2.

REVIEW OF LITERATURE

2.1.Urinary Tract Infection

Urinary tract infection (UTI) is a condition in which one or more parts of the urinary
system become infected. UTIs are the most common of all bacterial infections and can occur at
any time in the life of an individual. Nearly 95% of cases UTIs are caused by bacteria that
typically multiply at the opening of the urethra and travel up to the bladder. Much less often,
bacteria spread to the kidney from the bloodstream. The most members of these genera include
Escherichia, Klebsiella, Enterobacter and Proteus. Also gram positive Staphylococcus sp. plays
a role in the infection.

There are thousands of species of medicinal plants used globally for the cure of different
infections. These plants are used as antimicrobial agents and several works have been carried out
by scientists to find out its scientific basis. Celery (Apium graveolens) is a plant variety in the
family Apiaceae, commonly used as a vegetable. The major components of Apium graveolens
are alkaloids, glycosides, terpenoids, flavonoids, tannins and polyphenols.

2.2. Isolation and Identification of uropathogens

Janifer et al. (2009) studied prevalent UTI organisms from lower tract infected south
Indian Type 2 Diabetic patients. A total of 1157 (M: F 428: 729) type 2 diabetic subjects were
collected and subjected to antimicrobial susceptibility pattern by Kirby-Bauer method. This
study was reported that 533 pathogens of gram positive and gram negative bacilli were isolated
from 495 subjects. Escherichia coli (E. coli) was the most commonly found organisms and
highly sensitive to sulbactum, cefoperazone and pipercillin, tazobactum respectively. Finally
concluded that the prevalence of UTI was significantly higher in women than men with E. coli
being the major isolated pathogen.

Ejaz et al. (2011) determined the frequency and antimicrobial resistance of ESBL
producing E. coli and K. pneumoniae. A total of 13638 urine samples were processed for culture
and antimicrobial sensitivity testing. E. coli and K. pneumoniae were identified using API 20E.
A double disk synergy test (DDST) was performed to determine the ESBL production. ESBL
production was detected in 312 (57.4%) E. coli and 386 (71.7%) K. pneumoniae. A multidrug
resistance pattern was seen in ESBL producing E. coli and K. pneumoniae. ESBL producing E.
coli showed maximum resistance to cefotaxime (100%), ceftazidime (99.4%) and cefuroxime
(93.3%) while minimum resistance was seen with meropenem (1.3%), piperacillin/tazobactam
(10.3%) and nitrofurantoin (27.6%). ESBL producing K. pneumoniae showed maximum
resistance to ceftazidime (100%), cefotaxime (98.7%) and cefuroxime (98.1%) while minimum
resistance was seen with meropenem (3.6%), piperacillin, tazobactam (17.6%) and nitrofurantoin
(28.5%). In ESBL producing bacteria, high prevalence of antibacterial resistance of non-β-lactam
antibiotics is a serious matter of concern. Monitoring of ESBL production and antimicrobial
susceptibility testing are necessary to avoid treatment failure in patients with urinary tract
infection (UTI).

Shalini et al. (2011) studied the changing pattern of antibiotic sensitivity among
uropathogens causing urinary tract infections (UTI). A total of 170 urine culture sensitivity
reports were analyzed. The predominant growth of single bacteria was seen in 143 (84 .12%)
samples. The most common organisms isolated were Escherichia coli, Klebsiella, Pseudomonas
and Staphylococcus aureus. (These represented 64.33%, 92; 20.3%, 29: 9.1%, 13 and 6.30%, 9
of isolates respectively). More than 80% of the isolates were sensitive to Amikacin and
nitrofurantoin, while more than 70% were sensitive to norfloxacin, ciprofloxacin and
levofloxacin. Very high rate of resistance was seen against cotrimoxazole (81.82%), amoxicillin
(77.42%) and amoxi-clav (64.34%). E. coli showed high sensitivity to amikacin 98.91% (91),
Nitrofurantoin 93.48% (86). 75% of E. coli isolates were sensitive to minocycline, showing a
good utility of this drug for the treatment of outdoor patients with urinary tract infections.

Bahalo et al. (2013) determined the occurrence of some virulence gens among a hundred
Escherichia coli isolates obtained patients with urinary tract infection (UTI). A total of 100 E.
coli strains were isolated, the prevalence of genes such as: pyelonephritis associated pili (pap
genes), S-family adhesions (sfa gene), type fimbriae (fimHgene), aerobactin (aer gene) and
pyelonephritis isolates compared (pic gene) among the isolated strains was: 40%, 30%, 12%,
4%.
 Tarchouna et al. (2013) was determined the Escherichia coli is the predominant pathogen
causing urinary tract infection (UTI), the most common bacterial infectious disease encountered
in clinical practice, accounting for significant morbidity and high medical costs. The severity of
UTI produced by E. coli is due to the expression of a wide spectrum of virulence factors. In this
study was evaluated the role of E. coli virulence determinants in the pathogenesis of UTI. The
prevalence of genes coding for fimbrial adhesive systems was 68% for fimH, 41% for pap, and
34% for sfa/foc. The operons coding for afa fimbrial adhesins were identified in 20% of strains.
The hly and cnf genes coding for toxins were amplified in 19% and 3% of strains, respectively.
A prevalence of 52% was found for the gene. The various combinations of detected genes were
designated as virulence patterns. The strains isolated from hospitalized patients displayed a great
diversity of gene associations compared to those isolated from ambulatory patients.

Sangeetha et al. (2014) detected the current antibiotic resistance pattern of E. coli with a
special reference to fluroquinolone resistance. Among 311 culture positive urine samples, 203
were E. coli. High resistance rate to Ampicillin (81.3%), Co-trimoxazole (83.3%) and low
resistance rate to Nitrofurantoin (17%) were noted for E. coli. Among the 203 E.coli, 141 and
121 isolates showed a MIC of >4µg/ml for Ciprofloxacin and ≥ 8µg/ ml for levofloxacin
respectively. The resistance rate to other antibiotics and the MIC of Ciprofloxacin and
Levofloxacin increased as the patient’s age increases.

2.3. Molecular studies

Rahman and Deka (2012) reported E. coli isolates from patients with urinary tract
infection from northeastern India were investigated for detection and characterization of NTEC.
A total of 550 E. coli were isolated and tested for the presence of cnf genes. Of these, 84
(15.27%) belonged to NTEC. The cnf1 gene was present in 52 (61.9%) isolates, cnf2 in 23
(27.4%) and 9 (10.7%) carried both cnf1 and cnf2 genes. All the NTEC strains were found to
harbor the pap and aer genes. Sero group O4 was found to be the most common among the 12
sero groups identified amongst the NTEC isolates. Majority of the isolates (96.4%) were
sensitive to furazolidone and were highly resistant to ampicillin. NTEC were found to harbor
different numbers of plasmids (1 to 7). No association was observed between the number of
plasmids and the antibiotic resistance of the isolates.
 Foysal et al. (2012) determined the pathogenicity of any isolates because these genes act
multi-factorially. This study was forward and reverse primers with a length of 20bp for both
were used for detection of gyrB2 genes in clinical isolates of Pseudomonas sp. Collected from
patients suffering from urinary tract infection (UTI). A total of 12 isolates of Pseudomonas
sp.viz. Ps1,Ps2,Ps3,Ps4,Ps5,Ps6,Ps7,Ps8,Ps9,Ps10,Ps11 and Ps12 were used in this study in
which gyrB2 gene amplified in all 12 isolates and gave the expected 1130bp PCR product after
visualization under gel documentation system in 1.2% agarose gel. This PCR was outstanding in
the detection of gyrB2 gene in urinary tract infected patients caused by Pseudomonas sp.

2.4. Antibiotic susceptibility test

Mojtaba et al. (2008) evaluated the etiology, risk factors, and patterns of antimicrobial
resistance of intensive care unit (ICU)-acquired urinary tract infections (UTIs) in patients
admitted with sepsis. A UTI was present in 28 (28%) patients; the male to female ratio was 19:9
and the mean age of the patients was 58.71 19.45 years. From the total of 28 isolates, 27 were
resistant to ciprofloxacin, 23 to amikacin, 27 to meropenem, 28 to cefepime, 26 to ceftazidime,
and 27 to ceftriaxone

Mohandas et al. (2009) was determined the prevalence of asymptomatic bacteria,

bacterial agents and their antibiotic susceptibility pattern in pregnant women. 297 samples
10(3.3) were positive for asymptomatic urinary tract infection. The dominant bacterial isolate
was Escherichia coli (70%). The antibiotic susceptibility was observed to ciprofloxacin,
ceftazidime and cefotaxime (80%), the most resistance was amoxillin-clavulanic acid (90%).

Beyene et al. (2011) determined the type of antibiotic resistance pattern of the urinary
pathogens isolated from patients. Isolated bacteria were detected from 9.2% of the total patients.
The most common pathogens isolated were E. coli (33.3%), K. pneumonia (19%) and S.
saprophyticus (14.3%). E. coli and K. pneumonia showed the highest percentage of resistance to
ampicillin and amoxicillin (100%) however, all isolated of E. coli and K. pneumoniaewas
susceptible to ciprofloxacin. S. saprophyticus and S. aureus were resistant to ampicillin (100%)
and amoxicillin (66.7%). For all UTI isolated, least resistant was observed against drugs such as
ceftriaxone, gentamycin and chloramphenicol.
 Nerurkar et al. (2012) demonstrated the ethological bacterial pathogens of the UTI /and
to determine the antibiotic sensitivity pattern of pathogens isolated. E. coli was the most common
isolate (44.96%) followed by Enterobacter sp. (17.83) and Klebsieblla sp. (14.72%) amongst the
gram negative bacilli, gram positive bacteria Staphylococcus aureus (92.3%)was commonest E.
coli which was the main isolate identified was found to be highly susceptible to Amikacin
(82%)followed by Ciprofloxacin (78.2%). Gentamician (80.4%) Ampicillin (59%) and
Nitrofurantoin (57%) the type of Gram negative bacteria related to Enterobacteriaceae involved
in UTI and antimicrobial susceptibility pattern of the urinary pathogens. Total 48.48% urine
samples showed significant bacterial growth. The most common pathogens were Escherichia
coli (42.71%), Klebsiella pneumonia (23.96%), Proteus sp. Amikacin showed 69.57% resistance
in K. pneumonia, however, Levofloxacin showed 100% sensitivity, Nitrofurantoin showed
92.30% resistance in Enterobacter sp. and most quinolones and carbenicillins was susceptible to
Enterobacter sp. Proteus sp. was 100% resistant against Third generation cephalosporin,
however, Carbepenems was highly susceptible to isolated Proteus sp. Meropenem (90.63%) was
most sensitive among all isolated UTI pathogens and Nalidixic acid showed 67.71% sensitivity
among all isolates.

DevanandPrakash et al. (2013) was determined the distribution and antimicrobial

susceptibility of uropathogens in the Indian community as well as to determine the effect of
gender and age on the etiology of bacterial uropathogens. Clean catch midstream urine samples
were collected from 288 patients of the age ranging from 15 to 48 years Antimicrobial
susceptibility was performed on all isolated bacteria by Kirby Bauer’s disc diffusion method .The
multiple antibiotic resistance (MAR) index of each antibiotic was calculated. The UTI prevalence
was 53.82% in patients however, the prevalence was significantly higher in females than in males
(females: 73.57%; males: 35.14 %;). Females within the age group of 26-36 years and elderly
males of 48 years showed higher prevalence of UTI. Gram negative bacteria (90.32%) were found
in high prevalence than Gram positive (9.68%). Escherichia coli (42.58%) were the most
prevalent gram negative isolate. Nitrofurantoin (78.71%) was found the most resistant drug
among all uropathogens. Tested carbapenems were found the most susceptible drug against
isolated uropathogens which showed 92.36% and 84.52% susceptibility, respectively.
 Sharma et al. (2013) was identified the etiologic agents of UTI and to determine the
patterns of antimicrobial drug susceptibility among pregnant women attending antenatal clinic.
Isolated and diagnosed bacteria from pregnant women were found in Escherichia coli (43.47%),
Staphylococcus aureus (23.91%), Proteus vulgaris (19.56%), and Klebsiella species (5.43%),
Coagulase negative Staphylococci (7.6).The isolated uropathogens showed resistant to
ampicillin, co-trimoxazole ciprofloxacin, ceftazidime and sensitive to nitrofurantoin and
cephotaxime.

Chaudhary et al. (2014) was studied the prevalence of extended-spectrum β-lactamases

(ESBLs) among 663 clinical isolates obtained from various parts of India and to study the
occurrence of different variants of ESBLs among these isolates. Among the six hundred sixty
three isolates, the identified isolates were Acinetobacter baumannii (72), Escherichia coli (218),
Klebsiella pneumonia (30), Klebsiella oxymora (63), Pseudomonas aeruginosa (264) and
Staphylococcus aureus (16). PCR results revealed that approximately 89.0% of
Pseudomonasaeruginosa isolates were positive for ESBL followed by Escherichia coli (85.3%),
Klebsiella pneumonia(76.6%), Klebsiella oxymora (73.0%), Acinetobacter baumannii (72.2%)
and Staphylococcus aurous (31.2%). The overall prevalence of ESBL was 82.5%. The presence
of TEM type ESBLs were the predominant (in 186 isolates) followed by SHV (138), OXA (92),
CTX-M (65), AmpC (33), KPC (28) and blaZ (5). Of the drugs involved in the study, CSE1034
was found to be the most efficacious against all of ESBL positive clinical isolates showing
susceptibility approximately 95.7% with minimal inhibitory concentration values between 0.125
and 8.000 µg/mL for all strains tested. The susceptibilities of penmen (meropenem and
imipenem and cilastatin) ranged between 83% and 93% for all the isolates. The susceptibilities
of other drugs like piperacillin and tazobactam, amoxicillin and clavulanic acid, cefoperazone
and sulbactam were <45% for all the isolates.

2.5. Biological activity of plant extract

The antimicrobial activity of ethanolic extracts of pomegranates (punica granatium L.)

pricarps and celery (Apium graveolens L.), sweet clover (Melilotus officinalis L.), Alfalfa
(Medicago sativa L.), Fenugreek seed (Trigonellafeonum-graecum L.), leaves and carrot
(Daucus carota L.) seeds against some pathogenic fungi, gram negative and gram positive
bacteria were studied. All bacteria which were tested have shown susceptible to the P.granatium
and D. carota ethanolic extracts. Proteus vulgaris, Klebsiella aerogens and Streptococcus aureus
the most susceptible bacteria to the D. carota and P. granatium extracts successively reported by
RadhiMalih (2008).

Mahalingam et al. (2011) reported the antifungal properties of the leaf extracts of
Pandanus odoratissimum, Cedrusde odara, Capparis zeylanica, Mirabilis jalapa, Eicchorni
acrassipes, Gaedenia jasminoides, Spermaco cehispida, Nymphae anouchali, Melothria
heterophylla, Enicostem malittorale, against Setosphaeri arostrata isolated from sugarcane
fields. The pathogen Setosphaeri arostrata a commonly was causing the seedling blight disease
in sugar cane. The antifungal assay was determined by well diffusion method. Three different
solvents such as methanol, ethanol and aqueous were used. Among the three solvents methanolic
extracts of all the treated plant leaves showed maximum significant inhibitory activity followed
by ethanolic extract. There was no activity found with aqueous extract of all the plants tested.

Mahdi (2011) evaluated the antibacterial potential of three type’s plants essential oils
were extracted from the seeds of Apium graveolens, Coriandrum sativum and Cuminum
cyminum and assayed in vitro for antibacterial activity against most prevalent pathogens
Staphylococcus aureus, Salmonella sp. Escherichia coli and Pseudomonas aeruginosa. The
result shows that the extract of Apium graveolens, Coriandrum sativum and Cuminum cyminum
alcoholic extract at 25◦C exhibited an inhibition zone on S. aureus in the concentration
200mg/ml and had a exhibited significantly (p<0.05) greater than that produced by gentamicine
also the extract of Apium graveolens at 25◦C in the concentration 100 mg/ml had a significant
antibacterial activity on S. aureus, and the extract Apium graveolens alcoholic extract at 25◦C
and 37◦C pronounced antibacterial activity against Pseudomonas aeruginosa in the concentration
100 and 200 mg/ml respectively more ever the inhibition produced by gentamicine. While the
antibacterial effect Apium graveolens and Coriandrum sativum at 37◦C on S.aureus in the
concentration 200 mg/ml show very similar effect with gentamicine. The antibacterial activity
against Salmonella sp. and E. coli were moderate in action. It was found that the investigated
extracts of Apium graveolens, Coriandrum sativum and Cuminum cyminum were exhibited a
considerable inhibitory effect against S.aureus and extracts of Apium graveolens and
Coriandrum sativum against P.aeruginosa.
 Renisheya et al. (2011) reported the antimicrobial potential of five medicinally important
plants namely, Curcuma mangga (C. mangga) Valeton&Van Zip, Ficus racemosa (F. racemosa)
Roxb, Vitex negundo (V. nnegundo) L., Ocimum basilicum (O. basilicum) L., and Etling
eraelatior (E. elatior) K. Schum. Against the human pathogens. The results of this study showed
the presence of wide spectrum of antibacterial activities against all the above bacterial pathogens
studied. The maximum zone of inhibition observed for each bacterium was as follows: S. typhi
(12mm), K. pneumonia (13mm), P. vulgaris (20mm), P. aeroginosa (16mm) and S. aureus
(12mm).

Manzur et al. (2011) used 91 clinically important strains for the study which was both
clinical isolates as well as identified strains. The antimicrobial activity of all the extracts was
determined by agar disc diffusion method. The antibacterial activity was more pronounced
against bacteria than fungal strains. The Gram positive bacteria were more susceptible than Gram
negative bacteria. The methanol extract showed best antibacterial activity. T. catappa leaf
extracts showed better antibacterial activity than commercially used antibiotics.

Mogana et al. (2011) studied in six different extracts of Canarium patentinervium leaves
and barks were screened for their phytochemical composition, and antimicrobial and free radical
scavenging activities. Among the different extracts tested, the ethanol extract of leaves showed
significant antimicrobial and radical scavenging activities. The most susceptible microorganisms
were found to be Gram positive bacteria (Staphylococcus aureus, Methicillin resistant
Staphylococcus aureus or MRSA) and Gram negative bacteria (Pseudomonas aeruginosa).
Phytochemical analysis of the extracts revealed that the antimicrobial and the radical scavenging
activities are mainly due to the presence of tannins and flavonoids. The results obtained suggest
that Canarium patentinervium could be exploited in the management of various infectious
diseases.

Baby Joseph (2011) identified the bioactive compounds from the Sida cordifolia plant
and the antimicrobial, cytotoxic effect on HeLa cell lines. The GC-MS analysis results showed
mainly four different compounds such as Vasicinol, Ephedrine, Vasicinone and Hypaphorine in
various criteria like retention time and peak observation among the four phytochemical
compounds based upon in vivo study. The agar disk diffusion method was used to study the
antibacterial activity of S. cordifolia extracts against 7 bacterial strains. The minimum inhibitory
concentration (MIC) of the plant extracts were tested using two fold agar dilution method at
concentration ranging from 6 to 18µm/ml. The cytotoxicity results of S. cordifolia (L) extract on
HeLa cell lines clearly reflected the treated cell lines are fortunately all the uncontrolled growth
has been arrested there is declined level of cancerous cells observed. The methanol extract was
found to be an effective against all phytopathogens with low MIC of 6µm/mm and the methanol
extract exhibited a higher inhibition activity against Escherichia coli, Bacillus subtilis,
Enterobacter aerogens, Mycobacterium sp. and Micrococcus variance, Pseudomonas
aeruginosa and B. subtilis, Meanwhile, the results also indicate the presence of major
phytochemical compounds such as vasicinol, Ephedrine, vasicinone, Hypaphorine in the S.
cordifolia extracts. Hence, the isolation and purification of therapeutic potential compounds from
S. cordifolia could be used as an effective source against bacterial diseases in human and plants.

Gomathy et al. (2012) determined the qualitative analysis of phytochemical screening

and possible chemical components of Mukiamader aspatana (L) (family:Cucurbitaceae) leaves
GC-MS. The plant is an indigenous plant; traditionally it is uses as an ingredient of various
cocktail preparations and for the management of severe inflammatory disorders in Indian system
if medicine. GC-MS analysis of hydroalcoholic extracts leads to identification of 7 compounds.
This analysis revealed that contains Mukiamader aspatang (L) leaves mainly Dichloroacetic
acid,4-methylpentyl ester,2-Butyn-1-0l,4-methoxy and also showed the presence of other
constituents like flavonoids, saponins, carbohydrates, steroids, tannins and phenolic compounds.

Rana and Rana (2012) evaluated the larvicidal acticity of essential oils derived from ten
aromatic plants.With special reference to Foeniculum vulgare(F. vulgare) against early fourth
instar larvae of Culex quinquefasciatus (Cx. quinquefasciatus) say (Diptera:Culicidae). Essential
oils were extracted from plant materials through hydro distillation and efficacy was determined
through bioassay method. Two of the effective oils were evaluated further for the determination
of their LC50 and LT50 values based on profit analysis Essential oils of one plant was analyzed
through TLC and HPLC. Most of the essential oils caused 100% mortality within 120min at
250ppm.Oil of Tagetes patula (T. patula) and F. vulgare gave more than 90% mortality within
40 min. LC50 values calculated after 60 min of treatment were (84.80 and 7.65) and (24.69
,1.24) ppm for T. patula and F. vulgare respectively. At the same exposure period positive
control temperature (22.13, 2.39) ppm LC50 value. At 50ppm T. patula, F. vulgara and
exhibited LT50 values (113.71, 10.29), (11.02, 3.31) and (38.15, 5.90) min respectively.
Presence of high quantity of the essential oils of F. vulgare was confirmed by TLC and HPLC
results. This study was indicated that essential oils of spices/aromatic medicinal plants
particularly F. vulgare and T. patula carry huge potential as a mosquito larvicide. This potential
could be exploited for the development of safer and effective.

Iswantini et al. (2012) determined the inhibition effect of celery extracts toward xanthine
oxidase by in vitro method, and its active compounds. The ethanol extract of herbal at 1400
pomade the greatest inhibition effect with percent inhibition value at 91.64%. The fractionation
of the ethanol extract of herb yielded five fractions. The enzymatic test showed that fraction 4
was the fraction with the highest inhibition effect with percent inhibition of 88.62%, followed by
fraction 5 with percent inhibition at 85.44%. The UV and IR spectrum of both fraction 4 and 5
showed a very similar pattern. The analysis of UV spectrum of fraction 4 indicated the presence
of TT-TT transition which was resulted by the aromatic C=C,-OH, and C-O chromospheres.
While the n- transition was given by –C=O chromospheres. The IR spectrum analysis indicated
that the functional groups of aromatic –C=C, -OH, and C-O were presence. Based on
phytochemical assay and spectrum data, it was thought that fraction 4 and 5 were the flavonoid
group.
Senthamaraiselvi and Anushabaskar (2012) analysed the Sauropus androgynus was
carried out the phytochemical analysis of the leaves of this plant is reported for the first time.
The leaves indicated the presence of proteins, resins, steroids, tannins, glycosides, reducing
sugar, carbohydrates, saponins, sterols, terpenoids, acidic compounds, cardiac glycosides,
catechol, phenols, alkaloids, flavonoids. In the GC-MS analysis the Sauropus androgynus extract
result shows the presence of bioactive compounds which revealed a broad spectrum of many
medicinal property and antioxidant activity were identified. The functional group present in these
compounds was identified by IR spectral analysis. This study also helped to identify the formula
and structure of bimolecular which can be used as drugs.

Gnanavel and Mary saral (2013) studied the petroleum ether and ethanol leaf extracts
from Abrus precatorius were subjected to GC-MS analysis to study the important phytochemical
constituents responsible for the above reported pharmacological activities. The crude extracts of
petroleum ether and ethanol were obtained by immersion method. The preliminary
phytochemical analysis of various extracts confirmed the presence of secondary metabolites like
alkaloids, flavonoids, saponins, tannins, triterpenens, carbohydrates, proteins, glycosides and
steroids. The GC-MS analysis of petroleum ether leaf extract from Abrus precatorius revealed
the presence of nine phytocompounds: n-hexadecanoic acid,[ 1,1-bicyclopropyl]-2-octanoic acid,
2-hexyl-methyl ester, Isopropyl linoleate, 9,12-octadecadienoic acid, ethyl ester, Hexadecanoic
acid, 2-hydroxy-1-[hydroxymethyl]methyl ester,9,12,15-octadecadienoic acid (Z,Z,Z)-2,3-
dihydroxypropylester, 9,12- octadecadienoic acid (Z,Z)-2,3 dihydroxypropyle ester,
Octadecanoic acid, 2- hydroxyl-1-(hydroxymethyl)ethyl ester. The GC-MS analysis of ethanol
leaf extract from Abrus precatorius revealed the presence of ten phytocompounds: 1,2 Benzene
dicarboxylic acid, butyl cyclohexyl ester, Hexadecanoic acid, ethyl ester, 9,12, octadecadienoic
acid ethyl ester, Phytol, Oleic acid, 9,12- octadecadienoic acid (Z,Z)- 2,3- dihydroxypropyl ester,
9,12 – octadecadienoic acid (Z,Z)-2,3- dihydroxy propyl ester, Hexadecanoic acid, 2-hydroxy-1-
(hydroxymethyl) ethyl ester,9,12-octadecadienoic acid (Z,Z), 2- hydroxymethyl ethyl ester,
cyclopropyl methyl cyclopropy [methyl]- methyl ester.

Kalimuthu and Prabhakaran (2013) reported the preliminary phytochemical screening of

Cerope giapusilla a medicinal plant was carried out. Qualitative phytochemical analysis of these
plants confirms the presence of various secondary metabolites like saponins. Triterpenoids,
steroids, tannins, lignins, alkaloids, glycosides, flavonoids and phenols. The results suggest that
the phytochemical properties for curing various ailments and possess potential anti-
inflammatory, antimicrobial and antioxidant and leads to the isolation of new and novel
compounds. GC-MS analysis showed the existence of various compounds with different
chemical structures. The presence of various bioactive compounds confirms the application of
Cerope giapusilla for various ailments by traditional practitioners. However, isolation of
individual phytochemical constituents may proceed to find a novel drug.

Dhivya and Manimegalai (2013) determined the phytochemicals present in the ethanolic
flower extract of Calotropsis gigantean and further analysis of the components present in it by
GC-MS analysis. Ten grams of flower power was sequentially extracted by ethanol. The results
showed the presence of phytochemical compounds of alkaloids, tannins, phenol, flavoids, sterols,
antraquinones, proteins and quinonens in the flower extract. The GC-MS analysis of the
ethanolic extract revealed the presence of 14 major compounds. This study forms a basis for the
biological characterization and importance of the compounds identified and creates a platform to
screen many bioactive components to treat many diseases.
Ushadevi et al. (2013) evaluated the phytochemical constituents of Allium porrum leaf
extracts and its antibacterial potentialities were using different solvents. Phytochemical screening
of the different extracts showed that leaves contains important compounds such as amino acid
with sulphate, carbohydrates, Phenols, Proteins, Flavonoids, alkaloids, steroids, Tannins and
saponin. Aqueous, methanol and acetone extracts of plant leaves were usedfor assessing
antibacterial activity against Salmonella typhi, Klebsiella pneumonia, Staphylococcus aureus,
Pseudomonas aeruginosa and Proteus vulgaris by agar well diffusion method. This study was
demonstrated that the aqueous, methanolic, acetone extracts of A. porrum consists different
phytochemical components thus exhibit variations in the antibacterial activity. These promissory
extracts open the prospect of finding new clinically effective antibacterial compounds.

Manikandan and Prabhakaran (2014) the potential of an allopathic to extract direct and
indirect effect depends in large part on the chemistry of the plant and whether putative all
chemicals reach meaningful levels in the environment surrounding the plant. Cleome viscose L
(Capparidaceae) (synonym:C. icosandra L.) is a weed distributed throughout the tropics of the
world and the plains of India. The qualitative analysis on the root, stem and leaves of C.viscosa
showed that the presence of saponins and flavonoids in all their three organs.The presence of
alkaloids was noticed only in wagner’s test not in the mayor’s and Dragendorff’s test. GC-MS
result of whole plant of C. viscosa showed the presence 3-0-Methyl-d-glucose (73%),followed
by Benzofuran, 2,3-dihydro(9.844%) and n-Hexadecanoic acid (4.707%) of the total 32
compounds.
2.6. Antibacterial activities of plants

Abdul Ahameethunisa et al. (2010) reported the antibacterial activity of A. nilagirica

extracts against 15 bacterial strains. The minimum inhibitory concentration (MIC) of the plant
extracts were tested using two fold agar dilution method at concentrations ranging from 32 to
512 µg/ml. The phytochemical screening of extracts was carried out for major phytochemical
derivatives in A. nilagirica. All the extracts showed inhibitory activity for gram positive and
gram negative bacteria except for Klebsiella pneumonia, Enterococcus faecalis and
Staphylococcus aureus. The hexane extract was found to be effective against all phytopathogens
with low MIC of 32µg/ml and the methanol extract exhibited a higher inhibition activity against
Escherichia coli, Yersinia enterocolitica, Salmonella typhi, Enterobacter aerogens, Proteus
vulgaris, Pseudomonas aeruginosa (32µg/ml), Bacillus subtilis (64µg/ml) and Shigella flaxneri
(128µg/ml). The phytochemical screening of extracts answered for the major derivative of
alkaloids, amino acids,flavonoids, phenol, quinines, tannins and terpenoids.
Sunil (2010) The methanolic extract of this plant was found to have antimicrobial
activities against six species of pathogenic and non-pathogenic microorganisms: Bacillus
subtilis, Sataphylococcus aureus, Salmonella typhi, Escherichia coli, Pseudomonas aeruginosa
and Candida albicans. The maximum zone of inhibition (20mm) was found to be exhibited
against S. aureus. For this organism the minimum bactericidal concentration (MBC)of the crude
extract was 1,0001g/ml .The extract was found to be equally effective against gram positive and
gram negative bacteria. The antimicrobial activity of the extract was found to be decreased
during purification. The chemical constituents of organic plant extracts were separated by thin
layer chromatography (TLC) and the plant extracts were purified by column chromatography and
were further identified by Gas chromatography mass selection (GC-MS) analysis. The
composition of A. catechu extract had shown major components of terpene i.e. camphor
(76.40%) and phytol (27.56%) along with other terpenes in minor amounts which are related
with their high antibacterial and antifungal properties.

Vijayakumar et al. (2012) evaluated the antimicrobial activity of hexane; ethyl acetate
and methanol extract seeds and fruits of Illicum griffithi (I. griffithi) family: schisandraceae).
Ethyl acetate extract of fruits was effective against most of the tested reference cultures such as
Staphylococcus aureus, Yersinia enterocolitica, Vibrio parahaemolyticus, Bacillus subtilis,
Salmonella paratyphi, Enterococcus feacalis, Xanthomonas oryzae and Pseudomonas
aerugenosa, whereas methanol extract showed activity only against Staphylococcus aureus,
Bacillus subtilis and Xanthomonas oryzae. The hexane and ethyl acetate extracts of fruits were
more effective against most of the clinical isolates, whereas methanol extract was effective only
against Klebsiella pneumonia ESBL. The extracts of fruits and seeds did not show any
significant antifungal activity against tested fungi. The presence of phenols, tannins,
flavonoids,triterpenoids, steroids, alkaloids, saponins and carbohydrates in the different extracts
were established. Gas chromatography-mass spectrometry studies on hexane and ethyl acetate
extract of fruits resulted in the identification of 31 and 39 compounds respectively.
Radulovic et al. (2012) reported totally 304 constituents were identified, representing
more than 90% of the isolated oils. The volatiles of G. sanguineum have been studied for the first
time identified in this study. The antimicrobial assay showed a strong activity of the G.
robertinaum oil against Escherichia coli and Aspergillus fumigates. Micrococcus flavus and A.
fumigatus were the most susceptible strains to G. sanguineum oil.
Pathan et al. (2012) evaluated the antimicrobial efficacy of Citrus aurantifolia L (CA)
against some microorganisms bacteria and fungus were Staphylococcus aureus, Escherichia coli,
Klebsiella pneumonia, Pseudomonas sp. Aspergillus niger, Aspergillus fumigates and
Penicillium. The dried leaf extract of CA with chloroform, ethanol, acetone, petroleum ether and
aqueous ethanol was approximately 15%, 18%, 09%, 11%and 24% respectively. Due to its high
yield value hydroalcoholic extract of CA was used for estimating the antimicrobial activity and
its phytochemical screening. Phytochemical screening of CA plant reveals the presence of
Alkaloids, carbohydrates, flavonoids, steroids and tannins.
Govindasamy et al. (2012) identified the antibacterial activity and phytochemical
analysis of Catharanthus roseus (C. roseus) and also screening of leaf, stem, flower and root
extracts of the plant for its antibacterial activity. The ethanolic extract showed a maximum zone
of inhibition (21.15,1.64mm) against S. typhi minimum and zone of inhibition (06.24,0.69mm)
with ethanolic extract against S. aureus Further, the and minimum (05.20 0.8 6mm) zone of
inhibition against E. coli. In addition to the phytochemical analyses were showed the presences
of soluble sugar,reducing sugar, protein, methanolic extract observed in maximum (15.61
1.35mm) against S. typhi amino acid, lipids, total chlorophyll, phenol and ortho-
dihydroxyphenols in the ethonolic extract.

2.7. Antioxidant activities

Olayinka et al. (2010) evaluated invitro antioxidant activities and to screen for
phytochemical constituents of Helichrysum longifolium (family: Asteraceae) aqueous crude
extract. Phytochemical analysis revealed the presence of tannins, flavonoids, steroids and
saponins. The total phenolic content of the aqueous leaf extract was 0.499 mg gallic acid
equivalent/g of extract powder respectively. The percentage inhibition of lipid peroxide at the
initial stage of oxidation showed antioxidant activity of 87% compared to those of BHT (84.6%)
and gallic acid (96%). Also the percentage of inhibition of malondialdehyde by the extract
showed percentage inhibition of 78 % comparable to those of BHT (72.24%) and Gallic
(94.82%).

Kalucka (2010) The lipophilic and hydrophilic antioxidants were evaluated in eight
plants:safflower (Carthamus tinctorius )viper’s bugloss (Echium vulgare ) quince (Cydonia
vulgaris ) evenings primrose (Oenother abiennis ) rose mosqueta (Roseafinisru biginosa),black
seed (Nigella sativa) seabuckthorn (Hippophe arhamnoides) and borage (Borago officinales).
The highest amounts were contained in seed of borage and sea buckthorn (66.9mg/100gand
45.9mg/100g,respectively). The sea buckthorn seed lipids had the highest amounts of total
sterols (10.4mg/g of lipids). The predominant form was campesterol. Sitosterol was the major
sterol in the lipids of other tested seeds .The content of phenolic compounds ranged from
736.5mg/100g dry matter (d.m) (evening primrose) to 74.8 mg /100g d.m (safflower).The
highest antioxidant activity, expressed in % scavenged DPPH free radicals, was observed for
evening primrose (91.2%),while the lowest for safflower (36.2%). The correlation coefficient
between the level of phenolic compounds and antioxidant activity was 0.53.

Pracheta Sharna (2011) the antioxidant potential and phytochemical constituents of crude
hydro-alcoholic extract of Euphorbia neriifolia (EN) using tests involving inhibition of DPPH,
H2O2, superoxide anions, reducing power, FRAP and metal chelating activities. The phenolic,
flavonoids and tannin contents of the extracts were also determined using standard
phytochemical reaction methods. EN extract showed the presences of alkaloids, tannins,
saponins, flavonoids and cardiac glycosides. A positive correlation between the antioxidant
activities and physiochemical assays was observed and the highest scavenging activity of extract
was noticed at concentration of 1mg/ml. The percentage inhibition of lipids peroxide at the initial
stage of oxidation showed antioxidant activity of 76.15% compared to those of ascorbic acid
(75.6%), BHA (60.8%) and BHT (75.6%). The percentage inhibition of metal chelating capacity
of extract and standard was found to be 73.24%and 85.37% respectively. Our findings reveals
that the hydro-alcoholic extract of EN leaves possess antioxidant properties and could serve as
free radical inhibitors or scavenger or, acting possibly as natural antioxidants and this justified its
uses as anti-inflammatory, anti-analgesic, anti-anemic, anticancer in folkloric medicines

20
 Gul et al. (2011) were studied the antioxidant, antimicrobial and anti-proliferative
activities of A. moschatus extracts were evaluated in a series of invitro assay involving free
radicals, reactive oxygen species and their IC50 values were also determined. The results were
showed that the antioxidant activities of A. moschatus as determined by the total phenol,
flavonoids, and total antioxidant and FRAP methods were higher in leaf than that of the seed
extracts. The aqueous overnight seed extract (AMS-I) has shown significant radical scavenging
activity as in 1,1-Diphenyl-2-picrylhydrazyl (DPPH), hydrogen peroxide, hydroxyl radical,
superoxide and lipid peroxidation as compared to other seed and leaf extracts. The AMS-I and
AML-IV have shown activity against six and seven microorganisms respectively.
Simultaneously, AMS-IV and AML-IV have demonstrated potential antiproliferative activity
against two human cell lines- Colorectal adenocarcinoma (COLO-205) and retinoblastoma
(Y79).
VellingiriVadivel et al. (2011) was reported the methanolic extract of ox-eye bean
[Mucuna gigantea (Willd) DC.] contained total free phenolic content of 14.80±1.28 g catechin
equivalent/100 g extract dry matter. Encouraging levels of ferric reducing antioxidant power
(FRAP, 1,023mm Fe [II]/mg extract) inhibition of β-carotene degradation (59.35%) and radical
scavenging activity against DPPH (72.12%) and superoxide (43.11%) were exhibited by the raw
samples. Further, it also recorded 82.17% of α-amylase and 91.26% of α-glycosidase enzyme
inhibition characteristics. Sprouting oil-frying caused a apparent increase on the total free
phenolic content and also significant improvement on the antioxidant and free radical scavenging
capacity of methanolic extract, while soaking cooking as well as open-pan roasting treatments
showed diminishing effects. Moreover, inhibition of α- amylase and α-glycosidase enzyme
activities was declined to 22.82 and 45.47%, respectively during sprouting oil frying treatment,
which are more desirable for the dietary management of type II diabetic patients

Chew et al. (2012) was evaluated antioxidant, antimicrobial and cytotoxic activity of
different parts (root, flower, leaf and stem) of Lecuas aspera (L. aspera). Extracts showed
moderate to potent antioxidant activity, among which the root extract demonstrated the strongest
antioxidant activity with the IC50 value of 6552µg/ml. Methanol extract of root possessed
antioxidant activity near the range of vitamin E and thus could be a potential rich source of
natural antioxidant. In case of antimicrobial screening, crude extracts of root, flower, leaf and
stem showed notable antibacterial activity against tested microorganisms. The root extract

21
highest mean zone of inhibition ranging from 9.0-11.0mm against tested microorganisms, at a
concentration of 100 mg/ml. In the brine shrimp lethality bioassay, it was evident that the
methanol root extract did not show significant toxicity. The LC50 value for 12h and 24h
observation was 2.890 mg/ml and 1.417 mg/ml, respectively.

Jaures Noumedem et al. (2013) was designed the phytochemical composition,

antimicrobial and radical-scavenging activities of A. manniana methanol leaf extract and its
fractions. The results obtained showed that A. manniana contains alkaloids, tannins,
anthocyanins, flavonoids, phenols and steroids. The methanol extract as well as the hexane, ethyl
acetate and residual fractions exhibited both antibacterial and anti dermatophytic activities that
varied between the microbial species (MIC = 0.12 – 2.04 mg/mL). These tested samples also
showed high radical-scavenging activities (RaS50 = 3.34 – 4.80 μg/mL) when compared with
vitamin C used as reference antioxidant (RaS50 = 1.74 μg/mL).

Sohrevardi et al. (2013) was reported we have investigated essential oil constituents and
antioxidant property of Apium graveolens plant. Celery plant with scientific name Apium
graveolens is belongs to umbllifereae and consists of 114 genus and 420 species. For antioxidant
activity investigation, used of FTC, TBA, DPPH andTPC method. Then result by comparison of
Cvitamin antioxidant activity (naturalantioxidant) and BHT (synthetic antioxidant). Percentage
of antioxidant activity were determined for ethanolic extract of plant under study 37%, C vitamin
25.92%, BHT 32% by TBA method. 50.5% from leaf and stem of plant 22.5%, BHT 49.7%, C
vitamin 31.9% for FTC method. Absorption earn at phenolic compound method for this plant
0.41, amount of phenolic compound in this plant 312.3 mg/g. IC50 value of plant extract in
sDPPH assay was 8 μg/ml. Coumarone component at this plant is diluter and anticlotting and
cause of decrease of blood pressure and spathoulenol have antibacterial and pesticide properties.
By regards of this plant in natural, of this plantcan use as a natural antioxidant in industrial food
and drugs.

2.8. Synthesis of silver Nanoparticle suspension

Song et al. (2009) studied five plant leaf extracts (Pine, Persimmon,Ginkgo, Magnolia
and Platanus) were used and compared for their extracellular synthesis of metallic silver
nanoparticles. The synthesized silver nanoparticles were characterized with inductively coupled

22
plasma spectrometry (ICP), energy dispersive X-ray spectroscopy (EDS), scanning electron
microscopy (SEM), transmission electron microscopy (TEM), and particle analyzer. The average
particle size ranged from 15 to 500 nm. The particle size could be controlled by changing the
reaction temperature, leaf broth concentration and AgNO3 concentration. This environmentally
friendly method of biological silver nanoparticles production provides rates of synthesis faster or
comparable to those of chemical methods and can potentially be used in various human
contacting areas such as cosmetics, foods andMedical applications.
Inbaneson et al. (2011) reported the synthesized silver nanoparticles were investigated to
evaluate the antibacterial activity against urinary tract infections (UTIs) bacterial pathogens.
Escherichia coli was predominant (47%) followed by Pseudomonas aeruginosa (22%),
Klebsiella pneumonia (19%), Enterobacter sp, (6%). Proteus morganii (3%) and Staphylococcus
aureus (3%). The antibacterial activity of silver nanoparticles was evaluated by disc diffusion
assay. P. aeruginosa showed maximum sensitivity (11±0.58mm) followed by Enterobacter sp.
(8±0.49mm) at a concentration of 20µg d ¹ and the sensitivity was highly comparable with the
positive control kanamycin and tetracycline. K. pneumonia, E. coli, P. morganii and S. aureus
showed no sensitivity against all the tested concentrations of silver nanoparticles might indeed
the potential source to treat urinary tract infections caused by P. aeruginosa and Enterobacter sp.
Umamaheswari et al. (2012) evaluated the antioxidant activities of the extracts was
determining using various in vitro assays. The ACE inhibitor potency of the seed extract (IC
50=666.26±1.32µg/ml) was found to be significant (P<0.01) when compared with the standard
lisinopril (IC50=0.19±0.02ng/ml). The extract showed a strong antioxidant activity when
compared with the standard. The results of this study suggest that the methnolic seed extract of
A. graveolens has significant antioxidant and ACE inhibitor effect of antioxidant activities of the
methanolic seed extract of Apium graveolens.Traditionally the seeds of A. graveolens were used
as diuretic, anti-inflammatory.

Logeswari et al. (2012) determined the Plants extract from Ocimum tenuiflorum,
Solanum tricobatum, Syzygium cumini, Centella asiaticaand Citrus sinensis was used for the
synthesis of silver nanoparticles (Ag NPs) from silver nitrate solution. Ag NPs were
characterized by UV–Vis spectrophotometer, X-ray diffract meter (XRD), atomic force
microscope (AFM) and scanning electron microscope (SEM). Antimicrobial activity of the silver
bio-nanoparticles was performed by well diffusion method against Staphylococcus aureus,

23
Pseudomonas aeruginosa, Escherichia coli and Klebsiella pneumoniae. The highest
antimicrobial activity of silver nanoparticles synthesized by S. tricobatum, O. tenuiflorum
extracts was found against S. aureus (30 mm) and E. coli (30 mm) respectively. The AgNPs
synthesized in this process has the efficient antimicrobial activity against pathogenic bacteria. Of
these, silver nanoparticles are playing a major role in the field of nanotechnology and
nanomedicine.

Vanaja et al. (2012) The present study reported the plantmediated synthesis of silver
nanoparticles using the plant leaf extract of Coleus aromaticus The silver nanoparticles were
characterized by ultraviolet visible spectroscopy, X-ray diffraction, scanning electron
microscopy, energy-dispersive X-ray spectroscopy, Fourier transform infrared spectroscopy, and
the size of the silver nanoparticles is 44 nm. The bactericidal activity of the silver nanoparticles
was carried out by disc diffusion method that showed high toxicity against Bacillus subtilis and
Klebsiella planticola.
Anand and Gokulakrishnan (2012) reported the present study was carried out to
characterize the bioactive constituents present in ethanolic extracts of Hybanthus enneaspermus
using UV, FTIR and GC-MS. The FTIR spectrum confirmed the presence of phenols, alcohols,
alkanes, alkyl halides, carboxylic acids, aromatics, nitro compounds and amines in ethanolic
extract. The results of the GC-MS analysis provide different peaks determining the presence of
phytochemical compounds with different therapeutic activities. The major phytoconstituents
were (5E, 13E)-5, 13-Docosadienoic acid (20.90%) and Cedran-diol, 8S, 14- (13.02) which
possess many biological activities. hence this study creates a platform to screen many bioactive
components to treat many diseases.

Nagati et al. (2012) was reported that this research article our present a simple and eco-
friendly biosynthesis of silver nanoparticles using P. longifolia leaf extract as reducing agent.
TEM showed the formation of silver nanoparticles with an average size of 57 nm.

El-Sherbiny et al. (2013) was studied, green synthesis of densely dispersed and stable
silver nanoparticles (AgNPs) using myrrh extract as green-reducing and stabilizing agent was
investigated The resulting AgNPs were characterized by UV-Vis spectrophotometry,

24
transmission electron microscopy, Fourier transform infrared spectroscopy, and X-ray
diffraction. The developed NPs demonstrated a relatively high antibacterial activity against
Bacillus thuringiensis and Pseudomonas aeruginosa bacteria as compared to that of myrrh
extract. The antibacterial activity increased with increasing the AgNPs concentration.

2.9. Isolation of Phytocompounds in Apiu m graveolens

The primary phytochemical analysis on the seed extract of Apium graveolens indicates
the presence of carbohydrates, flavonoids, alkaloids, steroids and glycosides in the methanolic
extract (Khar, 2008) the plant included phenols and furocoumarins. Furocoumarins contained
celerin, bergapten, apiumoside, apiumetin, apigravrin, osthenol, isopimpinellin, isoimperatorin,
celereo side and 5 and 8-hydroxy methoxypsoralen. Phenols (155.41-177.23mg/100g) included
graveobioside A and B, apiin, apigenin, isoquercitrin, tannins (3.89-4.39 mg/100g) and phytic
acid (19.85-22.05 mg/g) (Khar, 2008).

Celery leaves and stem contain phenols. Flavonoids are the main component of apigenin
celery leaves whose amount is 202 mile-grams per kilo-gram. There are also luteolin and
chrysoeriol 7-glucosides with the amounts of 48 mg per kg and 27 mg kg respectively in celery
leaves. Celery leaves also contain furanocomarin, bergapten, xanthotoxin and isopimpinellin,
whose amounts vary from 12 to 50 mg kg in celeries grown in Florida in the United States
(Tahar et al., 2007). Among the 12 fragrant elements in the leaves and stem of celery, first 3-n
butyphthlide, sedanolide have the largest contribution to creating a fragrant aroma in the plant
(TamasBarberan, 2000).

Jung et al (2012) in a study examined the combined total phenolics and flavonoids
antioxidant properties of the leaves of celery using Folin-Ciocalteu method. The results of their
study showed the alcoholic extract of the leaves of celery contains the highest amount of
phenolics compounds and aqueous extract came in the second in this regard (Emertcan et al.,
2007). Celery root contains phenolics derivatives. Studies on the phenolics derivatives of the
celery root show that the amount of caffeic acid is at the highest level (89 to 168 milligrams per
kilogram, depending on the soil where it grows). The amount of ferulic acid is noticeable too (34
to 61 mg kg) while its p-coumaric acid derivatives content was very small(less than 5.0 mg/kg).
Celery root contains coumarin, scopoletin, and aesscoumlic acid. However the amount of these

25
elements is less than 5.0 milligrams per kilogram in the plant (Tahar et al., 2007). Celery seeds,
stems and leaves oil (2.5-3.5%) included volatile oils, sesquiterpene alcohols (1-3%) and fatty
acids. The derived major compounds were selenine (10-15%) limonene (60%), β-
pinene,camphene,cymene,limonene,α-thuyene,α-pinene,β-phellendrene, p cymene, γterinene,
sabineneterinolene, myristicic ,myristic, linoleic, petroselinic, palmitoleic, palmitic, oleic,
myristoleic, stearic acid,santalol,β-eudesmol,α-eudesmol,sedanenolide,3-n-butylphthalideand
phthalide. Celery tuber also contained methoxsalan (8-methoxypsoralen), 5- methoxypsoralen
and the allergen profiling (Apigl) (Khare 2008).

Celery seeds contain 2 to 3% essential oil. Its oil contains mostly limonene 9usually 60
percent), selenine (10%), furocoumarins and furocoumarins glycosides and their flavonoids.
Photochemistry tests of celery seeds approve the presence of flavonoids apigenin (as main
component), andvitamins A and C.A total of 16 combinations of seed extract have been
identified in celery which make up 98.7% of the whole extract whose main components are
D.limonene and myrcene. (Al-Snafi,2014).Celery seeds can be a potential candidate for the
production and development of commercial insect repellents and thus an alternative for the
common synthetic chemicals used in the community to control insects particularly the vector
ones.Recently a substance known as CAH has been found in the ethanol extract of celery seed
which can be investigated as an H. pylori infection (Haflmann, 1990).

2.9.1.Therapeutic Properties

A. graveolens seed on liver carcinoma, which was injected in Wister rats in the laboratory
(chemical), had inhibitive effect (Sah, 1995) A. graveolens can also stimulate pancreas to secrete
insulin to reduce blood glucose levels, so that it can be used to reduce or treat diabetes
complications (Thium et al., 2002). In another part of an invitro study, antioxidant systems in
homogenized liver of rats and their blood were examined. Evaluation of antioxidant systems
after applying extract of celery leaf, alone or in combination with carbon tetrachloride (it is
thought that free radicals derived from CCl4 due to lipid per oxidation have damaging effect on
liver)showed that the exerted extracts havea protective effect (Zidorn et al., 2005). Celery
contact with skin can cause acute skin inflammation (Tuetum et al., 2005). Celery seed is
effective on liver injuries in rats which were caused by a single dose of paracetamol. It was
noticed clear that celery has the protective activity against thioacetamide drugs (Hamza and

26
Amin, 2007). In a study by Kooti et al., 2014 it was shown that celery can protect spermatozoa
and tests of rats against the toxic effects of propylene glycol (Jung et al., 2012). The results in an
in vitro study evaluated the ability of methanol extract of celery to remove OH and DPPH and
inhibition of liposomal per oxidation showed that extract obtained from the removal of free
radicals and DPPH and reduces the intensity of LPX (liposomal per oxidation) indicating the
protective activity (antioxidants). Pretreatment with celery seed extract reduces most of the
effects caused by sodium evaporate which shows protective effect of celery extracts against
toxicity caused by VPA produced in laboratory.VPA is commonly used in the treatment of
epilepsy and other diseases and is thought to have a severe toxic effect on the tests in animals
and humans (Kooti et al., 2014). Celery can regulate heart function. Its levels and roots water has
effects on biochemical parameters such as GSH content, activity of catalase, peroxides’ and
intensity of lipid per oxidation in homogenized liver and hemolyzed blood (Hamza and Amin,
2007). Taher’s study in Isfahan showed that the ingredients of celery stabilizes liver cell
membranes and reduce release of AST and ALT enzymes into the blood (Khare, 2008). In a
study, Kootiet al. (2014) evaluated the effects of celery on serum lipids of mice fed high- fat
meals showed the plant causes a significant decrease in LDL and Cholesterol, However not on
VLDL and HDL (Teng et al., 1985). In a study Tsi et al. (1995, 2000), examined the attributes of
anti-hyperlipidemia of the celery in the rat. At the end of the experiment a significant reduction
was observed in the concentration of serum total cholesterol, triglyceride levels and hepatic
lipase tricycle glycerol in the treatment group (Korti et al., 2014). In a study conducted by
AbdElmageed (2011) about regulatory effects of celery on lipid metabolism and also prevention
from the fatty liver, biochemical analysis of serum liver enzymes and blood lipids showed that
eating 10% of celery alone lowers liver enzyme levels and blood fats (Tsi et al., 1995, 2000). In
another study, Cheng et al. (2008) showed that celery seed extract has a hyperlipidemia activity
and also has a role in cleanup of free radicals due to the antioxidant property (AbdElMagged,
2011). Long term consumption of aqua and butanol content celery extract primarily decreases
total cholesterol levels by increasing the excretion of bile acids and, rather than by the
modulating activity of limiting enzyme in cholesterol biosynthesis ( Cheng et al., 2008). Some of
the compounds in it seed show anti-inflammatory and analgesic effects (Atta, 1998; Tsi, 2000).
In a study affects of celery simultaneously paracetamol analgesia and aminopyrine by celery. It
was also shown that celery juice reduces cytochrome P450 in comparison to the control group
homogenized liver (Lewis et al., 1985). Zhang et al. (1999) showed protective effects of celery’s
active substances in ischemic brain (Jakovljevie et al., 2002). Kooti et al (2014) showed in a
study that celery can reduce sex hormone LH in rats however had no significant effect on
Testosterone and FSH (Zhang et al., 1999). Kooti et al. (2014) studies determined that celery
concurrent use of both male and female rats, cause an increase in number of newborns however,
there was no significant effect on birth rates (Ghasemiboron et al., 2014). GharibNaseri et al.,
2007 found that celery leaf extract have concentration-dependent inhibition on rat ileum
concentrations and concluded that probably voltage and ligand gated calcium channels involved
in the genesis of this antispasmodic effect (Kooti et al.,2014).

Celery considering pain relief as an important factor in the management of colicky pain.
This herb was developed in the form of soft capsules and subjected for multi centric clinical
trials. These trails included the evaluation of new herbal anti-spasmodic in patients with colicky
disorders like nonspecific abdominal coli, irritable bowel syndrome, infective diarrhea, acute
amoebic colitis, pain associated with lower urinary tract infection and painful menstruation
(Naseri et al., 2007). Celery has the ideal quantities of iron and magnesium to stop oncological
diseases from progressing. The juice extracted from the petioles can be used for oedema,
rheumatic tendencies gout, flatulence, chronic pulmonary catarrh, tendencies toward
overweightand lack of appetite (Harrisons, 1998). It is a strong diuretic, and is used as a urinary
antiseptic, mainly because of the volatile oil apiol. Celery seed has been found to help regulate
nervous system by producing a combing effect. It stimulates seeds drives and produce sedative
effect (Shalini et al., 2014). Its seeds also used as nervine, an agent that act on the nervous
system to reduce distress and irritation. Both stem and seed of celery have been reported to help
balance acidity in the body. Celery seed also promotes perspiration which may help with weight
loss when water retention is a problem (Ahmad et al., 1998). In the study of Naema et al. (2010)
was represented that the antiulcerogenic activity of celery extracts was evaluated in rats by the
HCL/ETOH method. Inhibition of gastric lesions by A. graveolens extracts was dose dependent
(Fazal et al., 2012). Wild celery is said to be useful in cases of hysteria, promoting restfulness
and sleep, and diffusing through the system a mild sustaining influence (Naema et al., 2010).
The potential of essential oil extracted from the seeds of celery evaluated as larvicidal and
repellent agent against dengue vector, Aedes aegypti. The investigations validated the significant
potential of essential oil as the probable agent for the control and management of rising
Ae.aegypti population (Tyagi et al., 2013). Celery apigenin relaxes rat isolated thoracic aorta by
reducing entry of calcium into the cell through voltage dependent and receptor-dependent
channels (Kumar et al., 2014). In the studywas shown that essential oil obtained from celery is
reported to have a calming effect on the central nervous system. Moreover, some of its
constituents showed antispasmodic, sedative and anticonvulsant actions. People with kidney
inflammation should refrain from excessive use of celery. In traditional medicine, the effects of
celery on abortion and the periods mentioned. Pregnant women should be careful in eating celery
seed because too high of its doses can stimulate the uterus (Kolarovic et al., 2009). Celery can
cause photo dermatitis and contact dermatitis. In a case-control study it was shown that it can
cause damage to the small intestine (Ashburn and staals., 1999). In 2002, Sausenthaler and
colleagues performed an experiment to evaluate the effect of maternal diet in pregnancy on
creating eczema and allergies in babies; in this study 2641 two years old children were
examined. The results showed that a high intake of celery (OR: 1.85; 95% Cl: 1.18, 2.89) during
the last 4 weeks of pregnancy may increase the risk of complications (Jones, 1950). In study of
Gadermaier et al. (2011) was shown that A. graveolens is one of the most important plant food
allergen sources in the adult Central European population. The most important celery allergens
are PR-10 protein (API g1), non-specific lipid transfer proteins- LTPI (API g2), pro-filing (API
g4) and flavor protein (API g5). API g2 and API g4 allergies are potentially dangerous for
allergic individuals because these allergens may induce an anaphylactic reaction (Sausenthaler et
al., 2002; Gadermaier et al., 2011). In a study Kooti et al. (2014) indicated that eating celery
alone in male rats is effective in reducing the number of newborns (Kooti et al., 2014). In
another study, this group demonstrated a certain dose of celery decreases the serum levels of T3
and T4 and increases TSH (Kooti et al., 2014).

2.9.2. Antimicrobial effects of A.graveolens

Essential oil and aqueous extract prepared from the aerial parts of A. graveolens were
tested to determine their antibacterial activity. Essential oil of A. graveolens was strongly
inhibitory against Escherichia coli and moderately inhibitory against Pseudomonas aeruginosa
and Staphylococcus aureus (Baananou et al., 2013) A.graveolens boiling water extract showed a
wide zone of inhibition E.coli growth in concentration of 5% (Naema et al., 2010). The
antimicrobial activity of the liquid carbon dioxide of A. graveolens were tested against E.coli,
Listeria monocytogenes, Citrobactor freundii, Hafni aalvei, Salmonella typhimurium, Bacillus
cereus, Enterococcus faecalis, Enterobactor aerogens, Staphylococcus aureus and Proteus
vulgaris. It was found that all the investigated leaf extracts were effective inhibitors of H. alvei,
S. aureus; E. coli, B. cereus and E. faecalis, C. freundii and P. vulgaris were resistant against
celery extracts isolated both from roots and leaves (Sipailien et al., 2003). The hexane,
chloroform, methanol and water extract showed various degrees of inhibition of bacterial growth
when tested against Escherichia coli, Bacillus subtilis, Shigella flexneri, Staphylococcus aureus,
Pseudomonas aeruginosa and Salmonella typhi, ranging between 6 to 14 mm (zone of
inhibition). The methanolic extract of all the examined celery showed positive antibacterial
activity all strains.