ABSTRACT
Nowadays, antibiotic resistance is a global public health threat. Bangladesh is accelerated to this owing to its sub-standards healthcare along with the self-medication and overuse of antibiotics. The study aimed to assess patterns of antibacterial resistance in the clinical samples. The study was carried out at Ibn Sina Diagnostic and Consultation Center Uttara, Dhaka, from January to December 2019. All cultures and antimicrobial susceptibility test results of patients were extracted from laboratory records, using a semi-structured checklist. Data were analyzed using Microsoft Excel and SPSS version 20.0. To ensure confidentiality coding was used instead of the patient’s identity. A total of 925 culture-positive results were analyzed, of which blood 620(65.0%) and urine 297(32.1%) samples were commonly diagnosed. The most frequently isolated bacterial were Salmonella spp. [601(65%)], Escherichia coli [244(26.4%)] and Klebsiella spp. [57(6.16%)]. The majority of the patients were females [540(58.4%)]. E. coli was found to be highly sensitive (>80%) to nitrofurantoin, meropenem, amikacin, amoxiclav, and imipenem; simultaneously, resistant (>45%) to cefixime, cephalexin, piperacillin, aztreonam, ampicillin, cefuroxime, and ciprofloxacin. S. typhi and S. paratyphi were sensitive (>80%) for cefepime, ceftriaxone, imipenem, tetracycline, cefixime, ceftazidime, cephalexin, cotrimoxazole, aztreonam, cefuroxime, and amoxiclav; concurrently, above 80% resistance for ciprofloxacin, azithromycin, gentamycin, and ampicillin. Overall, most of the isolates showed a significant rising rate of microbial resistance to ciprofloxacin, azithromycin, piperacillin, cephalexin, gentamycin, and ampicillin. The study findings revealed gradually rising rates of antibiotic resistance to commonly prescribed antibiotics. The study suggested the prescribers should be avoided overuse and irrational use of drugs to reduce antimicrobial resistance.
Key words: Bacteriology, antibiotic susceptibility, clinical samples, Dhaka.
Antibiotic resistance is a well-known public health concern at the community, national and global levels (Nordberg et al., 2004). Decreasing the effectiveness of antibiotics in treating bacterial common infections and a decline in the new drug development rate is a concerning issue (Kandelaki et al., 2015; Luepke and Mohr, 2017; Spellberg et al., 2004). Antibiotic resistance poses a significant risk of mortality and economic burden worldwide (Ahmed et al., 2019). The causes of antibiotic resistance are complex which include enzymatic degradation of antibacterial drugs, alteration of bacterial proteins that are antimicrobial targets, and changes in membrane permeability to antibiotics (Kandelaki et al., 2015). The low- and middle-income countries are more affected because of extensive misuse of antibiotics, non-human antibiotic use, poor quality of drugs, insufficient surveillance, and other factors associated with poor healthcare standards, malnutrition, chronic and repeated infection, unaffordability of more effective and costly drugs (Ayukekbong et al., 2017; Sosa et al., 2010). In 2014, the World Health Organization (WHO) reported on global surveillance of antimicrobial resistance, significant gaps prevail in surveillance, absence of standards methodology, data sharing and coordination. WHO identified the major gaps in the South-East Asia Region, the African Region, and the Eastern Mediterranean region (WHO, 2014).
Bangladesh is one of the South-East Asian developing countries and has a high rate of antibiotic resistance which poses a regional and global concern (Rahman and Huda, 2014). Enteric fever caused by salmonella spp. has been detected among children aged <5 years of age than the age group ≥5 years in the South-East Asian especially in India and Bangladesh. Though, there are no valid data regarding paratyphoid fever in Bangladesh (Naheed et al., 2010). Therapeutic failures in Bangladesh are not uncommon. Multiple studies have demonstrated irrational antibiotic prescribing by physicians, self-medication habits of patients, and indiscriminate use of antibiotics in agriculture and farming in different segments of the country (Biswas et al., 2014a; Biswas et al., 2014b; Sutradhar et al., 2014). Therefore, the prevalence of antibiotic resistance in Bangladesh is high, but no attempts have been undertaken to alleviate it. This study aims to serve as a reference for future works and to guide policymakers and prescribers to adopt the best strategy to lower the extent of antibiotic resistance as well as combat the problems following the expanding resistance.
Study design and setting
This retrospective cross-sectional study was executed from January to December 2019 at IbnSina Diagnostic and Consultation Center Uttara, Dhaka. The sample was collected by using a sterile ascetic technique. A total of 925 culture-positive test result samples were analyzed. All cultures and antimicrobial susceptibility test results of patients were extracted from laboratory records notebook by using a semi-structured checklist. The sample-set included blood, urine, stool and sputum samples as well as wound swabs.
Bacterial isolates and identification
All of the received clinical specimens were initially cultured and subcultures into brain heart infusion, blood agar, Salmonella-Shigella agar, Chocolate agar and Mac-Conkey agar as per need, and after overnight incubation at 37°C, the bacteria identification was completed by gram staining as well as standard biochemical tests (catalase, coagulase, oxidase). This was done by sub-culturing on mediums such as triple sugar iron agar (TSI), SIM medium, and Simmons’ citrate agar.
Antimicrobial susceptibility tests
The antimicrobial sensitivity tests of the isolated bacteria were performed by using the Kibry Bauer disk diffusion test on Mueller-Hinton agar (Bauer et al., 1966). The antibiotics agents used were: tetracycline (30 μg), nitrofurantoin (300 μg), azithromycin (15 μg), gentamicin (10 μg), ciprofloxacin (5 μg), doxycycline (30 μg), cotrimoxazole (25 μg), imipenem (10 μg), ceftriaxone (30 μg), ceftazidime (30μg), cefepime (30μg), meropenem (10μg), ampicillin (10 μg), penicillin (g) (10 μg). cefixime (5μg), cephalexin (30μg), piperacillin (75 μg), aztreonam (30 μg), cefuroxime (30 μg) amikacin (30μg), amoxiclav (30μg) vancomycin (30μg) fusidic acid (10μg) and cloxacillin (30 μg). The pattern of sensitivity and resistance was interpreted according to the guideline of the National Committee for Clinical Laboratory Standards (NCCLS) (National Committee for Clinical Laboratory Standards and Barry, 1999).
Statistical analysis
The data were entered into Microsoft Excel and analyzed by SPSS version 20. The results were presented as descriptive statistics in terms of relative frequency, percentage, mean ± standard deviation (SD) and to summarize patients’ attributes and other related information.
Ethical considerations
Ethical approval was obtained from the Institutional Review Board. Administrative authorization for this study was obtained from the Branch Manager of the Diagnostic Center. The researchers highly consider the human right of the participants. To ensure the confidentiality coding method was used instead of other identifiers of the patients.
A total of 925 samples were analyzed; 32.1% presented urine, followed by stool (0.5%), blood (67%), sputum (0.2%) and wound swab (0.1%) (Figure 1). Among them males were 385 (41.6%) and females were 540 (58.4%). The ages of the patients ranged from 3 months to 90 years with a mean age of 27 years. The infection was most common among age groups of above >40 years (22.4%) followed by age groups of 20-29 years (22.6%) (Table 1). The set of most frequently isolated organisms included Salmonella spp., 601(65.0%); Escherichia coli were,244(26.4%) and Klebsiella spp. and Staphylococcus aureus, 57(6.16%) and 15(1.62%) respectively (Figure 2 and Table 2).


The sensitivity pattern of E. coli shows that the microbes were highly (>80%) sensitive for imipenem, nitrofurantoin, gentamycin, meropenem, amikacin, and amoxiclav. E. coli is booming developed resistance (>45%) for some antibiotics such as cefixime, cephalexin, piperacillin, aztreonam, ampicillin, cefuroxime, and ciprofloxacin. The microbes Salmonella spp. (S. typhi and S. paratyphi) were more than 80% sensitive tocefepime, ceftriaxone, imipenem, tetracycline, cefixime, ceftazidime, cephalexin, gentamycin, cotrimoxazole, aztreonam, ampicillin, cefuroxime, amikacin, and amoxiclav. At the same time above 80% had developed resistance to ciprofloxacin and azithromycin. S. typhi has developed (>20%) resistant to several antibiotics like ampicillin, piperacillin, and cotrimoxazole.
Klebsiella spp. was above 80% sensitive tocefepime, ceftriaxone, imipenem, gentamycin, meropenem, amikacin, and amoxiclav. This bacteriumhas developed significant resistance (>40%) to some antibiotics such as ciprofloxacin, piperacillin, cephalexin, and cefepime. Staphylococcus aureus was highly (>80%) sensitive tocotrimoxazole, gentamycin, amikacin, doxycycline, vancomycin, fusidic acid, cloxacillin. S. aureus developed resistance for ciprofloxacin, penicillin, and azithromycin. Pseudomonas spp. was substantially sensitive to cefepime, imipenem, ceftazidime, piperacillin, gentamycin, and amikacin. These bacteria developed resistance to aztreonam, cefepime, and tetracycline. Proteus spp. was 100% sensitive tocefepime, imipenem, cotrimoxazole, piperacillin, amikacin, gentamycin, meropenem, amikacin, and amoxiclav. Of the resistance to ceftazidime and cefixime, throughout the study period, only two samples were positive Proteus spp. Enterococcus spp. was 100% resistance tocefepime, ceftriaxone, imipenem, cotrimoxazole, tetracycline, cefixime, ceftazidime, gentamycin, amikacin and meropenem and resistive for piperacillin, ampicillin, azithromycin. Though, throughout the study period, we have found a single culture positive of Enterococcus spp. (Table 3).


Bacterial infections are the predominant problem in developing countries like Bangladesh where water, sanitation, and hygiene (WASH) continue to be below international standard. The shortage of reliable microbial and antimicrobial data is also a problem in managing the physicians treating patients with a bacterial infection before the appropriate treatment is applied to get the best outcome (Tjaniadi et al., 2003). The major cause behind antibiotic resistance makes the bacteria to be smart. However, in Bangladesh, prescribers usually diagnose microbial infection based on clinical finding and choose antimicrobial drugs on an experiential basis (Faiz and Rahman, 2004), which critically distresses the sensitivity pattern of microorganisms. Besides, the unwillingness of the policymakers and officials to sanction law to overcome insufficient guidelines and instruction to control antimicrobial prescription and administration leads to the deteriorating of the circumstance.
In the present study, female patients were found to be higher than the males as found in other studies (Derbie et al., 2017; Tahira and Singh, 2017; Kolawole et al., 2009). The females were more infected than males due to their physiological and anatomical differences (Kibret and Abera, 2014). The current study revealed that Escherichia coli was highly sensitive (>80%) to imipenem, nitrofurantoin, gentamycin, meropenem, amikacin, and amoxiclav. The susceptibility pattern of E. coli for nitrofurantoin was (89.7%), which compares to a study done in London (Bean et al., 2008). E. coli is developing resistance (>45%) to some antibiotics such as cefixime, cephalexin, piperacillin, aztreonam, ampicillin, cefuroxime, and ciprofloxacin. Ahmed et al. (2019) showed that E. coli was highly resistance to commonly used antibiotics like ampicillin (94.6-100%), amoxiclav (67.1-85.5%), ciprofloxacin (65.2-80.5%) and cotrimoxazole (72-82.2%). Patil and Mule described the isolates of E. coli were resistant to ampicillin (96.6%), tetracycline (79%), ceftriaxone (62%) and gentamicin (51.7%) (Patil and Mule, 2019).
Salmonella spp. showed (>80%) sensitive to cefepime, ceftriaxone, imipenem, tetracycline, cefixime, ceftazidime, cephalexin, gentamycin, cotrimoxazole, aztreonam, ampicillin, cefuroxime, amikacin, and amoxiclav. This result is comparable with Ahmed et al. (2019) who demonstrated Salmonella spp. was highly sensitive to cefixime and ceftriaxone. Salmonella typhi developed resistance to ciprofloxacin and azithromycin. Ebrahim et al. (2016) showed that Salmonella typhi has developed resistance from 2003 to 2014 from 0% to 14% in Canada. The previous literature showed Salmonella isolates had 100% sensitivity to ceftriaxone and cefixime (Bhan et al., 2005; Bhetwal et al., 2017; and Mule, 2019).
Klebsiella spp. was above 80% sensitive to cefepime, ceftriaxone, imipenem, gentamycin, meropenem, amikacin, and amoxiclav. These bacteria developed significant resistance (>40%) to some antibiotics like ciprofloxacin, piperacillin, cephalexin, and cefepime. Klebsiellaspp.
Sensitive to Meropenem (100%), Nitrofurantoin (83.3%), whereas it is resistant to Penicillin (100%), Ampicillin (93.61%) and intermediate to Gentamicin (18.5%), Augmentin (17.4%) (Ahmed et al., 2019). Antimicrobial sensitivity pattern reveals the development of resistance to common antibiotics, which is comparable with that of previous studies (Akond et al., 2009; Rogers et al., 2011). However, antibiotics like ciprofloxacin, ceftazidime, ampicillin, piperacillin, cotrimoxazole, and ceftriaxone have become ineffective because of the beginning of the resistance. The microbes are still sensitive to imipenem, azithromycin, vancomycin and amikacin, nitrofurantoin, gentamycin, meropenem, and amoxiclav. This outline of sensitivity does not compare with previous findings (Hasan et al., 2011). Prescribers cited diagnostic insecurity and advent of resistance as pediment reasons for prescribing antimicrobials; nevertheless some of them revealed the possibility of losing patients as one reason (Rahman and Huda, 2014).In this study, we found a very small proportion of S. aureus 15(1.7%), Pseudomonas spp. 4(0.43%), Proteus spp. 2(0.21%), Enterococcus spp. 1(0.10%). Therefore, we did not compare it with other studies.
The present study revealed that E. coli, salmonella typhi, and salmonella paratypi were the most frequently isolated bacterial in the clinical samples. The majority of the isolated bacteria showed certain levels of antimicrobial resistance to commonly recommended drugs like ampicillin, norfloxacin, ciprofloxacin, azithromycin, cephalexin, piperacillin and cotrimoxazole. However, strict policy and appropriate use of antibiotics can assuage the burden of antimicrobial resistance. It is highly suggested to perform antimicrobial susceptibility testing before the administration of antibiotics and ensure the rational use of drugs to reduce antibiotic resistance.
The authors have not declared any conflict of interests.
REFERENCES
Ahmed I, Rabbi MB, Sultana S (2019). Antibiotic resistance in Bangladesh: A systematic review. International Journal of Infectious Diseases 80:54-61.
Crossref
|
|
Akond MA, Alam S, Hassan SMR, Shirin M (2009). Antibiotic resistance of Escherichia coli isolated from poultry and poultry environment of Bangladesh. Internet Journal of Food Safety 11:19-23.
|
|
|
Ayukekbong JA, Ntemgwa M, Atabe AN (2017). The threat of antimicrobial resistance in developing countries: causes and control strategies. Antimicrobial Resistance and Infection Control 6(1):47.
Crossref
|
|
|
Bauer HW, Kirby WM, Sherris JC, Turck M (1966). Antibiotic susceptibility testing by standard single disc method. American Journal of Clinical Pathology 45:494-549.
Crossref
|
|
|
Bean DC, Krahe D, Wareham DW (2008). Antimicrobial resistance in community and nosocomial Escherichia coli urinary tract isolates, London 2005-2006. Annals of Clinical Microbiology and Antimicrobials 7(1):13.
Crossref
|
|
|
Bhan MK, Bahl R, Bhatnagar S (2005). Typhoid and paratyphoid fever. The Lancet 366(9487):749-762.
Crossref
|
|
|
Bhetwal A, Maharjan A, Khanal PR, Parajuli NP (2017). Enteric fever caused by Salmonella entericaSerovars with reduced susceptibility of fluoroquinolones at a community based teaching Hospital of Nepal. International Journal of Microbiology 2017.
Crossref
|
|
|
Biswas M, Roy DN, Tajmim A, Rajib SS, Hossain M, Farzana F, Yasmen N (2014a). Prescription antibiotics for outpatients in Bangladesh: a cross-sectional health survey conducted in three cities. Annals of Clinical Microbiology and Antimicrobials 13(1):15.
Crossref
|
|
|
Biswas M, Roy MN, Manik MIN, Hossain MS, Tapu STA, Moniruzzaman M, Sultana S (2014b). Self medicated antibiotics in Bangladesh: a cross-sectional health survey conducted in the Rajshahi City. BMC Public Health 14(1):847.
Crossref
|
|
|
Derbie A, Hailu D, Mekonnen D, Abera B, Yitayew G (2017). Antibiogram profile of uropathogens isolated at Bahir Dar regional health research laboratory centre, northwest Ethiopia. The Pan African Medical Journal 26.
Crossref
|
|
|
Ebrahim M, Gravel D, Thabet C, Abdesselam K, Paramalingam S, Hyson C (2016). Anitmicrobial Resistance (AMR): Antimicrobial use and antimicrobial resistance trends in Canada: 2014. Canada Communicable Disease Report 42(11):227.
Crossref
|
|
|
Faiz MA, Rahman MR (2004). Rational antimicrobial use. Journal of Chittagong Medical College Teacher's Association 15:1-3.
|
|
|
Hasan B, Nahar SG, Akter L, Saleh AA (2011). Antimicrobial sensitivity pattern of Salmonella typhi isolated from blood culture in a referral hospital. Bangladesh Journal of Medical Microbiology 5(1):16-20.
Crossref
|
|
|
Kandelaki K, Lundborg CS, Marrone G (2015). Antibiotic use and resistance: a cross-sectional study exploring knowledge and attitudes among school and institution personnel in Tbilisi, Republic of Georgia. BMC Research Notes 8(1):495.
Crossref
|
|
|
Kibret M, Abera B (2014). Prevalence and antibiogram of bacterial isolates from urinary tract infections at Dessie Health Research Laboratory, Ethiopia. Asian Pacific Journal of Tropical Biomedicine 4(2):164-168.
Crossref
|
|
|
Kolawole AS, Kolawole OM, Kandaki-Olukemi YT, Babatunde SK, Durowade KA, Kolawole CF (2009). Prevalence of urinary tract infections (UTI) among patients attending DalhatuAraf Specialist Hospital, Lafia, Nasarawa state, Nigeria. International Journal of Medicine and Medical Sciences 1(5):163-167.
|
|
|
Luepke KH, Mohr III JF (2017). The antibiotic pipeline: reviving research and development and speeding drugs to market. Expert Review of Anti-Infective Therapy 15(5):425-433.
Crossref
|
|
|
Naheed A, Ram PK, Brooks WA, Hossain MA, Parsons MB, Talukder KA, Mintz E, Luby S, Breiman RF (2010). Burden of typhoid and paratyphoid fever in a densely populated urban community, Dhaka, Bangladesh. International Journal of Infectious Diseases 14:e93-e99.
Crossref
|
|
|
National Committee for Clinical Laboratory Standards, Barry AL (1999). Methods for determining bactericidal activity of antimicrobial agents: approved guideline (Vol. 19). Wayne, PA: National Committee for Clinical Laboratory Standards.
|
|
|
Nordberg P, Monnet DL, Cars O (2004). Antibacterial Drug Resistance (Background Document for the WHO Project: Priority Medicines for Europe and the World. A Public Health Approach to Innovation). WHO: Geneva, Switzerland.
|
|
|
Patil N, Mule P (2019). Sensitivity Pattern of Salmonella typhi and Paratyphi a Isolates to Chloramphenicol and other Anti-Typhoid Drugs: An in Vitro Study. Infection and Drug Resistance 12:3217.
Crossref
|
|
|
Rahman MS, Huda S (2014). Antimicrobial resistance and related issues: An overview of Bangladesh situation. Bangladesh Journal of Pharmacology 9(2):218-224.
Crossref
|
|
|
Rogers BA, Aminzadeh Z, Hayashi Y, Paterson DL (2011). Country-to-country transfer of patients and the risk of multi-resistant bacterial infection. Clinical Infectious Diseases 53(1):49-56.
Crossref
|
|
|
Spellberg B, Powers JH, Brass EP, Miller LG, Edwards Jr JE (2004). Trends in antimicrobial drug development: implications for the future. Clinical Infectious Diseases 38(9):1279-1286.
Crossref
|
|
|
Sosa ADJ, Byarugaba DK, Amábile-Cuevas CF, Hsueh PR, Kariuki S, Okeke IN (2010). Antimicrobial Resistance in Developing Countries (p. 554). New York: Springer.
Crossref
|
|
|
Sutradhar KB, Saha A, Huda NH, Uddin R (2014). Irrational use of antibiotics and antibiotic resistance in southern rural Bangladesh: perspectives from both the physicians and patients. Annual Research and Review in Biology, pp.1421-1430.
Crossref
|
|
|
Tahira F, Singh AK (2017). Bacterial and Antimicrobial Resistance Profle of Urinary Tract Infections among Women in Reproductive Age Group Attending the Tertiary Care Hospital Set Up. International Journal of Contemporary Medical Research 4:77-83.
|
|
|
Tjaniadi P, Lesmana M, Subekti D, Machpud N, Komalarini S, Santoso W, Simanjuntak CH, Punjabi N, Campbell JR, Alexander WK, BEECHAM III HJ (2003). Antimicrobial resistance of bacterial pathogens associated with diarrheal patients in Indonesia. The American Journal of Tropical Medicine and Hygiene 68(6):666-670.
Crossref
|
|
|
World Health Organization (WHO) (2014). Antimicrobial resistance: global report on surveillance. World Health Organization.
|
|