Antimicrobial Agents

Antimicrobial agent is a general term that is mainly concerned with antibiotics, anti-bacterial,

anti-fungal, antivirals and anti-protozoans. Antimicrobial agents are drugs, chemicals or
other substances that are capable of acting by two modes either kill (microbiocidal) or slow the
growth of microbes (micro-biostatic).

An antimicrobial is an agent that kills microorganisms or stops their growth. Antimicrobial

medicines can be grouped according to the microorganisms they act primarily against. For
example, antibiotics are used against bacteria, and antifungals are used against fungi.

They can also be classified according to their function. Agents that kill microbes
are microbicides, while those that merely inhibit their growth are called bacteriostatic agents.

The use of antimicrobial medicines to treat infection is known as antimicrobial chemotherapy,

while the use of antimicrobial medicines to prevent infection is known as antimicrobial
prophylaxis.

The main classes of antimicrobial agents are 

a) disinfectants (non-selective agents, such as bleach), which kill a wide range of microbes on

non-living surfaces to prevent the spread of illness, 

b) antiseptics (which are applied to living tissue and help reduce infection during surgery). E.g.
Dettol.

c) antibiotics (which destroy microorganisms within the body).

d) Sterilant Chemicals applied to inanimate objects to kill all micro-organisms as well as

spores. E.g. Peracetic acid – use to remove surface (countertops) contaminants. They are also
widely used in food industries, particularly during processing, as it does not contain toxic by-
products.
The term "antibiotic" originally described only those formulations derived from living
microorganisms but is now also applied to synthetic agents, such
as sulfonamides or fluoroquinolones. Though the term used to be restricted to antibacterials (and
is often used as a synonym for them by medical professionals and in medical literature), its
context has broadened to include all antimicrobials.

Antibacterial agents can be further subdivided into bactericidal agents, which kill bacteria,

and bacteriostatic agents, which slow down or stall bacterial growth. In response, further
advancements in antimicrobial technologies have resulted in solutions that can go beyond simply
inhibiting microbial growth. Instead, certain types of porous media have been developed to kill
microbes on contact.

Natural Antimicrobials, their Sources and Food Safety

By Muhammad Sajid Arshad and

Syeda Ayesha Batool
Submitted: September 18th 2016Reviewed: June 23rd 2017Published: September 6th 2017
DOI: 10.5772/intechopen.70197

Retrieved from https://www.intechopen.com/books/food-additives/natural-antimicrobials-their-

sources-and-food-safety

Antimicrobial agents and food safety

Traditional food preservation methods are less efficient in reducing the growth of
food-borne pathogens in food products, and the ever-increasing demand for
chemical-free food has paved the way for antimicrobials to be used in food
industry [7]. The use of antimicrobials is a new technology by the food industry to
increase the shelf life of food and overcome the issues of food quality and safety.
These antimicrobials could be of natural or synthetic type, but natural
antimicrobials are gaining much importance than synthetic ones. Even though
synthetic preservatives are approved by government agencies for human use, many
of these preservatives still threaten our health. Thus, researchers give more
importance toward the potential of natural products for their antimicrobial
activities [8–10].

3. Natural antimicrobial agents

Chemical compounds having pharmacological and biological activity and

produced by living organisms are called natural products. Living organisms
produce primary and secondary metabolites [11–13]. Primary metabolites are the
products that have essential function in the organism, while secondary metabolites
could simply be waste products or could have some important function in their
producers. Secondary metabolites can be used as drugs against diseases such as
cancer, inflammation (swelling), and so on and also have antimicrobial activity
[1, 14]. Secondary metabolites possessing antimicrobial activity are called the
natural antimicrobials and could be extracted from different sources like plants
(fruits, vegetables, seeds, herb, and spices), animals (eggs, milk, and tissues), and
microorganisms (fungi and bacteria) [15–17]. With special reference to plants,
secondary metabolites are found to be healthy ingredients that work as
antimicrobials or disease-controlling agents [4]. Owing to the potential of
antimicrobials against pathogenic and spoilage microorganisms, these secondary
metabolites gain much importance for the application in food products [18–20].
They contain the properties of antimicrobials and antioxidants at the same time and
so are considered as a better option for food preservation as compared to synthetic
preservatives [21].

Several researches have been conducted to find out the antimicrobial potential of
natural products, especially the plant sources like fruits, vegetables, herbs, and
spices because they are enriched with compounds having antimicrobial activity.
Nowadays, there are more than 1350 plants with antimicrobial activities and more
than 30,000 antimicrobial components have been extracted from plants [22].
However, many studies have also been conducted on antimicrobial potential of
microorganisms and animals. Food applications of antimicrobials have also been
investigated.

Nowadays, plant extracts and essential oils (EOs) have gained much importance
due to their flavoring as well as antimicrobial potential [23]. Research conducted
on the antimicrobial activity of the extracts from different fruit peels like banana,
apple, pomegranate, sweet lime, orange, mango, and papaya indicated that fruit
peel extracts have mild inhibitory effect against pathogenic bacteria [24–29].
Plants secondary metabolites contain many antimicrobial agents, so they have a
greater inhibitory effect against Gram-positive and Gram-negative bacteria
[14, 27, 30–32]. The chemical composition, concentration, and structure of the
antimicrobial component determine their efficacy. Antimicrobial components of
plant origin include flavonoids, thiosulfinates, glucosinolates, phenolics, organic
acids, flavonoids, and saponins [31, 33, 34]. However, the main compounds with
antimicrobial activity are phenols which include terpenes, aliphatic alcohols,
aldehydes, ketones, acids, and isoflavonoids [35–38].
Antimicrobial components in plant materials are commonly found in herbs and
spices (rosemary, sage, basil, oregano, thyme, cardamom, and clove), fruits and
vegetables (guava, pepper, cabbage, garlic, and onion, citrus), seeds and leaves
(grape seeds, fennel, nutmeg, parsley, and olive leaves) [39–42].

In this chapter, we discuss the role of antimicrobials from different sources with
special reference to meat and meat products. Consumption of meat is important for
the growth, development, and maintenance of health in human beings. Meat is an
animal origin food and is a rich source of proteins, vitamins, minerals, and so on
which is why the safety of meat and meat products is of much importance [43, 44].
Proteins of meat are of much importance with a high amount of essential amino
acids being available and of biological value. Meat and meat products are at a high
risk of microbial spoilage and also cause losses to economy [45]. Although food
industry has developed several new techniques for hygienic slaughtering and
production of meat products, a major concern related to meat consumption is the
presence of pathogenic microorganisms that cause food-borne diseases, for which
raw meat provides an ideal substrate
[46, 47]. Salmonella spp., Campylobacter spp., L. monocytogenes, E. coli, and S.
aureus are the most common meat spoilage agents that cause food-borne diseases
worldwide [48]. Synthetic preservatives are used to overcome this problem, but
their overuse leads to multidrug-resistant phenomenon in bacteria. Moreover, meat
industry is facing a new trend of developing all natural food products, where there
is no place for synthetic preservatives that could be the causative of food
sensitivities, toxicities, and allergies [49–51].

Essential oils, as plant extracts possessing antimicrobial agents and also

antioxidative and flavoring properties, can be considered as healthy ingredients to
be used in meat and meat products. If essential oils are used in meat products, they
can reduce the chances of food-borne diseases and can retard the oxidation of
lipids in meat [52–54].

4. Antimicrobials from plant sources

4.1. Herbs and spices

Herbs and spices have long been used by human beings for different reasons like
food additives, flavorings, and preservatives. They are considered the most
commonly used natural antimicrobials against different pathogens. The
antimicrobial activity of herbs and spices depends on the type of essential oil
present in it, food type in which it has to be used, and the type of microorganism
[11, 55–57].

The efficiency of essential oils from herbs and spices depends upon their chemical
structure, in particular to the presence of hydrophilic functional groups such as
hydroxyl groups [58]. Essential oils from clove, oregano, rosemary, thyme, sage,
and vanillin are the most effective containing the phenolic groups [58]. They
possess inhibitory activity against Gram-positive than Gram-negative bacteria
[59, 60]. Essential oils have high vapor pressure and are able to reach pathogenic
microorganism through gas or liquid phases. Many investigations have proved the
antimicrobial efficiency of essential oils against several pathogenic and spoilage
microflorae. However, the efficiency of essential oils depends upon the pH, storage
temperature, and concentration of oxygen [61].

Some of the antimicrobial compounds that are present in spices and herbs are
eugenol, thymol, thymol and carvacrol, vanillin, allicin, cinnamic aldehyde, and
allyl isothiocyanate that are, respectively, present in cloves, thyme, oregano,
vanilla, garlic, cinnamon, and mustard [26].
Essential oils possess antimicrobial activities against several pathogenic
microorganisms present in meat, including both Gram-positive and Gram-negative
bacteria [62]. Many studies have been conducted to analyze the effects of essential
oils extracted from sources such as oregano, rosemary, thyme, basil, garlic, and
clove, when used alone or in combination with other essential oils [4, 63].

Essential oils extracted from herbs and spices were found to be effective against
several pathogenic microorganisms. Studies showed the antimicrobial activities of
14 essential oils (clove, oregano, rosemary, pepper, nutmeg, liquorice, turmeric,
aniseed, cassia bark, fennel, prickly ash, round cardamom, dahurian angelipca root,
and angelica) against four meat spoilage and pathogenic bacteria (L.
monocytogenes, E. coli, Pseudomonas fluorescens and Lactobacillus sake), and the
results showed that extracts of clove, rosemary, and cassia bark contained strong
antimicrobial activity against these bacteria but a combination of rosemary and
liquorice extracts was the best inhibitor against all four types of bacteria.
Antimicrobials from herbs and spices are widely used by the industry, and
government agencies have approved them to be safe
[64, 65]. Pseudomonas bacteria are responsible for the unacceptability of meat
sausages. The use of thymol extracted from thyme and oregano as an antimicrobial
inhibits the growth of Pseudomonas in sausages [53]. Researches show that
Marjoram, mustard, cinnamon, lemon grass, and rosemary extracts have inhibitory
effects against E. coli O157:H7, S. typhi, and Listeria [66, 67].

The oregano essential oils have antibacterial activities against E. coli, S. aureus, B.
subtilis, and Saccharomyces cerevisiae. The main component in the essential oil of
oregano is carvacrol (80.5%). Some other studies show that S. typhimurium is more
sensitive to oregano essential oils than S. enteritidis [68].
Sodium nitrite has been used as a preservative in meat and meat products, but
researches showed that if it is used in combination with oregano essential oils, it
will slow down the growth of bacteria more efficiently than sodium nitrite alone
[14]. The amount of EOs used in meat and meat products should be higher than the
dose used in in vitro conditions because of the interaction with components of
meat. Antimicrobial essential oils can be used directly or as polyethylene oxide
(PEO)-based antimicrobial packaging [69].

Another research showed that the addition of oregano essential oils at a

concentration of 0.7% will provide antimicrobial activity in minced sheep meat
against S. enteritidis. In vitro tests detected the antibacterial activity of oregano oil
against S. enteritidis in foods such as traditional salted fish and cod fish [70].

Mustard and horseradish essential oils also have antimicrobial activities against
Gram-negative bacteria. Major antimicrobial agent in both is allyl-isothiocyanate
[71–73].

When applying the antimicrobials in meat or meat products, depending upon the
properties and type of pathogen, some EOs are more effective than others.
Eugenol, coriander, clove, oregano, and thyme oils were found to be effective at
levels of 5–20 μl/g in inhibiting L. monocytogenes in meat products, while
mustard, mint, and sage oils were less effective or ineffective [74].

Rosmarinus officinalis L. commonly called rosemary is cultivated in southern

Europe and is used as a flavoring agent due to its better flavor, high antioxidant,
and antibacterial capacity [74, 75]. CARNOSIC ACID AND CARNOSOL are the
major antimicrobial components of rosemary and are effective against both Gram-
negative and Gram-positive bacteria. In meat and meat products, rosemary oil has
high antibacterial activity against L. monocytogenes [76].

Thyme essential oils have high antimicrobial activity owing to the presence of
different compounds. The most prominent of all identified compounds of thyme
essential oils were thymol (50%), followed by p-cymene (24%), linalool (4.6%), γ-
terpinene (4.1%), and 1,8-cineole (4.3%). Thyme oils are effective against L.
monocytogenes at a dose level of 5–20 μl/g. When added at a dose level of 0.3–
0.9%, they are very effective against E. coli in beef. In vitro antimicrobial activity
of thyme essential oil has been tested against E. coli at a temperature higher than
that of refrigeration [77, 78].

Extensive research has been conducted to analyze the efficiency of essential oils
against Salmonella, and results showed that oils extracted from thyme and oregano
reduce the growth of Salmonella up to many folds of colony-forming unit (CFU)
levels, while cinnamon oils at a rate of 7000 mg/kg of meat have strong
antibacterial activity against Salmonella [62, 77].

Research has been conducted to find out the antimicrobial activity of clove oil
against L. monocytogenes in minced mutton. Thymol essential oil from thyme at a
concentration of 250–750 mg is used in fresh minced beef in combination with
modified atmosphere packaging against different microorganisms and also
increases the shelf life of beef [26, 29].

Sage essential oil is used at a concentration of 0.3% in minced beef in combination

with soy protein. Rosemary or Chinese mahogany (500, 1000, and 1500 ppm) is
used to increase fresh chicken sausage [14, 69, 79].

4.1.1. Safety aspect of essential oils

Antimicrobial agents, though very effective against microbial population and able
to extend the quality and shelf life of meat and meat products, should be added
with care because they can cause some side effects. Many essential oils like thymol
and eugenol can cause mucous membrane irritation, if used in higher
concentrations. In vitro studies of various essential oils like carvacrol, carvone,
thymol, and so on show a mild to moderate toxic effects [30]. Some essential oils
can cause allergy or some can have photoactive molecules which can cause
phototoxic reactions [80, 81].

4.2. Fruits and vegetables

Many fruits and vegetables are nowadays well known to have antimicrobial effect
against different pathogenic and spoilage microbes due to their contents of
phenolic and organic acids. Fruit peels that are mostly discarded also contain
antimicrobial compounds [30, 82].

Research showed that the antimicrobial activity of orange peel and capsicum was
due to the presence of phenolic compound (coumaric acid) [83]. In minced beef,
the extracts of capsicum annum have inhibitory effect against S.
typhimurium and Pseudomonas. The minimum dose level of capsicum extract was
1.5 ml/100 g of minced beef to inhibit the growth of S. typhimurium, while a dose
of 3 ml/100 g was required for a bactericidal effect against P. aeruginosa [83].

Pomegranate extract reduces the growth of E. coli. The peel of pomegranate
contains different phenols and flavonoids that have great antimicrobial activity
against Gram-positive bacteria. Peel extracts have inhibitory effects against S.
aureus and B. cereus at a concentration of 0.01%. The addition of pomegranate
peel extract to chicken meat products increases its shelf stability by 2–3 weeks
during chilled storage and its extract is also effective in controlling oxidative
rancidity in these chicken products [7, 62].

Citrus peel extract, lemon grass, and lime peel extracts were investigated for their
antimicrobial activities in meat and meat products. The extracts showed high
potential of antibacterial activity against B. cereus, S. typhimurium, and S. aureus.
Hot water extract of lemon fruit peels, seeds, and juices displayed promising
evidence of antibacterial activity against bacteria E. coli, P. aeruginosa,
and S. aureus [84, 85].

Garlic is a potential inhibitor for food pathogens. Foods contaminated with

pathogens pose a potential danger to the consumer’s health. The use of garlic can
increase the shelf life and decrease the possibilities of food poisoning and spoilage
in processed foods. Garlic extract has antimicrobial activity due to the presence of
an organic sulfur compound allicin, which acts as a growth inhibitor for both
Gram-positive and Gram-negative bacteria
-including E. coli, Salmonella, Streptococcus, Staphylococcus, Klebsiella, Proteus, 
and Helicobacter pylori. Garlic aqueous extract has antibacterial properties
against S. aureus present in hamburger. Freshly ground garlic in combination with
lean camel meat at a concentration of 10, 15, and 25% was used to increase the
shelf life of meat at different temperatures (rooms, incubators, refrigerators). After
4 days of storage at room temperature, 12 days of incubation, and 28 days of
refrigeration, it was found that treatments with 15 and 25% garlic resulted in
complete inhibition of microbial growth with no sign of any organoleptic spoilage
of the meat [11].

The antimicrobial effect of onion extract on the fresh beef fillet meat was
investigated. Beef fillet samples were cut into pieces and treated with 5, 10, 20,
and 50% onion-water extract (v/v) and stored in refrigeration conditions at 4°C.
Microbiological quality of the samples was investigated during storage for 9 days.
Increasing concentrations of onion extract significantly affected E. coli and yeast-
mold counts, but Pseudomonas spp., aerobic mesophilic bacteria, and total coli
forms were not affected significantly for some concentrations and days.

Antimicrobial efficacy of curcumin, one of the active components of the Curcuma

longa (turmeric) plant, was evaluated against food pathogens in a minced meat
medium. S. typhimurium, L. monocytogenes, E. coli O157:H7,
and S. aureus strains were used as food pathogens [86].

5. Antimicrobial edible coatings

Today, many fresh products are available commercially with best nutritional
profile and low cost of production. Consumers also prefer consuming fresh meat
and meat products, but a limit for the commercial availability of fresh meat is its
low storage life because of high moisture contents that cause the growth of
pathogenic and spoilage microorganisms [87].

To avoid this, the spoilage use of antimicrobials is one of the best ways to increase
the shelf life of these perishable food products especially meat and meat products.
The use of antimicrobial films and coatings dates back to twelfth century. The only
difference between film and coating is its thickness. There are many ways of
applying these antimicrobials on food products to enhance the natural appearance
and safety of fresh meat and meat products like spray or spread of antimicrobials
on meat [88, 89].

By the combination of different preservation techniques, researchers have been

successful in achieving the objectives related to microbial quality storage life of
perishable products. The addition of natural antimicrobials in combination with
modified atmosphere packaging and refrigeration has proven to show the best
results. Antimicrobials can also be added in coatings and films to be used in meat
and meat products [88, 90].

The use of antimicrobials in edible films and coatings is an emergent technique

that is helpful in enhancing the quality and safety aspect of food. This technique
includes a control release of antimicrobial agents in effective concentration in the
food product, when required.

The use of oregano essential oil (EO) as natural antimicrobial in combination with
modified atmosphere packaging and refrigeration highly enhances the storage life
of fresh beef or chicken during storage. Whey protein isolate coatings containing
antimicrobial agents like oregano EO, 3-polylysine, or sodium lactate were used on
fresh beef under refrigeration, which was evaluated against the progression of
microflora like Pseudomonas bacteria [91]. By using 1.5% of oregano EO or
0.75% of 3-poly-lysine, the growth of Pseudomonas spp. was reduced and the
development of lactic acid bacteria was completely inhibited.
Both Pseudomonas spp. and total viable microorganisms were completely
inhibited with 2% sodium lactate, even though the effect on LAB was less intense
[89].

The effect of soy protein isolate films containing up to 5% of oregano and/or

thyme EO was evaluated to be effective against coliform and Pseudomonas spp.,
but not significantly effective against total viable microorganisms, LAB,
or Staphylococcus spp. in vacuum-packaged minced beef burgers for a 12-day
period of cold storage at 4°C. CARVACOL AND CINNAMON ALDEHYDES,
the main active compounds of oregano and cinnamon essential oils, were evaluated
for their antimicrobial activity; they were incorporated in edible films based on
apple puree containing 1.5 and 3% of carvacrol or cinnamaldehyde over chicken
breast under refrigeration. These films inactivated the autochthonous spoilage
microflora of chicken [89].

Whey protein isolates-based edible films were evaluated for antimicrobial

activities with different essential oils. These films showed high effectiveness
against L. monocytogenes, E. coli O157:H7, and S. enterica Typhimurium, when
used in combination with 1% sorbic acid in meat sausages. Oregano containing
carvacrol as antimicrobial agent and clove containing eugenol EOs were highly
effective against S. aureus, Salmonella, and L. innocula. Coatings act as barrier
against oxygen transfer leading to growth inhibition of aerobic bacteria.
CHITOSAN has been used as an antimicrobial agent and also as a coating and
wrapper in salami and film and coating combined with lauric arginate and nisin to
reduce L. monocytogenes population in sliced turkey deli meat and also in seafood
and fish [63, 64].

Similarly, milk protein coatings are used in beef in combination with oregano
essential oils against E. coli and Pseudomonas. Chistosan coatings dissolved in
lactic acid in combination with 1% acetic acid are used in roasted beef products
against L. monocytogenes. Similarly, chitosan coatings in combination with
oregano oil at a concentration of 0.7% are used against Pseudomonas spp.
and Brochothrix thermosphacta [91]. Gelatin films are used in Turkey bologna in
combination with nisaplin-based films (GNF) (0.025–0.5%; w/v nisin)
against L. monocytogenes bacteria [89].

Chitosan coatings in different molecular weights and viscosities (14, 57, or 360
mPa) were used in Atlantic cod fish against psychotropic bacteria. Whey protein
coatings were used in smoked fish in combination with Lactoperoxidase system
(0–0.5%, w/v) against L. monocytogenes. Gelatin films were used in sardine
pilchardus in combination with oregano extracts against Enterobacter bacteria.
Alginate, carrageenan, pectin, gelatin, or starch coatings were also used in smoked
salmons in combination with sodium lactate against a mixture
of L. monocytogenes [92].

6. Conclusion

All the researches and studies conducted till now have proved that the use of
synthetic preservatives to increase the shelf life of food and food products is in any
way harmful for the human health, so there is a call for the use of natural products
as preservatives to increase the quality and shelf stability of the food and food
products. Natural antimicrobials contain all the qualities to be used as preservatives
especially in meat and meat products, and plants are the main source of these
antimicrobials.

Plant essential oils have great antimicrobial activity against Gram-positive and
Gram-negative bacteria owing to the potential of phenolic compounds. Essential
oils from herbs and spices like clove, oregano, rosemary, thyme, sage, and vanillin
are the most effective against spoilage and pathogenic microorganisms
like L. monocytogenes, E. coli, P. fluorescens, L. sake, S. aureus, and B. subtilis.
Mustard and horseradish essential oils also have antimicrobial activities against
Gram-negative bacteria. Major antimicrobial agent in both is allyl-isothiocyanate.
Antimicrobial agents, though very effective as antimicrobial agents, should be used
with care because they can cause side effects like irritation. Many fruits and
vegetables also contain antimicrobial activity against pathogenic and spoilage
microbes. Extracts of capsicum annum showed antimicrobial effects
against S. typhimurium in minced beef; similarly, pomegranate extracts reduced the
growth of E. coli. Citrus peel extract, lemon grass, and lime peel extracts showed
great antimicrobial effect against B. cereus, S. typhimurium, and S. aureus. Garlic
is a potential inhibitor for food pathogens. Garlic aqueous extract has antibacterial
properties against S. aureus present in hamburger.

To increase the shelf life of meat and meat products, a new trend is the use of
antimicrobial in edible films and coatings in combination with different packaging
techniques. Oregano essential oils in combination with modified atmosphere
packaging highly increase the shelf life of chicken and beef. Whey protein isolate
coatings added with oregano essential oils in combination with refrigeration were
very effective against Pseudomonas in beef and beef products. Whey protein
isolate-based edible films were evaluated for antimicrobial activities with different
essential oils and were very effective against S. aureus, Salmonella,
and L. innocula. Antimicrobials can be sprayed upon meat and meat products or
meat can be dipped into them. They are completely harmless to human health
owing to the potential of all natural compounds, so there is an increasing market
for the natural antimicrobials to be used as preservatives.