CHAPTER ONE

1.0 INTRODUCTION

Moringa oleifera is a plant native to Northern India that can also grow in other tropical and sub-

tropical places, with flowers, seeds, and roots at 25-35 oc temperature (Saini et al., 2016). This

species has been found suitable for human as well as animal consumption and it has proved to

show anti-lipidemic anti-inflammatory, antioxidants and anti-diabetic properties (Paliwal et al.,

2011). M. oleifera is rich in phytochemicals like alkaloids, saponin, tannins, steroids, phenolic

acids, glucosinolates, flavonoids and terpens (Rani et al., 2018).

M. oleifera is one of the most useful tropical trees. The relatives ease with which it

propagate through both sexual and asexual means and its low demand for soil nutrients and water

after been planted make its production and management easy. Introduction of these plants into a

farm which has bio-diverse environments can be beneficial for both the owner of the farm and

the surrounding ecosystem (Fugile, 2009). The great social economic importance of Moringa has

been demonstrated by Tomiro et al. (2009), Madi et al. (2012), Animashaun and Toye (2013),

and Azeez et al. (2013). For instance in western Nigeria, the analyzes of the profitability of a

leaves-based Moringa production indicated and average net profit of 59.81% Hectares and per

year from the scale of leaves (Animashaun and Toye, 2013).

Otherwise, the majority (51.66%) of sellers and retailers believe that the scale of Moringa

leaves is profitable and even 26.66% of them claim it is twice as profitable as other leafy

vegetable in northern Cameroon (Madi et al., 2012).

Antibiotics are mainly used both in human and veterinary medicine to insure human

and animal health worldwide. Beside medical therapy, antibiotics have also been used to

1
improve aquaculture and agricultural production. However, the emergence of resistant

bacteria to commonly used effective antibiotics, resulted in the need for stronger drugs and

more costly therapy. New forms of antibiotic resistance easily crossing with remarkable

speed across international boundaries and spread between continents. World health leaders

have described antibiotic resistant microorganisms as “nightmare bacteria” that “pose a

catastrophic threat” to people in every country in the world (Wang et al., 2012).

Some studies on bacterial resistance have shown that there is a huge diversity of

resistance mechanisms, in which the distribution and interaction is mostly complex and

unknown. However, there are varieties of biochemical and physiological mechanisms that

are responsible for the development of antibiotic resistance. The mechanism of resistance

may be evolution of either genetically inherent or the result of the microorganism being

exposed to antibiotics. Most of the antibiotic resistance has emerged as a result of mutation

or through transfer of genetic materials between microorganisms. Several of various recent

studies revealed that almost 400 different bacteria have demonstrated about 20,000 possible

resistant genes (Davies, 2010).

The resistances that evolve within bacteria that affect animals have the potential to affect

humans. Zoonosis of the resistant strains is able to occur, posing a risk to human health.

People who are employed at farms or food animal production facilities are at a higher risk of

infection with a resistant strain of bacteria (Sarmah et al., 2006).

Antibiotic resistant infections occur too often and with increasing frequency, interfering

with the effective treatment of people and animals. Antibiotic resistance has increased due to

the introduction of antibiotics into an environment. In general practice, there are concerns

about some common infections which are becoming difficult to treat as illness with

2
antibiotics due to the presence of antibiotic resistant bacteria which may take longer to

resolve (Butler et al., 2006). To preserve the effectiveness of antibiotics, it is critical to

examine the uses of these drugs, in both humans and animals. Several new initiatives are

being put in place to halt the alarming trend of resistance to antibiotics and to deal with the

ever-increasing number of infections caused by resistant bacteria (CDC) (2008).

1.2 Objective of the study

The aim of this study is to Assess the antimicrobial effects of Mornings oleifera leaves extract on

selected bacterial isolates

The specific objectives are ;

1) to determine the susceptibility of selected organisms

2)to the leaf extracts of Moringa oleifera,

3) to determine the effectiveness of extraction solvent and their different concentrations on the selected

microorganisms and

4) to compare the susceptibility of selected organisms to some antibiotics

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 CHAPTER TWO

2.0 LITERATURE REVIEW

2.1 MORINGA OLEIFERA

Moringa oleifera is the most widely cultivated species of the tropical flowering plant

family moringaceae containing thirteen diverse species (Shazad et al., 2013). M. oleifera is

indigenous to south Asia where it grows in the Himalayan foothills from north-eastern Pakistan

to north-western Bengal, India (Sharma et al., 2011).

M. oleifera contains known 92 nutrients includes 46 antioxidants which also includes

Quercetin and chlorogenic acids, 36 anti-inflammatory agents, 18 amino acids and 9 essential

amino acids. It also contains “Niazimicin” which helps to fight against cancer cells (Paliwal et

al., 2011).

2.2 BRIEF HISTORY OF M. OLEIFERA AS MEDICINAL PLANTS

The history dates back to 150 B.C. Historical proofs reveal that ancient kings and queens

used Moringa leaves and fruits in their diet to maintain mental alertness and healthy skin.

Ancient maurian warriors of India were feed with Moringa leaf extract in the warfront (Jahn,

2006). The Moringa drink was believed to add them extra energy and relieved them of stress and

pain incurred during war. These brave soldiers were the ones who defeated “Alexander” the

great (Shazad et al., 2013). Inspired by the news of this Moringa drinks, three young

4
professional started analyzing the nutritional benefits of Moringa tree (Rani et al., 2018). As

their research deepened, they were analyzed by the huge amount of nutritional contents of these

small leaves and fruits bears (Makkar et al., 2007). Traditionally, besides been a daily used

vegetable among people of these regions. Moringa is widely known and used for health benefits.

Among commoners, it has earned its name as ‘the miracle tree’ due to its analyzing healing

abilities for various ailments and even some chronic diseases several investigations were carried

outs to isolate bioactive compounds from various part of the plants due to its various applications

(Guevara et al., 2009). Therefore, herbal plants in medicine or known as phytomedicine are still

trustworthy and widely applied as one of the alternative ways in medicinal field due to its

affordable cost (Abalaa et al., 2009).

2.3 TAXONOMY AND DISTRIBUTION OF M. OLEIFERA

Moringa oleifera belongs to the family Moringaceae, order capparales, class

magnoliopsida and division magnoliophta. The family includes 12 other species of shrubs and

trees (Verdcourt, 1985; Olson, 2002). The 13 species of the genus Moringa are indigeneous to

Arabia, Africa and Asia (Olson, 2002). Among all these species, M. oleifera (hereafter referred

to as Moringa ) has so far become the most used and studied (Neo et al., 2015). M. oleifera was

utilized by the ancient Romans, Greek and Egyptians, and is now widely cultivated throughout

the tropical and subtropical regions of the world (Fahey, 2005).

5
Figure 1.1: A typical Moringa oleifera (Source: Emmanuel et al., (2011)

M. oleifera has been widely naturalized in other tropical regions of the world and has

been reported from large parts of southern and eastern Asia (Verdcourt, 2005; Lu and Olson,

6
2001). It has been also widely naturalized in Africa mainly in sub-Sahara Africa (Verdcourt,

2005).

2.4 SIGNIFICANCE OF MORINGA

Moringa oleifera is one of the most widely used plants globally (Palada and Chang,

2003). Studies have shown that all part of the species are traditionally used for different

purposes, such as medicine and food, with the leaves most generally used (Popoola and Obenbe,

2013; Sivasankari et al., 2014; Popoola and Obenbe, 2013; Agoyi et al., 2014).

2.5 SOCIO-ECONOMIC IMPORTANCE

M. oleifera is one of the most useful tropical trees. The relatives ease with which it

propagate through both sexual and asexual means and its low demand for soil nutrients and water

after been planted make its production and management easy. Introduction of these plants into a

farm which has bio-diverse environments can be beneficial for both the owner of the farm and

the surrounding ecosystem (Fugile, 2009). The great social economic importance of Moringa has

been demonstrated by Tomiro et al. (2009), Madi et al. (2012), Animashaun and Toye (2013),

and Azeez et al. (2013). For instance in western Nigeria, the analyzes of the profitability of a

leaves-based Moringa production indicated and average net profit of 59.81% Hectares and per

year from the scale of leaves (Animashaun and Toye, 2013).

Otherwise, the majority (51.66%) of sellers and retailers believe that the scale of Moringa

leaves is profitable and even 26.66% of them claim it is twice as profitable as other leafy

vegetable in northern Cameroon (Madi et al., 2012).

2.6 USES OF MORINGA OLEIFERA

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2.6.1 Human Consumption of Moringa Oleifera

Moringa oleifera leaves are highly nutritious. The young leaves are edible and are

commonly cooked and eaten like spinach or used to make soup and salad. The leaves can be

consumed either in raw, cooked or dried over a screen for several days and grind into fine

powder that can be added to almost any food as a nutrients supplements (Makkar and Beca,

2006), such as pap, cereal and drinks to improve their nutritive value (Gardener and Ellen,

2002). The leaves which were boiled resulted in three times move bio-available iron than the raw

leaves. This result were also seen in the powdered Moringa leaves, the protein quality of

Moringa leaves compared with that of milk and egg (Gardener and Ellen, 2002). On the other

hand, in 100g dry matter; they contain 29+6g of protein, 28+ 6mg of Iron, 1924+ 288mg of

calcium, 15620+ 6475 IU of Vitamin A and 773+91mg of Vitamin C (Gardener and Ellen,

2002). This is at least twice the protein in milk and half the protein in egg, and has more iron

than in beef, more calcium than in milk, equal Vitamin A to carrots and more Vitamin C than in

orange (Wangchareon and Gomlmanec, 2015).

2.6.2 INDUSTRIAL USES OF M. OLEIFERA OIL

The oil content of de-hulled seed (kernel) is approximately 42%. The oil is brilliant

yellow and is used as a lubricant for fine machinery such as time pieces because it has little

tendency to deteriorate and become rancid and sticky as well as vegetables cooking and frying

oil and valuables in the perfume industry for stabilizing scents (Ramachandran et al., 2000). The

free fatty acid contents varies from 0.5-3%; the seed oil of Moringa contains approximately 13%

saturated fatty acids and 82% unsaturated fatty acids (Ferrao and Mendez, 2000). It has a

particularly high level of oleic acid (70%). Other vegetables oils normally contain only 40%

oleic acid (Ferrao and Mendez, 2000).

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2.6.3 M. OLEIFERA AS PLANT GROWTH ENHANCERS

The extract obtained from the leaves of Moringa in 80% ethanol contains growth

enhancing principles (hormones of cytokinine type) and can be used in the form of a folia spray

to accelerate growth of young plants, which enables the plants to be firmed and more resistant to

pests and disease (Al-kharusi et al., 2000; (Makkar and Becker, 2006).

Spraying the leaves of plant with Moringa extract prepared in 80% ethanol and then

diluted with water produced some notable effects such as a longer, more vigorous life span,

heavier roots stems and leaves, bigger fruits and higher sugar levels etc. (Makkar and Becker,

2006).

2.7 MORINGA OLEIFERA NUTRITIONAL VALUES.

Through research, Moringa was found to contain many essential nutrients, for instance

vitamins (Fahey, 2005; Hsu et al., 2006, Kasolo et al., 2010). Nutritional content of plant plays

essential function in medicinal, nutritional, and theraupetic properties (Al kharusi et al., 2009).

Moringa leaves consists high source of vitamin C, calcium, potassium as well as protein and as

an effective source of antioxidants Sddhuraju and Becker, 2003). Due to the presence of several

sorts of antioxidants compounds such as flavonoids, ascorbic acid, carotenoids, and phenolics.

Moringa is able to extend the period of food containing fats (Dillard and German, 2000). It was

also found that each different part of the Moringa tree which was studied, be it fruits, seeds,

leaves, flowers, bar and roots, all resulted in the discovery of at least one, or in most studies, a

number of beneficial nutrients (Brunelli et al., 2010). It was similarly mentioned in an article by

tress, for life organization that every part of the Moringa tree is said to have beneficial properties

9
that can serve humanity, in addition, the Moringa was found to have a group of compounds

containing sugar and rhamnose, which are uncommon sugar-modified glucosinolates (Fahey et

al., 2010; Fahey, 2005; Amaglo et al., 2010). These compounds were reported to demonstrate

certain chemopreventive activity by reducing apoptosis (Brunelli et al., 2010).

2.7.1 Table 1. Analysis of dried leaf powder contains the following per 100g of edible

portion

Nutritional value per 100g Values

Moisture (%) 7.5

Calories (kcal) 205

Protein (g) 2.7.1

Fat (g) 2.3

Carbohydrate 38.2

Fibre (g) 19.2

Trace minerals

Calcium (mg) 2,003

Magnesium (mg) 368

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Phosphorus 204

Potassium 1,324

Iron 28.72

Sodium 870

Vitamins

Vitamin A- Betacarotene (mg) 16.3

Vitamin B1- Thiamine (mg) 2.64

Vitamin B2- Riboflavin (mg) 20.5

Vitamin B3- Nicotinic acid (mg) 8.2

Vitamin C- Ascorbic Acid (mg) 17.3

Vitamin E- Tocopherol acetate (mg) 113

Amino Acids

Arginine (g/16gµ) 1.33%

Histidine (g/16gµ) 0.16%

Lysine (g/16gµ) 1.32%

Tryptophan (g/16gµ) 0.43%

Phenyalanine (g/16gµ) 1.39%

Threonine (g/16gµ) 1.19%

Leucine (g/16gµ) 1.95%

Isoleucine (g/16gµ) 0.83%

Valine (g/16gµ) 1.06%

Other Constituents

Oxalic acid (mg) 1.6%

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2.8 Medicinal uses of M. Oleifera

Moringa oleifera is considered rich with several medicinal properties, as well as all parts have

been reportedly singly or with other plants for treating diverse illness and disease (Fugile, 2009;

Pamok et al., 2011). In traditional medicine, the leaves are used to treat ailments including

malaria, typhoid fever, parasitic disease, arthritis, swellings, cuts, and disease of the skin, genito-

urinary ailments, hypertension and diabetes (Leone et al., 2015). They are also used to elicit

lactation and boost the immune system (to treat HIV/AIDS related symptoms) (Anwar et al.,

2007; Popoola and Obenbe, 2013; Sivasankari et al., 2014), as well as cardiac stimulants and

contraceptive remedy for the treatments of these ailments, one can directly consume either raw

and dried leaves, or the extract of an aqueous infusion (Leone et al., 2015).

2.9 MORINGA AS A TREATMENT FOR DIABETIC NEPHROPATHY

M. oleifera is used in traditional folktore for treating many ailments such as asthma,

spasm, enlarged liver and spleen, infection and nervous debility, ulcer, inflammation and for

wound healing (Mishra et al., 2011; Promkun et al., 2010). This tree has in recent times been

advocated as an outstanding indigenous sources of highly digestible protein, calcium, iron,

vitamin C, and carotenoid suitable for utilization in many so-called "developing" regions of the

world where under nourishment is a major concern studies have shown that the extract of M.

Oleifera leaves also has anti-diabetic and antioxidants activities (Pari et al., 2007; Jaiswal et al.,

2009). Khongrun et al. (2012) investigated the activity of leaf extract of M. Oleifera in

improving neuropathic pain induced by diabetic condition. Experimental rats administered with

extracts of M. Oleifera leaves at different doses once daily in a period of 21 days shows the

12
analgesic efficacy of the plant extract was evaluated using Von Frey filament and hot plate tests

every 3 days.

2.9.1 ANTIOXIDANT ACTIVITY OF MORINGA OLEIFERA

Free radical and reactive oxygen species are well known as inducers of cellular and tissue

pathogenesis which is causing some diseases like diabetes, cancer, inflammatory and also

cardiovascular diseases also free radical reactions taking place in the human body and food

systems can cause injury and death (Halliwell, 2008). Free radicals are one of the main factors

which necessarily cause DNA mutation and involved in the initiation stage of carcinogenesis

(Verma et al., 2009). Reactive Oxygen Species (ROS) are constantly produced in human body

by normal metabolic system. An over-production of reactive oxygen can occur as imbalance of

defense system. Therefore, investigations of antioxidants are needed which focused on natural

compounds from natural sources (Fahey et al., 2005).

The most widely used synthetic antioxidants in food are butylated hydroxytoluene (BHT)

or butylated hydroxyanisole (BHA). Both of them are very effective as antioxidants but their use

in food products is not popular anymore due to their instability and also due to a suspected action

as promoters of carcinogenesis. For this reason, there is another interest in the studies of natural

healthy (non-toxic) additives as potential antioxidants (Tomaino et al., 2005). The total

antioxidant values should include methods applicable to both lipophilic and hydrophilic

antioxidants, with regards to similarity and differences of both hydrogen atom transfer and

electron transfer mechanism (Hamza et al., 2010). Some methods have been used to evaluate

antioxidant activities of natural compounds by using stable free radical DPPH and ABTS

(Tomaino et al., 2005). Most of the antioxidant compounds derived from plant source have wide

13
variety and chemical properties. The antioxidant characteristic is based on its ability to trap free

radicals.

In Asia and Africa, the leaves is recommended as a supplement because of rich in

nutrients for breastfeeding mothers and infant because it contains compounds like Nitrile,

mustard oil glycosides, benzyl glycosides, phenolic glycosides, flavonoid glycosides,

thiocarbamate glycosides and amino acids had been isolated from this plant (Hallliwell, 2008).

M. oleifera leaves contain of natural source of polyphenol that potential to have antioxidant

(Santos et al., 2012).

2.10 ANTIBIOTICS

Antibiotics are chemical agents that prevent bacterial growth by stopping the bacterial cell

from dividing (bacteriostatic) or by killing them (bactericidal). The terms antibiotic and

antimicrobial are often used interchangeably but are not synonymous. Antibiotics are substances

of microbial origin (such as penicillin) while “antimicrobial” refers to any substance including

synthetic compounds which destroys microbes (Guardabasse and Courvalin, 2006).

Antibiotics are used to treat and or prevent disease in human and animals. The reductions in

death afforded by effective antibiotics for bacterial infections of all types, ranging from simple

skin infections to infections of the bloodstream, lung, abdomen, as well as brain, so enormous

14
that the lives of both human and animals are saved due to treatment by using antibiotics

(Spellberg, 2011).

2.10.1. Mechanism of action of antibiotics

In order to appreciate the mechanisms of resistance, it is important to understand how

antimicrobial agents act. One of the most common mechanisms of action is targeting the cell

wall, which is present in bacteria (prokaryotic cells) but absent in humans (eukaryotic cells).

Thus, antimicrobial agents act selectively on vital microbial functions with minimal effects or

without affecting host functions. Different classes of antibiotics possess specific modes of action

by which they inhibit the growth or kill bacteria (Young, 2011).

Table 1: A list of antimicrobial agents and their modes of action

S/N Antimicrobial agents Group Mode of action

1 Ampicillin , Penicillins Inhibitor of cell wall

Augmentin synthesis

2 Amoxycillin Cephalosporins Inhibitor of cell wall

Ceftriaxone synthesis

3 Chloramphenicol Chloramphenicol Inhibitor of protein

15
 synthesis.

4 Erythromycin Macrolides Inhibitor of protein synthesis

5 Azithromycin, Aminoglycosides Inhibitor of protein synthesis

Gentamycin,

streptomycin

6 Oxytetracycine, Quinolones Inhibitors of DNA synthesis

Tetracyclines, Ciprofloxacin

Nalidixic acid

7 Sulfamethazine Sulfonamides Competitive inhibitors of

folic

8 Trimethopim acid synthesis

Source: (Young, 2011).

2.10.2. Antibiotic Resistance

Antibiotic resistance is the ability of a bacterium or other microorganisms to survive and

reproduce in the presence of antibiotic doses that were previously thought effective against them

(WHO, 2011). The origin of antibiotic resistance genes are unclear; however, studies using

clinical isolates collected before the introduction of antibiotics demonstrated susceptibility,

although, conjugative plasmids were present (Denyer et al., 2011).

Normally, most cells in a naive, susceptible bacterial population which can cause an

infection are susceptible to particular antibiotic upon exposure. However, there is always a

16
minute subpopulation of resistant bacterial cells that will be able to multiply at higher rate in

sufficient antibiotic concentration (Smith et al., 2005). Resistance is often associated with

reduced bacterial fitness, and it has been proposed that a reduction in antibiotic use will pose

selective pressure to acquire resistance would benefit the fitter susceptible bacteria, enabling

them to outcompete resistant strains over time.

Antibiotic resistant bacteria are growing public health emergency since infections from

resistant bacteria are more hard and costly to treat. For instance, since the 1990s, some strains of

Salmonella became resistant to a range of antibiotics. The major problem in the clinical practice

today is the emergence of multiple-drug resistance, which is resistance to several types of

antimicrobial agent (Amenu, 2014).

Resistance to an antibiotic may be an inherent property of the infecting organism or it may

be acquired. Acquired resistance may result from mutation or from transfer of an extra

chromosomal genetic material followed by selection of resistant organisms during therapy

(Courvalin, 2005). There is a range of mechanisms by which an organism can acquire resistance,

the simplest being genetic mutation. Resistant mutants will have a strong survival advantage in

the face of antibiotic exposures, giving rise to the total usage of antibacterial agents in a

population and the increased proportion of isolates that exhibit resistance to those agents

(Hegstad et al., 2010).

2.10.3. Mechanism of antibiotic resistance

As there are many different ways in which antibiotics can kill or inhibit the growth and

multiplication of microorganisms, there are also many mechanisms of resistance that

microorganisms innately possess or have developed over time through exposure of antibiotics. It

is possible that through one mechanism, an organism can become resistant to many different

17
classes of antibiotics, especially if the modes of action are similar. Sometimes resistance can be

shared between individual bacteria through the production of “resistance plasmids,” the pieces of

DNA capable of being transferred from one cell to another (Clewell, 2014).

Resistance genes transferred between organisms via these mobile genetic elements (MGEs)

is the most common and clinically more important in multi-drug resistance (MDR) of Gram-

negative bacteria than resistance arises through mutation. There is ample evidence that MGEs are

able to transfer resistance mechanisms between genera; for example, MGEs of enterococci being

transferred to Staphylococcus aureus (Hegstad et al., 2010).

A microorganism is resistant if it exhibits “significantly reduced susceptibility” when

compared with that of the “original isolate” or a group of sensitive strains. Resistance can result

from mutations in housekeeping structural or regulatory genes, or alternatively, horizontal

acquisition of foreign genetic information (Courvalin, 2005).

Resistance can be described in two ways: intrinsic or natural whereby microorganisms

naturally do not possess target sites for the drugs and therefore the drug does not affect them

(e.g. Mycoplasma species resistant to penicillins) or they naturally have low permeability to

those agents because of the differences in the chemical nature of the drug and the microbial

membrane structures especially for those that require entry into the microbial cell in order to

affect their action (e.g. many enterobacteriaceae). The other is acquired resistance whereby a

naturally susceptible microorganism acquires ways of not being affected by the drug (Fluit et al.,

2001).

2.10.4. Antibiotic inactivation:

On some occasions cell may gain resistance to antibiotics by making an enzyme that renders

the drug inactive, or that decreases the functionality of the antibiotics. The best example is beta

18
lactamases which is capable of breaking the beta-lactam rings of beta lactam antibiotics such as

penicillin. In such manner, the breakage of the beta-lactam ring stops the antibiotic from being

able to attach to the peptidoglycan precursors. But it will be less likely that penicillin or other

similar drugs will be able to disrupt the integrity of the cell wall, as long as the organism

produces beta lactamases (Sageman, 2015). This method of resistance can be transferred from

one bacterium to another through the production of the R-plasmids, and is common in strains of

methicillin resistant such as Staphylococcus aureus (MRSA) (Holcomb et al., 2008).

2.10.5. Reduced membrane permeability:

Another common way of interfering with antibiotics is through the prevention of entrance of

the drug into the cell. Gram negative bacteria have an outer cell membrane, and drugs must pass

through the cell pores, which are channels that span the outer membrane and allow the entry and

exit of materials into or out of the cell. In order to enter the cell or interact with the cell wall, the

drugs must be able to pass through the pores (Willey et al., 2013).

A gene mutation can result in altered pores, usually by changing the electrical charge or the

physical structure which can make it more difficult for antibiotics to enter the cell. The antibiotic

is still functionally active, but it will fail to reach its target site. A microorganism can develop

resistance to multiple drug classes at once in this manner. But some gram negative bacteria are

innately resistant to large drugs like vancomycin, which is too large to pass through the pores

even before a mutation occurs (Galdiero et al., 2012).

2.10.6. Modification of target site:

Many antibiotics act by binding to a target molecular component of the microorganism. A

microorganism can decrease the effectiveness of a drug if the target molecule changes slightly its

structure so that antibiotic may no longer be able to bind to the target molecule. For example,

19
tetracyclines block the transfer RNA access site by binding to it. In turn slight changes in the

access site may result in microbial resistance to tetracyclines (Denyer et al., 2011).

2.10.7. Efflux or transport of antibiotic:

Another mechanism by which microorganisms can become resistant to antibiotics is by

utilizing an efflux pump. An efflux pump is a biological pump that can force the antibiotic out of

the cell, so that it cannot reach or stay in contact with its target. This method of antimicrobial

resistance may often create resistance to more than one class of antibiotics, especially the

macrolides, tetracyclines, and fluoroquinolones because these antibiotics inhibit different aspects

of protein and DNA biosynthesis and therefore must be intracellular to exert their effect (Willey

et al., 2013).

The genetic alterations in bacteria cause resistant to antibiotics in one or more of four

principle ways, as shown in Figure 2; the target molecules are structurally altered to prevent

antibiotic binding; reduce membrane permeability (antibiotics are excluded from cell entry);

antibiotics are inactivated through enzymatic degradation; or they are pumped out of the cell by

efflux pump .

2.10.8. Multiple antibiotic resistances

R-plasmids possess regions with the resistance genes and resistance to a number of different

antibiotics that can be mediated by the same R-factor and is known as multiple antibiotic

resistances (Dessen et al., 2001). The prevalence of multiple drug resistance bacteria itself is a

serious problem, but transfer of multiple drug resistance to other members of the family

Enterobacteriaceae, particularly E. coli, Salmonella and Shigella makes it even greater concern

to clinicians in curbing infections in medical and veterinary practice (Levy, 2002).

20
 An example of MDR pathogen is vancomycin resistant Enterococcus (VRE), which can

cause infection in many parts of the body. Researcher Jane Siegel found that VRE “was

associated with increased mortality, length of hospital stay, admission to the intensive care unit,

surgical procedures, and costs when VRE patients were compared with a matched hospital

population” (Siegel et al., 2006).

Another example of MDR pathogen is MRSA. Staphylococcus aureus is a common

bacterium that causes the urinary tract, wound, skin infections, and other complications. It is also

one of the most common nosocomial infections, and one in twenty health care workers are

colonized with MRSA and has a higher rate of causing symptomatic or fatal infections than

methicillin susceptible S. aureus (Smith, 2008).

21