REPUBLIC OF IRAQMINISTRY OF HIGHER EDUCATION AND SCIENTIFIC RESEARCH

UNIVERSITY OF AL-QASSIM GREEN

COLLEGE OF VETERINARY MEDICINE

Studying Antibiotics Sensitivity Patterns of

Pseudomonas aeroginosa

Isolated From Patient With Otitis Media

ـــــراقــجمهوريـة العـوزارة التعليم العالي والبحث العلمي

القاسم الخضراء-كلية الطبجامعة البيطري

:دراسة عن

Pseudomonas) بكتيريا حساسية انماطaeroginosaاالذن التهاب عزالت ( عن الحيوية المضادات لبعض الوسطى

اشراف: د.زينة خضير ضياء : احمد اعداد

م2015هـ1436

وبشر الصابرين

العليَصدَقَ اللُّه العظيم

اإلهداء... إلى صاحب السكينة ... المدفون

بالمدينة ... نبي الرحمةورسول االنسانية ... المصطفى )ص(.

إلى صاحب القبة البيضاء في ارض الغري ... امام الهدى امير المؤمنين

علي بن ابي طالب )ع(. إلى من سهرَت عيونُه لكي تغمض

عيناي ...

إلى َمثــــلَي االعــــلى وقـــدوتي .)أبي العزيز(.فـــــي الحــــياة

إلى من حملتني وربّتني وسقتني بدموع عينيها ...

إلى من وِضعت الجنان تحت قدميها ...

... )اميإلى رمز الوفاء والحنان الغالية (.

إلى اخوتي ... واخواتي ... واصدقائي جميعا .

إلى كل شهداء العراق ... الذين سقطوا لكي نبقى نحن على قيد

الحياة ...

Abstract :

Otitis Media(OM) is one of the common hearing problems

which can cause many complications if not treated properly.

Hence, the aim of this study is to identify the bacterial isolates

of Pseudomonas aeroginosa as one of the causing agents of OM

and to study their antimicrobial susceptibility patterns. The

study was carried out in Al- Hillah Teaching hospital from

December 2014 to March 2015.

Twenty swabs from patients of OM were included in this study.

Out of 20 ear discharge samples cultured, 17(85%) samples

have bacterial growth, P. aeroginosa isolates represented

8(47%) of the bacterial growth.

Regarding antibiotic sensitivity test, all the isolates were highly

sensitive to cefotaxim and imipenem (100), while

chloramphenicol was not effective against tested isolates.

الخالصة: األذن التهاب من يعتبر واحدة السمع الوسطى مشاكل

تسبب أن يمكن التي و كثيرة الشائعة يعالج إذا مضاعفات لمصحيح الدراسة ،إن. بشكل هذه من على الهدف التعرف هوالعوامل Pseudomonas aeroginosaبكتيريا عزالت من كواحدة

األذن ل المسببة الحساسية التهاب فحص واجراء الوسطى. وقد للعالج فاعلية األكثر المضادات لتحديد الحياتية للمضادات

الدراسة من التعليمي الحلة مستشفى في أجريت الفترة2015مارس الى 2014 ديسمبر

األذن 20جمعت بالتهاب مصابين مرضى من مسحةفي بكتيريا نموا النتائج أظهرت ، ،%) 85مسحة (17الوسطى

عزالت .Pشكلت aeroginosa 8 (47%). النموالبكتيري منيتعلق فقد باختبار وفيما الحيوية للمضادات كانتالحساسية( العزالتجميع االميبنيم و للييفوتاكسيم ,100حساسة (% الكلورامفينيكول ضد بينما فعالية ذا يكن المختبرة لم .العزالت

Introduction:

Otitis media is inflammation of the middle ear. This is most

commonly caused by the buildup of fluid behind the ear drum, as a

result of a blockage to the Eustachian tube. Otitis media is more

common in children, as their Eustachian tube is shorter and more

horizontal than

adults and is made up of more flaccid cartilage, which can impair its

opening. (Bluestone and KLien, 2001). Otitis media can cause a mild

to moderate hearing loss, due to the fluid interfering with the

transmission of sound through to the inner ear. There are three

common types of otitis media, acute purulent otitis media, otitis

media with effusion and chronic suppurative otitis media (Berman,

1997).

Aetiologic agents of otitis media include bacteria, fungi and

viruses (Li et al., 2001), though bacterial agents of otitis media

depends on age of individuals, geographical location and whether

the infection is acute and chronic ( Herzon, 1992).

Treatment is very necessary and urgent to prevent complications

such as meningitis, septicaemia, amongst others ( Herzon, 1992).

Increased antimicrobial resistant bacteria in chronic otitis media can

lead to the development of complications of chronic otitis media like

meningitis and brain abscess (Agrawal et al, 2005). Studies have

indicated that otomycosis, which results from prolonged bacterial

treatment of chronic otitis media can complicate the management of

chronic otitis media (Ahmed et al, 2010).

Materials and Methods:

1 -Isolation and identification of bacteria: Atotal of 20 swabs were collected from patients with OM admitted to

Al- Hillah Teaching hospital in Babylonprovince, from December 2014

to March 2015. All the specimens were isolated and identified using

standard procedures. Culture media used for isolation and purification

of bacteria included: (Nutrient agar medium, Blood agar base,

MacConkey agar medium, EMB medium) were prepared according to

the manufacturer's instructions. Each swab was cultured

immediately and incubated aerobically at 37 ºC for 24 hours. After

positive results of growth were appear, bacterial samples were

identified with Gram stain and Biochemical tests .

2 -Biochemical tests:Bacterial isolates were identified by standard biochemical tests

according to (MacFaddin, 2000)by :1 .IMVIC test (indol production, methyl red, vogas-proskauer and

citrate utilization) .2 .TSI (triple sugar iron).

3 .Motility test.4 .Oxidase test .

5 .Mannitol salt agar (for Staphylococcusaureus) .

3-Antimicrobial susceptibility testing:Susceptibility to some antimicrobial agents for all P. aeroginosa was determined by the standard disk diffusion test (DDT) on Muller- Hinton agar (Oxoid, UK). Plates were incubated for 18 hours at 37 ºC. The selection of antibiotic discs was performed according to the guideline recommended by the Clinical and Laboratory Standards Institute (CLSI, 2010). After incubation, the diameter of each inhibition zone was measured and recorded in (mm). The results

then interpreted according to CLSI documentation (CLSI, 2010).

Results and discussion:

1- Isolation of bacteria:

In this study a total of (20) samples from patients with otitis media

were collected. The results showed that 17 (85%) of the swabs were

positive on culture, while 3 (15%) has no growth as shown in Table

(1). This is in agreement with a study reported by Ogbogu et al

(2013), who found that (84. 55%) of their specimens were culture-

positive otitis media, while (15.45%) of the specimens were culture-

negative.

Table (1): Percentage of otitis media swabs.

From the 20 samples only 8(47%) isolates were belonged to P.

aeruginosa and 9(52.9%) were belonged to the other genera as

shown in fig (1). Similar results have been reported by Kumar et al

(2014) who found that 42.22% of isolated bacteria were belonged to

P. aeruginosa .

Figure (1)

Type of swab No(%) .

Positive 17) 85%(

No Growth 3) 15%(

Total No. 20) 100%(

2- The sensitivity of isolates to antibiotics by using DDT: The effects of different antibiotics on P. aeruginosa isolates were

investigated (Fig. 2). The overall antimicrobial susceptibility

demonstrated that cefotaxime and imipenem revealed high level of

sensitivity (100%).

Figure (2)

Antibiotic disc Symbol Potency Total no. (%) of susceptible

isolates

Cefotaxime 20 µg 8 (100%)

Cefoxitin FOX 30 µg 5 (62.5%)

Gentamicin CN 10 µg 4 (50%)

Amikacin AK 30 µg 2 (25%)

Ciprofloxacin CIP 5 µg 5 (62.5%)

Chloramphenicol C 30 µg 0 (100%)

Imipemem IMP 10 µg 8 (100%)

However, the isolates were sensitive to cefoxitin and ciprofloxacin

(62.5%), similar to reports of findings in Brazil (Weckwerth et al,

2009).

Results also revealed that P. aeruginosa isolates were highly

resistant to chloramphenicol (100%). This is comparable with results

from Ethiopia (Yismaw et al, 2010) and Iraq (Ihsan et al, 2010).

However, the study also showed that 50% and 25% of isolates were

sensitive to gentamicin and amikacin, respectively. Similar results

have been reported by Ihsan et al (2010) who found that 50%

resistance rate for gentamicin.

References

- Agrawal, S.; Hussein, M. & MacRae, D. (2005) Complication

of otitis media: an evolving state. J. Otolaryngol. 34(1): 33-39.

- Ahmed, Z., Hafeez, A., Zahid, T., Jawaid ,M. A., Matiullah,

S.& Marfani, M. S. (2010) Otomycosis: Clinical Presentation

and Management, Pakistan J. Otolaryngol. 26:78-80.

- Berman, S. (1997). Classification and criteria of Otitis Media. Clin.Microbiol. Infect (Suppl)., 3: 1-4.

- Bluestone, C. D., Klein, J. O. (2001). Microbiology. In: Bluestone CD, Klein JO, eds. Otitis Media in Infants and Children. 3rd ed. Philadelphia, PA: W. B. Saunders., pp. 79-1014.

-Clinical & Laboratory Standards Institute (CLSI, formerly NCCLS). (2010): performance standareds for antimicrobial susceptibility testing, 7th informational supplement.

- Herzon, F.S. (1992). Ear and Sinus infections. In: Brillnan, J.C. and Quenzer, R.N. (ed). Infectious diseases in emergency medicine. Little Brown and Company, Boston. pp. 867 – 885.

-Ihsan, E,. Alsaimary, Ahmed, M,. Alabbasi, Jassim, & Najim, M,. Antibiotics susceptibility of bacterial pathogens associated with otitis media. J Bacteriol Res 2010;2(4):41-50.

- Li, W. C., Chiu, N.C., Hsu, C. H., Lee, K.S., Hwang, H.K.& Huang, F.Y. (2001). Pathogens in the middle ear effusion of children with persistent otitis media; implications of drug resistance and complication J. Microbiol. Immunol. Infect. 34(3):190–194.

-MacFaddin, J. F. 2000. Biochemical tests for identification of medical bacteria. Lippincott Williams and Wilkins. Philadelphia, USA.

-Ogbogu, P. I. , Eghafona, N. O. & Ogbogu, M. I. (2013) . Microbiology of otitis media among children attending a

tertiary hospital in Benin City, Nigeria. J. Public Health Epidemiol.5 (7): 280-284 .

-Raghu ,K,G,. Kumar, K. R., Navya, S & Basavarajappa, K. G. (2014). A Study of Bacterial Profile and Antibiotic Susceptibility Pattern of Chronic Suppurative Otitis Media among Patients attending a Tertiary Care Centre, Davangere. Sch. J. App. Med. Sci. 2(5B):1606-1612.

-Weckwerth, P,H,. de Magalhães Lopes, C,A,. Duarte, M,A, .Weckwerth A,C,. Martins, C,H, .Neto, D,L,. de Aguiar, H,F,. Chronic suppurative otitis media in cleft palate: microorganism etiology and susceptibilities.Cleft Palate Craniofac J 2009;46(5):461-467.

-Yismaw, G,. Abay, S,. Asrat, D,. Yifru, S,. Kassu, A,. Bacteriological profile and resistance patterns of clinical isolates from pediatric patients, Gondar University teaching hospital. Gondar, Ethiopia. Ethiop Med J 2010;48 (4):293-299.