Volume 4, Issue 8, August – 2019 International Journal of Innovative Science and Research Technology

ISSN No:-2456-2165

Nutritional Studies and Antimicrobial Activities of

Jatropha tanjorensis Leaves Extracts against
Escherichia coli Isolates
1
Ebana, R. U. B., 2Andy, I. E., 1Edet, U. O., 1Anosike, I. K. 2Mbim, E. N. & 1Benjamin, A. U.
1
Obong University, Faculty of Natural and Applied Sciences, Department of Microbiology, Obong Ntak, Etim Ekpo,
Akwa Ibom State, Nigeria.
2
Department of Microbiology, Faculty of Biological Sciences, University of Calabar, PMB 1115, Calabar,
Cross River State, Nigeria.

Abstract:- This study was aimed at evaluating the I. INTRODUCTION

extract of Jatropha tanjorensis leaves for bioactive
components, proximate composition and antimicrobial The discovery of the drug Penicillin by Alexander
activity. The collection of the plant leaves, processing of Flemmings from the Fungus: Penicillium notatum was
the leaves and extracts were all done using standard timely since it significantly reduced the mortality and
methodologies. Qualitative screening and gas morbidity of infectious diseases globally.01 Following its
chromatography (GC-MS) were used to screen for discovery, several other antibiotics have successfully been
phytochemicals. Proximate composition, vitamins, discovered and are currently in the global market where they
minerals and anti-nutrients were all carried out using have been mass produced. Besides the significant global
standard techniques. Standard microbiological reduction in morbidities and mortalities associated with
techniques were used to isolate, characterize and identify antibiotics, they have been found to increase global life
Escherichia coli isolates from dumpsite soil samples. The expectancy by atleast two decades.01,02 Studies revealed that
standard disc method was used to assay the antibiotics the advent of antibiotics paved way for advances in surgery,
sensitivity of the test antibiotics and that of the extracts. cancer treatment, and organ transplant, as well as influenced
The proximate nutrients and their respective amounts in the expansion of livestock production amongst other uses. 02,
03
g/100g per dry matter were moisture (80.23±1.53),
protein (5.01±0.17), ash (2.91±0.31), crude fiber
(3.21±0.17) fat (1.94±0.11) and carbohydrate Sadly, the very microorganisms and their associated
(86.93±1.47). Mineral analysis revealed the presence of infections that were effectively controlled and treated by
sodium, potassium, calcium, magnesium, zinc, copper, these antibiotics are now becoming resistant to them. The
and iron. Furthermore, vitamin analysis showed the challenge of antibiotics resistance has now assumed a global
presence of vitamins A, B, C and E. Anti-nutrient dimension as resistance to antibiotics has been reported in
analysis revealed the presence of permissible levels of all continents of the world.03 Even more worrisome, is the
hydrocyanic acid, soluble oxalate, total oxalate, and fact that resistance has completely outpaced the search and
phytate. Phytochemical analysis via GC-MS revealed a development of newer and more effective antibiotics.04
total of 16 compounds belonging to five different Furthermore, microbes have the evolutionary capacity to
categories of phytochemical namely: alkanes and acquire and spread resistance genes even via plasmids and
associated hydrocarbons, organic acid and fatty acid, transposons. 05, 06 Studies revealed that these genes abound
flavonoids, alkaloids and amines. Organic and fatty in diverse environments including in the hospital samples,
acids had the highest concentration of 80.52% followed effluents and as well as other environments. 05, 07
by flavonoids with a concentration of 18.44%. Alkaloids,
amines and alkanes and associated hydrocarbons had Despite advances in research development especially
concentrations of 0.18, 0.692 and 0.113, respectively. The in the areas of medicine, lead compound discovery,
test antibiotics showed zones that ranged from 12.00 to molecular docking and combinatorial chemistry, only few
23.00mm. The highest and least zones of inhibition were antibiotics have made it to the market since the 1970s.04
28.00 and 10.00 mm for aqueous extract while it was Given the gravity of this scourge and the global paucity of
17.30 and 10.00 mm for ethanolic extract, respectively. newer antibiotics to stem the tide of antibiotics resistance,
Jatropha tanjorensis could be rich source of nutrients, alternative medicine has been one of the major interests of
vitamins, essential minerals as well as phytochemicals researchers currently. 08
with effective antimicrobial potential against Escherichia
coli. One of such alternatives is the use of ethno-medicinal
plants. Since time immemorial, medicinal plants and its
Keywords:- Nutrition, Phytochemicals, Jatropha tanjorensis products have been utilized traditionally in the treatment of
Leaf Extract, Escherichia coli, Antimicrobial Susceptibility. common ailments.08,09 These plants and their products are
currently gaining more attention because they have been
reported to possess phytochemicals which exhibit noble
antimicrobial properties against even the multidrug-resistant

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Volume 4, Issue 8, August – 2019 International Journal of Innovative Science and Research Technology
ISSN No:-2456-2165
microorganisms with little or no side effects to the host. 09, 10  Processing of Leaves Sample
One of such medicinal plants is Jatropha tanjorensis which The collected leaves were processed following
belongs to the family Euphorbiacea and grows very well in procedures previously described.13 Briefly, the freshly
the tropics.11 collected leaves were first washed and allowed to air dried.
There were then chopped into tiny pieces using a clean
Jatropha tanjorensis commonly called “When hospital stainless kitchen knife and dried using an electric oven
is too far” or “When the doctor is not near” in Cross River maintained at 60oC for atleast 1 hour. Afterwards, the dried
and Akwa Ibom States is used as a source of medicine and leaves were then grinded into powder using a clean Mortar
as a vegetable substitute for fluted pumpkin in these areas. and pestle and the powdered leaves were then stored in a
In Nigeria, this plant has been utilized by the locals in the clean, dry sample bottle and kept in a dry place till required.
treatment of malaria and hypertension and has been reported
to contain alkaloids, flavonoids, tannins, cardiac glycosides,  Preparation of Extracts
saponins, and anthraquinones.12 However, there is a dearth Both aqueous and ethanolic extracts were prepared as
of information on the nutritional content and antimicrobial previously reported.09 Briefly, 20g each of the powdered
potential of this plant against microorganisms including the leaves was weighed out and dissolved separately into beaker
multidrug-resistant isolates. Therefore, this study was aimed containing 200ml each of distilled water and freshly
at evaluating the nutritional and antimicrobial potentials of prepared 75% ethanol. The mixtures were then stirred gently
Jatropha tanjorensis extracts against Escherichia coli to ensure proper mixing. The flasks were sealed and
isolates. incubated for 24 hours. Following incubation, the content of
the flasks were then filtered into separate conical flasks
II. MATERIALS AND METHODS using Whitman No.1 filter papers neatly folded into glass
funnels. The filtrates from both flasks were allowed time to
A. Materials: filter properly. At the end of the filtration, the residues were
discarded while some portions of both filtrates were heated
 Soil Samples and Plant Parts using an electric water bath maintained at 1000C till slurries
Soil samples from a dumpsite and plant parts of of both filtrates were obtained. The slurries were then stored
Jatropha tanjorensis were obtained and utilized for this away at room temperature till further use.
work.
 Qualitative Screening for Phytochemical
 Antibiotics discs employed were commercially sourced Qualitatively, the remaining filtrates (extracts) were
for and included; gentamycin (CN) (10μg), ofloxacin screened for crude phytochemicals using slightly modified
(OFX) (10μg), ceporex (CEP) (10μg), amplicin (PN) methods previously described.10, 14, 15, 16, 17 The
(10μg),ciprofloxacin (CPX) (10μg), reflacine (PEF) phytochemicals screened for were alkaloids, tannins,
(10μg), amoxicillin+clavulanic acid (AU) (30μg), saponins, flavonoids, glycosides and polyphenol.
streptomycin (S) (30μg), sulfamethoxazole+
trimethoprim (SXT) (30μg), nalidixic acid (NA) (30μg),  Screening for Alkaloids
(Optun laboratories, Nigeria). Briefly, 5ml of 1 % HCl was added to exactly 2ml of
each extract and gently stirred after which they were placed
 Media and Reagents employed included Eosin on steam bath. Then, a few drops of Mayer’s reagent and
Methylene blue medium, Muller Hinton Agar, Nutrient 1ml of Dragendorff’s reagent were added to the mixtures.
agar, Citrate agar, Urease agar, Grams reagents, indole, The presence of a precipitate with either of these reagents
H2O2, oxidase reagent, methyl red, alcohol, among others. was taken as a positive result.

 Other Materials  Screening for Tannins

Other materials employed for this study included To the extracts, exactly 2mls of 5% ferric chloride was
incubator, GC-MS, rotary evaporator, petri dishes, among added to 1ml of each of the extracts. The formation of a
others. dark blue or greenish black coloration on addition of ferric
chloride indicated the presence of tannins.
B. Methodology:
 Screening for Saponins
 Collection of Plant Parts To each of the extracts, 2ml of distilled water was
The leaves of the plant employed for this study were added and the resulting aqueous mixtures shaken vigorously
freshly collected from Obong University Community with for about 5 – 10minutes. The formation of a stable and
the help of locals. The plant parts were identified persistent froth of about 1cm in length was taken as positive
scientifically at the Botany Department, University of result.
Calabar, Cross River State as Jatropha tanjorensis. The
freshly collected leaves were immediately taken to the
Microbiology laboratory for further analysis.

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Volume 4, Issue 8, August – 2019 International Journal of Innovative Science and Research Technology
ISSN No:-2456-2165
 Screening for Flavonoids  Collection and Processing of Soil Samples
To exactly 2ml of both extracts, a few pieces of This was done following methods previously
aluminum metal and concentrated HCl were added. described.24 Briefly, soil samples from three different
Following addition, the formation of an orange or red or locations in Uyo Village dumpsite were collected into sterile
crimson or magnetic coloration was regarded as a positive nylon bags and transported immediately to the laboratory
result for flavonoids. for further analysis. Approximately, 1g of each soil sample
was introduced into test tubes containing 10mL of distilled
 Screening for Glycosides water. Then, ten-fold serial dilutions were carried out on the
Exactly 2ml of chloroform with 10% ammonia soil samples.
solution was added to 2ml of each of the extracts. Formation
of a pink color was regarded as positive test for glycosides.  Isolation and Characterization of E. Coli Isolates
This was performed following methods previously
 Screening for Polyphenol described.25 Briefly, the 10-3 dilutions were then plated out
Briefly, 2ml of both extracts were treated with 5ml of on freshly prepared Eosin Methylene Blue (EMB) Agar and
distilled water and heated for 30min in a water bath. After incubated at room temperature overnight. Following
heating, 1ml of 1% potassium ferrocyanide solution was incubation, the plates were examined for growth. Following
added to it. The formation of green-blue coloration indicated purification of test isolates, they were characterized
the presence of polyphenol. following methods described previously.26

 GC-MS Analysis  Antimicrobial Susceptibility Testing

This was done at Mifor Consult Laboratory located at
Marian Calabar, Cross River State. Briefly, exactly 10g of  Antibiotics Susceptibility against Test Isolates
the powdered leaves was weighed and introduced into an This was performed using the Agar Disc diffusion
extraction thimble of a soxhlet extractor. Then, 100ml of method previously described.27, 28,29 Briefly, exactly, 3-5
methanol was measured and poured into the round bottom colonies of each test organism was selected using a sterile
flask attached to the soxhlet extractor. This was refluxed for inoculating loop and suspended in saline after which the
at least three times. The extract was concentrated to about inoculum was adjusted to a turbidity equivalent to a 0.5
2mL using a rotary evaporator after which it was then McFarland standard (corresponds to approximately 1.5 x
transferred into a Teflon screw-cap vial and appropriately 108 CFU/ml). The suspension was then vortexed to make
labeled. Subsequently, the extract was cleaned up with 3g of sure it was well-mixed. Then a fresh, sterile cotton-tipped
anhydrous sodium sulfate in a well packed column, swab was dipped into the suspension, the excess liquid from
conditioned with methanol to form slurry after which it was the swab removed by pressing it against the side of the tube.
injected into GC-MS for qualitative and quantitative Subsequently, the swab was inoculated unto a plate
analyses. containing freshly prepared Muller Hinton Agar (MHA)
starting at the top, the surface was inoculated with the swab
 Proximate Composition and Mineral Analysis covering the entire plate by spreading back and forth from
The leaves were also evaluated for proximate nutrients edge to edge, rotating the plate approximately 60° and
and mineral elements. These were all done following the repeating the swabbing procedure thrice ensuring that the
methods of the Official Analytical Chemists (A.O.A.C). The entire surface was properly covered. Then the disc
analysis included moisture content, ash content, crude containing the antimicrobial agents was applied using a
protein, lipid, crude fiber, carbohydrate, vitamins, calorific sterile pair of forceps within 15 minutes of inoculating the
value and mineral elements (Ca, Fe, Mg, Zn, Cu, Na, K, and MHA plate and then pressed down firmly to ensure firm,
P). leveled contact with the agar. The plate was inverted and
incubated at 350C for 16-18 hours. After incubation, the
 Vitamin Analysis clear zone around each disc was measured and interpreted
Vitamins A, B, C and E were also evaluated for in the using procedures already described (CLSI, 2014). Isolates
leaves of our study plant. These were done using methods showing resistance to atleast two antibiotics were regarded
previously described.18, 19, 20, 21 as multidrug-resistant E. coli isolates. This procedure was
carried out on all bacterial isolates.
 Anti -Nutrients
The anti-nutrients examined were hydrocyanic acid,
soluble oxalate, phytate and total oxalate. These were
estimated using procedures previously described.22, 23, 24

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 Antimicrobial Susceptibility Test of Extracts against Test present in excess while the remaining phytochemicals were
Isolates just present but not in excess as presented in Table 1.
This was carried out using agar disk diffusion methods Similarly, the results of the GC-MS as presented in Tables 2
previously described.27,28 Briefly, the test isolates were first and 3 as well as in Fig. 1 further confirm the presence of
inoculated on nutrient broth and allowed to stand overnight. different bioactive compounds in the leaf extracts. The 16
After overnight incubation, 0.1ml (a loopful) of each test compounds observed via GC-MS belong to five different
isolate was inoculated unto plates containing freshly categories of phytochemicals namely alkanes and associated
prepared MHA. Then, a manual borer was used to obtain hydrocarbons, organic acid and fatty acid, flavonoids,
discs of about 6mm from Whitman filter paper No1. The alkaloids and amines. Amongst these categories of
discs were sterilized in an oven for 30minutes at 60oC. phytochemicals, the most diverse was the organic and fatty
Following sterilization, these discs were soaked in different acid family which had a total of 13 compounds namely
concentrations (0.1, 0.3, 0.5mg/ml) of each extract for thiazole, 2-amino-4-(p-methoxyphenyl), 1,16-
15minutes and were placed gently on the Mueller Hinton Cyclocorynan-17-oic acid, 19,20-didehydro, methyl ester,
Agar (MHA) plates pre-inoculated with the test organisms. 2-(Acridin-9-ylamino)-3-(3H-imidazol-4-yl)-propionic acid,
This was done in duplicates for each isolate and 2-Hydroxy-2-methylbutyric acid, Butanoic acid, 4-[(2,4-
concentration. The plates were then incubated at 37oC dichlorophenyl)oxy]-, butyl ester, 2-propenoic acid, 2-
overnight. Following incubation, zone diameter were methyl-, 3-[(phenylsulfonyl)amino]phenyl ester, Heptyl
obtained and compared with appropriate standards. isobutyl carbonate, Octadecanoic acid, 2-hydroxy-1-
(hydroxymethyl)ethyl ester, n-Hexadecanoic acid, n-
 Statistical Analysis Hexadecanoic acid, Octadecanoic acid, Propanoic acid, 2-
Replicate readings obtained from this study were (3-acetoxy-4,4,14-trimethylandrost-8-en-17-yl), and 3-
analyzed using Analysis of Variance (ANOVA) at 95% pyridinecarboxylic acid, 2,7,10-tris(acetyloxy)-
confidence level using Microsoft Excel version 2007. 1,1a,2,3,4,6,7,10,11,11a-decahydro-1,1,3,6,9-pentamethyl-
Results analyzed using mean ± Standard deviation. 4-ox. In the organic and fatty acid family of phytochemicals,
thiazole, 2-amino-4-(p-methoxyphenyl) was the most
III. RESULTS abundant with a concentration of 41.04%. Flavonoids and
amines had 3 compounds each while alkaloids, and the
 Phytochemical Analysis alkanes and associated hydrocarbons had 2 compounds
The leaves when subjected to phytochemical screening each. As shown in Fig. 2, the combined concentration of
using crude means as well as via GC-MS revealed that the various phytochemicals revealed that the organic and
Jatropha tanjorensis leaves contain various phytochemicals. fatty acids had the highest abundance of 80.52% followed
The results of the crude qualitative screening revealed the by flavonoids (18.44%). Alkaloids, amines, alkanes and
presence of alkaloids, tannins, flavonoids, saponins, associated hydrocarbons had concentrations of 0.18, 0.692
glycosides, glycosides and polyphenol. Among the and 0.113, respectively.
phytochemicals, alkaloids, flavonoids and polyphenol were

Phytochemicals Aqueous extract Ethanolic extract

Alkaloids ++ ++
Tannins + +
Flavonoids ++ ++
Saponins + +
Glycosides + +
Polyphenol ++ ++
Table 1:- Qualitative screening for phytochemicals in Jatropha tanjorensis
Key:- + = present and ++ = present in excess.

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Volume 4, Issue 8, August – 2019 International Journal of Innovative Science and Research Technology
ISSN No:-2456-2165
S/N Phytochemical Compounds Concentration (%)
groups
1 Alkanes and Cyclohexane, 1,1'-[4-(3-cyclohexylpropyl)-1,7-heptanediyl]bis- 0.053
2 associated 2-Isopentyl-N-(1-naphthyl)-1-cyclopropanecarboxamide 0.060
hydrocarbons
3 Organic acid and Thiazole, 2-amino-4-(p-methoxyphenyl)- 41.040
4 Fatty acid 1,16-Cyclocorynan-17-oic acid, 19,20-didehydro-, methyl ester, 7.281
5 2-(Acridin-9-ylamino)-3-(3H-imidazol-4-yl)-propionic acid 0.061
6 2-Hydroxy-2-methylbutyric acid 0.361

7 Butanoic acid, 4-[(2,4-dichlorophenyl)oxy]-, butyl ester 0.113

8 2-propenoic acid, 2-methyl-, 3-[(phenylsulfonyl)amino]phenyl ester 0.057
9 Heptyl isobutyl carbonate 0.057
10 Octadecanoic acid, 2-hydroxy-1-(hydroxymethyl)ethyl ester 0.056
11 n-Hexadecanoic acid 0.059
12 n-Hexadecanoic acid 9.476
13 Octadecanoic acid 10.474
14 Propanoic acid, 2-(3-acetoxy-4,4,14-trimethylandrost-8-en-17-yl)- 8.138
15 3-Pyridinecarboxylic acid, 2,7,10-tris(acetyloxy)-1,1a,2,3,4,6,7,10,11,11a- 3.347
decahydro-1,1,3,6,9-pentamethyl-4-ox
15 Flavonoids 2-Pentanone, 4-hydroxy-4-methyl- 18.380
17 4''-Dehydroxy-2'',3',3'',4',5,6'',7-hepta-O-methylisoorientin 0.062
18 1-Dimethyl(chloromethyl)silyloxydecane 0.055
19 Alkaloid 2,16-Dihydro-iboxyphylline 0.072
20 3,5-Dimethyl-1-dodecylpyrazole 0.105
21 Amine 5-benzoxazolamine, 2-[4-[2-[4-(2-benzoxazolyl)phenyl]ethyl]phenyl]- 0.070
22 Heptadecanenitrile, 6-aza-2-(4-hydroxy-3-methoxyphenyl)-2-propyl 0.561
23 Cyclohexanecarbonitrile, 4-[4'-(nonyloxy)[1,1'-biphenyl]-4-yl]-1-octyl- 0.061
Table 2:- GC-MS analysis of Jatropha tanjorensis leaves

Fig 1:- Chromatogram of the various phytochemicals in the leave extract using GCMS

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Table 3:- Peaks properties of the various phytochemicals

Fig 2:- Combined concentrations of the various phytochemical categories

 Nutrients and Anti-Nutrient Analyses ash, crude fiber and fat. The least abundant nutrient was fat.
The results of the nutrient analyses are presented in The leaves employed in this study were also analyzed for
Tables 4-7 as mean ± SD (standard deviation). The results the presence of mineral elements and the results indicate the
of the proximate composition presented in Table 4 indicate presence of sodium, potassium, calcium, magnesium, zinc,
that the leaves of the study were rich in nutrients. The copper, and iron. The most abundant mineral element was
proximate nutrients and their respective amounts in g/100g magnesium which had an abundance of 19.23±1.71
per dry matter were moisture (80.23±1.53), protein mg/100g per dry matter. The second most abundant mineral
(5.01±0.17), ash (2.91±0.31), crude fiber (3.21±0.17) fat element was calcium with an abundance of 18.64±1.33
(1.94±0.11) and carbohydrate (86.93±1.47). It can be seen mg/100g per dry matter. These were followed by potassium
that the carbohydrate was the most abundant nutrient in the and sodium that had abundance of 16.22±1.57 and
study plant part. This was followed by moisture, protein,

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14.14±2.14 mg/100g per dry matter. The least abundant Anti-nutrients Amount (mg/100mL)
element was copper with a value of 1.21±0.10 ±0.10. Hydrocyanic acid 4.07±0.14a
Soluble oxalate 8.23±0.17
Table 5 shows the presence of vitamins. From the Total oxalate 9.27±2.11
results, the leaves had vitamins A, B, C and E in varying Phytate 7.18±0.10
amounts. The most abundant vitamin was B with a mean Table 7:- Anti-nutrients levels in the leaves
value of 102.81 mg/100mL followed by vitamin A with a Key:- aSuperscript represents significant Mean±SD (p <
mean value of 81.09 mg/100mL. Vitamins C and E had 0.05) following analysis of variance.
mean values of 6.13 and 3.05 mg/100mL, respectively. In
addition to the nutrient analysis presented in Tables 4-7, the  Microbiological Analysis
leaves of the study plant were also evaluated for anti- A total of ten (10) test isolates were recovered from
nutrients. The anti-nutrients evaluated were hydrocyanic soil samples and characterized as presented in Table 8. In
acid, soluble oxalate, total oxalate, and phytate. The results addition to showing a green metallic sheen on EMB plates,
indicate that total oxalate was the most abundant anti- the isolates were subjected to a battery of various
nutrients with a mean value of 9.27 followed by soluble biochemical tests. The isolates were positive for catalase,
oxalate with a mean value of 8.23. The least abundant was indole and H2S/Gas, in addition to appearing as rods and
hydrocyanic acid which had a mean value of 4.07. showing motility. However, they were negative for oxidase,
MR-VP, citrate and urease tests.
Proximate nutrients Amount (g/100g per dry
matter)  Antimicrobial Analysis of Antibiotics and Extracts
Moisture 80.23±1.53a against Test Isolates
Protein 5.01±0.17 Table 9 shows the antibiotics sensitivity profile against
Ash 2.91±0.31 test isolates. The isolates showed varying zones of
Crude fiber 3.21±0.17 inhibition against the various antibiotics. For Ciprofloxacin,
Fat 1.94±0.11 the zones ranged from 12.00-21.50 mm, 8.50-20.50mm for
Carbohydrate 86.93±1.47 OFX and 15.50-21.10mm for NA. CN and PEF recorded
Table 4:- Proximate composition of the Jatropha zones of inhibition ranging from 14.00-23.00mm and 12.50-
tanjorensis 20.00, respectively. 9.50-21.20mm and 10.50-19.50mm
Key:- aSuperscript represents significant Mean±SD (p < were the zones of inhibition recorded for AU and CPX,
0.05) following analysis of variance respectively. For SXT, S and PN, the zones were 13.50-
23.00, 13.20-22.00mm and 12.00-21.50 mm, respectively.
Elements Amount (mg/100g per dry matter)
Sodium 14.14±2.14a Similarly, Tables 10 and 11 show the antimicrobial
Potassium 16.22±1.57 activities of the aqueous and ethanolic extracts of our study
Calcium 18.64±1.33 plant, against the test isolates. Generally, as the
Magnesium 19.23±1.71 concentration of the extract increased, there was a gradual
Zinc 3.64±0.17 increase in the zone inhibition. Isolate 1 showed no
Copper 1.21±0.10 susceptibility to the aqueous extracts at concentration of 0.1
Iron 3.80±0.20 mg/ml as shown in Table 10. Isolate 7 also showed resistant
at 0.1 and 0.2 mg/ml. The highest zone of inhibition
Table 5:- Mineral levels in the leaves
obtained was 28.00mm against isolate 10 while the least
Key:- aSuperscript represents significant Mean±SD (p <
was 10.00mm. Furthermore, only isolate 10 showed
0.05) following analysis of variance.
complete resistances to all the concentrations of the
ethanolic extract used in our study as shown in Table 11.
Vitamins Amount (mg/100mL)
The highest susceptibility observed was 17.30 mm while the
A 81.09±2.14a least was 10.00 against isolate 2 and 1, respectively. Apart
B 102.81±2.89 from isolate 10, all other isolates gave zones that increased
C 6.13±1.41 with increasing concentrations of the ethanolic extract.
E 3.05±0.17
Table 6:- Estimation of vitamin contents
Key: - aSuperscript represents significant Mean±SD (p <
0.05) following analysis of variance.

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Isolates Grams Shape Motility Catalase Oxidase Methyl Voges Indole Citrate Urease H2S/Gas
Reaction Red Proskauer
Isolate - Rods Yes NA - - - + - - -/+
1
Isolate - Rods Yes NA - - - + - - -/+
2
Isolate - Rods Yes NA - - - + - - -/+
3
Isolate - Rods Yes NA - - - + - - -/+
4
Isolate - Rods Yes NA - - - + - - -/+
5
Isolate - Rods Yes NA - - - + - - -/+
6
Isolate - Rods Yes NA - - - + - - -/+
7
Isolate - Rods Yes NA - - - + - - -/+
8
Isolate - Rods Yes NA - - - + - - -/+
9
Isolate - Rods Yes NA - - - + - - -/+
10
Table 8:- Biochemical characterization of the microbial isolates
Keys: + = Positive, - = Negative, NA=Not applicable

E. coli isolates CEP OFX NA PEF CN AU CPX SXT S PN

Isolate 1 18.00 17.20 20.30 12.50 14.00 15.20 17.50 14.30 15.00 16.00
Isolate 2 - - - - - - - - - 12.00
Isolate 3 16.00 18.30 15.50 13.00 17.30 18.30 15.50 14.30 13.20 16.00
Isolate 4 12.00 16.50 17.30 14.00 15.50 16.20 17.30 16.30 22.00 17.20
Isolate 5 19.00 20.50 18.10 18.00 23.00 21.20 19.50 23.00 22.00 21.50
Isolate 6 20.00 8.50 19.30 18.50 18.30 9.50 10.50 13.50 17.30 10.80
Isolate 7 21.50 15.30 20.50 20.00 22.00 21.50 19.50 18.00 15.00 12.00
Isolate 8 - - - - - - - - - -
Isolate 9 15.30 13.20 21.10 20.00 17.50 18.00 19.00 14.50 16.00 17.30
Isolate 10 14.20 16.40 15.20 19.00 22.00 19.30 18.30 21.00 19.00 18.50
Table 9:- Antibiotics sensitivity of the various E. coli isolates
Key: - = No zone of inhibition, Key: - = No zone of inhibition, CPX, SXT, S, PN, CEP, OFX, NA, PEF, CN, and AU
respectively.

Isolates Concentration (mg/ml)

0.1 0.2 0.3
1 - 15.00 16.00
2 13.10 14.00 15.20
3 12.20 14.00 15.50
4 14.30 15.00 16.70
5 11.00 12.99 14.78
6 11.19 12.00 13.89
7 - - 10.00
8 12.11 13.28 15.56
9 12.50 13.10 14.00
10 - 10.00 28.00
Table 10:-Antimicrobial sensitivity (mm) of aqueous extracts of Jatropha tanjorensis against E. coli
Key: - = No zone of inhibition.

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Volume 4, Issue 8, August – 2019 International Journal of Innovative Science and Research Technology
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Isolates Concentration (mg/ml)
0.1 0.2 0.3
1 10.00 11.00 12.00
2 11.50 12.00 17.30
3 12.50 13.00 14.60
4 13.50 16.40 20.00
5 12.50 15.00 17.00
6 11.10 12.00 13.90
7 12.00 14.11 15.00
8 12.00 12.50 13.00
9 13.00 13.90 14.00
10 - - -
Table 11:- Antimicrobial sensitivity (mm) of ethanolic extracts of Jatropha tanjorensis against test isolates
Key: - = No zone of inhibition.

IV. DISCUSSION vegetable was low in minerals analyzed in this study. The
highest was magnesium with a value of 19.23 and this was
Jatropha tanjorensis is popular plant that is well used followed by calcium with a value of 18.64 mg/100g per dry
as a leafy vegetable and as a medicinal plant. Analysis of matter. Compared to earlier findings, all the observed
the plant parts for nutrients indicate that the leaves have a minerals were slightly higher.31 However; minerals
very high moisture content of 80.23%. This slightly higher observed in this study were somewhat lower than previously
than the 78.77% previously reported.30 The moisture reported.11
contents of various edible leafy vegetables such as cassava,
cabbage, cowpea and sweet potato have been observed to The presence of essential vitamins such as A, B, C and
range from 79.00-85.00 with cabbage having the least and E in this study is a further indication of the nutritional
cowpea with the highest amounts of moisture.30 Compared capacity of the plant. The level of vitamins in this study was
to other common tropical edible leafy vegetables including agreeable to those of mushroom in our earlier study.24 Since
Lasianthera africana and Dennettia tripetala, which gave Jatropha tanjorensis is widely eaten in the Southern part of
moisture contents of 78.95 and 76.62 %, respectively, the the country, the anti-nutrient levels present in the leaves
moisture content of our sample plant was only slightly were consistent with anti-nutrients in other widely eaten
higher.09 Similarly, the carbohydrate content was observed leafy vegetables such as Lasianthera Africana, Dennettia
to be 86.93 g/100g per dry matter. This is somewhat higher tripetala , cassava, water leaf, bitter leaf, Ugu leaf and green
than the carbohydrate contents of Lasianthera africana vegetable. 09, 32
(73.04g/100g) and Dennettia tripetala (49.70g/100g).09
Jatropha tanjorensis has been adjudged to have atleast
The values for the protein, ash, crude fiber and fat eight benefits ranging from increasing men vitality
were observed to be 5.01, 2.91, 3.21 and 1.94, respectively. (increasing blood circulation and size of the penis during
The 5.01% protein content observed in this study although sex), placing slices of the leaves on the forehead of children
lower than the values of moisture and that of carbohydrate cures fever, and treatment of vaginal discharge.33 Other
recorded, it is slightly higher than the 2.04% previously beneficial uses include curing skin problems and
reported from Benin City, Nigeria. Generally, this low rheumatism, relieve for inflammation, smoothing defecation
protein value may be indicative of the fact that leafy and curing of the scratch and skin blisters.31, 33
vegetables are usually low in protein.30 The protein content
of this leaf extract has also been observed similar to other The medicinal properties of all ethno-medicinal plants
leafy vegetables such as cassava, cowpea and sweet potato have been associated with the presence of phytochemicals in
which have reported protein contents of 6.00, 4.70 and 4.60, various parts of the plants.04, 31 Phytochemicals including
respectively.30 The protein content observed in this study is alkaloids, tannins, flavonoids, saponins, glycosides and
consistent with earlier findings where protein content was polyphenols observed in this study are similar to those
reported to be 4.75 %. 11 earlier reported. 31, 34 A total of 23 individual
phytochemicals classified into five categories namely
The low amount of fiber in this study correlates with alkanes and associated hydrocarbons, organic acid and fatty
those of earlier studies and this nutrient is usually an acid, flavonoids, alkaloids and amines were quantified. The
indication of low mineral content.11, 30 The presence of little organic and fatty acid class were the most abundant with a
fat observed in this study agrees with previous findings.11 total of 13 compounds and a combined total of 80.52%.
This makes the leafy vegetable a good option for those that This was followed by flavonoids, amines and alkaloids with
are dieting and/or diabetic. Studies revealed that the combined concentrations of 18.44, 0.692 and 0.18%,
presence of low fiber in leafy vegetables is always an respectively. This observation is contrary to an earlier
indication of low mineral content.09,30 In line with this report.31 Flavonoids is a diverse group of compounds have
assertion, it is therefore not surprising that our leafy

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Volume 4, Issue 8, August – 2019 International Journal of Innovative Science and Research Technology
ISSN No:-2456-2165
been shown to possess anti-oxidant and anti-inflammatory [6]. Mbim, E. N., Mboto, C. I., Edet, U. O., Umego, C. F.,
properties in humans. George, U. E. &Temidayo, I.. Comparative Evaluation
of Methicillin-resistant Staphylococcus aureus
The antimicrobial susceptibility profile of the plant (MRSA) Isolates from Hospital and Community
extracts revealed consistent and increasing inhibitions Settings in Nigeria. Journal of Advances in
against test isolates as the concentrations increased from Microbiology, 2017, 4(4), 1-9.
0.10-0.30mg/ml and none of the isolates showed absolute [7]. George U. E., Mboto, C. I., Mbim, E. N., Edet, U. O.,
resistance to the extracts. However, isolate 10 showed mild Umego C. F. & Sunday, H.. Comparative Evaluation
resistances to all concentration of the ethanolic extract. The of the Effect of Annona muricata (Graviola) Leaves
highest and least inhibitions recorded were 28.00 and Extracts and Cefoxitin on Staphylococcus aureus.
10.00mm against isolates 10 and 7 at a concentration of 0.30 Asian Journal of Biology 2017, 3(3): 1-7.
mg/ml for aqueous extract and 10.00-20.00mm for ethanolic [8]. Kållberg C, Årdal C, Salvesen Blix H, Klein E, M.
extract. Similarly, test isolates showed complete resistance Martinez E, Lindbæk M, (2018) Introduction and
to the antibiotics used and these were isolates 2 and 8. These geographic availability of new antibiotics approved
two isolates were however, sensitive to plant extracts and between 1999 and 2014. PLoS ONE 13(10):
gave zones of 13.10-15.20 and 12.11-15.56mm, e0205166.
respectively. The highest zone of inhibition for the https://doi.org/10.1371/journal.pone.0205166.
antibiotics was 23.00mm against isolate 5 with gentamycin [9]. Ebana, R. U. B., Asamudo, N. U., Etok, C. A., Edet U.
and sulfamethoxazole+ trimethoprim. The aqueous extract O. & Onyebuisi, C. S. Phytochemical screening,
was earlier reported to inhibit the growth of both Gram nutrient analysis and antimicrobial activity of the
negative and positive bacteria (Staphylococcus aureus and leaves of Lasianthera africana and Dennettia tripetala
E. coli) with inhibition zones of 16.00 and 11.30 mm, on clinical isolates. Journal of Advances in Biology
respectively.31 and Biotechnology. 2016, 8(4):1-9.
[10]. Ebana, R. U. B., Etok, C. A. & Edet, U. O. Chemical
V. CONCLUSION Composition and Antimicrobial Analysis of the Pods
and Seeds of Cola rostada and Cola nitida. Intl. J.
This study shows that Jatropha tanjorensis is indeed a App. Studies. 2015, 10 (4):1245-1249.
rich source of nutrients with features comparable to other [11]. Egbon, E. E., Ize-Iyamu, O. K., Okojie, V. U., Egbon,
known edible leafy vegetables. Further evaluations revealed I. E. Proximate and Mineral Composition of Jathropha
that they are a rich source of vitamins and essential minerals tanjorensis. Chemical and Process Engineering
as well as low in anti-nutrients. This plant has been Research, 2013, 17, 21-24.
observed to possess phytochemicals and possess effective [12]. Falodun, A., Udu-Cosi, A. A., Erharuyi, O., Imieje,
antimicrobial potential comparable to those of routinely V., Falodun, E. J., Agbolahor, O., Hamann, M.
used antibiotics against Escherichia coli in our study. Jatropha tanjorensis – Review of phytochemistry,
pharmacology and pharmacotherapy. Journal
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