32

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

32

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

32

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

32

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) .

32

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

22

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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