See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/331546079

Evaluation of Antimicrobial Activity of the Aquatic Extracts of Zea Mays,

Cranberries and Raisins against Bacterial isolates from Urinary Tract Infection
in Babylon Province, Iraq

Article  in  Indian Journal of Public Health Research and Development · March 2019

DOI: 10.5958/0976-5506.2019.00402.9

CITATION READS
1 224

3 authors:

Assist.Prof.Dr. Mays Hadi Jebur Nada Khazal K. Hindi

University of Babylon Nursing of college\University of Babylon
20 PUBLICATIONS   24 CITATIONS    105 PUBLICATIONS   356 CITATIONS   

SEE PROFILE SEE PROFILE

Mohammed Abdul-Hassan Jabarah Al-Zobaidy

University of Baghdad
20 PUBLICATIONS   28 CITATIONS   

SEE PROFILE

Some of the authors of this publication are also working on these related projects:

microbiology researches View project

Psychological support of COVID-19 patients. View project

All content following this page was uploaded by Assist.Prof.Dr. Mays Hadi Jebur on 17 May 2019.

The user has requested enhancement of the downloaded file.

 DOI Number: 10.5958/0976-5506.2019.00402.9

Evaluation of Antimicrobial Activity of the Aquatic Extracts of

Zea Mays, Cranberries and Raisins against Bacterial isolates
from Urinary Tract Infection in Babylon Province, Iraq

Mohammed Abdul-Hassan AL-Zobaidy1, Mays Hadi Jebur2, Nada Khazal Kadhim Hindi3
1
Department of Basic Medical Sciences/ College of Nursing/ University of AL-Qadisiyah, Iraq, 2Basic and Medical
science Department, College of Nursing, University of Babylon, 3Basic and Medical science Department, College
of Nursing, University of Babylon

ABSTRACT

Background: Urinary tract infection costs hundreds millions of dollars every year. In addition, causative
bacterial species are diverse and some of them are multi-drug resistant. Objectives: evaluating the effects
of aqueous extracts of Zea Mays, Cranberry and Raisins on growth, adherence and biofilm formation of
bacterial uropathogens isolated from clinical urinary tract infections. Method: The extracts of Zea Mays,
Cranberry and Raisins were prepared as 30 gram of powder soaked in 100 ml distillated water, allowed
to stand for 72 hr and sterilized by filtration. Bacterial isolates were obtained from urinary tract infection
clinical samples. Agar well diffusion assay, tissue culture plate method assay and bacterial ability to adhere to
oral epithelial cells used to evaluate effects of these extracts on growth, motility, adhesion and biofilm formation
in bacterial isolates. Results: the aqueous extracts of Zea Mays, Cranberry and Raisins, individually and
in combination, have significant inhibitory effects on growth, motility, adherence and biofilm formation in
the bacterial isolates considered in current study. In addition, combinations that contain Zea Mays exerted
more powerful antibacterial effects than other combinations. Conclusions: plant extracts considered in
current study may provide useful substitutes for commonly utilised antibiotics for treatmnet of urinary tract
infections, especially recurrent cases. However, the exact mechanisms and the active ingredients responsible
for the reported antibacterial effects need further exploration and specification. In addition, further studies at
the molecular level and/ or in vivo studies involving humans and animals are needed to confirm the observed
effects of these plant extracts.

Keywords: Zea Mays, Cranberry, Raisins, adherence, biofilm, uropathogens.

INTRODUCTION Moreover, uropathogens may implement phenotypic

Urinary tract infection (UTI) is a prevalent bacterial
disease as it is associated with approximately two
million of cases worldwide (1). It mostly affects male
infants, women of various age groups and elderly men
(2).
Also, inadequate treatment of UTI results in serious
complications like recurrent infection, septicemia and
renal dysfunction.
Causative micro-organisms of UTI (uropathogenes)
include both Gram-positive and Gram-negative bacteria
and some fungal species (3,4). The initial step in the
pathogenesis of UTI is adherence of uropathogenes to
lining epithelium of urinary tract (uroepithelium) via
flegallae and pili (2).
alterations such as “filamentation” to resist killing by
host neutrophiles and achieve progressive colonization
of uroepithelium (5).
It is well-recognized that UTI can cause significant
financial and health troubles for the patient, family and
the community, therefore, effective treatment is a priority
at the individual, organizational and governmental
levels (6). Commonly prescribed antibiotics for treatment
of UTI include trimethoprim, sulfamethoxazole,
ciprofloxacin and ampicillin (7). However, the demand
for alternative therapeutics is increasing. The latter must
be inexpensive, effective against resistant uropathogens
and without affecting vaginal and intestinal flora
 Indian Journal of Public Health Research & Development, February 2019 2018, Vol. 10, No. 02
(8).
Also, these potential therapies should target the
virulence elements of concerned uropathogens. In this
regard, numerous studies have evaluated the potential
antimicrobial properties of plants against a wide range
of bacterial isolates, especially those responsible for
urinary tract infections. These plants have been shown to
be inexpensive, have wide therapeutic window and have
few unwanted effects compared to standard antibiotics(9).
One of these plants is Zea Mays. Its aqueous
extract contains phenols, flavanoids, tannins, saponnins,
alkaloids and cardiac glycosides (10) whereas its
alcoholic (methanol) extract contains also terpenoids
and anthraquinones. Also, Zea Mays hairs have
been successfully employed in the treatment of DM,
immunological, proliferative and renal diseases (10).
Previous studies reported antibacterial properties
of raisins and raisin products. For example, there was
99.99% inhibition of Salmonella typhi, Escherichia coli
O137:H7 and Listeria monocytogenes growth following
18 and 8 days incubation with finished raisins and raisin
juice, respectively. Also, raisins contain phytochemicals
with antibacterial properties (11-16).
The objective of current in vitro study was to
evaluate antibacterial effects of aqueous extracts of Zea
Mays, Cranberry and Raisins on bacterial uropathogens,
individually and in combination.
MATERIAL AND METHOD
1. Aqueous extracts: Preparation of Aqueous
Extract of plants according to (17).
2. Bacterial Isolates
A total of 8 Gram negative, and 3 Gram positive
isolates were isolated from UTI samples were used
in this study. The bacterial isolates represented by; S.
aureus, S.epidermidis, S. saprophyticus, P.aeruginosa,P.
fluresence, E. coli, E.aerugenes, K. pneumoniae, Proteus
mirabilis, P.vulgaris, Acinetobacter. These bacteria
were activated and cloned three successive times in
nutrient agar and stored on nutrient agar slant at 4 ºC.
The identification of these organisms was confirmed by
using conventional biochemical tests (18).
3- Antimicrobial activity test by Agar-well
diffusion assay (In vitro)
According to Forbes (17), the antimicrobial activity of
855
ciprofloxacin was determined by agar disc diffusion (the
plates were performed in triplicates).
4-Biofilm Formation Assay: Semi quantitative
micro-titer plate test or Tissue culture plate method assay
(TCP) designated by Christensen et al. (19) was assumed
as the gold standard test to detect biofilm formation
(table 1).
5-Adherence test: Bacterial adherence to oral
epithelial cell can be identified using method designated
by (20) (table 1).
6-Inhibition of motility (swarming) by plant
extract:
Plant extract was added separately in concentrations
of (10%, 20%, 30%).
All the extracts involved in this study were used
in each tests to detect the antibacterial properties of
these extracts separately (the plates were performed in
triplicates).
Table (1) Classification of bacterial adherence
and biofilm formation by TCP method
Mean of OD value at Biofilms
Adherence
630nm formation
<0.120 None None
0.120-0.240 Moderate Moderate
>0.240 Strong High
DATA ANALYSIS
Data were expressed as Mean±SE and presented
using descriptive statistics. Independent t-tests and
one-way Analyses of Variance (ANOVA), followed
by Bonferroni multiple comparison test, were used for
comparisons between variables. Probability values less
than .05 were considered statistically significant.
RESULTS
Current study showed that the individual aqueous
extracts of Zea Mays, Cranberry and Raisins exerted
powerful anti-growth effects on Gram-positive and
Gram-negative bacterial uropathogens as compared
to the standard antibiotic; Ciprofloxacin (Figures 1-3).
Also, compared to Ciprofloxacin, Gram-negative isolates
were more sensitive to current extracts than were Gram-
 856 Indian Journal of Public Health Research & Development, February 2019 2018, Vol. 10, No. 02
positive isolates (P<0.001 and <0.05, respectively; Table
1).
The salient finding of current study was that
combinations of two or more extracts produced
more significant anti-growth activities than those of
corresponding single exracts (Figures 1-4; Tables 1-2)
and those of Ciprofloxacin (Figures 4-6). Interestingly,
combinations contained Zea Mays produced more
powerful anti-growth effects than other combinations
(Table 2).
Moreover, aqueous extracts of Zea Mays, Cranberry
and Raisins exerted powerful anti-adherence and anti-
biofilm properties on Gram-negative isolates of interest.
The former two extracts were more powerful than
Raisins (Table 3).
DISCUSSION
Previous studies reported that cranberry extracts
exerted significant inhibitory effects on the growth of
specific isolated uropathogens (21). For example, results
of a retrospective study (22) reported that cranberries
exerted significant antibacterial effects againt
antibiotic-responsive and antibiotic-resistant bacterial
uropathogens. In addition, (23) reported that cranberries
contain two components that inhibited adhesion of
E. coli; one of them inhibited mannose-susceptible
fimbrial component and the other blocked mannose-
resistant element of adhesion in E. coli. Also, (24) reported
effective inhibition of biofilmn formation in bacterial
uropathogens by cranberries.
The reported antibacterial activities of cranberries
could be attributed to their contents of hipauric acid
and phytochemicals like flavanoids, production of
nitric oxide and/ or presence of specific anti-adhesion
ingredients (25-28).
On the other hand, other researchers (29) stated that
Zea Mays was useful for treatment of recurrent nephritis
and cystitis. The reported antibacterial activities of Zea
Mays could be attributed to its contents of flavonoids,
tannins, saponnins, phenols and cyclic hydroxamic acids
(30).
The latter are lethal for bacteria, fungi and insects (31).
Furthermore, previous studies revealed reported
powerful antimicrobial activities of raisins against a wide
spectrum of pathogenic micro-organisms. For example,
adding raisin products to meats and bread had prevented
their rancidity without having unwanted sensory effects
(32)
. Also, both skin and meat of raisins exhibit effective
antibacterial properties whether independently or in
combination (33).
The documented antibacterial properties of raisins
could be attributed to their acidic pH (3.5-4), their
dryness and presence of phytochemicals like acid-active
phenolic acid and oleanolic acid (34,35). The latter has anti-
adherence and anti-biofilm formation against cariogenic
Staphylococcus mutans (36,37).
The highly significant inhibition of bacterial
growth reported in current study, following the use
of combinations of two or more extracts, could be
attributed to the possibility that these extracts have
different mechanisms of actions against the selected
uropathogens (38).
CONCLUSIONS
Plant extracts studied in current study, individually
or in combinations, may provide useful substitutes
for synthetic antibiotics for treatmnet of urinary tract
infections, especially recurrent cases. However, further
studies are required to explore the exact pharmacokinetics
and pharmacodynamics of these plants, maybe at the
molecular level and/ or in vivo studies involving humans
and animals.
Ethical Clearance : This study was approved by
the ethical committee of scientific research according to
ministry higher edcation and sintifc research in Iraq,
also according to biosafty low of good lab.potection .
Source of Funding; self-fund
Conflict of Interest; the authors declare that they
have no conflict of interest.
Indian Journal of Public Health Research & Development, February 2019 2018, Vol. 10, No. 02 857

Figure (1): Antibacterial activity of white Raisins against bacterial isolates

Figure (2): Antibacterial activity of Cranberry against bacterial isolates

Figure (3): Antibacterial activity of Zea Mays against bacterial isolates

858 Indian Journal of Public Health Research & Development, February 2019 2018, Vol. 10, No. 02

Figure (4): Antibacterial activity of white Raisins and Cranberry against bacterial isolates

Figure (5): Antibacterial activity of Zea Mays and Cranberry against bacterial isolates

Figure (6): Antibacterial activity of Zea Mays, white Raisin and against bacterial isolates
 Indian Journal of Public Health Research & Development, February 2019 2018, Vol. 10, No. 02 859

Table (1) Effects of individual extracts of Raisins, Cranberry and Zea Mays on growth of G+ve and G-ve
bacterial uropathogenes

Zone of inhibition (Mean±SE) / mm

Extracts G+ve (n=3) G-ve (n=8)
Raisins 22.3 ± 1.5* 21.6 ± 0.8**
Cranberry 24.0 ± 1* 24.4 ± 1.1**
Zea Mays 25.3 ± 2.6* 26.4 ± 0.7**
Control (Ciprofloxacin) 17.3 ± 0.7 8.8 ± 1.9
* and ** Significant difference from ciprofloxacin (P≤0.05) and (P≤0.001), respectively.

Table (2) Effects of combinations of Raisins, Cranberry and Zea Mays extracts on growth of G+ve and G-ve
bacterial uropathogenes

Zone of inhibition (Mean±SE) / mm

Extracts G+ve (n=3) G-ve (n=8)

Raisins + Cranberry 30 ± 0.0***† 30.3 ± 0.6***†

Raisins + Zea Mays 34 ± 0.6***†† 34.5 ± 0.7***††

Cranberry + Zea Mays 35.3 ± 0.3***†† 35.6 ± 0.6***††

Raisins + Cranberry + Zea Mays 40.± 0.0*** 39.8 ± 0.2***

Control (Ciprofloxacin) 17.3 ± 0.7 8.8 ± 1.9

*** Significant difference from ciprofloxacin (P≤0.001).† Significant difference from other combinations (P<0.001).††
Significant difference from the combination of Raisins, Cranberry and Zea Mays (P<0.001).

Table (3) Anti-biofilm and anti-adherence activities of Zea Mays, white Raisin and Cranberry against G-ve
bacteria

Biofilm formation Adherence

Bacteria Aquatic Aquatic Aquatic

Aquatic extract Aquatic extract Aquatic extract
extract of extract of extract of
of Cranberry of Cranberry of Zea Mays
white Raisin Zea Mays white Raisin

P. aeroginosa Moderate* High** High Moderate High High

P.fluresence Moderate High High Moderate High High

P.vulgaris Moderate High High Moderate High High

P.mirabilis Moderate High High Moderate High High

K. pneumoniae Moderate High High Moderate High High

E. aerugenes Moderate High High Moderate High High

Acinetobacter Moderate High High Moderate High High

E. coli Moderate High High Moderate High High

*Moderately (0.120-0.240) **High (>0.240)

 860 Indian Journal of Public Health Research & Development, February 2019 2018, Vol. 10, No. 02
REFERENCES
1. Stamm WE, Norrby SR. Urinary tract infections:
disease panorama and challenges. J Infect Dis.
2001; 183 (Suppl 1):S1–S4.
2. Flores-Mireles AL, Walker JN, Caparon M,
Hultgren SJ . Urinary tract infections: epidemiology,
mechanisms of infection and treatment options. Nat
Rev Microbiol. 2015 ; 13(5): 269–284.
3. Nielubowicz GR, Mobley HL. Host–pathogen
interactions in urinary tract infection. Nature Rev
Urol. 2010; 7:430–441. Ronald A. The etiology of
urinary tract infection: traditional and emerging
pathogens. Am J Med. 2002; 113 (Suppl 1A):14S–
19S.
4. Kostakioti M, Hadjifrangiskou M, Hultgren
SJ. Bacterial biofilms: development, dispersal,
and therapeutic strategies in the dawn of the
postantibiotic era.Cold Spring Harb Perspect Med.
2013 ;1;3(4):a010306.
5. Danese PN, Pratt LA, Dove SL, Kolter R. The outer
membrane protein, antigen 43, mediates cell-to-cell
interactions within Escherichia coli biofilms. Mol
Microbiol. 2000; 37:424–432.
6. Mittal R, Aggarwal S, Sharma S, Chhibber S, Harjai
K. Urinary tract infections caused by Pseudomonas
aeruginosa: a mini review. J Infect Publ Health.
2009; 2:101–111.
7. Justice SS, Hunstad DA, Seed PC, Hultgren SJ.
Filamentation by Escherichia coli subverts innate
defenses during urinary tract infection. Proc Natl
Acad Sci USA. 2006; 103:19884–19889.
8. Nagamatsu K, et al. Dysregulation of Escherichia
coli α-hemolysin expression alters the course of
acute and persistent urinary tract infection. Proc
Natl Acad Sci USA. 2015; 112:E871–E880.
9. Kostakioti M, Hultgren SJ, Hadjifrangiskou M.
Molecular blueprint of uropathogenic Escherichia
coli virulence provides clues toward the development
of anti-virulence therapeutics. Virulence. 2012;
3:592–594.
10. Khadim WA, Khadim MJ, Hameed IH. Antibacterial
activity of several plant extracts against proteus
species. International Journal of Pharmaceutical and
Clinical Research 2016; 8(12): 1673-1684.
11. Solihah, M.A., Wan Rosli, W.I. and Nurhanan, A.R.
Phytochemicals screening and total phenolic content
of Malaysian Zea mays hair extracts. International
Food Research Journal 2012; 19(4): 1533-1538.
12. Elben BS. Science Base to Support the Antimicrobial
Action of Raisins. Center for Chemical Regulation
and Food Safety, Exponent, Inc.September 30, 2009.
13. Bower C. K., Schilke, K. F., and M. A. Daeschel
. Antimicrobial properties of raisins in beef jerky
preservation. J. Food Sci. 2003; 68: 1484-1489.
14. Bolin, H., Petrucci, V., Amino acids in raisins. J.
Food Sci. 1985; 50, 1507.
15. Palma, M., Taylor, L.T., Supercritical fluid
extraction of 5-hydroxymethyl-2- furaldehyde from
raisins. J. Agric. Food Chem. 2001; 49, 628–632.
16. Rahbar M, Diba K . In vitro activity of cranberry
extract against etiological agents of urinary tract
infections. African Journal of Pharmacy and
Pharmacology 2010; 4(5), 286-288, May 2010.
17. Gunther NW, Lockatell V, Johnson DE, Mobley HL
. In vivo dynamics of type 1 fimbria regulation in
uropathogenic Escherichia coli during experimental
urinary tract infection. Infect. Immun. 2001; 69:
2838–2846
18. Hindi NKK. In vitro Antibacterial
activity of Plants extracts against
Porphyromonasgingivalis, Prevotellaintermedia
and Aggregatibacteractinomycetemcomitans
Streptococcus mutanus, Isolated from Periodontitis
Patients in Babylon province, Iraq. International
Journal of PharmTech Research, 2016; 9(11):84-93.
19. Forbes BA, Sahm DF, Weissfeld AS. Bailey and
Scotts’ Diagnostic microbiology. 12th ed. Elsevier.
China.2007.
20. Christensen GD, Simpson WA, Younger JA,
Baddour LM, Barrett FF, Melton DM. Adherence
of cogulase negative Staphylococi to plastic tissue
cultures:a quantitative model for the adherence
of staphylococci to medical devices. J Clin
Microbiol.1985; 22:996–06.
21. Avila-Campos MJ, Simionato MR, Cai S, Mayer
MP, Delorenzo JL, Zelant F. Virulence Factors
of Actinobacillus actinomycetecomitans: other
Putative Factors. Pesq.Odont. Bras.2000; 14(1): 05-
11
22. Harkins KJ . what’s the use of cranberry juice? Age
and Ageing, 2000; 29: 9- 12.
23. Mulvey MA.Adhesion and entry of uropathogenic
 Indian Journal of Public Health Research & Development, February 2019 2018, Vol. 10, No. 02
Escherichia coli. Cell Microbiol. 2002; 4(5):257–
271.
24. Dugoua JJ, Seely D, Perri D, Mills E, Koren
G.Safety and efficacy of cranberry (Vaccinium
macrocarpon) during pregnancy and lactation. Can.
J. Clin. Pharmacol., 2008; 15: 80-86.
25. Cimolai N, Cimolai T The cranberry and the urinary
tract. Eur. J. Clin. Microbiol. 2007; 26: 767-776.
26. Johnson BJ, Lin B, Rubin RA, Malanoski AP Media
acidification by Escherichia coli in the presence of
cranberry juice., 2009; 2(226): 1-4.
27. Wang SY, Stretch AW. Antioxidant capacity in
cranberry is influenced by cultivar and storage
temperature. J Agric Food Chem 2001; 49: 969–974.
28. MacMicking J, Xie QW, Nathan C. Nitric oxide and
macrophage function. Annu Rev Immunol 1997; 15:
323–350.
29. Howell AB, Leahy M, Kurowska E, Guthrie N. In
vivo evidence that cranberry proanthocyanidins
inhibit adherence of p-fimbriated E. coli bacteria to
uroepithelial cells. Fed Am Soc Exp Biol J 2001;
15: A284.
30. Yue, Q.; Bacon, C.W.; Richardson, M.D.
Biotransformation of BOA and 6-methoxy-
benzoxazolinone by Fusarium moniliforme.
Phytochemistry 1998, 48, 451–454. (CrossRef).
31. Frey, M.; Chomet, P.; Glawischnig, E.; Stettner, C.;
View publication stats
861
Grün, S.; Winklmair, A.; Eisenreich, W.; Bacher,
A.; Meeley, R.B.; Briggs, S.P.; et al. Analysis of
a chemical plant defense mechanism in grasses.
Science 1997, 277, 696–699.
32. Niemeyer, H.M. Hydroxamic acids derived from
2-hydroxy-2H-1,4-benzoxazin-3(4H)-one: Key
defense chemicals of cereals. J. Agric. Food Chem.
2009, 57, 1677–1696.
33. Wei, Q., Wolf-Hall, C., and C. A. Hall III. Application
of raisin extracts as preservatives in liquid bread
systems. J. Food Sci. 2009; 74: M177-M184.
34. United States Department of Agriculture (USDA).
2009. Pathogen Modeling Program Version 7.0.
Available at http://pmp.arserrc.gov/PMPOnline.
aspx
35. Aruscavage, D., Lee, K., Miller, S., and J. T. LeJune.
Interactions affecting the proliferation and control
of human pathogens on edible plants. J. Food
Sci.2006; 71: 89-99.
36. Vasavada, M. N., and D. P. Cornforth. Evaluation
of antioxidant effects of raisin paste in cooked
ground beef, pork, and chicken. J. Food Sci. 2006;
71: C242-C245.
37. Tamma PD, Cosgrove SE, Maragakis LL
Combination Therapy for Treatment of Infections
with Gram-Negative Bacteria. J Clinical
Microbiology Reviews; 2012; 25(3): 450–470.