BACKGROUND for their major health needs (S. Indradevi, et


It is estimated by The World
Health Organization (WHO) that 80% of the individuals of emerging countries
depend on traditional medicines, usually 
plant-originated drugs, for their major health needs (S. Indradevi, et al. 2012)because fruits and juices are
nutrient rich foods which provide vitamins, minerals and other bioactive
compounds with few calories (Ribeiro C,et al.2017). In past few
years, interest in using natural products for pharmacological purpose has
tremendously increased (Silmara Baroni,et al.2016).

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Citrus fruits belong to kingdom
Plantae, the Rutaceae family and Citrus genera and are grown all over the
world. These fruits include, lemon (Citrus limonum),  grapefruit (Citrus vitis), lime (Citrus
aurantifulia), tangerine (Citrus reticulata) and sweet orange (Citrus
sinensis)(Anthony Cemaluk C. Egbuonu et al.2016) . Citrus fruits are highly nutritious, medicinal and constitute
0.9% of daily calorie intake and about 1.7% of carbohydrate intake. The peel
waste of citrus fruit is highly fresh and seasonal in terms of wealth for the
farmers if the industries bring useful products from them by evolving
different  methodologies. (Suja D1,et al, 2017).



An orange fruit namely Citrus sinensis  belongs to genus Citrus and Rutaceae family
is among the most important crops grown throughout the world. (Ngele, K.K. ,et al. 2014) with about 60% of the total citrus world production. Antioxidant and
antibacterial activity of orange juice and its edible portion have been
reported in many studies. (A.E.
Hegazy,et al. 2012).



The peel of Citrus sinensis is rich in flavonoids including methylated
derivatives such as polymethoxyflavones (PMFs) Alexander Gosslaua, et
al. 2014) which are rarely found in other
plants. The peel also contain
other constituents such as carotenoids, ascorbic acid, hydroxycinnamic acids
and anthocyanins (Roberta Fusco,et al. 2016). The evaluation of its ethanolic peel extract
indicated the presence of carbohydrates, saponins, alkaloids, tannins, fixed
oils, phenols and steroids while the inner white layer of orange peel contain
hesperidin which releases its aglycone portion that is hesperetin on
indigestion (Sharma pradeep kumar, et al. 2014).



Phytochemicals not only play an
important in ecological and physiological functions but are also used
commercially because of their multiple applications in food and pharmaceutical
industries (F. GÜLAY KIRBA?LAR, et al.
2009).  These compounds boost immunity, aids
digestion, promote healthy skin and are used as vitamin-packed flavouring
agents (J.O. Arawande, et al. 2015. They are highly potent in decreasing plasma cholesterol level so have
an inverse relation with coronary heart disease (Nesrin M. Fayek, et al 2017). Orange
peel extract may function as surfactant by decreasing the surface tension of
liquids in the stomach, decreasing the potential for the
fluid to splash up into the oesophagus. Researches show that orange peel
extract inhibits the growth and division of cancer cells (Narjis
Hadi Mansoor Al-Saadi, et al 2009).  Orange peel oil has lethal
effect on fleas, fire ants, houseflies due to its 90-95% limonene while the
roasted pulp and fresh peel are used for skin dressing and acne treatment
respectively. Oranges are eaten to relieve fever and potion of the immature
fruit is taken to relieve gastrointestinal complaints. Plants containing protoberberines,
oligosaccharides, saponin, flavonoid, alkaloid, and phylates are used in the
African traditional system and show antimicrobial activity (Uchechi N. Ekwenye,
et al. 2010).



Bacteria have the inborn ability to
transfer and gain resistance against the drugs which are used in therapeutics.
It’s been a long time that plants have been a good source of natural products
for conserving human health therefore the use of plant extracts and phytochemicals
which are synthesized in secondary metabolism of the plant having well-known
antimicrobial properties, can be of great importance in therapeutics or
alternative remedies. The active substances in these products include phenolic
compounds which are a part of the essential oils, as well as tannin. Essential
oils are quite more effective in controlling biofilm cultures due to their
better diffusing ability and interaction. (Maruti J.
Dhanavade, et al. 2011).
 Screening of crude extract shows that
although the compounds may not have their own string antibacterial activity but
can be used in combination with antibiotics increase the efficacy of the drug
by different modes of actions. (IqbalAhmad, et al. 2007).













E. coli are rod shaped gram negative bacteria
usually found in warm blooded organisms (E.V.Bulychevaa.
et al.2014).


Eschericrhia coli may be considered as most prevailing
opportunistic enterobacteria (Lilian
AparecidaSanches, et al. 2017). Along with living organisms e. coli also
lives in water where it hits household and agricultural effluents. They are facultative anaerobes which
form lactate, ethanol, succinate, carbon dioxide and acetate as their metabolic
end products. (E.V.Bulychevaa. et al. 2014).


This bacterium is a main model organism in
microbiology. They are also used in biological engineering. Most of E. coli
strains are harmless, but some serotypes can cause serious food poisoning in
their hosts.. According to the United States Environmental Protection Agency
(USEPA) E. coli is the best indicator of health risk from water contact in
recreational waters (E.V.Bulychevaa. et
al. 2014). E. coli are transmitted by fecal-oral route, food-borne,
environmental or from person to person. They have ability to release
shiga-toxin which destroys host cells in intestine and through bloodstream move
to other body parts such as kidneys and brain and affect them too(EricaKintz,
et al. 2017). E.
coli has been a common
infection pathogen associated with both community acquired and nosocomial
infections. Antibiotic resistance among E coli isolates
continues to increase, limiting the choices of antibiotics available (Hui-HsiuWu, et al. 2016). The Extraintestinal
pathogenic E. coli (ExPEC) is the most common cause of extraintestinal
infections such as urinary tract infections, bloodstream, kidney and other
infections. The prevalence of extraintestinal infections is thought to exceed 7
million medical visits, 1 million emergency visits and 100 000
hospitalizations every year in the USA. (A.R.Manges, 2016).









Pseudomonas aeruginosa is a gram-negative bacillus (Alice
S. Prince, 2012) and encompasses a high colonization
and transmission capacity (FatemehNanvazadeh,
et al. 2013). The characteristic blue color of the organism is due to the production
of fluorescent siderophores pyoverdin, pyochelin and pyocyanin which has iron
scavenging and oxidant ability respectively (Alice S. Prince, 2012).


P. aeruginosa is found widely in nature i.e in soil and water. It has
few nutritional requirements and can adapt to intolerable conditions. It
oxidizes glucose and xylose but cannot ferment lactose or other carbohydrates.
If nitrate is available as an inorganic electron acceptot then P. aeruginosa
can grow aerobically or anaerobically as in the lungs of entities with cystic
fibrosis (Alice S. Prince, 2012).


P. aeruginosa is a common multidrug-resistant (MDR) gram-negative
pathogen which may cause pneumonia in hospitalized individuals. P. aeruginosa infections have a high rate of
mortality (ranging from 10% to 70%) particularly in patients given
inappropriate empirical therapy, immunocompromised patients, ICU patients and
drug-resistant P. aeruginosa infections (O.Asuphon, et al. 2016). Pseudomonas aeruginosa is a common lung pathogen and a main
cause of morbidity and mortality in patients with cystic fibrosis (Alaya Mikalauskas, et al.2017). Data from developing countries indicates
that P. aeruginosa is the most
common cause of pneumonia in hospital (29%), and is the third most common cause
of Intensive Care Unit (ICU)-acquired infections (17%). Antimicrobial
susceptibility and pathogenesis mechanisms of P. aeruginosa are
poorly understood (AliBadamchi, et al.










K. pneumoniae is a Gram negative, encapsulated, non-motile,
lactose-fermenting, facultative anaerobic, bacillus (FahmiYousefKhan, et al. 2014) which belongs to Enterobacteriaceae family (HacerAkturk, et al. 2016).


K. pneumonia inhabits a wide variety of hosts ranging from
plants to mammals, but can also be found in the soil and surface water (Dennis J.Doorduijn, et al. 2016). It is found as the normal flora of the
skin, mouth and intestine (FahmiYousefKhan,
et al. 2014).


A wide range of hospital acquired infections
are caused by  K. pneumoniae  which include primary bacteremia, pneumonia,
urinary tract infections, wound infections and intra-abdominal infections(HacerAkturk, et al. 2016) particularly
in susceptible individuals such as newborns, the elderly, or the
chronically ill (Sonal P.Henson, et al.
2017). Recently, in both community and nosocomial settings, it has gained
an increasingly important role in adult meningitis with substantial
geographical diversity in its clinical patterns (FahmiYousefKhan, et al. 2014). In the WHO report of 2014  Klebsiella pneumoniae
is  considered as one of the top three
bacteria of international concern on the global status
of antibacterial resistance (Sonal
P.Henson, et al. 2017) causing high numbers of both healthcare- and
community-acquired infections(C.Pichler,
et al. 2017). K. pneumoniae have innate
resistance to penicillins such as ampicillin but are prone
to third generation cephalosporins such as ceftriaxone (Sonal P.Henson, et al. 2017). The
limitation in the treatment options for K. pneumoniae infections
is due to the  acquisition of antibiotic
resistance genes and intrinsic resistance to several classes of antibiotics
Currently,  strains of K. pneumoniae producing
Extended Spectrum Beta-Lactamases (ESBLs) and carbapenemases have spread worldwide
(Dennis J.Doorduijn, et al. 2016).







Proteus mirabilis is a Gram negative motile
bacillus which belongs to Enterobacteriaceae family (Rakesh D.MistryMD, et al. 2010) (Chi-YuChen , et al. 2012)  


Proteus mirabilis inhabits the skin blisters
of the axilla (Rakesh D.MistryMD, et al.


Proteus mirabilis causes urinary
tract infections (AnaUmpiérrez, er al.
2013) and wound infections in humans (Chun-HaoLiu,
et al. 2015). Proteus mirabilis is also a common cause of other types
of hospital-acquired infections, including bacteraemia, meningitis, empyema and
osteomyelitis. Since 1970s along with sporadic infections, sporadic infections
have also been reported (A.Adler, et al.
2013). Proteus
mirabilis has been model organism
for urease-producing uropathogens (Allison
N. Norsworthy, et al. 2017)
which generates
ammonia and elevates the pH of the urine to >7.2(Chi-YuChen , et al. 2012). The
generated ammonia causes severe metabolic disorders and damage of GIT
epithelium by its interaction with the immune system of human (A.Adler, et al. 2013). Infection stones
in the urinary tract and crystalline biofilms on indwelling urinary catheters,
or in patients with urolithiasis are formed by these diverse bacteria (Chi-YuChen , et al. 2012)  frequently
leading to polymicrobial infection(Allison
N. Norsworthy, et al. 2017). Biofilm formation, toxins, adhesins, motility, immunoavoidance
and nutrient gaining  are the virulence
factors of Proteus mirabilis for UTIs (Sandra
M. Fox-Moon, et al. 2015).The increased resistance to antimicrobial agents has led not
only to a changes in antimicrobial therapies, but also to poor prognoses and
an increase in the mortality rate of hospitalized patients ( Chi-YuChen , et al. 2012).








Klebsiella oxytoca is one of four members of the genus Klebsiella
(Kazutoshi HoRi, et al. 1998). It is a rod-shaped, non-motile, Gram
negative bacterium encapsulated in polysaccharide capsule, which provides a
resistance against host immune system (Mahendra
Kumar Trivedi , et al. 2015).


Klebsiella oxytoca is found in patients with antibiotic associated
hemorrhagic colitis as well as in the stools of healthy individuals. (Christoph
Högenauer, et al. 2006).


Klebsiella oxytoca is understood to be an opportunistic pathogen that
usually causes hospital acquired infections, frequently involving immunosuppressed
individuals or those patients who require intensive care (Christopher Lowe, et al. 2012). Patients having neurosurgical procedures,
prostatectomies, urinary tract infections (UTI), intravascular catheters,
colonoscopies, platelet transfusions, and pre-existing viral or antibiotic
induced colitis are more susceptible to K. oxytoca infection (Mahendra Kumar Trivedi , et al. 2015). Current evidences show Klebsiella
oxytoca as a possible cause of Antibiotic-associated hemorrhagic colitis (AAHC)
(Stephen A Smith, et al. 2009) Klebsiella oxytoca isolated from
patients with antibiotic associated haemorrhagic enterocolitis had been shown
to produce a cytotoxin which exerts  a
cytotoxic effect, example include, cell-rounding followed by cell death, on
many tissue culture cells (JUNZABURO
MINAMI, et al. 1992). K. oxytoca constitutively produce
chromosomal –lactamases which provides resistance to ampicillin and amoxicillin
(LAURENT BEAUGERIE. Et al. 2003). K.
oxytoca is typically resistant to some antibiotics like cefotaxime, ceftazidime
and aztreonam (Mahendra Kumar Trivedi , et al. 2015).



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