer, etc. protein powder, eggs, fruits, yogurt, avocado,
cooked quinoa, olive oil, seeds, and nuts are most essential Probiotic foods.
Commonly, food industries were using the chemicals for food saving processing. Chemicals were used to increase
the shelf life of food material for later use. Chemicals are manufactured hazardous effects on the human physical
condition and effect on the dietary significance of foodstuff. Nitrates & sulfur dioxide such type of chemical were
used. The term antimicrobial peptide was explained that how to food preserved without any altering of dietary
value and quality of food and its aroma. Chemicals are unhealthy for a consumer that's why we prefer the best
naturally occurring preservation method held to recognize. Hence, there is necessary or significant require to
establish the best methodology and other way to preserve the foodstuff. The food item can be preserved by
microbes & microorganisms & bacteria to strengthening the shelf life of food. Some technique is using nowadays
for the preservation of dairy products like peptide synthesis, chemical synthesis, enzymatic synthesis, and
recombinant synthesis.[2]
The overweight population has been beloved to reduce their weight through do exercises and diet planning, there
are big population who has most wanted to weight loss through in well and sustainable way & that's why dairy
products help them to control their overweight by checking and balanced way their weight. The obese person wants
to become healthy through different types of exercise and diet roles. The micro-nutrients and macro-nutrients occur
in the preserved food and dairy fermented foodstuff. The most advantageous dietary nutrient composition to
achieve weight reduces remains a controversy. The nutrition is allowing to patients, the take proteins lower
carbohydrates, lipid in their diets for a healthy lifestyle.
Middle East Journal of Applied Science & Technology (MEJAST)
(Peer Reviewed International Journal) Volume 2, Issue 4, Pages 72-79, October-December 2019
74 | P a g e ISSN (Online): 2582- 0974 Website: www.mejast.com
When overweight people eating higher proteins than they should go for exercise & workout. While those one who
intakes higher proteins for weight maintainer are the concern and other one intake higher proteins in that case
compared who intake low proteins. Higher proteins with a combination of hard work are better than take lower
calories with no workout. Humans have linked the consumption of dairy food materials and calcium gain from
dietary food to faster weight reduction and loss’s fat molecules from the body. In this study, we expected to get the
weight loss with a high value of fats and introduced the prototype which is sufficient for the long term metabolic &
and short term metabolic activities that are obligatory for body figure maintain. In one program, we should collect
the steps that should follow such as the energy limit, protein with low carbohydrates, highly nutritional value &
increase the dose of dietary fibers and aerobic exercise.
Nowadays, we are facing the major problems that are food losses in the worldwide level, and one-third part around
destroys from the human consumption food, such products include dairy items, fish, vegetables, seafood, meat, and
some other cereal crops are wasted in huge amount. Fungi and some type microorganisms or the main reason of
disturbing the food chain because they have the ability to grow in every environment, fungal infection have ability
produced to the toxic effect on human health, animals and plants. Fungi be heterotopy and food absorbing
microorganisms from moist areas and either they produce from the sexual or asexual process, they can move easily
from one to another place by spores and wind and rapidly mature and start lifecycle again and again and cause the
infection. These foods are tested by thermal treatment such as sterilization, pasteurization, & water removal by
drying & freezing-drying & high titer of high sugar or salt [3]. The major issue of loss disease free years in Nordic
countries today is diseases like type 2 diabetes, cardiovascular diseases, and cancers. Moreover, the increasing
occurrence of obesity greatly increases the risk of these chronic diseases. The increasing prevalence of different
chronic diseases, it is most important to understand the health effect of milk and dairy products in the diet. In earlier
times, milk was used for a longer time due to its vast process of preservation as it was completed in three steps:
Pasteurization, Sterilization, Dehydration.
Figure 2: Overall association between dairy product intake and health outcomes
Middle East Journal of Applied Science & Technology (MEJAST)
(Peer Reviewed International Journal) Volume 2, Issue 4, Pages 72-79, October-December 2019
75 | P a g e ISSN (Online): 2582- 0974 Website: www.mejast.com
These processes convert raw milk into other forms of dairy products for the consumption of mankind. This method
was adopted on a small scale level and other level for milk processing. The main purpose for the preservation is that
to control the growth of spoilage microorganisms that cause contamination in storage and to increase the shelf-life
of products. [4]
Favorable involvement conditions
Two types of bacteria can grow on milk and its products: Spoilage bacteria, Pathogenic bacteria.
Spoilage bacteria cause unpleasant flavor, odor, and taste while pathogenic bacteria not show these symptoms and
cause illness.
The main principles of preservation are to control microorganisms by killing them and their spores, by control their
enzymes and control all physical sources that provide deterioration in milk.
Table 1
Raw side Processing Packaging and distribution
Receiving Pasteurization By filling machines
Storage Sterilization Delivered to market
Dehydration Homes and refrigerator
As this process is time-consuming and laborious so chemicals also added to preserve milk products, for example,
acids organic & inorganic acids, antimicrobial agents, antioxidants, and other compounds like these are added. Due
to their adverse effect on human health restricted the use of chemical method.[5]
Methods for preservation of milk and dairy products
Biopreservatives play an important role due to their advantageous effects on health as compared to chemical
preservatives. Different types of microorganisms are classified and studied by researchers to estimate beneficial
uses for preservation.[6], [7]
The role of lactic acid bacteria in preservation
Lactic acid bacteria (LAB) LAB is generally included Lactococcus, Streptococcus, Lactobacillus, Pediococcus,
Leuconostoc, Enterococcus, Carnobacterium, Aerococcus, Oenococcus, Tetragenococcus, and Vagococcus. All
these micro-organisms are Gram-positive, nonmotile, non-spore forming, rod- and coccus-shaped organisms and
are able for fermentation of carbohydrates. The main product obtained as a result of fermentation is lactic acid. The
composition of the LAB contains fewer proportions of G+C contents in their DNA. The basic function of Lactic
Acid Bacteria in food fermentation and processing is like GRAS (generally recognized safe). The LAB can be used
for the production of different fermented products of dairy Major compounds produced by LAB for fermentation
are organic acids, hydrogen peroxide, and bacteriocins. Additionally, Lactic Acid Bacteria can be used for the
production of probiotics. The application of antimicrobial producing lactic acid bacteria (LAB)[8] for the
manufacture of dairy products, which can be used into fermented or non fermented milk and dairy products, help
for a processing advantage to improve safety and increase the quality of dairy products, providing an additional
Middle East Journal of Applied Science & Technology (MEJAST)
(Peer Reviewed International Journal) Volume 2, Issue 4, Pages 72-79, October-December 2019
76 | P a g e ISSN (Online): 2582- 0974 Website: www.mejast.com
hurdle to less the likelihood of food-borne diseases. These peptides are considered natural biopreservatives and
their potential application in the food industry provides great interest. [15] as biopreservative: Bacteriocins defined
as peptides that are synthesized from bacteria that slow down & destroy other related or unrelated microorganisms
to save the Food. Bacteriocins are classified into 3 major classes on the base of their biosynthetic action and
activity.
Class-1 tiny post-translationally customized peptides
Class-2 unmodified bacteriocins9
Class-3 large peptides
Bacteriocins have been used as biopreservative by different methods either alone or combine with other methods
like in hurdle technique. The recent application of bacteriocins into dairy food to control the pathogens includes the
food with lactic acid bacteria that cause to produce bacteriocins. Mostly dairy products which made from
pasteurized milk, conserved at refrigerator temperature allow the enlargement of bacteria such as L.
monocytogenes. So infectivity can happen in later stages of processing of dairy products. Bacteriocins are also used
for other purposes like food fermentation, improve flavors and cheese ripening.[9] These also help to improve the
safety of dairy products by using different Strains combinations of different preservation methods that provide high
protection than the single strain. These bacteriocins helpful for the prevention and reduction of intestinal pathogens
produce probiotics that have the ability to survive in GIT may be advantageous. [18]
There are some types of bacteriocin that are used for the prevention of milk-borne pathogens like
Nisin
The bacteriocin nisin can be classified as lantibiotic and the most utilized and commercially available bacteriocins.
These are approved as a food preservative and are recognized the safest by the World Health Organization (WHO)
Experts Committee on Food preservatives. Nisin has an antimicrobial activity which is next to for Gram-positive
bacteria as well as Lactic Acid Bacteria, pathogens like Listeria and Staphylococcus, and the spore-forming
bacteria, Bacillus, and others. One of the most important applications of nisin was to prevent the late blowing in
dairy product cheese caused due to gas-producing Clostridium spp. It has been of great interest to use new
bacteriocins with a wide range of antibacterial activity, stable in different food varieties, tolerable to high
temperature and resistant to proteolytic enzymes.[2]
Pediocin
These classified as ll. Bacteriocins and commercially available for the preservation of dairy products these
bacteriocins are more effective than other classes. They are stable in aqueous solutions, pH resistant and have little
impact on freezing and boiling[10].
Lacticins
lacticins are formed from convinced strains of Lc. Latics and consist of lacticins 3147 and lactic in 481. A lacticins
3147 powder preparation was effective for control of Listeria and Bacillus in infant milk formulation, natural
Middle East Journal of Applied Science & Technology (MEJAST)
(Peer Reviewed International Journal) Volume 2, Issue 4, Pages 72-79, October-December 2019
77 | P a g e ISSN (Online): 2582- 0974 Website: www.mejast.com
yogurt, cottage cheese, and other dairy products. However, the application of lactic in food not likes to ensure the
complete eradication of pathogens such as L. monocytogenes and others.[8], [11]
Enterocins
Enterocins are strains that evolve from enterococci and are safe to use in food applications, but these are associated
with infectious microorganisms in humans and other living organisms. To use enterococci for food purposes their
purified form is available and suitable for food intake. They become active against foodborne pathogens and help to
prevent the growth of other microorganisms.[17]
The elevated cost of bacteriocin separation and distillation also confines the commercial discovery of new
bacteriocins. In adding, the restrictive food legislation of the physical condition regulatory authorities limit the
support of new bacteriocins as food preservatives and, as a consequence, only two bacteriocins (nisin and pediocin)
are commercially accessible[12]
Utility of enterococci to preserve dairy products
Enterococci were described for the first time from scientist Thiercelin in 1899 and genus Enterococcus was
proposed by Thiercelin and Jouhaud in 1903. Presence of enterococci in milk and dairy products are beneficial by
contributing to produce unique and emerging by-products, by protecting against diverse spoilers, and as probiotics.
Due to their adverse nature, heat resistance and adaptation to the different substrate and environmental conditions,
they can grow during freezing and survive after the pasteurization of milk and its products. So, enterococci are a
part of raw and pasteurized milk. More importantly, in cheeses, such as Mozzarella, Cheddar, enterococci are the
major microorganisms in the completely ripened product High levels of contaminating enterococci could produce
to deterioration of sensory products in some cheeses but they also play the important role in cheese ripening and
aroma development. Safe and commercially available strains of enterococci are use in food and as probiotics and
they should possess neither any virulence factors, nor should be able to obtain an antibiotic resistance gene. [16]
BACTERIOPHAGE AS BIOPRESERVATIVE
Bacteriophages or phages are the most widely found microorganisms on planet earth and widespread on foods of
different origins. These are viruses that specialize to infect and multiply in bacteria and are harmless due to host
specific to humans and other living organisms. In bio-preservation, bacteriophages can work as symbiotic an
association with fermenting organisms during fermentation. It is a very effective method for the preparation of food
processing and reduces effectively the like Salmonella in cheddar production Endolysins. Endolysins are called as
phage lysins which are referred to as phage-encoded peptidoglycan hydrolases and employed from majority of
bacteriophages to enzymatically degrade the peptidoglycan layer of the host bacterium. Bacteriophages are also
utilizing the endolysins at each end of their replication cycle for degradation, of the peptidoglycan of the bacterial
host. Endolysins are good antimicrobials due to the absence of an outer membrane covering in the Gram-positive
bacterial cell wall. In bio-preservation, bacteriophages work symbiotically with fermentative microorganisms
during fermentation. It is the most effective method to prepare food processing and fewer effects like Salmonella in
cheddar cheese, S. aureus growth in curd manufacturing and complete eradication of Listeria monocytogenes
Middle East Journal of Applied Science & Technology (MEJAST)
(Peer Reviewed International Journal) Volume 2, Issue 4, Pages 72-79, October-December 2019
78 | P a g e ISSN (Online): 2582- 0974 Website: www.mejast.com
during ripened formation and storage of acid coagulation and semi-hard cheeses. It was also inhibition of
Enterobacter sakazakii for reconstituted infant formula milk. [14]
Endolysins
Endolysins are also good antimicrobial agents due to the absence of an outer membrane in the Gram-positive
bacterial cell wall. It is easily accessible to peptidoglycan. Endolysins usually have narrow spectrum lytic activity
and less restricted to phage host bacterial activity. The role of endolysins in milk bio-preservation is still in
seniority. But there are many endolysins are isolated that work against different foodborne pathogens. The good
thing in this regard is that no-existence of resistance against endolysins now date. It is suggested by several
researchers now that transgenic cows expressing more endolysins that have low chances of mastitis and S. aureus
milk contamination. Endolysins are also having a role in dairy processing, particularly in endolysins to destroy S.
aureus in pasteurized milk. [14]
CONCLUDING REMARKS
Major, aim of dairy trade is to produce dairy foodstuffs with more nutrition and fitness enhancing property. This
review explains all the aspects related to the recent trends for using bacteriocins producing bacteria and
bacteriocins in dairy products. Pasteurization and refrigeration are important to control pathogens but restricted due
to the presence of species that survive after this process so it is also the need of time to discover novel bacteriocins
that having unique properties for application in dairy foods. Concentrated culture of bacteriocin is more efficient as
compared with the direct addition of cultures. The starter culture should be able to produce bioactive metabolites.
The foremost imperfection linked with this function that the lack of compatibility of strains and other cultures for
the fermentation of dairy products. But biopreservative have a promising role in food preservation and safety.
Genatically modified Bacteriophages helpful for preservation, but their safety should be proper for selected as
biopreservative agent. Some lactic acid bacteria are of great importance in traditional cheese formation from raw
milk. The use of lactic acid bacteria is functional starter culture in dairy products like in yogurt and ice-cream.
These lactic acid bacteria have positive effect like anti obesity and anti diabetes. Bacteriocins, enterococci,
Bacteriophages, endolysins, LAB these all biopreservative have major role and applications in improving food
safety.
REFERENCES
[1] S. E. Evivie, G. C. Huo, J. O. Igene, and X. Bian, ―Some current applications, limitations and future
perspectives of lactic acid bacteria as probiotics,‖ Food Nutr. Res., vol. 61, no. 1, 2017.
[2] M. Rai, R. Pandit, S. Gaikwad, and G. Kövics, ―Antimicrobial peptides as natural bio-preservative to enhance
the shelf-life of food,‖ J. Food Sci. Technol., vol. 53, no. 9, pp. 3381–3394, 2016.
[3] M. Leyva Salas, J. Mounier, F. Valence, M. Coton, A. Thierry, and E. Coton, ―Antifungal Microbial Agents for
Food Biopreservation—A Review,‖ Microorganisms, vol. 5, no. 3, p. 37, 2017.
[4] T. K. Thorning, A. Raben, T. Tholstrup, S. S. Soedamah-muthu, I. Givens, and A. Astrup, ―Milk and dairy
science,‖ vol. 1, pp. 1–11, 2016.
Middle East Journal of Applied Science & Technology (MEJAST)
(Peer Reviewed International Journal) Volume 2, Issue 4, Pages 72-79, October-December 2019
79 | P a g e ISSN (Online): 2582- 0974 Website: www.mejast.com
[5] J. D. Murray, N. Mohamad-Fauzi, C. A. Cooper, and E. A. Maga, ―Current status of transgenic animal research
for human health applications.‖
[6] D. E. Kerr and O. Wellnitz, ―Mammary expression of new genes to combat mastitis,‖ J. Anim. Sci., vol. 81, no.
15_suppl_3, pp. 38–47, Mar. 2003.
[7] D. M. Donovan, D. E. Kerr, and R. J. Wall, ―Engineering disease resistant cattle,‖ Transgenic Res., vol. 14, no.
5, pp. 563–567, 2005.
[8] D. E. Kerr et al., ―Lysostaphin expression in mammary glands confers protection against staphylococcal
infection in transgenic mice,‖ Nat. Biotechnol., vol. 19, no. 1, pp. 66–70, 2001.
[9] L. M. Houdebine, ―Transgenic animal production,‖ Biotechnol. Sustain. Agric. Emerg. Approaches Strateg.,
vol. 61, pp. 141–184, 2017.
[10] S. Mishra et al., ―Effects of nutraceuticals on genetic expressions.,‖ Open Nutraceuticals J., vol. 2, pp. 70–80,
2009.
[11] L. Luo et al., ―Fabrication of Synthetic Mesenchymal Stem Cells for the Treatment of Acute Myocardial
Infarction in Mice,‖ Circ. Res., vol. 120, no. 11, pp. 1768–1775, 2017.
[12] D. R. Brundige, E. A. Maga, K. C. Klasing, and J. D. Murray, ―Lysozyme Transgenic Goats’ Milk Influences
Gastrointestinal Morphology in Young Pigs,‖ J. Nutr., vol. 138, no. 5, pp. 921–926, 2018.
[13] Rai, M., Pandit, R., Gaikwad, S. & Kövics, G. (2016). Antimicrobial peptides as natural bio-preservatives to
enhance the shelf-life of food. Journal of Food Science and Technology, 53, 3381–3394.
[14] Singh, V.P. (2018). Recent approaches to food bio-preservation – a review. Open Veterinary Journal, 8, 104.
[15] Arqués, J.L., Rodríguez, E., Langa, S., Landete, J.M. & Medina, M. (2015). Antimicrobial activity of lactic
acid bacteria in dairy products and gut: Effect on pathogens. BioMed Research International, 2015.
[16] Bhardwaj, A., Malik, R.K. & Chauhan, P. (2008). Functional and safety aspects of enterococci in dairy foods.
Indian Journal of Microbiology, 48, 317–325.
[17] Gautam, N. & Sharma, N. (2009). Bacteriocin: Safest approach to preserve food products. Indian Journal of
Microbiology, 49, 204–211.
[18] Silva, C.C.G., Silva, S.P.M. & Ribeiro, S.C. (2018). Application of bacteriocins and protective cultures in
dairy food preservation. Frontiers in Microbiology, 9.
[19] Leyva Salas, Morphology in Young Pigs. The Journal of Nutrition 138:921–926. doi: 10.1093/jn/138.5.921
[20] Donovan DM, Kerr DE, Wall RJ (2005) Engineering disease resistant cattle. Transgenic Research 14:563–
567. doi: 10.1007/s11248-005-0670-8
[21] Hickey, C.D., Sheehan, J.J., Wilkinson, M.G. & Auty, M.A.E. (2015). Growth and location of bacterial
colonies within dairy foods using microscopy techniques: A review. Frontiers in Microbiology, 6, 1–8.
[22] Hill, D., Sugrue, I., Arendt, E., Hill, C., Stanton, C. & Ross, R.P. (2017). Recent advances in microbial
fermentation for dairy and health. F1000Research, 6, 751.

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Preservation of Milk and Dairy Products by Using Biopreservatives

  • 1. Middle East Journal of Applied Science & Technology (MEJAST) (Peer Reviewed International Journal) Volume 2, Issue 4, Pages 72-79, October-December 2019 72 | P a g e ISSN (Online): 2582- 0974 Website: www.mejast.com Preservation of Milk and Dairy Products by Using Biopreservatives Saba Ameer1 *, Soha Aslam1 & Muhammad Saeed1 1 *Institute of Molecular Biology & Biotechnology, Bahauddin Zakariya University, Multan-60800, Pakistan. Article Received: 29 June 2019 Article Accepted: 30 October 2019 Article Published: 25 December 2019 INTRODUCTION Some compounds having medical properties like spices, for example, black pepper, coriander, cassia bark, cinnamon, cumin, and cardamom. In the earliest time, the human has used spices for a valuable purpose. A hundred varieties of spices were used to create the preserved food. In different industries, some spices are used for the wide applications which are beneficial for living organisms such as the type of spices are good for digestion and metabolism similar to cinnamon and ginger. Hence, material are using for production of perfumes, a color of food, and its flavor. Turmeric, ginger, red chili, garlic and small cardamom are antimicrobial effects, not all types of spices & herbs are safe for us. [22] Figure 1 Bacteria are unsurprisingly occurring in the sealed food items and fermented dairy food. Bacteria held with dairy fermentation can grow over a wide temp interval from 4 to 50c. Mesophilic bacteria approximately have a growth assortment from 25-35c another side, thermophilic variety contain range 37-45c. (Johnson Steele, 2013). Such types of parameters like temperature, pH, and as well as water activity growth of bacterial cells start within dairy items including the above parameters. In the fermentation process, lactic-acid bacteria are the most important ABSTRACT Dairy products help to meet the need of essential nutrients that are difficult to obtain in daily life without milk and its products like yogurt, cheese, butter. Milk is almost sterile when secreted from a healthy udder. Lower temperature and boiling the milk help in retarding the growth of spoilage organisms. Biopreservation is a method in which natural microbiota or antimicrobials are used in food to enhance the shelf-life. For this purpose affordable microorganisms are selected to control the pathogenic activity. The main organism which is used is lactic acid bacteria and their metabolites. They have the ability to show the antimicrobial features and maintain the sole flavor of food. Lactic acid bacteria are considered as great biopreservatives. Other LAB such as nisin has broad range of application in food industry and approved by food and drug administration (FDA). Other metabolites such as enterococci, Bacteriophages and endolysins used as bio-preservation and have promising role in milk storage. As these are economically important due to their drastic advantages like non-toxic, availability, non-immunogenic and broad activity so, they are considered to be good agent for biopreservation. This review will focus on application of biopreservatives to reduce the spoilage and increase the safety of milk products. Keywords: Bacteriocins, Lactic Acid Bacteria (LAB), Enterococci, Milk, Cheese, Cream. ButterCream Yogurt Milk Cheese Ice cream Main dairy foods
  • 2. Middle East Journal of Applied Science & Technology (MEJAST) (Peer Reviewed International Journal) Volume 2, Issue 4, Pages 72-79, October-December 2019 73 | P a g e ISSN (Online): 2582- 0974 Website: www.mejast.com starter culture. Lactic acid bacteria cooperate an important role and used to control the pH of dairy objects & foodstuffs & glycolysis process in milk. Another important role of lactic Bacillus is augmentation flavor in dairy products. [21] Such types of dairy foods (include cheese & sour cream & yogurt) are formed through lactic acid bacteria fermentation process. Such type of starter culture was used in the dairy industry which has the following of this lactococcus lactic sp, L. lactic sp, and Lactobacillus Helveticas. Probiotics help to increase & addition the nutritional value of milk and its dairy material. Some lactic acid bacteria have same nature as Probiotics. In the food material, using are Probiotics strains & gram positive Lactobacillus sp & Bifidobacterium. In review paper, we discuss live bacteria that occur in the gut & are vital for Probiotics that’s why due to its quantifying and envision the colonies of bacteria in food items.[22] In the field of food processing, different species have been utilized for the purpose of preservation milk and dairy products in different countries through various methodologies for preserved food & milk. There are use Probiotics for humanizing the dietary value of food and enhance the flavor and health. Prebiotics are also used for the superior performance of Probiotics. [1] Basically, Probiotics are living microorganisms include yeast & bacteria which is preferred to use in food due to increasing the shelf life of food and better quality of food. A few important Probiotics are used for the management of health issues such as food allergic reaction & colon cancer, etc. protein powder, eggs, fruits, yogurt, avocado, cooked quinoa, olive oil, seeds, and nuts are most essential Probiotic foods. Commonly, food industries were using the chemicals for food saving processing. Chemicals were used to increase the shelf life of food material for later use. Chemicals are manufactured hazardous effects on the human physical condition and effect on the dietary significance of foodstuff. Nitrates & sulfur dioxide such type of chemical were used. The term antimicrobial peptide was explained that how to food preserved without any altering of dietary value and quality of food and its aroma. Chemicals are unhealthy for a consumer that's why we prefer the best naturally occurring preservation method held to recognize. Hence, there is necessary or significant require to establish the best methodology and other way to preserve the foodstuff. The food item can be preserved by microbes & microorganisms & bacteria to strengthening the shelf life of food. Some technique is using nowadays for the preservation of dairy products like peptide synthesis, chemical synthesis, enzymatic synthesis, and recombinant synthesis.[2] The overweight population has been beloved to reduce their weight through do exercises and diet planning, there are big population who has most wanted to weight loss through in well and sustainable way & that's why dairy products help them to control their overweight by checking and balanced way their weight. The obese person wants to become healthy through different types of exercise and diet roles. The micro-nutrients and macro-nutrients occur in the preserved food and dairy fermented foodstuff. The most advantageous dietary nutrient composition to achieve weight reduces remains a controversy. The nutrition is allowing to patients, the take proteins lower carbohydrates, lipid in their diets for a healthy lifestyle.
  • 3. Middle East Journal of Applied Science & Technology (MEJAST) (Peer Reviewed International Journal) Volume 2, Issue 4, Pages 72-79, October-December 2019 74 | P a g e ISSN (Online): 2582- 0974 Website: www.mejast.com When overweight people eating higher proteins than they should go for exercise & workout. While those one who intakes higher proteins for weight maintainer are the concern and other one intake higher proteins in that case compared who intake low proteins. Higher proteins with a combination of hard work are better than take lower calories with no workout. Humans have linked the consumption of dairy food materials and calcium gain from dietary food to faster weight reduction and loss’s fat molecules from the body. In this study, we expected to get the weight loss with a high value of fats and introduced the prototype which is sufficient for the long term metabolic & and short term metabolic activities that are obligatory for body figure maintain. In one program, we should collect the steps that should follow such as the energy limit, protein with low carbohydrates, highly nutritional value & increase the dose of dietary fibers and aerobic exercise. Nowadays, we are facing the major problems that are food losses in the worldwide level, and one-third part around destroys from the human consumption food, such products include dairy items, fish, vegetables, seafood, meat, and some other cereal crops are wasted in huge amount. Fungi and some type microorganisms or the main reason of disturbing the food chain because they have the ability to grow in every environment, fungal infection have ability produced to the toxic effect on human health, animals and plants. Fungi be heterotopy and food absorbing microorganisms from moist areas and either they produce from the sexual or asexual process, they can move easily from one to another place by spores and wind and rapidly mature and start lifecycle again and again and cause the infection. These foods are tested by thermal treatment such as sterilization, pasteurization, & water removal by drying & freezing-drying & high titer of high sugar or salt [3]. The major issue of loss disease free years in Nordic countries today is diseases like type 2 diabetes, cardiovascular diseases, and cancers. Moreover, the increasing occurrence of obesity greatly increases the risk of these chronic diseases. The increasing prevalence of different chronic diseases, it is most important to understand the health effect of milk and dairy products in the diet. In earlier times, milk was used for a longer time due to its vast process of preservation as it was completed in three steps: Pasteurization, Sterilization, Dehydration. Figure 2: Overall association between dairy product intake and health outcomes
  • 4. Middle East Journal of Applied Science & Technology (MEJAST) (Peer Reviewed International Journal) Volume 2, Issue 4, Pages 72-79, October-December 2019 75 | P a g e ISSN (Online): 2582- 0974 Website: www.mejast.com These processes convert raw milk into other forms of dairy products for the consumption of mankind. This method was adopted on a small scale level and other level for milk processing. The main purpose for the preservation is that to control the growth of spoilage microorganisms that cause contamination in storage and to increase the shelf-life of products. [4] Favorable involvement conditions Two types of bacteria can grow on milk and its products: Spoilage bacteria, Pathogenic bacteria. Spoilage bacteria cause unpleasant flavor, odor, and taste while pathogenic bacteria not show these symptoms and cause illness. The main principles of preservation are to control microorganisms by killing them and their spores, by control their enzymes and control all physical sources that provide deterioration in milk. Table 1 Raw side Processing Packaging and distribution Receiving Pasteurization By filling machines Storage Sterilization Delivered to market Dehydration Homes and refrigerator As this process is time-consuming and laborious so chemicals also added to preserve milk products, for example, acids organic & inorganic acids, antimicrobial agents, antioxidants, and other compounds like these are added. Due to their adverse effect on human health restricted the use of chemical method.[5] Methods for preservation of milk and dairy products Biopreservatives play an important role due to their advantageous effects on health as compared to chemical preservatives. Different types of microorganisms are classified and studied by researchers to estimate beneficial uses for preservation.[6], [7] The role of lactic acid bacteria in preservation Lactic acid bacteria (LAB) LAB is generally included Lactococcus, Streptococcus, Lactobacillus, Pediococcus, Leuconostoc, Enterococcus, Carnobacterium, Aerococcus, Oenococcus, Tetragenococcus, and Vagococcus. All these micro-organisms are Gram-positive, nonmotile, non-spore forming, rod- and coccus-shaped organisms and are able for fermentation of carbohydrates. The main product obtained as a result of fermentation is lactic acid. The composition of the LAB contains fewer proportions of G+C contents in their DNA. The basic function of Lactic Acid Bacteria in food fermentation and processing is like GRAS (generally recognized safe). The LAB can be used for the production of different fermented products of dairy Major compounds produced by LAB for fermentation are organic acids, hydrogen peroxide, and bacteriocins. Additionally, Lactic Acid Bacteria can be used for the production of probiotics. The application of antimicrobial producing lactic acid bacteria (LAB)[8] for the manufacture of dairy products, which can be used into fermented or non fermented milk and dairy products, help for a processing advantage to improve safety and increase the quality of dairy products, providing an additional
  • 5. Middle East Journal of Applied Science & Technology (MEJAST) (Peer Reviewed International Journal) Volume 2, Issue 4, Pages 72-79, October-December 2019 76 | P a g e ISSN (Online): 2582- 0974 Website: www.mejast.com hurdle to less the likelihood of food-borne diseases. These peptides are considered natural biopreservatives and their potential application in the food industry provides great interest. [15] as biopreservative: Bacteriocins defined as peptides that are synthesized from bacteria that slow down & destroy other related or unrelated microorganisms to save the Food. Bacteriocins are classified into 3 major classes on the base of their biosynthetic action and activity. Class-1 tiny post-translationally customized peptides Class-2 unmodified bacteriocins9 Class-3 large peptides Bacteriocins have been used as biopreservative by different methods either alone or combine with other methods like in hurdle technique. The recent application of bacteriocins into dairy food to control the pathogens includes the food with lactic acid bacteria that cause to produce bacteriocins. Mostly dairy products which made from pasteurized milk, conserved at refrigerator temperature allow the enlargement of bacteria such as L. monocytogenes. So infectivity can happen in later stages of processing of dairy products. Bacteriocins are also used for other purposes like food fermentation, improve flavors and cheese ripening.[9] These also help to improve the safety of dairy products by using different Strains combinations of different preservation methods that provide high protection than the single strain. These bacteriocins helpful for the prevention and reduction of intestinal pathogens produce probiotics that have the ability to survive in GIT may be advantageous. [18] There are some types of bacteriocin that are used for the prevention of milk-borne pathogens like Nisin The bacteriocin nisin can be classified as lantibiotic and the most utilized and commercially available bacteriocins. These are approved as a food preservative and are recognized the safest by the World Health Organization (WHO) Experts Committee on Food preservatives. Nisin has an antimicrobial activity which is next to for Gram-positive bacteria as well as Lactic Acid Bacteria, pathogens like Listeria and Staphylococcus, and the spore-forming bacteria, Bacillus, and others. One of the most important applications of nisin was to prevent the late blowing in dairy product cheese caused due to gas-producing Clostridium spp. It has been of great interest to use new bacteriocins with a wide range of antibacterial activity, stable in different food varieties, tolerable to high temperature and resistant to proteolytic enzymes.[2] Pediocin These classified as ll. Bacteriocins and commercially available for the preservation of dairy products these bacteriocins are more effective than other classes. They are stable in aqueous solutions, pH resistant and have little impact on freezing and boiling[10]. Lacticins lacticins are formed from convinced strains of Lc. Latics and consist of lacticins 3147 and lactic in 481. A lacticins 3147 powder preparation was effective for control of Listeria and Bacillus in infant milk formulation, natural
  • 6. Middle East Journal of Applied Science & Technology (MEJAST) (Peer Reviewed International Journal) Volume 2, Issue 4, Pages 72-79, October-December 2019 77 | P a g e ISSN (Online): 2582- 0974 Website: www.mejast.com yogurt, cottage cheese, and other dairy products. However, the application of lactic in food not likes to ensure the complete eradication of pathogens such as L. monocytogenes and others.[8], [11] Enterocins Enterocins are strains that evolve from enterococci and are safe to use in food applications, but these are associated with infectious microorganisms in humans and other living organisms. To use enterococci for food purposes their purified form is available and suitable for food intake. They become active against foodborne pathogens and help to prevent the growth of other microorganisms.[17] The elevated cost of bacteriocin separation and distillation also confines the commercial discovery of new bacteriocins. In adding, the restrictive food legislation of the physical condition regulatory authorities limit the support of new bacteriocins as food preservatives and, as a consequence, only two bacteriocins (nisin and pediocin) are commercially accessible[12] Utility of enterococci to preserve dairy products Enterococci were described for the first time from scientist Thiercelin in 1899 and genus Enterococcus was proposed by Thiercelin and Jouhaud in 1903. Presence of enterococci in milk and dairy products are beneficial by contributing to produce unique and emerging by-products, by protecting against diverse spoilers, and as probiotics. Due to their adverse nature, heat resistance and adaptation to the different substrate and environmental conditions, they can grow during freezing and survive after the pasteurization of milk and its products. So, enterococci are a part of raw and pasteurized milk. More importantly, in cheeses, such as Mozzarella, Cheddar, enterococci are the major microorganisms in the completely ripened product High levels of contaminating enterococci could produce to deterioration of sensory products in some cheeses but they also play the important role in cheese ripening and aroma development. Safe and commercially available strains of enterococci are use in food and as probiotics and they should possess neither any virulence factors, nor should be able to obtain an antibiotic resistance gene. [16] BACTERIOPHAGE AS BIOPRESERVATIVE Bacteriophages or phages are the most widely found microorganisms on planet earth and widespread on foods of different origins. These are viruses that specialize to infect and multiply in bacteria and are harmless due to host specific to humans and other living organisms. In bio-preservation, bacteriophages can work as symbiotic an association with fermenting organisms during fermentation. It is a very effective method for the preparation of food processing and reduces effectively the like Salmonella in cheddar production Endolysins. Endolysins are called as phage lysins which are referred to as phage-encoded peptidoglycan hydrolases and employed from majority of bacteriophages to enzymatically degrade the peptidoglycan layer of the host bacterium. Bacteriophages are also utilizing the endolysins at each end of their replication cycle for degradation, of the peptidoglycan of the bacterial host. Endolysins are good antimicrobials due to the absence of an outer membrane covering in the Gram-positive bacterial cell wall. In bio-preservation, bacteriophages work symbiotically with fermentative microorganisms during fermentation. It is the most effective method to prepare food processing and fewer effects like Salmonella in cheddar cheese, S. aureus growth in curd manufacturing and complete eradication of Listeria monocytogenes
  • 7. Middle East Journal of Applied Science & Technology (MEJAST) (Peer Reviewed International Journal) Volume 2, Issue 4, Pages 72-79, October-December 2019 78 | P a g e ISSN (Online): 2582- 0974 Website: www.mejast.com during ripened formation and storage of acid coagulation and semi-hard cheeses. It was also inhibition of Enterobacter sakazakii for reconstituted infant formula milk. [14] Endolysins Endolysins are also good antimicrobial agents due to the absence of an outer membrane in the Gram-positive bacterial cell wall. It is easily accessible to peptidoglycan. Endolysins usually have narrow spectrum lytic activity and less restricted to phage host bacterial activity. The role of endolysins in milk bio-preservation is still in seniority. But there are many endolysins are isolated that work against different foodborne pathogens. The good thing in this regard is that no-existence of resistance against endolysins now date. It is suggested by several researchers now that transgenic cows expressing more endolysins that have low chances of mastitis and S. aureus milk contamination. Endolysins are also having a role in dairy processing, particularly in endolysins to destroy S. aureus in pasteurized milk. [14] CONCLUDING REMARKS Major, aim of dairy trade is to produce dairy foodstuffs with more nutrition and fitness enhancing property. This review explains all the aspects related to the recent trends for using bacteriocins producing bacteria and bacteriocins in dairy products. Pasteurization and refrigeration are important to control pathogens but restricted due to the presence of species that survive after this process so it is also the need of time to discover novel bacteriocins that having unique properties for application in dairy foods. Concentrated culture of bacteriocin is more efficient as compared with the direct addition of cultures. The starter culture should be able to produce bioactive metabolites. The foremost imperfection linked with this function that the lack of compatibility of strains and other cultures for the fermentation of dairy products. But biopreservative have a promising role in food preservation and safety. Genatically modified Bacteriophages helpful for preservation, but their safety should be proper for selected as biopreservative agent. Some lactic acid bacteria are of great importance in traditional cheese formation from raw milk. The use of lactic acid bacteria is functional starter culture in dairy products like in yogurt and ice-cream. These lactic acid bacteria have positive effect like anti obesity and anti diabetes. Bacteriocins, enterococci, Bacteriophages, endolysins, LAB these all biopreservative have major role and applications in improving food safety. REFERENCES [1] S. E. Evivie, G. C. Huo, J. O. Igene, and X. Bian, ―Some current applications, limitations and future perspectives of lactic acid bacteria as probiotics,‖ Food Nutr. Res., vol. 61, no. 1, 2017. [2] M. Rai, R. Pandit, S. Gaikwad, and G. Kövics, ―Antimicrobial peptides as natural bio-preservative to enhance the shelf-life of food,‖ J. Food Sci. Technol., vol. 53, no. 9, pp. 3381–3394, 2016. [3] M. Leyva Salas, J. Mounier, F. Valence, M. Coton, A. Thierry, and E. Coton, ―Antifungal Microbial Agents for Food Biopreservation—A Review,‖ Microorganisms, vol. 5, no. 3, p. 37, 2017. [4] T. K. Thorning, A. Raben, T. Tholstrup, S. S. Soedamah-muthu, I. Givens, and A. Astrup, ―Milk and dairy science,‖ vol. 1, pp. 1–11, 2016.
  • 8. Middle East Journal of Applied Science & Technology (MEJAST) (Peer Reviewed International Journal) Volume 2, Issue 4, Pages 72-79, October-December 2019 79 | P a g e ISSN (Online): 2582- 0974 Website: www.mejast.com [5] J. D. Murray, N. Mohamad-Fauzi, C. A. Cooper, and E. A. Maga, ―Current status of transgenic animal research for human health applications.‖ [6] D. E. Kerr and O. Wellnitz, ―Mammary expression of new genes to combat mastitis,‖ J. Anim. Sci., vol. 81, no. 15_suppl_3, pp. 38–47, Mar. 2003. [7] D. M. Donovan, D. E. Kerr, and R. J. Wall, ―Engineering disease resistant cattle,‖ Transgenic Res., vol. 14, no. 5, pp. 563–567, 2005. [8] D. E. Kerr et al., ―Lysostaphin expression in mammary glands confers protection against staphylococcal infection in transgenic mice,‖ Nat. Biotechnol., vol. 19, no. 1, pp. 66–70, 2001. [9] L. M. Houdebine, ―Transgenic animal production,‖ Biotechnol. Sustain. Agric. Emerg. Approaches Strateg., vol. 61, pp. 141–184, 2017. [10] S. Mishra et al., ―Effects of nutraceuticals on genetic expressions.,‖ Open Nutraceuticals J., vol. 2, pp. 70–80, 2009. [11] L. Luo et al., ―Fabrication of Synthetic Mesenchymal Stem Cells for the Treatment of Acute Myocardial Infarction in Mice,‖ Circ. Res., vol. 120, no. 11, pp. 1768–1775, 2017. [12] D. R. Brundige, E. A. Maga, K. C. Klasing, and J. D. Murray, ―Lysozyme Transgenic Goats’ Milk Influences Gastrointestinal Morphology in Young Pigs,‖ J. Nutr., vol. 138, no. 5, pp. 921–926, 2018. [13] Rai, M., Pandit, R., Gaikwad, S. & Kövics, G. (2016). Antimicrobial peptides as natural bio-preservatives to enhance the shelf-life of food. Journal of Food Science and Technology, 53, 3381–3394. [14] Singh, V.P. (2018). Recent approaches to food bio-preservation – a review. Open Veterinary Journal, 8, 104. [15] Arqués, J.L., Rodríguez, E., Langa, S., Landete, J.M. & Medina, M. (2015). Antimicrobial activity of lactic acid bacteria in dairy products and gut: Effect on pathogens. BioMed Research International, 2015. [16] Bhardwaj, A., Malik, R.K. & Chauhan, P. (2008). Functional and safety aspects of enterococci in dairy foods. Indian Journal of Microbiology, 48, 317–325. [17] Gautam, N. & Sharma, N. (2009). Bacteriocin: Safest approach to preserve food products. Indian Journal of Microbiology, 49, 204–211. [18] Silva, C.C.G., Silva, S.P.M. & Ribeiro, S.C. (2018). Application of bacteriocins and protective cultures in dairy food preservation. Frontiers in Microbiology, 9. [19] Leyva Salas, Morphology in Young Pigs. The Journal of Nutrition 138:921–926. doi: 10.1093/jn/138.5.921 [20] Donovan DM, Kerr DE, Wall RJ (2005) Engineering disease resistant cattle. Transgenic Research 14:563– 567. doi: 10.1007/s11248-005-0670-8 [21] Hickey, C.D., Sheehan, J.J., Wilkinson, M.G. & Auty, M.A.E. (2015). Growth and location of bacterial colonies within dairy foods using microscopy techniques: A review. Frontiers in Microbiology, 6, 1–8. [22] Hill, D., Sugrue, I., Arendt, E., Hill, C., Stanton, C. & Ross, R.P. (2017). Recent advances in microbial fermentation for dairy and health. F1000Research, 6, 751.