CHAPTER ONE

1.0 INTRODUCTION

The deterioration of food products is one of the main problems of people. People apply many

different methods to prolong the process of food spoilage and increase shelf life. It is estimated

that almost half of food losses at the retail and consumer level in developed countries are lost

(Amit et al., 2017). Food spoilage is caused by Physical changes (Light, temperature, mechanical

damage, etc.), Chemical changes (Enzymatic reactions, non-enzymatic reactions, chemical

interactions, etc.), Food decay, Microorganism proliferation and other elements (Insect, animal,

bird, rodent etc.) (Sahu et al., 2017). Food spoilage can result in changes in color, taste and

nutritional value. There are many different methods to prevent food spoilage. These are Adding

sugar and salt, Food preservatives, Packaging and Processes such as freezing are applied

(Lorenzo et al., 2018). Microorganisms in food can lead to food borne illness. Foodborne illness

is referred to as illness caused by ingesting food that is deteriorated and contaminated by

microorganisms, such as bacteria, fungi, and viruses or parasites, as well as natural toxins,

chemicals, and physical agents. Foodborne illness is a serious and common threat to community

health all over the world and is the main cause of morbidity (CDC, 2020). Both industrialized

and developing countries suffer large numbers of illnesses and the incidence, on a global basis,

appears to be increasing. Mostly, foodborne illnesses are mild and are linked with acute

gastrointestinal (GI) symptoms, such as vomiting and diarrhea. Sometimes these illnesses

become a serious threat to life mostly in children of developing nations, and followed by

disability and chronic sequelae or disability. Foodborne illness may be caused by different

pathogens while harmful bacteria are the leading cause. These pathogens and their toxins enter
into body through different modes and cause foodborne illness. Contamination may occur during

growing, harvesting, preparation, storage, processing, and distribution. Soil, handling techniques,

food preparation, preservation, and storage conditions are the different sources of contamination.

The most common symptoms of foodborne illness include intestinal flu that may last a few hours

or even several days. Symptoms of foodborne illness range from mild to serious that may include

abdominal cramps, nausea, vomiting, dehydration, diarrhea, and fever (CDC, 2020).

1.1 Background of Study

Although processes of food spoilage and methods of food preservation and food fermentation

have been recognized since ancient times. It was not until the 1800s that the relationship between

foods and micro-organisms was established. In 1837, Schwann proposed that the yeast which

appeared during alcoholic fermentation was a microscopic plant, and between 1857 and 1876,

Pasteur showed that micro-organisms were responsible for the chemical changes that take place

in foods and beverages. The observations laid the foundation for the development of food

microbiology. Knowledge in terms of the role that micro-organisms play in food preservation,

food spoilage and food preservation, food spoilage and food poisoning accelerated rapidly until

food microbiology gradually emerged as a discipline. Not all groups of micro-organisms are of

equal interest to the food microbiologist. Bacteria come on top of the list with molds and yeasts

also of considerable importance and viruses. (CDC, 2020).

1.2 Significance of Study

Food spoilage results when microbiological, chemical, or physical changes occur, rendering the

food product unacceptable to the consumer. Microbiological food spoilage is caused by the

growth of microorganisms which produce enzymes that lead to objectionable by-products in the

food.
 CHAPTER TWO

2.0 LITERATURE REVIEW

2.1 Factors affecting Organisms in Food Spoilage

2.1.1 Intrinsic parameters

Intrinsic factors affecting the proliferation rate are more related to the internal characteristics of

food products, and the way in which these characteristics maintain or affect the growth of

microorganisms; these factors include water activity, pH, oxidation-reduction potential, content

and type of nutrients, inhibiting substances, and biological structures (Dashdorj et al., 2017).

2.1.1.1 Water activity

It is defined as the amount of water available for the growth of microorganisms; microbial

proliferation decreases when water availability also decreases (Akhtar et al., 2018). The water

available for met activity determines the degree of microbial growth instead of the total moisture

content (Akhtar et al., 2018). The unit of measurement for the water that microorganisms require

is usually expressed as water activity (Aw), which is defined as the water vapor pressure of food

substrate, divided by the water vapor pressure of pure water, at the same temperature. This

concept is related to relative humidity (RH), thus: RH = 100 × Aw (Akhtar et al., 2018). The

approximate optimal Aw for the growth of most microorganisms is 0.99; most bacteria require

an Aw greater than 0.91 to grow. Gram-negative bacteria require higher values than Gram-

positive bacteria. Most of the natural food products have an Aw of 0.99 or more. Generally,
bacteria have the highest requirements of water activity, fungi have the lowest, and yeasts have

intermediate requirements. Most bacteria that decompose food do not grow with an Aw less than

0.91, but fungi and yeasts can grow with values of 0.80 or less, including surfaces partially

dehydrated. The lowest value reported for bacteria in food is 0.75 for halophytes, while

xerophilic fungi and osmophilic yeasts have shown growth at Aw values of 0.65 and 0.61,

respectively (Akhtar et al., 2018).

2.1.1.2. pH

The pH is defined as the negative logarithm of hydronium ions concentration; it is considered as

a unit of measure to establish acidity or alkalinity levels of a substance, in this case food, and it is

determined by the number of free hydrogen ions (H+) (Akhtar et al., 2018). The effects of

adverse pH affect at least two aspects of the microbial cell-functioning of its enzymes and

nutrients transportation to the cell. The cytoplasmic membrane of microorganisms is relatively

impermeable to H+ and OH− ions; its concentration in the cytoplasm remains reasonably

constant, despite the wide variations that may occur in the pH of the surrounding medium. When

microorganisms are in an environment below or above the neutral level, their ability to

proliferate depends on their ability to change the environmental pH to a more appropriate range,

since key components like DNA or ATP require a neutral medium (Akhtar et al., 2018). The pH

for the optimal growth of most microorganisms is close to neutrality (pH = 6.6– 7.5). Yeasts can

grow in an acid environment and thrive in an intermediate range (4.0–4.5), although they survive

in values between 1.5 and 8.5. Fungi tolerate a wide range (0.5–11.0), but their growth is

generally higher in an acid pH (too acid for bacteria and yeast). Bacterial growth is usually

favored by pH values closer to the neutral level. Nevertheless, acidophilic bacteria grow on

substrates with a pH of up to 5.2 and below that point the growth reduces dramatically (Akhtar et
al., 2018). In general, fruits, vinegars, and wines have pH values lower than those required for

bacterial growth, so they can usually be decomposed by fungi and yeasts. Most vegetables have

pH values lower than those from fruits, and consequently, vegetables are more exposed to

bacterial or fungi decomposition. In contrast, most meats and sea products have pH values equal

or greater than 5.6, making them susceptible to decomposition by bacteria, fungi, and yeasts

(Carrasco et al., 2017).

2.1.1.3. Oxidation-reduction potential

The oxidation-reduction potential (O/R) is an indicator of the oxidizing and reducing power of a

substrate; that is, the O/R potential of a substrate can be generally defined as the ease with which

a substrate loses or gains electrons (when a food product loses electrons, it oxidizes, whereas,

when it gains electrons it is reduced; thus, a food product that easily gives electrons is a good

reducing agent and the one that receives electrons is a good oxidizing agent) (Carrasco et al.,

2017). To achieve optimum growth, some microorganisms require reducing conditions and

others require oxidizing conditions. The O/R potential of a system is expressed with the Eh

symbol (when electrons are transferred from one compound to another, a potential difference is

created between the two compounds; this difference can be measured and expressed as millivolts

[mV]). The more oxidized a substance is, the more positive the electrical potential will be; and

the more reduced a substance is, the more negative the electrical potential will be. When the

concentration of oxidant and reducer is equal, there is an electrical potential of zero (Carrasco et

al., 2017). Saprophytes that are capable of transferring hydrogen as H+ and e− (electrons) to

molecular oxygen are aerobic; that is, aerobic microorganisms require positive Eh values

(oxidized) for their growth, whereas anaerobic microorganisms require negative values of Eh

(reduced). Facultative microorganisms can grow under any of the conditions (Carrasco et al.,
2017). It has to be considered that maximum and minimum Eh values (in mV) necessary for

aerobic and anaerobic growth could be lethal to the other group. Among food substances that

help to maintain reducing conditions are the –SH groups in meats and the ascorbic acid, as well

as, reducing sugars in fruits and vegetables. Some aerobic bacteria grow better under slightly

reducing conditions being known as microaerophiles such as Lactobacillus and Campylobacter

(Carrasco et al., 2017). Most of fungi and yeasts found in food are aerobic, although a few tend

to be facultative anaerobes. Regarding the Eh value of food, vegetables, especially juices, tend to

have Eh values of +300 to +400 mV; so, it is not surprising to find that aerobic bacteria and

fungi are the common cause of decomposition in this type of products. Meats have Eh values

around −200 mV; in ground meats, Eh is usually around +200 mV. Various types of cheese show

Eh values between −20 and −200 mV (Carrasco et al., 2017).

2.1.1.4. Content of nutrients

Microorganisms have nutritional requirements, most of them need external sources of nitrogen,

energy, minerals, as well as vitamins, and related growth factors; these requirements are found in

our food, so if they have the right conditions to develop, they will. In general, fungi have the

lowest nutrient requirement, followed by Gram-negative bacteria, then yeasts and finally, Gram-

positive bacteria, which have the highest requirements (Carrasco et al., 2017). The primary

sources of nitrogen used by heterotrophic microorganisms are amino acids. A great number of

other nitrogen compounds may serve for this function for several types of organisms. For

example, some of them can use free nucleotides and amino acids, while others can be capable of

using peptides and proteins. In general, simple compounds like amino acids will be used by

almost all of the organisms before attacking more complex compounds such as high molecular

weight proteins. The same applies to polysaccharides and lipids (Wildman, 2016).
Microorganisms in food tend to use as energy sources, sugars, alcohols, and amino acids. Fungi

are the most efficient in the use of proteins, complex carbohydrates, and lipids because they

contain enzymes capable of hydrolyzing these molecules into simpler components; many

bacteria have a similar capacity, but most yeasts require simpler molecules (Wildman, 2016). All

microorganisms need minerals, although vitamin requirements vary. Fungi and some bacteria

can synthesize enough B vitamins to meet their needs, while others need to have a source of

vitamins, food products being an excellent source of them (Wildman, 2016). Gram-positive

bacteria are the ones that have lower synthesized capacity, so they need one or more of these

components to grow. In contrast, Gram-negative bacteria and fungi are capable of synthesizing

the most, if not all, of their requirements and consequently, these two groups of organisms can

grow in food products with low content of B vitamins (Wildman, 2016).

2.1.2. Extrinsic parameters

Food factors are very important for the development of microorganisms; there are external or

extrinsic factors. This term refers to environmental factors that affect the growth rate of

microorganisms; these factors include temperature, oxygen availability, and relative humidity, as

well as, the presence and activities of other microorganisms (Carrasco et al., 2017).

2.1.2.1 Storage temperature

Microorganisms have an optimal range, as well as a minimum and maximum temperature to

grow (Carrasco et al., 2017). Therefore, ambient temperature determines not only the

proliferation rate, but also the genera of microorganisms that are going to be developed, along

with the microbial activity degree that is registered. The change in only a few degrees in

temperature will favor the growth of completely different organisms, and it will result in a

different type of food decomposition and/or foodborne disease. Due to these characteristics,
thermal treatment is employed as a method to control microbial activity (Carrasco et al., 2017).

The optimal temperature for the proliferation of most microorganisms ranges from 14 to 40°C,

although some genera develop below 0°C, and other genera grow at temperatures above 100°C.

Nevertheless, food quality must be taken into account when selecting storage temperature.

Although it can be desirable to storage all food products at temperatures equal or less to those of

refrigeration, this is not the best thing to do to maintain a desirable quality in some food products

such as banana, whose quality is best maintained in storage at 13–17°C than at 5–7°C. Similarly,

many vegetables are favored at temperatures near 10°C such as potatoes, celery, cabbage, and

many others. In each case, the success of storage temperature depends, to a large extent, on the

relative humidity and the presence or absence of gases such as carbon dioxide and ozone

(Carrasco et al., 2017).

2.1.2.2. Oxygen availability and presence of other gases in the environment

Like temperature, the oxygen availability determines the microorganisms that will be active.

Some have an absolute requirement for oxygen, while others grow in total absence of it, and

others may grow with or without oxygen. Microorganisms that require free oxygen are called

aerobic microorganisms, while those that thrive in the absence of oxygen are called anaerobic;

and those that grow both in presence or absence of free oxygen are known as facultative

microorganisms (Peelman et al., 2017). Carbon dioxide is the most important atmospheric gas

that is used to control food microorganisms. Along with oxygen, it is used in packaged food with

modified atmosphere.

Ozone is another atmospheric gas with antimicrobial properties, and for decades, it has been used

as an agent to lengthen shelf life of certain types of food. Although being effective against a

variety of microorganisms, it is a highly oxidizing agent; thus, it cannot be used in food products
with high lipid content, as it could accelerate rancidity. Normally, ozone levels of 0.15–5.00 ppm

in the air inhibit the growth of some bacteria that decompose food as well as yeast growth

(Cheng et al., 2017).

2.1.2.3. Relative humidity in the environment

Relative humidity in the environment Relative humidity (RH) of the environment is important

from the point of view of water activity within food and the growth of microorganisms on

surfaces. This extrinsic factor affects microbial growth and can be influenced by temperature

(Lee et al., 2020). All microorganisms have a highwater requirement, this being needed for their

growth and activity. When the Aw of a food product is set at 0.60, it is important that this food is

stored under RH conditions that do not allow food to draw humidity from the air and, therefore,

it increases its own Aw from the surface and subsurface to an extent where microbial growth can

occur. A high relative humidity can cause humidity condensation in food, equipment, walls, and

ceilings (Lee et al., 2020). Condensation causes wet surfaces, which lead to microbial growth

and decomposition. Microbial growth is inhibited by a low relative humidity. When food

products with low Aw values are placed in high RH environments, food takes in moisture until

they reach balance. Similarly, food products with high Aw lose moisture when placed in an

environment with low RH (Lee et al., 2020). There is a relationship between RH and temperature

that must be taken into account when selecting the appropriate storage environments for food

products. Overall, the higher the temperature, the less the RH, and vice versa (Lee et al., 2020).

Bacteria require higher humidity than yeasts and fungi. The optimal relative humidity for

bacteria is 92% or higher, while yeasts prefer 90% or higher, and fungi thrive if the relative

humidity is between 85 and 90%. Food products suffering superficial decomposition by fungi,

yeasts, and specific bacteria, should be stored under low RH conditions. Poorly packed meats
such as whole chickens and beef cuts, tend to suffer a lot of superficial decomposition inside the

refrigerator before internal decomposition occurs, usually, due to high RH in refrigerators, and to

the fact that the biota decomposing meat is essentially aerobic in nature (Lee et al., 2020).

Although it is possible to decrease the possibility of superficial decomposition in certain food

products by storing them in low RH conditions, it should be remembered that the food itself will

lose moisture into the atmosphere under such conditions, and thus, it will become undesirable.

When selecting appropriate RH conditions, there should be taken into account both the

possibility of superficial microbial growth and the quality that the food product needs to have.

By altering the gas atmosphere, it is possible to delay superficial decomposition without

lowering the relative humidity (Lee et al., 2020).

2.1.2.4. Presence and activities of other microorganisms

Some food origin organisms produce substances that can inhibit or be lethal for other organisms;

these include antibiotics, bacteriocins, hydrogen peroxide, and organic acids. Bacteriocins

produced by lactic acid-producing bacteria originated in various food products such as meat, are

of high interest (Lee et al., 2020). Bacteriocins produced by Gram-positive bacteria are

biologically active proteins with bactericidal action. Some bacteriocins produced by these

bacteria inhibit a variety of food pathogens including, B. cereus, C. perfringens, Listeria spp., A.

hydrophila, and S. aureus, among others (Lee et al., 2020). Normally food products can reach the

final consumer at home, in community dining rooms, or restaurants. Measures to prevent food

poisoning should be implemented at these locations, particularly in areas where large volumes of

food are distributed such as cold chain, frozen chain, hot chain, and vacuum cooking. Likewise,

in the frozen chain, food temperature is gradually lowered to −18°C and defrosted at

temperatures higher than 65°C at the time it will be served to the costumer (not before); while in
the hot chain, for example, in a buffet, food is kept at temperatures higher than 65°C and it

should be consumed within 12 h maximum (Lee et al., 2020). Other important measures are the

use of food preservation methods, which can be physical or chemical. Within the physical

methods, there are the traditional or industrial pasteurization, dehydration, preservation in

modified atmosphere, and irradiation. In order to maintain an adequate quality control and to

minimize the risk of food poisoning, microbial markers can be used; these markers do not

represent a potential health risk, however, a large number of them indicate deficiencies in

hygiene and sanitary quality of food products; it also leads to a decrease in the shelf-life and

could be related to the presence of pathogenic microorganisms. The main microbial markers are

aerobic mesophilic, total coliforms, fecal coliforms, Enterococci, E. coli, S. aureus, and lactic

acid bacteria (Lee et al., 2020). Once the risk factors are identified, it is necessary to establish a

system that allows to prevent and decrease all of them; to do this, a method with scientific basis

and systematic profile has been established, this is known as Hazard Analysis and Critical

Control Point (HACCP). A microbiological approach should consider the type of microorganism

or metabolite (toxins) that threatens human health; the analytical methods for its detection and

quantification; the number of samples to be taken and the size of the analytical unit; and the

microbiological limits considered to be adequate at specific points in the food chain (Lee et al.,

2020).

2.2 Food Spoilage Organisms

2.2.1 Bacteria

Bacteria belong to the prokaryotic kingdom Monera. These are unicellular and simple in

structure, rod-like, spherical, or curved in shape and may carry out all of their activities within

one cell. These are the smallest living bodies ranging in size from 0.1 to 10 microns (Karlsson et
al., 2021). Unlike other complex cells, bacteria do not have any membrane-bounded nucleus or

any other intracellular organelles. Bacterium genetic material is organized into a single circular

chromosome containing deoxyribonucleic acid (DNA) that contains all the genes that encode the

bacterium traits. Bacteria multiply in number as a result of rapid cell division and acquire

survival via altering its own genetic material by different methods, such as conjugation,

transduction, and transformation (Karlsson et al., 2021). Each method involves the transfer of

small or large blocks of DNA, which allows the bacteria to acquire new traits that are beneficial

to their survival. This simplicity of bacterial genetic material allows them to thrive in many

different types of environments. These are ubiquitous and found in the depths of the ocean, air,

and high in the atmosphere, soils, contact surfaces, and inside our bodies and hence are the chief

source of food contamination. Although most of bacteria are beneficial for us, other remains a

major source of disease. Campylobacter is unique in that even though it is the number one cause

of foodborne illness, we are still discovering new sources of infection. Campylobacteriosis is

mainly associated with poultry products but research is indicating that pets and cattle (beef) may

also be its major source. Though the versatility of Campylobacter allows it to thrive as an

infectious agent, this trait of bacteria as a whole also allows other bacteria to be beneficial, such

as the bacteria we use to make yogurt or those that maintain balance in the earth’s environment

(Karlsson et al., 2021).

2.2.1.1 Campylobacter

An individual suffers from this illness when a food or drink contaminated with bacteria

Campylobacter is ingested. According to the CDC (Centers for Disease Control and Prevention)

in the USA, Campylobacteriosis is the second most common foodborne illness in the country.

Several types of Campylobacter, including C. jejuni, C. coli, and C. fetus. C. jejuni is responsible
for most cases of human foodborne disease. Infections mostly arise from raw poultry, eating

improperly cooked poultry, drinking raw milk and nonchlorinated or untreated water, or handling

of infected human or animal feces (Law et al., 2019). Most common sources are wastes of

poultry and cattle, but feces of birds, pups, and kittens may also be the source of bacteria.

Symptoms normally occur in 2–5 days after ingestion of contaminated food or organism and

persist for 7 days. Symptoms mostly include abdominal pain and discomfort, nausea, bloody

diarrhea, vomiting, fever, and fatigue (Riddle et al., 2018). Use of antibiotics may be

recommended for people with severe attack. In chronic conditions Campylobacteriosis can leads

to arthritis and Guillain–Barré syndrome (GBS). Individuals with GBS suffer from paralysis of

the limbs and gait disturbances, which can continue for some weeks. In most severe cases, GBS

can lead to breathing problems leading to long stays in hospitals (Nyati and Nyati, 2017).

2.2.1.2 Shigella

Shigellosis is an infectious disease of the intestine that is caused by Shigella bacteria and spreads

mainly through ingestion of contaminated food and water (CDC, 2020). It can be transmitted

from person to person through contaminated tops. It can be transferred by consuming food or

drinks that are contaminated by food managers, who are infected with Shigella. Infections can

also spread through vegetables grown in fields irrigated with dirty sewage, by consuming food

polluted by flies, by drinking or swimming in water containing Shigella, or by the ingestion of an

extremely low number of bacteria, that is, 10 to 100 (CDC, 2020). Four different types of

Shigella-causing illness in humans are Shigella sonnei, S. flexneri, S. dysenteriae, and S. boydii.

In USA and many other developed countries, S. sonnei is the most common form that causes

illness (CDC, 2020). Epidemics of shigellosis mostly occur in the hot and humid climates,

especially in crowded and poor hygiene areas (CDC, 2020). Symptoms usually initiate within 2
days after consumption of contaminated food. Symptoms usually include nausea, fatigue, watery

and sometimes bloody stools, fever, vomiting, and stomachaches and usually resolves in 5–7

days (CDC, 2020). Children less than 5 years of age are more vulnerable for developing

infection. Mostly antibiotics are used for the treatment. Ampicillin or ciprofloxacin are well-

known antibiotics. Sometimes, to treat diarrhea, antidiarrheal medicines are given that further

worsen the situation because these don’t have the capability to kill intestinal bacteria. The most

complicated illnesses include Reiter’s syndrome and hemolytic uremic syndrome (Navaneethan

and Giannella, 2020).

2.2.1.3 Salmonella

Salmonella bacteria are the main cause of food poisoning in human beings. It causes illness when

contaminated meat, poultry, and eggs are eaten raw or undercooked or when foods to be eaten

come in contact with contaminated foods during preparation (Townes, 2020). Symptoms usually

begin from 12 h to 3 days and may include fever, nausea, cramps, headache, diarrhea, and

vomiting, which may persist from 2 to 7 days (CDC, 2020). Salmonella may arise in small

limited epidemics as well as large outbreaks in the general population or in a hospital,

institutions for children and elderly, or restaurants, respectively. Children are the most vulnerable

group to get infected by Salmonella bacteria. Infants, the elderly, and those with weak immune

systems are more likely to get severe illness. Salmonellosis is caused by many types of

Salmonella bacteria, while in the United States S. typhimurium and S. enteritidis are mainly

responsible (Townes, 2020).

The consumer can get these bacteria from different food products, such as poultry, beef, raw

eggs, and sometimes from unwashed fruits and vegetable. We also can get this organism after

handling pets (Townes, 2020). Symptoms resolved mostly in 5–7 days and typically needs no
treatment other than oral fluids. Patients who have severe diarrhea may need intravenous (IV)

fluids. Antibiotic therapy may be needed if the disease persists longer and spreads from the

intestines into the blood circulation. Some types of Salmonella become resistant to antibiotics

and pose serious threats to public health. Most victims improve successfully from Salmonella,

those who cannot, develop a condition called Reiter’s syndrome that results in arthritis. It

includes painful swollen joints, annoyed eyes, and irritated urination. If left untreated bacteria

can move from the intestine to blood and other organs and leads to death (Townes, 2020).

2.2.1.4 Clostridium Botulinum

Clostridium botulinum bacteria causes Botulism. It is most frequently related to inadequately

cooked foods or improperly handled home-canned foods (Caya et al., 2018). Symptoms may

develop within 4–72 h after ingesting contaminated food and may include nervous system (NS)

instabilities, such as double vision, exhausted eyes, pharyngitis, dysphagia, or laryngitis.

Untreated botulism can be life threatening. The three main clinical presentations of botulism

include foodborne botulism (FBB), wound botulism (WB), and infant botulism (IB). When

contaminated soil gets into an open wound, wound botulism results. Less than 5% of all botulism

cases are of this type and it occurs mostly in young boys (Caya et al., 2018). The soil is naturally

contaminated with the botulism organisms. Infant botulism is considered the most common and

may be one cause of sudden infant death (Caya et al., 2018). Most infants are infected less than 6

months of age. The most common source of infection is contaminated honey, which is used to

sweeten milk, because honey is often contaminated with botulinum. Among all of them, the best-

known type of the disease is foodborne botulism, where individuals ingest the toxins present in

food. People of all ages are vulnerable to this type of infection. Foodborne botulism is related to

foods that are not cooked or improperly cooked because the toxin is deactivated and destroyed
by proper cooking. In chronic conditions when a large number of the botulism species grow and

replicate in the intestines, the condition is called intestinal colonization. Organisms grow within

pouches of the intestine where oxygen concentration is low and release their toxins that are

directly absorbed in bloodstream and cause adult infectious botulism. People only acquire this

type of botulism by eating contaminated foods (Argudín et al., 2018).

2.2.1.5 Escherichia coli

Escherichia coli is also responsible for foodborne infection. Many strains of E. coli are harmless

and thrive in the GI tract of Homo sapiens and other warm-blooded animals (Wassenaar, 2016).

These are normally a part of intestinal microflora and are essential for GI health. E. coli serotype

O157:H7 results in serious poisoning in humans. If a person eats food contaminated from this

strain, it will definitely lead to illness. Symptoms include severe abdominal cramps, watery or

bloody diarrhea, and fatigue (CDC, 2020). Shiga toxin-producing E. coli (STEC) can also cause

nausea, vomiting, and low-grade fever. Symptoms usually begin from 2 to 5 days after ingestion

of contaminated food or drinks and may last for 1 week. Individuals can pass STEC infections to

one another during the meetings and interactions so hand-washing practice is considered the best

protective measure to away from this infection (Montealegre et al., 2018).

2.2.1.6 Listeria monocytogenes

Listeria monocytogenes bacteria causes Listeriosis. It can spread through untreated water,

unpasteurized milk and dairy products, raw or uncooked seafood, and vegetables infected by

dung fertilizer (Allerberger and Wagner, 2019). A person may develop symptoms in 2–30 days

after ingestion of contaminated food. Fever, intestinal flu, and chills are common symptoms in

adults while infants may vomit and refuse to drink or may suffer breathing difficulties. Two of

the most common forms of illnesses caused by listeria include noninvasive and invasive GI
listertiosis (Paras et al., 2022). People with strong immune systems develop noninvasive type

febrile gastroenteritis. While immune compromised individuals bear invasive diseases that can

cause meningoencephlitis (Allerberger and Wagner, 2019). Vulnerable groups are pregnant

ladies, newborns, elderly, and people with weakened immune systems. Listeria can be

transmitted from mother to child in uterus. Severe complications in pregnant ladies leads to

meningitis or encephalitis, blood poisonings, spontaneous abortions or still births (Janakiraman,

2018).

2.2.1.7 Staphylococcus aureus

Staphylococcal foodborne illness is one of the most common diseases acquired by S. aureus

(Xue et al., 2020). It is one of the most concerned diseases by public health programs in the

world; it is due to the production of one or more toxins by the bacteria during their growth at

permissive temperatures; however, the incubation period of the disease depends on the amount of

ingested toxin. Small doses of enterotoxins can cause the disease; for example, a concentration of

0.5 ng/mL in contaminated chocolate milk has been reported to cause large outbreaks (Xue et

al., 2020). S. aureus produces various toxins. Staphylococcal enterotoxins are a family of nine

thermostable enterotoxin serotypes belonging to a large family of pyrogenic toxins

(superantigens). Pyrogenic toxins can cause immunosuppression and nonspecific T cell

proliferation (Kim et al., 2018). Enterotoxins are highly stable and they resist high temperatures

(which makes them suitable for industrial use) and environmental conditions of drying and

freezing. They are also resistant to proteolytic enzymes (pepsin and trypsin) at low pH, enabling

them to be fully functional in the digestive tract after infection (Kim et al., 2018). The

mechanism by which poisoning is caused is not entirely clear yet. However, enterotoxins have

been observed to directly affect the intestinal epithelium and the vagus nerve causing stimulation
of the emetic center. It is estimated that 0.1 μg of enterotoxin can cause staphylococcal poisoning

in humans. Apart from causing poisoning, S. aureus can also cause toxic shock syndrome due to

the production of the Toxic Shock Syndrome Toxin 1 (TSST-1) and Enterotoxin Type B (Nassar

et al., 2021). Symptoms include nausea, vomit, abdominal cramps, salivation, diarrhea could be

present or absent. The first three symptoms are the most common ones. Usually, it is a self-

limiting disease and can be cured in 24–48 h, but it can become severe, especially in children, the

elderly, and immunocompromised people. Toxic shock syndrome is characterized by high fever,

hypotension, erythematous rash (similar to scarlet fever, peeling of the skin during recovery, flu-

like symptoms, vomiting, and diarrhea) (Tong et al., 2019). The diagnosis of the disease is

carried out by detecting the staphylococcal enterotoxin in the food or by recovering at least 105

S. aureus/g from food leftovers. The enterotoxin can be detected by several methods: bioassays,

molecular biology, and immunological techniques. The isolated strains can be genetically

characterized by multilocus sequences from the spa or SCCmec gene, and pulsed-field

electrophoresis (Tong et al., 2019). The mainly involved food products in outbreaks and where

S. aureus can grow optimally, since they are stored at room temperature, are meat and its derived

products, poultry and eggs, milk and its derived products, salads, and bakery products (cream-

filled cakes and stuffed sandwiches) (Tong et al., 2019). Other factors that must be taken into

account are the emergence of methicillin resistant strains, which may be found in food (mainly in

meat and milk). It is important to note that many of the isolates obtained from outbreaks are not

tested for antimicrobial susceptibility; due to the various problems that these strains can create,

the antimicrobial susceptibility test should be performed. They have been reported to be

causative agents of outbreaks in blood infections and wounds in immunocompromised patients in

hospitals (Ha et al., 2020). Foodborne illness due to S. aureus may be preventable. It is known
that the permissible temperature for the growth and production of the enzyme is between 6 and

46°C; thus, food products could be cooked above 60°C and refrigerated below 5°C. Therefore,

maintaining the cold chain of food can prevent the growth of the microorganism. By using good

manufacturing practices and good hygiene practices, the contamination by S. aureus can be

prevented (Ha et al., 2020).

2.2.1.8 Bacillus Cereus

Bacillus cereus is a ubiquitous microorganism in the environment, and it can easily contaminate

any food production and processing system, due to the formation of endospores. The bacterium

can survive pasteurization and cooking processes (Paul et al., 2021). It has been demonstrated

that this microorganism produces, cereulide or emetic toxin; three enterotoxins, hemolysin BL

(HBL), nonhemolytic (NHE), cytotoxin K (CytK), which are responsible for the emetic

syndrome and diarrhea; and three phospholipases, phosphatidylinositol hydrolase,

phosphatidylcholine hydrolase, and hemolytic sphingomyelinase (Tuipulotu et al., 2021).

Cereulide is a thermostable cyclic peptide that causes emesis by stimulating the afferent vagal

pathway through its bond to the serotonin receptor. The toxin is produced during the stationary

phase of growth of the microorganism and it accumulates in food over time. The structure of the

toxin explains its resistance to food processing methods (Bremer, 2018). In contrast, inside the

small intestine of the host, the thermolabile enterotoxins, HBL and NHE, produced during the

exponential phase of the vegetative growth of the bacterium are the cause of diarrheal syndrome;

the proteins that form enterotoxins (binding and lithic factors) are unable to traverse intact the

gastric barrier; that is why it is considered that preformed or extracellular enterotoxins in food

are not involved in the pathogenesis of the bacterium (Bremer, 2018). It is believed that the spore

germination that reaches the small intestine, the growth, and the simultaneous production of the
enterotoxin are the ones that cause diarrhea. HBL is a hemolysin formed by three components,

two protein subunits (L2 and L1), and one B protein; it has hemolytic, cytotoxic, and

dermonecrotic effect, and it induces vascular permeability. NHE also consists of three

components: NheA, NheB, and NheC. It has been demonstrated that strains producing emetic

toxin do not produce enterotoxin. The cytotoxin K is similar to the Alpha-toxin of S. aureus and

the Beta-toxin of C. perfringens (Bremer, 2018). Furthermore, the enterotoxin FM (EntFM) has

been described; it is a 45 kDa polypeptide encoded by the entFM gene, located in the bacterial

chromosome. It has not been directly involved in food poisoning; however, the presence of the

gene in strains that cause diarrheal outbreaks has been detected; in experiments with mice and

rabbits, it causes vascular permeability (Bremer, 2018). The emetic syndrome is characterized by

nausea and vomit similar to those produced by S. aureus poisoning. Symptoms appear soon after

consuming food contaminated with the preformed toxin. Generally, poisoning develops with

mild symptoms, usually lasting no more than 1 day, but severe cases require hospitalization. The

diarrhea that is caused belongs to the secretory type, similar to the one produced by V. cholerae.

Colic pain occurs similar to that of C. perfringens poisoning. Both syndromes are self-limiting

(Wu et al., 2019). Enterotoxins can be detected by immunoassays or molecular biology

(conventional PCR and multiple PCR) by looking for the ces gene (nonribosomal production of

cereulide); by detecting the hblD, hblC, and hblA genes encoding the L1, L2, and B protein

components of the HBL toxin, respectively; or the nheA, nheB, and nheC genes of the NHE

toxin components. The 16S ribosomal gene can be looked for by real-time PCR (Wu et al.,

2019). Apart from causing food poisoning, B. cereus can also cause local and systemic infections

in immunocompromised patients, neonates, people taking drugs, and patients with surgical or

traumatic wounds, or catheters (Cheng et al., 2021). The most susceptible food products to be
contaminated include flours, meats, milk, cheese, vegetables, fish, rice and its derived products;

generally, in food with high content of starch. The strains produced by the emetic toxin grow

well in rice dishes (fried and cooked) and other starchy products; although, there have been

studies where it has been demonstrated that the toxin can be in different types of food products;

while strains producing diarrheagenic toxins grow in a wide variety of food products, from

vegetables to sauces and stews (Cheng et al., 2021). Strains isolated from infections have been

shown to be sensitive to chloramphenicol, clindamycin, vancomycin, gentamicin, streptomycin,

and erythromycin; they are resistant to β-lactam antibiotics, including third-generation

cephalosporins (Cheng et al., 2021). Inadequate cooking temperatures, contaminated equipment,

and poor hygiene conditions at the food processing and preparation sites are the major factors

that contribute to food poisoning by B. cereus and its toxins; that is why, it is suggested to store

food at temperatures lower than 4°C or to cook them at temperatures higher than 100°C, and to

reheat or cool food rapidly, to avoid prolonged exposure to temperatures that allow spore

germination and to diminish the risks of a possible poisoning (Griffiths and Schraf, 2017).

2.2.2 Fungi

Fungi include all organisms in the Kingdom Fungi, such as yeasts, molds, mushrooms, and

mildews. These are eukaryotic (have a clearly defined nucleus) multicellular organisms in which

chromosomes contain the genetic material (DNA). Cytoplasm contains various membrane-bound

organelles, such as mitochondria (create and convert energy), Golgi apparatus (modifies and

transports proteins), and lysosome (digest polysaccharides or old cell parts). Fungi are similar to

plants but are unable to do photosynthesis. Approximately 70,000 recognized species of fungi

are known whereas only 300 of them cause human infections mostly in immunocompromised

people. Fungi in the form of mycotoxins cause diseases in humans and animals (Wu et al., 2019).
Histoplasmosis is an infectious disease caused by the fungus Histoplasma capsulatum. There are

also many beneficial fungi. For example, Penicillium is the source of antibiotic “penicillin”

commonly used to treat bacterial infections (Law et al., 2019).

2.2.2.1 Mycotoxins and Alimentary Mycotoxicoses

2.2.2.1.1 Aflatoxins

Aflatoxins originated from Aspergillus parasiticus and Aspergillus flavus. They are frequently

found in tree nuts, peanuts, maize, other oil seeds, including corn and cotton seeds. Aflatoxins

cause necrosis, cirrhosis and carcinoma of the liver (Li et al., 2020).

2.2.2.1.2 Altertoxins

Altertoxins originated from Alternaria spp. Some of the toxins can be present in wheat and

tomatoes. The toxins can be easily cross-contaminated between grain commodities, suggesting

that manufacturing and storage of grain commodities is a critical practice (Li et al., 2020).

2.2.2.1.3 Fumonisins

Crop corn can be easily contaminated by Fusarium moniliforme and its Fumonisin B1 causing

oesophageal cancer in humans (Adejumo et al., 2017).

2.2.2.1.4 Amanita phalloides

Amanita phalloides contains 2–3 mg of amatoxins per gram of dry tissue. A single mushroom

can kill an adult human. Amatoxins cause inhibition of RNApolymerase leading to inhibition of

protein synthesis and cell death of the intestinal epithelium and liver tissue and liver failure

follows (Adejumo et al., 2017). Ingestion of the amatoxincontaining mushroom results in

vomiting, nausea, abdominal pain and bloody diarrhea that develop within 6–24 hours. Within 3–
4 days, patients develop icterus (yellow skin), hypoglycemia, bleeding, renal failure and sepsis

(bacterial infections). The mortality rate in humans is 10–40% (Adejumo et al., 2017).

2.2.2.1.5 Amanita muscarina

Amanita muscarina is characterized by muscarine, which affects the autonomic nervous system.

Within a few minutes to a few hours of consumption of mushrooms containing these toxins, the

patient will experience perspiration, salivation and lacrimation syndrome, blurred vision,

abdominal cramps, watery diarrhea, constriction of the pupils, hypotension and a slowed pulse

(Adejumo et al., 2017).

2.2.2.1.6 Marine Bio Toxins

In many countries of the world this type of poisoning is a major public health problem, affecting

many thousands of people. The most common type is ciguatera, which is associated with

consumption of a variety of tropic and subtropical fish, mainly coral fish, feeding on toxin-

producing dinoflagellates, or predatory fish containing coral fish (CDC, 2020).

2.2.3 Viruses

Viruses are extremely small and very simple in composition. These range in size from 0.02 to 0.4

microns, even smaller than bacteria (Law et al., 2019). Viruses are unique organisms belonging

to their own kingdom and cannot truly be classified as a living entity because these remain

dormant until they find an appropriate host cell where these are multiplied or replicated. No free-

living forms of viruses have been found (Law et al., 2019). Viruses are not composed of cells,

which are considered to be the smallest structural unit of living matter that is able to function

independently. Viruses have the genetic material but absolutely need to borrow another cell’s

machinery to replicate. Viruses consist of genetic material and a protein shell called a capsid.
Some viruses have an additional outer envelope, rod, and/or tail. Once inside a host, they utilize

the host cell’s machinery to create new viruses. Eventually, new viruses accumulate in the host

cell leading to cell rupture. Afterward, viruses move to another cell and repeat the process,

causing more cell deaths and spread infectious diseases. Viruses vary in their degree of virulence

and ability to cause illnesses, such as the common cold, caused by the influenza virus, to fatal

diseases, such as AIDS (acquired immune deficiency syndrome) caused by HIV (human

immunodeficiency virus) (Law et al., 2019).

2.2.3.1 Norwalk-Like Virus

A well-known foodborne disease is viral gastroenteritis caused by group of viruses called

Norwalk-like virus (Nguyen et al., 2017). Sources are raw or partially cooked polluted shellfish

and transfer of infections from infected people involved in food handling and preparation.

Symptoms include nausea, vomiting, fever, chills, abdominal pain, and body aches. In a majority

of the cases Norwalk-like viruse-induced gastroenteritis is a self-controlled disease enduring for

24–60 h. Though after recovery from symptoms, the infected person may continue excreting

high loads of Noroviruses in feces. If the disease persists longer, it worsens the condition and

leads to dehydration, kidney failure, sudden weight loss, chronic diarrhea, malnutrition, and

ultimately death (Atmar et al., 2018).

2.2.3.2 Rotavirus

Rotavirus is one of several viruses that cause infections commonly known as stomach flu,

despite having no relation to influenza (Hallowell et al., 2022). By the age of five, nearly every

child in the world has been infected with rotavirus at least once and adults are rarely affected.

Janko et al. (2022) demonstrated the occurrence of Rotavirus in meat products (minced meat,

beef burger and sausage) as well as in milk and its products (ice-cream and cottage cheese)
intended for human consumption in different localities in Egypt. Rotavirus is transmitted by the

fecal-oral route. It infects cells that line the small intestine and produces an enterotoxin, which

induces gastroenteritis, leading to severe diarrhea and sometimes death through dehydration

(Janko et al., 2022).

2.2.3.3 Hepatitis A

Hepatitis A is an infectious disease of the liver caused by Hepatovirus A (HAV); it is a type

of viral hepatitis. Many cases have few or no symptoms, especially in the young (Matheny and

Kingery, 2019). The time between infection and symptoms, in those who develop them, is 2–6

weeks. When symptoms occur, they typically last 8 weeks and may include nausea, vomiting,

diarrhea, jaundice, fever, and abdominal pain young (Matheny and Kingery, 2019). Around 10–

15% of people experience a recurrence of symptoms during the 6 months after the initial

infection. Acute liver failure may rarely occur, with this being more common in the elderly

young (Matheny and Kingery, 2019).

It is usually spread by eating food or drinking water contaminated with infected feces young

(Matheny and Kingery, 2019). Undercooked or raw shellfish are relatively common sources

(Bellou et al., 2018). It may also be spread through close contact with an infectious person

(Matheny and Kingery, 2019). While children often do not have symptoms when infected, they

are still able to infect others (Matheny and Kingery, 2019). After a single infection, a person is

immune for the rest of his or her life. Diagnosis requires blood testing, as the symptoms are

similar to those of a number of other diseases. It is one of five known hepatitis viruses:

A, B, C, D, and E (Matheny and Kingery, 2019).

The hepatitis A vaccine is effective for prevention. Some countries recommend it routinely for

children and those at higher risk who have not previously been vaccinated (Matheny and

Kingery, 2019). It appears to be effective for life. Other preventive measures include hand

washing and properly cooking food. No specific treatment is available, with rest and medications

for nausea or diarrhea recommended on an as-needed basis. Infections usually resolve

completely and without ongoing liver disease. Treatment of acute liver failure, if it occurs, is

with liver transplantation (Matheny and Kingery, 2019).

2.2.3.4 Hepatitis E

Hepatitis E is a “self-limiting” disease but occasionally develops into an acute severe liver

disease and is fatal in about 2% of all cases. The disease in pregnant women is more often severe

and is associated with a clinical syndrome called "fulminant hepatic failure" with an elevated

mortality rate of around 20%. It spreads mainly through fecal contamination of water supplies or

food; person-to-person transmission is uncommon (Matheny and Kingery, 2019).

2.2.4 Parasites

These are organisms that live on or within the host and gain sustenance from it. Three major

groups of parasites that cause human infections include the parasitic protozoa, the parasitic

helminthes or worms, and certain arthropods that either directly cause disease or proceed as

vectors (Law et al., 2019). Parasitic infection means infection caused by one of these three

groups, such as protozoa, worms, and insects. Protozoa are unicellular eukaryotes ranging from

microscopic to several millimeters in length. The protozoa differ significantly in their physiology

and life cycles and can be found in soils and natural waters (Law et al., 2019).
2.2.4.1 Helminths

Roundworms, tapeworms and flukes are transmitted to humans via food and water in many parts

of the world. Nonhuman hosts play a vital role in the life cycles of many of these parasites.

Foods can be made safe by cooking, but not all foods are customarily cooked (Garcia et al.,

2018).

2.2.4.1.1 Tapeworms

Taenia solium

Taenia solium in the intestine and passing eggs through feces, sometimes a cysticercus (a larva

sometimes called a "bladder worm") develops in the human and the human acts like an

intermediate host. This happens if eggs get to the stomach, usually as a result of contaminated

hands, but also of vomiting. Cysticerci often occur in the central nervous system, which can

cause major neurological problems like epilepsy and even death. The condition of having

cysticerci in one's body is called cysticercosis (Garcia et al., 2018).

Taenia saginata

T. saginata, also known as the beef tapeworm is a parasite of both cattle and humans. T. saginata

occurs where cattle are raised by infected humans maintaining poor hygiene, human feces is

improperly disposed off, meat inspection programs are poor and where meat is eaten without

proper cooking (Hernández et al., 2017).

Echinococcus spp
Echinococcus spp lives in dogs, foxes and other canids and intermediate stages normally infect

sheep, goats, pigs, horses and cattle. Humans can also serve as an intermediate host if they ingest

tapeworm eggs in contaminated water or on raw contaminated vegetables. The larval tapeworms

form fluid-filled cysts (called hydatid cysts) in the liver, lungs and other organs of intermediate

hosts. Tissue damage may be severe in some cases (Hernández et al., 2017).

2.2.4.1.2 Roundworms

Anisakis

Anisakis is a genus of parasitic nematodes, which have a life cycle involving fish and marine

mammals (Ángeles et al., 2020. They are infective to humans and cause Anisakiasis and fish

which have been infected with Anisakis spp. (A. simplex) can produce apain, nausea and

vomiting may occur. Occasionally then anaphylactic reaction in people who have become

sensitized to immuno-globulin E (Ángeles et al., 2020). Within hours after ingestion of fish meal

containg infective larvae, violent abdominal larvae are coughed up. The larvae may burrow into

the walls of the bowel and produce severe eosinophilic granulomatous response which may

occur 1 to 2 weeks following infection, causing symptoms mimicking Crohn's disease (Ángeles

et al., 2020).

Trichinella spiralis 

Trichinella species, the smallest nematode parasite of humans, has an unusual lifecycle, and are

one of the most widespread and clinically important parasites in the world (Xiaolei et al.,

2019). The small adult worms mature in the small intestine of a definitive host, such as a pig.

Each adult female produces batches of live larvae, which bore through the intestinal wall, enters

the blood (to feed on it) and lymphatic system, and are carried to striated muscle. Once in the
muscle, they encyst, or become enclosed in a capsule. Humans can become infected by eating

infected pork, horsemeat, or wild carnivores such as fox, cat, hyena or bear (Xiaolei et al., 2019).

Trichinella spiralis is a viviparous nematode parasite, occurring in rodents, pigs, bears, hyenas

and humans, and is responsible for the disease trichinosis. It is sometimes referred to as the "pork

worm" due to it being typically encountered in undercooked pork products. It should not be

confused with the distantly related pork tapeworm (Xiaolei et al., 2019).

Trichinosis, also called trichinellosis, or trichiniasis, are C. parvum and C. hominis (previously

C. parvum is a parasitic disease caused by eating raw or genotype 1). C. canis, C. felis, C.

meleagridis and C. undercooked pork and wild game infected with the larvae muris can also

cause disease in humans (Gao et al., 2018). Infection is of a species of roundworm T. spiralis.

Only three through contaminated material such as water, uncooked Trichinella species are

known to cause trichinosis; or cross-contaminated food that has been in contact with T. spiralis,

T. nativa and T. britovi (Gao et al., 2018). The great majority the feces of an infected individual

or animal. The most of trichinosis infections have either minor or no symptoms important

zoonotic reservoirs are cattle, sheep and goats and no complications. Trichinosis initially

involves the (Gao et al., 2018). The parasite is transmitted by microbial cysts intestines. Within

1-2 days of contagion, symptoms such as nausea, heartburn, dyspepsia and diarrhea may appear.

Later on, as the worms encyst in different parts of the human body, other manifestations of the

disease may acute diarrhea or persistent diarrhea that can last for few appear, such as headache,

fever, chills, cough, eye weeks. The most dangerous case is worms entering the central nervous

system. They cannot survive there, but they may cause enough damage to produce serious

neurological deficits (such as ataxia or respiratory paralysis) and even death (Gao et al., 2018).
2.2.4.1.3 Trematodes

Fasciola hepatica

Fasciola hepatica (sheep liver fluke) and F. gigantica (cattle liver fluke) live in the bile

ducts of man and herbivorous animals. The eggs must find its way to fresh water to

complete its life cycle and develop into the infective stage, encysted metacercariae (Bargues

et al., 2018). Human infection occurs by ingestion of the encysted metacercariae on raw

vegetables or in water. Disease symptoms include fever, abdominal pain, weight loss and

enlarged liver. Some evidence suggests that heavy or chronic infections of this parasite are

associated with liver tumors (Bargues et al., 2018).

Paragonimus (Lung Fluke)

Lung Fluke reside in many parts of the world, including North America and infect the human

lung and occasionally other tissues, including the brain. The most common species is P.

westermani. Reservoirs are cats and monkeys. In fresh water, eggs develop into microcercous

cercariae in snail and then into the infective stage (encysted metcercariae) in fresh water crabs,

cray fish and shrimps (Bargues et al., 2018). Human infection occurs from ingestion of raw or

incompletely cooked crabs, cray fish or shrimps. Symptoms of diarrhea, abdominal pain and

fever may occur early after infection and progress to coughing and thoracic pain as the worms

settle in the lungs. Infections are sometimes initially misdiagnosed as tuberculosis, which can

delay effective treatment (Bargues et al., 2018).

2.2.5 Protozoa

E. histolytica is an anaerobic parasitic protozoan predominantly infecting humans and other

primates. E. histolytica is estimated to infect about 50 million people worldwide (Rawat et al.,
2020). When cysts are swallowed they cause infection by excysting (releasing the trophozoite

stage) in the digestive tract. E. histolytica infection in human can lead to amoebic dysentery or

amoebic liver abscess. Symptoms can include fulminating dysentery, bloody diarrhea, weight

loss, fatigue and abdominal pain. The amoeba can actually 'bore' into the intestinal wall, causing

lesions and intestinal symptoms and it may reach the blood stream. From there, it can reach

different vital organs of the human body, usually the liver and cause liver abscess, which can be

fatal if untreated (Rawat et al., 2020).

2.2.5.1 Toxoplasma gondii

T. gondii is a species of parasitic protozoa in the genus Toxoplasma. The definitive host of T.

gondii is the cat, but the parasite can be carried by all known mammals. Humans infection

(toxoplasmosis) occurs by ingestion of the infecive stage; oocysts (e.g., by eating unwashed

vegetables) or tissue cysts in improperly cooked meat (Rawat et al., 2020).

Symptoms of toxoplasmosis are flu-like and selflimiting. However, in case of

immunocompromised patients, the most notable manifestation of toxoplasmosis is

encephalitis, which can be deadly. If infection with T. gondii occurs for the first time during

pregnancy, the parasite can cross the placenta, possibly leading to hydrocephalus or

microcephaly, intracranial calcification and chorioretinitis, with the possibility of spontaneous

abortion (miscarriage) or intrauterine death (Rawat et al., 2020).

2.3 Functional Properties of Food Spoilage Microorganisms

2.3.1 Probiotic Microorganisms

Probiotics are defined as live microorganisms that, when administered in adequate amounts,
confer a health benefit on the host (Hill et al., 2020). Probiotic organisms used in foods must

have the ability to resist gastric juices, exposure to bile, and be able to proliferate and colonize

the digestive tract (Hill et al., 2020). The beneficial effects of probiotic foods on human health

and nutrition are constantly increasing, and probiotics are popularly using bio-ingredients in

many functional fermented foods (Hill et al., 2020). The most commonly used probiotic bacteria

belong to the heterogeneous group of LAB (Lactobacillus, Enterococcus) and to the genus

Bifidobacterium, however, yeasts and other microbes have also been developed as potential

probiotics during recent years (Varankovich et al., 2021). Products containing probiotic bacteria

generally include foods and supplements (Varankovich et al., 2021). Fermented milk products

are the most traditional source of probiotic strains of lactobacilli. However, commercial probiotic

lactobacilli have also been added to meat products, snacks, fruit juice, etc. (Shah, 2021).

Probiotic properties of Lactobacillus plantarum isolated from kimchi, Korean fermented

vegetable product, has been reported and is also found to prevent the growth of Helicobacter

pylori (Shah, 2021).

2.3.2 Antimicrobial Properties

Many species of LAB isolated from fermented vegetable and milk products have antimicrobial

activities due to production of antimicrobial compounds such as bacteriocin and nisin (Khan et

al., 2020). Many strains of LAB isolated from kimchi produce antimicrobial compounds such as

bacteriocin by L. lactis BH5 and L. citreum GJ7 and pediocin by P. pentosaceus (Khan et al.,

2020). Species of LAB isolated from kimchi show strong antimicrobial activity against Listeria

monocytogenes, Staphylococcus aureus, E. coli, and Salmonella typhimurium (Lee et al., 2009).

Weissella cibaria isolated from fermented cabbage product shows antimicrobial activity against

Gram-positive and Gram-negative pathogens (Lee et al., 2018). Lactococcus lactis isolated from
dahi, Indian curd, produces nisin Z that inhibits L. monocytogenes and S. aureus (Lee et al.,

2019). Several LAB species isolated from Romanian traditional fermented fruits and vegetables

have antimicrobial activity against L. monocytogenes, E. coli, Salmonella, and Bacillus (Lee et

al., 2021). Microorganisms as protective cultures, e.g., bacteriocin producers, may have several

advantages, as they can contribute to the flavor, texture and nutritional value of the product

besides the production of bacteriocin (Lee et al., 2022).

2.3.3 Antioxidant Activity

Antioxidant activities in fermented foods include 1,1-diphenyl 2-picryl hydrazyl (DPPH) radical

scavenging activity, 2,20 -azinobis (3-ethylbenzo-thiazoline-6-sulfonic acid; ABTS) radical

scavenging activity, total phenol content (TPC) estimation, and reducing power assay (Wang et

al., 2019). Many Asian fermented soybean foods have antioxidant properties, e.g., natto,

Bacillus-fermented soybean food of Japan, chungkokjang and jang, fermented soybean foods of

Korea, douchi, a fermented soybean food of China, kinema, Bacillus-fermented soybean food of

India and Nepal, bekang and tungrymbai, Bacillus-fermented soybean foods of India, thua nao,

Bacillus-fermented soybean food of Thailand, and tempe mold-fermented soybean food of

Indonesia (Wang et al., 2020). Antioxidant activityes have also been observed in kimchi and

yogurt (Tamang, 2020).

2.3.4 Peptide Production

Bioactive peptides are formed during food fermentation by proteolytic microorganisms (Singh et

al., 2018). In fermented foods peptides have some functional properties such as

immunomodulatory, antithrombic, and antihypertensive properties (Singh et al., 2018). Species

of Bacillus are involved in enzymatic hydrolysis of protein producing peptides and amino acids,

which claim to have health benefits (Tamang, 2020). Inhibitory properties of Angiotensin
converting enzyme (ACE) have been studied in various fermented milk products such as kefir,

koumiss, yogurt, fermented camel milk, cheese, and fermented fish products (Tamang, 2020).

2.3.5 Production of Enzymes by Microorganisms

Another important reason to ferment foods is to coax microorganisms into producing enzymes

that also provide very useful services. During food fermentation microorganisms produce

enzymes to break down complex compounds to simple bio-molecules for several biological

activities such as proteinase, amylase, mannase, cellulase, and catalase in many Asian fermented

soybean foods by Bacillus spp. (Tamang, 2020). Common genera of mycelial fungi in fermented

foods and beverages such as Actinomucor, Amylomyces, Aspergillus, Monascus, Mucor,

Neurospora, and Rhizopus produce various carbohydrases such as α- amylase, amyloglucosidase,

maltase, invertase, pectinase, ß-galactosidase, cellulase, hemi-cellulase; acid and alkaline

proteases; and lipases (Tamang, 2020). Taka-amylase A (TAA), an enzyme produced by

Aspergillus oryzae in koji has many uses in industry (Tamang, 2020). Dry, solid, cake-like mixed

amylolytic starters used for alcohol production in the Himalayas have yeasts Saccharomycopsis

fibuligera, S. capsularis and Pichia burtonii with high amylase activities (Singh et al., 2018).

2.3.6 Degradation of Anti-nutritive Compounds

Some microorganisms present in fermented foods may degrade anti-nutritive substances and

thereby convert the substrates into consumable products (Tamang, 2020). Various steps

employed during the processing of gari and fufu, fermented cassava products of Africa, such as

peeling, washing, grating, fermentation, dewatering and roasting minimizes the residual cyanide

contents of the product (Babalola, 2018). Bitter varieties of cassava tubers contain the

cyanogenic glycoside linamarin and lotaustralin, which are detoxified by species of Leuconostoc,

Lactobacillus, and Streptococcus during traditional method to gari and fufu productions to yield
hydrocyanic acid (HCN) which has low boiling point and escapes from the dewatered pulp

during toasting rendering the product safe for human consumption (Babalola, 2018). In tempe,

Rhizopus oligosporus eliminates the flatulence causing indigestible oligosaccharides such as

stachyose and verbascose into the absorbable monosaccharides and disaccharides (Babalola,

2018). Degradation of anti-nutritive compounds by B. subtilis has been reported in kinema.

Phytic acid is reduced during fermentation of idli and rabadi, a fermented cereal food of India

(Babalola, 2018).

2.4 Food Preservation

Food preservation includes food processing practices which prevent the growth

of microorganisms, such as yeasts and slow the oxidation of fats that cause rancidity. Food

preservation may also include processes that inhibit visual deterioration, such as the enzymatic

browning reaction in apples after they are cut during food preparation. By preserving food, food

waste can be reduced, which is an important way to decrease production costs and increase the

efficiency of food systems, improve food security and nutrition and contribute towards

environmental sustainability (Abakarov and Nunes, 2021).

2.4.1 Conventional Technologies of Food Preservation

2.4.1.1 Thermal Processing

The thermal process is one of the most widely used methods for food preservation, even

considering its drawbacks and the development of further non-thermal technologies (Augusto et

al., 2017). It is a unit operation where the food is heated to a certain temperature, maintained for

a certain time in order to promote the required microbial and/or enzymatic inactivation, and then
cooled. The food preservation by thermal processing is based on the use of thermal energy

(heat) for microbial and enzymatic inactivation, obtained due to protein denaturation and melting

of lipid components, among other effects. However, although microbial and enzymatic

inactivation is desirable, the thermal processing also results in other reactions (generally)

undesirable, such as sensorial changes and nutrient destruction. The challenge, therefore, is to

guarantee a safe and stable product (with the desired microbial and enzymatic inactivation), but

with better sensorial and nutritional properties, with lower costs and energy saving consumption

(Augusto et al., 2017).

2.4.1.2 Cooling

The cooling/chilling process is widely used in the food industry for ensuring the shelf life

extension of products with lower sensory and nutritional changes (Augusto, 2017). It is a unit

operation where the food is cooled to a certain temperature and maintained under these

conditions throughout its storage, generally used in association with other preservation processes

(with the exception of raw materials) (Augusto et al., 2017).

2.4.1.3 Freezing

The freezing process is a unit operation where latent heat is removed from the food, with

consequent solidification of the water (Augusto et al., 2017). The change in the physical state of

the water defines freezing as a very effective method of conservation, being more than a “super

cooling.” When food is frozen, the microorganisms enter into latency state, not, as a general rule,

inactivated. Thus, when the microbial load reduction is required, another method of preservation

should be used for safety assurance in consumption. Once again the thermal pasteurization
processes are frequently used in conjunction with freezing, since it results in microbial

inactivation (Augusto et al., 2017).

2.4.1.4 Hurdle Technology

Hurdle technology is a method of ensuring that pathogens in food products can be eliminated or

controlled by combining more than one approach. These approaches can be thought of as

"hurdles" the pathogen has to overcome if it is to remain active in the food. The right

combination of hurdles can ensure all pathogens are eliminated or rendered harmless in the final

product (Pereira and Vicente, 2020).

2.5 Innovative Technologies for Food Preservation

In recent years, the food industry has shown increased interest in innovative approaches, which

can provide products of proven quality and with a reduced environmental cost, increasing the

products’ added value (Pereira and Vicente, 2020).

2.5.1 Electromagnetic Technologies

This part of the chapter will focus on the basic principles and mechanisms of action of several

emerging electromagnetic-based technologies, such as UV-light, Pulsed Electric Field on food

preservation. All those technologies could be divided under the two main categories of thermal

and nonthermal electromagnetic technologies. Thermal technologies for food preservation rely

on heat to kill food-borne pathogens, while the nonthermal technologies are related to other

mechanisms of microbial inactivation rather than the temperature increase (Pereira and Vicente,

2020).
2.5.2 Pulsed Electric Fields

Pulse electric field (PEF) is a technology that has attracted the interest of the researchers over

almost the last century but only recently started to be exploited by the industrial sector (Li and

Farid, 2016). Most of the PEF research has been focused on decontamination of foods but its

effect on biological cells has also found applicability in other areas of food processing such as

extraction of bioactive compounds or as a pretreatment prior to freezing, canning, and drying (Li

and Farid, 2016). Thus, PEF has been applied for the processing of liquid and semiliquid food

matrices, as well as the treatment of selected solid foods (Li and Farid, 2016).

2.5.3 Ohmic Heating

Ohmic Heating (OH) can be described as a process in which electrical current passes through a

food product having an electrical resistance, also described as “resistance heating,”

“electroconductive heating,” or “Joule heating” OH has found application in several food

processes, such as cooking, blanching, pasteurization, sterilization, and extraction, specifically in

high viscous or particulate foods (Zhang, 2019).

2.5.4 Microwaves

Microwaves radiation has been applied to various heating applications related to foods

Microwaves energy has been used in pasteurization and sterilization as well as in drying,

thawing, tempering, blanching, and even combined to freezing (Zhang, 2019).

2.5.5 UV-Light
UV radiation is involved in food preservation technologies, which is considered as germicidal

range as it can effectively inactivate bacterial, fungi, viral, and protozoan microorganisms

(Koutchma, and Linden, 2015). The main factors and parameters for the efficiency of the

microbial inactivation by UV radiation are mainly related to the transmitted energy form, to the

food product specifications, and the microbial entity (Koutchma, and Linden, 2015).

CHAPTER THREE

3.0 CONCLUSION

One of the major revolutionary inventions of human civilization was acquiring the knowledge to

preserve foods as it was the precondition to man to settle down in one place and to develop a

society. However, increasing shelf lives of food items without compromising original food

properties is still critical and challenging. Food is an organic perishable substance, which is

susceptible to spoilage due to microbial, chemical, or physical activities. Different traditional

techniques, such as drying, chilling, freezing, and fermentation, had been evolved in the past to

preserve foods and to maintain their nutrition value and texture. With time and growing

demands, preservation techniques have been improved and modernized. Irradiation, highpressure

food preservation, and pulsed electric field effect are the latest innovations used to increase shelf

life of foods. Different chemical reagents have also been introduced as food additives and

preservatives. However, there are growing concerns of using chemical additives and

preservatives in food items because of possible health hazards.

3.1 RECOMMENDATION

To meet the growing demand of consumers, food preservation and processing sector has been

expanding in a rapid manner. To ensure food safety and long shelf life of foods, it is important to

understand food spoilage mechanisms and food preservation techniques. This review has

compiled and discussed different food categories, different food spoilage mechanisms, and

mechanisms and applications of traditional and advanced food preservation techniques. This

write up will be useful for the professionals and researchers working on food processing and

food safety to develop effective and integrated methods to preserve foods.

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