antimicrobial susceptibility of isolated e.coli from different water sources in nyala town. (sudan)
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Antimicrobial susceptibility of isolated E.coli from different water sources in Nyala Town. (Sudan)TRANSCRIPT
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Antimicrobial susceptibility of isolated E.coli from different
water sources in Nyala Town. (Sudan) Nada A. Elamin
1, Mustafa S. koya
2 Zeinab A.Elrabei
3
1department of Biology, faculty of education, University of Nyala, Sudan
2department of Chemistry, faculty of education, university of Nyala Sudan
3department of Chemistry, faculty of education, University of Nyala, Sudan
اختبار حساسية بكتيريا القولون المعزولة من المصادر المختلفة
لمياه مدينة نياال للمضادات الحيوية
:مستخلص الدراسة
اجريت هذه الدراسة بهدد اتباد ر اس سدوة اوم اودة بابايد المالداو ا الدة
disk)تمةودة ا اسدخدوت . ون ا ص در ا خبلفة ود ه وديةدة اود ب لدا ض ا تد داو ا وايدة
diffusion ) كثدر اسدبخداو ون ا ت داو ا واية األ 23بتبا راس سوة ا وم اوة عدد
Cefotaxime (30 mcg) , Ampicillin / Sulbactam( 20 االيت اشبملت علد يف ال الج
mcg ) , Co.Trimxazole(25mcg) , Piperacillin / Tazobatam(100/10 mcg) ,
Chloramphenicol(30 mcg) . Ciprofloxacin(5 mcg). Tetracycline(30 mcg) ,
Ofloxacin(5 mcg) , Gentamicin(10mcg) , Amikacin(30 mcg) , Pefloxcin (5
mcg) and Ceftizoxcime ( 30 mcg ).
و مجوع سدالبو بابايد المالداو ا الدة لل وةد و ددت الدراسدة أاضحت الةب ئج أ
و Chloramphenicol، Co-Trimoxazole ، (Ciprofloxacin)اس سددة لادد وددن
اس سددوبه بوةمدد (Tetracyclineا Ampicillin / Sulbactam)اوم اوددة لادد وددن
. ترى ت د وباسطةاوم اوبه للمت داو األ
Abstract
The prevalence of antimicrobial resistant among the faecal
bacteria found in different water sources in Nyala town South Darfur
was tested. Isolation and identification of E.coli was done by using
standard methods. Presumptive isolates obtained were confirmed by
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culturing in Brilliant Green Bile lactose media (BGB) and IMVC test.
Antimicrobial susceptibility testing by the disk diffusion method was
conducted for 12 antimicrobial agents: Cefotaxime(30mcg) ,
Ampicillin / Sulbactam(20mcg) , Co-Trimxazole (25mcg), Piperacillin
/ Tazobatam(100/10mcg) Chloramphenicol(30mcg) .
Ciprofloxacin(5mcg) . Tetracycline(30mcg) , Ofloxacin(5mcg) ,
Gentamicin(10mcg) , Amikacin(30mcg) , Pefloxcin(5mcg0 and
Ceftizoxcime(30mcg) .
The results indicate that all the studied E .coli strains were
sensitive to Ciprofloxacin, Co-Trimoxazole and Chloramphenicol and
resistant to Tetracycline and Ampicillin / Sulbactam . Key words:- Antimicrobial, susceptibility, Escherichia coli, Nyala, water sources
Introduction
Public and environmental health protection requires safe drinking
water, which means that it must be free of pathogenic bacteria. E .coli
is an organism that occurs universally in sewage and because it is a
faecal coliform, it plays an important role in the sanitary analysis of
water. (Zamxaka et al., 2004). According to the World Health
Organization (WHO, 1981), 80% of all diseases are attributed to
unsafe water.
About one quarter of billion people in the world are suffering
from major related diseases at any time (Odeyemi, 1998).
Antibiotic resistance in E.coli has been globally identified in
isolates from environmental, aninmal and human sources (Heike and
Reinhard,2005) . This is a consequence of the use of antimicrobials in
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medicine and their application in animal husbandry , which have
brought about phenotypic changes , often due to chromosomal
mutations .(Walsh et.al .2005) .
E.coli is resistant to a number of antibiotics. (Robert2003). In
addition to the consequences of microbial resistance to antibiotics on
human health, contamination of surface water bodies with resistant
bacterial strains from human activities and livestock operations has
also been reported (Harakeh et al., 2004).
However, bacteria have developed strategies for survival within
the host during an infection and one of these strategies is the
resistance of isolates to the antimicrobial drugs. Antibiotic resistance
is a serious problem because it limits the therapeutic possibilities in
the treatment of bacterial diseases in domestic animal species in
general and poultry in particular (Nicole et al., 2000). According to
Hussain et al., 1982, the numbers of multi-drug resistant E. coli are
continuously increasing although various antimicrobial agents are
being used. Considering the above facts, the present research work
was undertaken to determine the current status of drug sensitivity and
resistance pattern of the E. coli isolated from 10 different sources to
select the drugs of choice for therapeutic use against various infections
of man and animals caused by the organism.
The aim of this study was to isolate E.coli organisms from water
collected from different sources in Nyala town with the intention of
testing their resistance to commonly used antibiotics
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Materials and Methods:
Water samples were collected during the rainy season of 2009
from 50 sampling sites such as dug wells, boreholes along wadi Nyala
and tap water in 250 ml bottles, according to procedure mentioned by
WHO,1984.
Samples were kept at 4ᵒC from different sites until their arrival to
laboratory. 50 isolates of E.coli were collected and tested against 12
commonly used antimicrobial agents. Standard methods were used for
the enrichment, isolation, identification and biochemical confirmation
(indole,methyl, red , Voges - proskauer and Simmon citrate tests ) of
E.coli (Drangan .2000)
The bacterial isolates that were confirmed to be E.coli based on
the results of the biochemical tests were selected for antimicrobial
agent sensitivity testing. The antibiotic resistance was determined by a
standard disc diffusion technique using Muller-Hinton agar (Difco)
according to the recommendation of National Committee for Clinical
Laboratory Standards NCCLs 2008.
The fifty samples of E.coli mentioned above were cultured into
peptone water medium incubated at 37ᵒC for 24 hours following
according to methods mentioned by Nada, (2010), with a slight
modification. In exposed to light the growth turbidity was matched
with turbidity of a standard (Macforland 0.5). Small amount of the
cultured liquid were transferred into Muller Hinton agar medium
plates. The plates were left for 15 minuts to be absorbed, multi
antimicrobial susceptibility disks of Gram negative bacteria were
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placed and the plates then incubated at 37ᵒC for 24hr and examined
for evaluation of the inhibition zones. The names and concentrations
of the used antibiotics is shown in (Table 1)
Antimicrobial agent Code Concentration
Amikacin AK 30 mcg
As 20 mcg
CF 30 mcg
Ceftizoxime CL 30 mcg
Chloramphenicol CH 30 mcg
Ciprofloxacin CP 5 mcg
Co-trimxazole BA 25 mcg
Gentamicin GM 10 mcg
Oflxacin OF 5 mcg
Pefloxacin PF 5 mcg
Piperacillin/Tazobatam TzP 100/10 mcg
Teteacycline TE 30 mcg
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Results:
Disk diffusion zones were used to show the difference among
types of samples collected from several water sources.
Significant differences were seen inhibition diffusion zones for all
antimicrobial agents (Table 2) .
Multi disk zones of E.coli isolated from water were examined to
show the difference among types of antimicrobial agents. The largest
zones (a) indicates greater susceptibility were shown by
(Ciprofloxacin, 5mcg), but the smallest zones (f) indicating greater
resistance to the antibiotic were shown by (Ampicillin/ Sulbatctam, 20
mcg). That means E.coli isolates were sensitive to Ciprofloxacin (CP),
CO-Trimoxazole (BA, 25mcg) and chloramphenicol (CH 30 mcg).
The superscript (a) is common and there were no significant
differences among them, but (CP, 5 mcg) shows the largest inhibition
zone Fig(1) .
There were no significant differences among Co-Trimoxazole
(BA, 25mcg) , Chloramphenicol (CH,30 mcg) , Ofloxacin (OF) and
Pefloxacin (PF, 5mcg) (the superscript b is common ) .
Tetracyclline (TE 30 mcg) , Ampicilln / sulbactam (AS,20 mcg)
showed the smallest zones but these showed significant difference
among CP, BA , CH, OF , PF , CL , AK , TZP , Gm and As . (The
superscript is not common) .
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Antibiotic Code Susceptibility
according to
manufacture
standard
(NCCLS)
Statistical
analysis
Mean SD
(mm)
Superscript
significant
Amikacin AK I 15.88C as
2.07 c
Ampicillin /Sulbactam AS R 9.98f as 2.14 f
Cefotaxime CF R 15.56C as
3.67 c
Ceftizoxime CL I 16.44cd
as
2.90 c
Chloramphenicol CH I 17.92ab
as
3.25 ab
Ciprofloxacin CP S 21.02a as
2.65 a
Co-Trimoxazole BA S 18.02ab
as
4.02 ab
Gentamycin GM S 15.78C as
1.66 c
Ofloxacin OF S 17.60bc
as
3.19 bc
Pefloxacin PF I 17.48bc
as
3.03 cd
Piperacillin/Tazobactam TzP R 15.88C as
2.98 c
Tetracycline TE R 8.94e as
3.24 e
S: Sensitive R: Resistant I: Intermediate
*: means with the same superscript are not significantly different.
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antibiotic
Discussion :
The Enterobacteiaceae family has been linked to well known as
antibiotic gene pools. These genes are transferred into the normal flora
of humans and animals (Lin and Biyela,2005) .
In this study all strains of E.coli showed resistance against the
activity of Tetracycline and Ampicillin / Sulbactam . The highest
inhibition zones were found with Ciprofloxacin followed by co-
Trimxazole and chloramphenicol . These findings are in accordance
with (Erwa.1975; Hassan , 1985, Omer,1986; Shers et.al;1988
Kawjok,1992;Musa ,1994; Altaib,1997 and Raida etal,2005), who
Antibiotic
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found that the highest levels of resistance were observed for
tetracycline in E.coli isolated from different sources of water.
Angela (2004) reported that the highest rates of resistance were
towards Ampicillin (21.8%) and tetracycline (14.1%) The results of
the present study confirm the work done by.
Conclusion:
On the basis of the observations recorded and findings obtained
on various biochemical properties of the microorganisms, it is clearly
concluded that the water bodies of the study area revealed the
occurrence of multiple antibiotic resistant E.coli in the water sources
which in turn contributes in making it unsafe for drinking. Good
hygienic practices including sanitary water handling, safe use of
antibiotics, adequate water source protection mechanisms should be
implemented in the area of water points in order to protect the
surrounding community and livestock in general from water borne
diseases.
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