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)
Nada A. Elamin1, Mustafa S. koya2 Zeinab A.Elrabei3
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 نيالا للمضادات الحيوية
مستخلص الدراسة :
اجريت هذه الدراسة به دد اتبا د ر اس سدوة اوم او دة باباي د المال داو ا ال دة
ون ا ص در ا خبلفة ود ه وديةدة اود ب لدا ض ا تد داو ا وايدة . اسدخدوت تمةودة ا ) disk diffusion ( بتبا راس سوة ا وم اوة عدد 23 ون ا ت داو ا واية الأكثدر اسدبخداو
في ال لاج االتي اشبملت علد Cefotaxime (30 mcg) , Ampicillin / Sulbactam( 20 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 ).
أاضحت الةب ئج أو جموع سدلابو باباي د المال داو ا ال دة لل وة د و ددت الدراسدة
اس سددة لادد وددن ) Chloramphenicol ، Co-Trimoxazole ، (Ciprofloxacin و اوم اودة لاد ودن ) Ampicillin / Sulbactam ا Tetracycline ) بوةمد اس سدوبه
اوم اوبه للمت داو الأترى ت د وباسطة.
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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