Subscribe Now Subscribe Today
Abstract
Fulltext PDF
References
Research Article
 
Antibiotic Susceptibility Pattern of Salmonella and Shiegella Isolates Among Diarrheal Patients in Gedo Hospital, West Shoa Zone, Oromia State, Ethiopia



Thirumurugan Gunasekaran, Yonas Haileselassie and Befikadu Asefa
 
ABSTRACT

To evaluate the antimicrobial resistance profiling of bacterial strains from clinical isolates in Gedo Hospital, West Shoa Region, Oromia State, Ethiopia to create awareness of periodic antibiotic susceptibility testing, antimicrobial resistance among the health care providers. A convenience sample of patients who visited the adult outpatient department of Gedo hospital for diarrheal complaint was incorporated into the study. Isolation, identification of bacterial strains and antibiotic susceptibility tests were performed by the disk diffusion method recommended by the Clinical and Laboratory Standards Institute using commercial antibiotic discs. Out of 200 samples, 40 diarrheagenic bacteria were isolated and identified as Shiegella (22 = N) and Salmonella strains (18 = N). Shiegella and Salmonella strains were susceptible to gentamicin and ciprofloxacin, respectively but a high rate of resistance were observed to amoxicillin, chloramphenicol and tetracycline for both Shiegella and Salmonella strains. Notably, both bacteria have developed complete resistance to amoxicillin. Gentamicin may be the drug choice for Shiegella stains caused diarrhea and ciprofloxacin for the diarrhea caused by Salmonella in the study area. Antibiotics selection and treatment of infections based on bacterial strains identification and in vitro susceptibility testing rather than current empirical treatment.

Services
Related Articles in ASCI
Similar Articles in this Journal
Search in Google Scholar
View Citation
Report Citation

 
  How to cite this article:

Thirumurugan Gunasekaran, Yonas Haileselassie and Befikadu Asefa, 2014. Antibiotic Susceptibility Pattern of Salmonella and Shiegella Isolates Among Diarrheal Patients in Gedo Hospital, West Shoa Zone, Oromia State, Ethiopia. American Journal of Drug Discovery and Development, 4: 194-201.

DOI: 10.3923/ajdd.2014.194.201

URL: https://scialert.net/abstract/?doi=ajdd.2014.194.201
 
Received: May 07, 2014; Accepted: May 15, 2014; Published: August 23, 2014

INTRODUCTION

Diarrheal disease amounts to an estimated 4.1% of the total global burden of disease and is responsible for the deaths of 1.7 million people every year. Among the microbial pathogens Vibrio cholera, Salmonella and Shiegella strain are the most serious as they cause severe illness and as they are associated with outbreaks. Among the enteric bacterial pathogens Salmonella and Shiegella are of particular concern as causes of enteric fevers, food poisoning and gastroenteritis. The retrospective studies carried out in Ethiopia showed the prevalence of Salmonella (5.3-15.4%) and Shiegella (5-7.5%) (Mengistu et al., 2014). Antibiotic resistance is a drug resistance where a microorganism has developed the ability to survive exposure to an antibiotic. The volume of antibiotic prescribed is the major cause in increasing rates of microbial resistance rather than compliance with antibiotics (Thirumurugan and Dhanaraju, 2011). Antimicrobial resistance is a global concern causes infections by resistant microorganism resulting in prolonged illness and death and reduces effectiveness of treatment, thus patient’s remains infectious, increasing risk of spreading resistant to others pathogens and many infectious diseases become untreatable and increases the cost of therapy. Also the achievements of modern medicine are put at risk by antimicrobial resistance because lack of effective antimicrobials the successive treatments such as organ transplantation, cancer chemotherapy, major surgery would be compromised and possibilities of spreading antimicrobial resistance rapidly from countries to countries and continents through humans and food. Resistance has spread worldwide, antimicrobial resistance gonorrhoea emerged in Vietnam in 1967 and then spread to the Philippines and finally the USA (Thirumurugan et al., 2009; Holmes et al., 1967).

Appropriate antimicrobial drug use has unquestionable benefit, but physicians and the public often utilize these agents inappropriately. Inappropriate use results from physicians providing antimicrobial drugs to treat viral infections, using inadequate criteria for diagnosis of infections that potentially have a microbial etiology, unnecessarily prescribing expensive, broad-spectrum agents and not following established recommendations for using common prophylaxis. The availability of antibiotics over the counter, despite regulations to the contrary, also fuel inappropriate use of antimicrobial drugs in developing countries. The epidemiological and prescriptions pattern studies conducted in Ethiopia showed that a higher percentage and irrational use of antibiotics. Antibiotic prescribing pattern at Bishoftu hospital shows 28.3% of polypharmacy (Feleke et al., 2013), Hawassa University Referral hospital 58% (Desalegn, 2013) and average number of antibiotics per patient was 2.17 for prophylaxis and 2.18 for treatment at northwest Ethiopia (Abula and Kedir, 2004).

In most developing countries, laboratory investigations of resistant microbes are diagnostic challenges due to lack of adequate facilities that enable culture and antimicrobial susceptibility testing (Collee et al., 1996). As a result, there is restricted awareness of the prevalence of infections and antimicrobial resistance (Cook and Zumla, 2003; Sharma et al., 2005). Also, the injudicious use of antibiotics by patients and physicians alike in many developing countries such as Ethiopia has led to an increased antimicrobial resistance and in turn reduced therapeutic efficacy in these countries (Asrat, 2008). In Harar, Eastern Ethiopia, Salmonella and Shiegella have been reported to be resistant to first line antibiotics such as ampicillin, tetracycline and chloramphenicol and also resistant Shiegella serogroups identified from adult diarrhoeal out patients in Addis Ababa, Ethiopia (Aseffa et al., 1997; Mache et al., 1997).

The objective of this study is to evaluate the antibiogram and antibiotic resistance of bacterial strains from clinical isolates in Gedo Hospital, West Shoa Region, Oromia State, Ethiopia to create the awareness of periodic antibiotic susceptibility testing, antimicrobial resistance among the health care providers.

MATERIALS AND METHODS

Study design and specimen collection: The study was conducted in Gedo Hospital, West Shoa, Oromia state, Ethiopia. A convenience sample of patients who visited the adult outpatient department of Gedo hospital for diarrheal complaint was incorporated into the study. The convenient sampling technique employed to collect 200 samples and the patients who took any antibiotics for treatment was not included in the study. About 1.5 g of diarrheal stool was collected randomly using clean, dry, non-leak wide mouth stool containers from patients with different socio-demographic status.

Bacteriological investigation: The specimens collected were precisely transported to the Ambo University Microbiology Laboratory using Cary Blair transport media for isolation and identification of bacteria (NCCLS, 2004). Using sterile swabs, the samples were directly inoculated onto plates of Deoxycholate Citrate Agar and Xylose Lysine Deoxycholate Agar and the plates were incubated aerobically at 37°C for 24 h. The same samples were plated onto Selenite F broth and incubated as a fore stated for enrichment. Following the incubation of Selenite F broth, a loop (0.001 mL) were streaked onto both Deoxycholate Citrate Agar and Xylose Lysine Deoxycholate plates and incubated at 37°C for 24 h. Two to three colonies suspected were selected, purified by streaking on to nutrient agar plates and characterized biochemically using Klingler Iron Agar, Urease tests, motility and indole test (NCCLS, 2004).

Antibiotic susceptibility test: Antibiotic susceptibility tests were performed by disk diffusion method using guidelines established by Bauer et al. (1966) and recommended by the Clinical and Laboratory Standards Institute utilizing commercial antibiotic discs. Characterization of the resistance or susceptibility profile of the isolates were determined by measuring the inhibitory zone and then compared with the interpretative chart to determine the susceptibility of the isolates to the antibiotics (Thirumurugan et al., 2009). The antibiotics tested include Amoxicillin 20 μg (AMX), Chloramphenicol 30 μg (CHP), Tetracycline 30 μg (TEC), Gentamicin 30 μg (GEM), Co-trimaxazole 10 μg (COT) and Ciprofloxacin 30 μg (CIF) were purchased local distributers (Liverpool, UK).

Quality control: Culture media were checked for sterility and performance. Standard strains of E. coli ATCC 25922 and Staphylococcus aureus ATCC 25923 were employed for culture and antimicrobial susceptibility testing.

Data analysis: Tables and percentages were used to describe findings. Cross tabulations were utilized to examine the levels of antimicrobial resistance to the respective antibiotics.

Ethical considerations: Written consent was obtained from every study participant. The study will not cause any harm to the respondent, sample collectors and supervisors. The privacy was maintained for each procedure and confidentiality has also been kept. Ethical clearance was sought from the College of Medicine and Health Sciences and Ethical Review Committee, Ambo University, Ambo, Ethiopia.

RESULTS

In this study, 40 stool samples were detected with the diarrheagenic enterobacteriaceae from 200 diarrheal samples. Of 200 participants, 93 were male and 107 were female. Eighteen of the patients who became positive were male and 22 positive patients were female. Age distribution of diarrheagenic enterobacteriaceae positive patients ranged from 0-64 years. Nine (22.5%) diarrheagenic enterobacteriaceae positive patients were pediatric age group (up to 16 years) and out of nine pediatric patients, 4 was male and 5 were female. Fourteen (35%) positive samples were detected from the age group of 17-32 years and out of 14, 4 was male and 10 were female. From the age group 33-48, 12(30%) stool samples detected were diarrhegenic and 7 were male, 5 were female. Only 5 positive samples with 3 males and 2 females were detected in the age group of 49-64 years (Table 1).

Table 1: Distribution of Shiegella and Salmonella strains isolated from diarrhea stool by age and sex in Gedo hospital, Western shoa, Oromia state

Table 2: Percentage of stool consistency in patients with Shiegella and Salmonella infection in Gedo hospital, Western shoa, Oromia state

Table 3: Comparison of stool consistency results for age and sex groups of patients in Gedo hospital

Of a 40 enterobacteriaceae infected stool with various consistencies, 22 were Shiegella strains and 18 were Salmonella strains. Diarrheal stool samples were classified as watery diarrhea, mucoid diarrhea, bloody diarrhea and bloody mucoid diarrhea. Totally 4(10%) has been identified as watery diarrhea, Shiegella sp. were detected from 3(13.6) samples and Salmonella sp. were detected from one (5.6%) sample. Mucoid stools were 18(45%) in number, 12(54.6%) samples showed the presence of Shiegella sp. and 6(33.3%) showed the presence of Salmonella. Seven (17.5%) samples were bloody diarrhea and Shiegella, Salmonella detected were 2(9.1%), 5(27.8%), respectively. Among 11(27.5%) bloody mucoid diarrhea samples, 5(22.7%) were Shiegella and 6(33.3%) were Salmonella (Table 2 and 3).

Antimicrobial susceptibility test was carried out using the disc diffusion method against the Shiegella and Salmonella strains.

Table 4:Antimicrobial susceptibility profiling of Shiegella and Salmonella isolates among patients with diarrhea stool in Gedo hospital, Western shoa, Oromia state

Antimicrobial resistance pattern results were categorized as resistance, intermediate and susceptible for all antibiotics. Among all antibiotics tested, of a 22 Shiegella, 21(95.5%) Shiegella strain showed the resistance, 1(4.5%) intermediate to Amoxicillin (AMX) and 17(77.3%) Shiegella strain showed resistance, 5(22.7%) intermediate against TEC. Gentamicin (GEM) showed activity against Shiegella strain compared with other antibiotics tested, 16(72.7%) were susceptible and 6(27.3%) were intermediate. The CIF 11(50%) and COT 10(45.5%) showed more or less equal susceptibility pattern, 2(9.1%) were resistance, 9(40.9%) were intermediate for CIF and 3(13.6%) were resistance, 9(40.9%) were intermediate for COT. CHP showed 8(36.4%) resistance, 9(40.9%) intermediate and 5(22.7%) were susceptible against Shiegella.

AMX showed poor antibacterial activity against Salmonella compared with the Shiegella strain. Of 18 diarrheagenic Salmonella isolates, total 18(100%) were resistance for AMX but CIF showed good antibacterial spectrum, 17(94.4%) were susceptible and 1(5.6%) were intermediate. After CIF, COT showed 9(50%) susceptibility, 5(27.8%) were resistance and 4(22.2%) were intermediate. CHP, TEC and GEM showed more or less equal resistance pattern against Salmonella strain. CHP showed 13(72.2%) resistance, 2(11.1%) intermediate and 3(16.7%) were susceptible and GEM showed 13(72.2%) resistance, 3(16.7%) intermediate and 2(11.1%) was susceptible. Also TEC showed weak antibacterial activity, 15(83.3%) were resistance, 2(11.1%) were intermediate and 1(5.6%) were susceptible (Table 4).

DISCUSSION

The present study provides results of antibiotics resistance pattern of diarrheagenic Shiegella and Salmonella strains isolates conducted in Gedo hospital, West Shoa, Oromia, Ethiopia. Out of 200 samples, 40 diarrheagenic bacteria were isolated and identified as Shiegella (22 = N) and Salmonella strains (18 = n) (Table 1). Similarly, the study conducted at Harar, Estern Ethiopia reveals the presence of Salmonella (28 = n) and Shiegella (17 = n) isolates from 244 stool samples Reda et al. (2011), Asrat (2008) and Aseffa et al. (1997) also reported the similar percentage of presence of Salmonella and Shiegella strains in the diarrheal stool, but Roma et al. (2000) reported 34.6% prevalence of Shiegella isolated from a study done in Awassa, Southern Ethiopia. Out of 40 diarrheagenic isolates, 18(45%) were isolated from the mucoid diahrrea, 7(17.5%) from bloody diarrhea and 11(27.5%) from the bloody mucoid diarrhea sample (Table 2 and 3). The previous literature showed the many controversy in the presence of Shiegella and Salmonella strains, Asrat (2008) reported that 82.4% of Shiegella and Salmonella strains were isolated from watery diarrhea stool but Reda et al. (2011) reported that mucoid were 46.8%, mucoid and bloody were 42.8%. This may be because of variations in the strain from place to place and our findings were similar compared to the study done in Harar, Eastern Ethiopia (Reda et al., 2011).

Antimicrobial resistance was high among the Shiegella strains isolated in the study were resistant to AMX (95.5%), TEC (77.3%) and CHP (36.4%) and these findings were comparable with previous studies carried out in Ethiopia (Asrat, 2008; Mache et al., 1997; Roma et al., 2000). The Shiegella strains showed no resistance to GEM, but six had intermediary susceptibility and 72.7% were susceptible to this antibiotic tested. Similarly, the study done in harar, Estern Ethiopia showed that Shiegella strains were 94.1% of susceptibility, 0% of resistance to GEM but the study conducted in Gondar Teaching hospital, Northwest Ethiopia showed that Shiegella strains were 10% resistant to GEM (Huruy et al., 2011) and similar data has been reported in other African countries, such as Nigeria (Iwalokun et al., 2001) (Table 4). Resistance to CIF (9.1%) was observed in our study comparable with previous study in which 3.1% of were resistant against CIF (MoezArdalan et al., 2003) but 8.3% of resistance pattern of Shiegella strains to CIF were reported in the study conducted in the Gondar Teaching hospital, Northwest Ethiopia (Huruy et al., 2011). COT showed less susceptible (45.5%), 13.6% resistances compared with the CIF but the previous study showed 91.7% of resistance to COT (Jafari et al., 2008). The pattern of resistance to Salmonella strains isolates were 100% resistant to AMX, 83.3% to TEC followed by CHP (72.2%) and GEM (72.2%). The CIF showed 94.4% susceptibility, 0% resistance and 5.6% intermediate against Salmonella strains. COT showed 50% susceptibility and 27.8% resistance against Salmonella strains isolated in our study. The similar results were observed in the previous studies conducted in Ethiopia (Asrat, 2008; Huruy et al., 2011; Amabile-Cuevas, 2010). The high resistant to AMX might be due to high rate of AMX prescription in the study area and CHP, TEC resistance might reflect the indiscriminate and widespread uses of antibiotics in public health practices since the society in the setting have easy access to different antibiotics without prescription. Also, an antibiotic has been prescribed irrationally without susceptibility test.

CONCLUSION AND RECOMMENDATION

Shiegella and Salmonella strains were susceptible to gentamicin and ciprofloxaxin, respectively but a high rate of resistance were observed to AMX, CHP, TEC for both Shiegella and Salmonella strains. Notably, both bacteria have developed complete resistance to AMX. We suggest that gentamicin may be the drug choice for Shiegella strains caused diarrhea and ciprofloxacin for the diarrhea caused by Salmonella strains in the study area. However, there is required of extensive prevalence, antibacterial susceptibility profiling and underlying mechanisms of antimicrobial resistance by Shiegella and Salmonella isolates to be conducted. Moreover, selection of antibiotics and treatment of infections based on bacterial species identification and in vitro susceptibility testing should be implemented rather than current empirical treatment.

ACKNOWLEDGMENTS

This study was carried out with the financial support of the Ambo University. We appreciate the assistance of Gedo Hospital medical directors for their kindly support during data collection. We would like to acknowledge also College of Medicine and Health Sciences for consistent support of materials and appropriate guidance.

REFERENCES
Abula, T. and M. Kedir, 2004. The pattern of antibiotic usage in surgical in-patients of a teaching hospital, northwest Ethiopia. Ethiop. J. Health Dev., 18: 35-38.
Direct Link  |  

Amabile-Cuevas, C., 2010. Antibiotic resistance in Mexico: A brief overview of the current status and its causes. J. Infect. Dev. Ctries, 29: 126-131.
PubMed  |  

Aseffa, A., E. Gedlu and T. Asmelash, 1997. Antibiotic resistance of prevalent salmonella and shigella strains in northwest Ethiopia. East. Afr. Med. J., 74: 708-713.
PubMed  |  

Asrat, D., 2008. Shigella and Salmonella serogroups and their antibiotic susceptibility patterns in Ethiopia. East. Mediterr. Health J., 14: 760-767.
PubMed  |  

Bauer, A.W., W.M. Kirby, J.C. Sherris and M. Turck, 1966. Antibiotic susceptibility testing by a standardized single disk method. Am. J. Clin. Pathol., 45: 493-496.
PubMed  |  

Collee, J.G., R.S. Miles and B. Watt, 1996. Tests for Identification of Bacteria. In: Mackie and McCartney Practical Medical Microbiology, Collee, J.G., A.G. Fraser, B.P. Marmion and A. Simmons (Eds.). 14th Edn., Elsevier India Pvt. Ltd., India, pp: 131-149.

Cook, G.C. and A.I. Zumla, 2003. Malabsorption in the Tropics. In: Manson's Tropical Diseases, Cook, G.C. and A.I. Zumla (Eds.). 21st Edn., W.B. Saunders Company, USA., ISBN-13: 978-0702026409, pp: 121-126.

Desalegn, A.S., 2013. Assessment of drug use pattern using WHO prescribing indicators at Hawassa University teaching and referral hospital, south Ethiopia: A cross-sectional study. BMC Health Services Res., Vol. 13. 10.1186/1472-6963-13-170

Feleke, M., W. Yenet and J.L. Lenjisa, 2013. Prescribing pattern of antibiotics in pediatric wards of Bishoftu Hospital, East Ethiopia. Int. J. Basic. Clin. Pharmacol., 2: 718-722.
CrossRef  |  Direct Link  |  

Holmes, K.K., D.W. Johnson and T.M. Floyd, 1967. Studies of venereal disease. I. Probenecid-procaine penicillin G combination and tetracycline hydrochloride in the treatment of penicillin-resistant gonorrhea in men. J. Am. Med. Assoc., 202: 461-473.
CrossRef  |  Direct Link  |  

Huruy, K., A. Kassu, A. Mulu, N. Worku and T. Fetene et al., 2011. Intestinal parasitosis and shigellosis among diarrheal patients in gondar teaching hospital, Northwest Ethiopia. BMC Res. Notes, Vol. 4. 10.1186/1756-0500-4-472

Iwalokun, B.A., G.O. Gbenle, S.I. Smith, A. Ogunledun, K.A. Akinsinde and E.A. Omonigbehin, 2001. Epidemiology of shigellosis in Lagos, Nigeria: Trends in antimicrobial resistance. J. Health Popul. Nutr., 19: 183-190.
PubMed  |  

Jafari, F., M. Hamidian, M. Rezadehbashi, M. Doyle, S. Salmanzadeh-Ahrabi, F. Derakhshan and M.R. Zali, 2008. Prevalence and antimicrobial resistance of diarrheagenic Escherichia coli and Shigella species associated with acute diarrhea in Tehran, Iran. Can. J. Infect. Dis. Med. Microbiol., 19: e56-e62.
Direct Link  |  

Mache, A., Y. Mengistu and S. Cowley, 1997. Salmonella serogroups identified from adult diarrhoeal out-patients in Addis Ababa, Ethiopia: Antibiotic resistance and plasmid profile analysis. East Afr. Med. J., 74: 183-186.
PubMed  |  

Mengistu, G., G. Mulugeta, T. Lema and A. Aseffa, 2014. Prevalence and antimicrobial susceptibility patterns of Salmonella serovars and Shigella species. J. Microb. Biochem. Technol.,

MoezArdalan, K., M.R. Zali, M.M. Soltan-Dallal, M.R. Hemami and S. Salmanzadeh-Ahrabi, 2003. Prevalence and pattern of antimicrobial resistance of Shigella species among patients with acute diarrhoea in Karaj, Tehran, Iran. J. Health Popul. Nutr., 21: 96-102.
PubMed  |  Direct Link  |  

NCCLS., 2004. Performance standards for antimicrobial susceptibility testing. National Committee for Clinical Laboratory Standards (NCCLS) Approved Standard M100-S14, Wayne, PA., USA.

Reda, A.A., B. Seyoum, J. Yimam, G. Andualem, S. Fiseha and J.M. Vandeweerd, 2011. Antibiotic susceptibility patterns of Salmonella and Shigella isolates in Harar, Eastern Ethiopia. Infect. Dis. Immun., 3: 134-139.
Direct Link  |  

Roma, B., S. Worku, S.T. Mariam and N. Langeland, 2000. Antimicrobial susceptibility pattern of Shigella isolates in Awassa. Ethiop. J. Health Dev., 14: 149-154.
Direct Link  |  

Sharma, R., C.L. Sharma and B. Kapoor, 2005. Antibacterial resistance: Current problems and possible solutions. Indian J. Med. Sci., 59: 120-129.
PubMed  |  

Thirumurugan, G. and M.D. Dhanaraju, 2011. Novel biogenic metal nanoparticles for pharmaceutical applications. Adv. Sci. Lett., 4: 339-348.
CrossRef  |  

Thirumurugan, G., S.M. Shaheedha and M.D. Dhanaraju, 2009. In-vitro evaluation of anti-bacterial activity of silver nanoparticles synthesised by using phytophthora infestans. Int. J. ChemTech Res., 1: 714-716.

©  2019 Science Alert. All Rights Reserved
Fulltext PDF References Abstract