Get Permission Kammineni, Yajamanam, Reddy, Shafi, and Basireddy: Antimicrobial resistance surveillance of various clinical isolates in a diagnostic laboratory- An observational study


Introduction

Infectious diseases are among the top most killers with great mortality and morbidity. Antibiotic prescription is a very common entity in a clinical setup. Because of the inadvertent and irrational use of antibiotics, increasingly drug resistance is being observed among the clinical isolates, which takes a great toll on the health, economical and social status of the affected. Drug resistance is observed among all the major bacterial species with Multidrug resistance commonly observed among nosocomial infections.1,2

India is one of the largest consumer of antibiotics in the world and also carries highest burden of drug resistance pathogens. Irrational and over use of antibiotics have been the culprits for drug resistance in India. With the emergence and rapid spread of metallobetalactamase producers like NDM-1 in India, only very few alternative antibiotics are available for the treatment. As there is scarcity of the national data, Indian council of medical research initiated Antimicrobial Resistance Surveillance & Research Network (AMRSN) in the year 2013 which covers antibiotic resistance surveillance of major bacterial groups. Increasing resistance of gram negative isolates has been observed by this network. Alarming rates of resistance to fluoroquinolones, third generation cephalosporins was observed. Increased carbapenem resistance is particularly worrisome.2

Antimicrobial resistance surveillance should be done at local, regional, and national level in order to formulate the antibiotic policies which helps in guiding the clinicians in rational use of antibiotics which decreases the mortality and morbidity and in turn the emergence of drug resistance. Knowledge about the local drug resistance pattern is extremely important for the clinician in order to choose the appropriate empirical antibiotic. The present study emphasizes on systematic analysis of the Antimicrobial resistance surveillance of various clinical isolates in and around Kurnool which helps the local clinicians in the better management of the patients.

Aims and Objectives

The present study aims in identifying the etiological agents from various clinical samples and detecting their resistance pattern.

Materials and Methods

All the samples sent to the Microbiology department in Bharathi Diagnostics, Kurnool, Andhra Pradesh, during the study period of January 2018 to December 2018, were processed according to the standard protocols. Antimicrobial susceptibility testing was done for all the significant isolates by using Kirby Bauer disc diffusion method and the interpretation was carried out as per the CLSI guidelines.3 The most common clinical samples obtained were Urine, Blood, Pus/ Wound swab, Sputum and other Body fluids.

Results

A total of 5687 samples were processed during study period out of which 1696 samples were culture positive. Most common sample was Urine, followed by Pus/ Wound swab, Blood, Sputum and Body fluids.

Urinary tract infections

Out of 3,164 urine samples obtained 941 have shown significant growth. The distribution of the various bacterial pathogens among the urine clinical isolates were as follows

Figure 1

Distribution of the various bacterial pathogens among the urine clinical isolates

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Table 1
Urine E.coli Klebsiella spp. Proteus spp. Pseudomonas
Amoxyclav 81.86 89.64 52.94 -
Ceftriaxone 69.87 67.12 29.41 -
Ciprofloxacin 67.61 63.06 23.53 25
Co-trimoxazole 60.39 63.2 76.47 -
CFS 32.89 42.73 5.88 40
Gentamycin 29.34 37.98 29.41 22.5
Fosfomycin 23.02 34.24 29.41 -
Nitrofurantoin 14.76 36.65 - -
PIT 19.69 23.32 5.88 25
Amikacin 12.45 18.5 5.88 22.5
Imipenem 4.12 7.23 0 2.5
Meropenem 3.39 5.85 0 2.5
Tigecycline 0 0 - -
Colistin 0 0 100 0

Resistant pattern of common urinary pathogens isolated during the study period

Figure 2

Resistant pattern of Enterococcus spp. and Staphylococcus spp. isolated from the urinary tract infections

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Wound infections

Out of 573 pus/ wound swab samples obtained 235 were culture positive. The distribution of the various bacterial pathogens among the clinical isolates were as follows

Figure 3

Distribution of the various bacterial pathogens among the pus specimens

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Table 2
PUS E.coli Klebsiella spp. Pseudomonas Proteus spp. Others
Amoxyclav 86.79 91.3 NA 77.78 81.82
Ceftriaxone 71.7 71.74 NA 66.67 81.82
Ciprofloxacin 73.58 60.87 23.08 44.44 36.36
Co-trimoxazole 73.58 67.39 NA 66.67 54.55
CFS 30.19 36.96 42.31 11.11 9.09
Gentamycin 35.85 41.3 26.92 55.56 54.55
Fosfomycin 44 48 NA - -
Aztreonam 73.58 60.87 34.62 55.56 81.82
PIT 30.19 39.13 34.62 11.11 18.18
Amikacin 16.98 23.91 19.23 22.22 36.36
Imipenem 1.89 10.87 7.69 0 0
Meropenem 1.89 10.87 3.85 0 0
Tigecycline 0 0 - 100 0
Colistin 0 0 0 100 0

Resistant pattern of common pathogens isolated from the wound infections during the study period

Figure 4

Resistant pattern of S.aureus and Enterococcus spp. isolated from the wound infections

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Blood cultures

A total of 821 blood cultures were obtained during the study period out of which 166 samples have shown positive blood culture.

Table 3
Type of pathogen From Blood Cultures (N=166) No.of isolates % isolated
E.coli 31 18.67
CONS 30 18.07
Klebsiella 28 16.81
Pseudomonas 22 13.25
S.aureus 16 9.63
NFGNB 09 5.42
Acinetobacter 08 4.81
Candida 06 3.41
Enterococcus 05 3.01
Salmonella Typhi 02 1.20
Proteus spp. 02 1.20
Others 07 4.21

Distribution of the various bacterial pathogens among the blood clinical isolates

Table 4
Blood E.coli Klebsiella spp. Pseudomonas
Amoxyclav 87 82 -
Ceftriaxone 81 71 -
Ciprofloxacin 74 57 14
Co-trimoxazole 58 64 -
CFS 10 36 -
Gentamicin 39 43 45
Ceftazidime - - 68
PIT 13 32 9
Amikacin 19 43 45
Imipenem 7 21 9
Meropenem 7 14 9
Tigecycline 0 0 0
Colistin 0 0 0
For Gram Positive Organisms
BLOOD S.aureus CONS
Ciprofloxacin 81 50
Levofloxacin 69 40
Gentamycin 40 17
Amoxyclav 56 43
Clindamycin 31 43
Cefoxitin 36 43
Teicoplanin 0 0
Amikacin 25 7
Vancomycin 0 0
Linezolid 0 0

Resistant pattern of common blood pathogens isolated during the study period

Sputum

Figure 5

Distribution of the various bacterial pathogens among the Sputum specimens

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Table 5
Sputum Klebsiella spp. Pseudomonas Acinetobacter sp.
Amoxyclav 87.5 NA 100
Ceftriaxone 62.5 NA 100
Co-trimoxazole 64.3 NA 88.9
Ciprofloxacin 48.2 23.8 100
Gentamycin 44.6 19.1 88.9
CFS 41.1 9.5 77.8
PIT 41.1 19.1 88.9
Amikacin 32.1 19.1 88.9
Imipenem 12.5 3.2 55.7
Meropenem 8.7 3.2 55.7
Tigecycline 0 - 0
Colistin 0 0 0

Resistant pattern of common pathogens isolated from sputum specimens during the study period

Discussion

A phenomenal increase in the drug resistance has been observed recently among the clinical isolates which ranged from resistance to a few antibiotics to almost all the available antibiotics (MDR, XDR, PDR ). In the present scenario of evidence based medicine the role of microbiology services have gained much importance compared to the earlier days. Better understanding of the microbial aspects along with sound knowledge of pharmacokinetics and pharmacodynamics of the antibiotics helps the clinicians in choosing the wide variety of antibiotics appropriately for the better management of patients. Local antibiotic resistance patterns play a crucial role in this.4,5

In the present study, majority of the isolates were obtained from urine samples followed pus/ wound swab and sputum. It is a known fact that E.coli is the most common causative agent of UTI, the present study E.coli constituted half of the total urinary isolates followed by Klebsiella sp. & Enterococcus sp. E.coli was highly resistant to Amoxyclav followed by 3rd gen Cephalosporins (cefixime, ceftri axone, cefotaxime), Fluoroquinolones (Ciprofloxacin, Norfloxacin, Levofloxacin) and Co-trimoxazole making majority of the orally available antibiotics ineffective for treatment. The best oral antibiotics with least resistance in the present study were Fosfomycin (23.02%) & Nitrofurantoin(14.76%). In a study conducted by Sardar et al similar findings were observed where all the isolates were susceptible to fosfomycin on par with the carbapenems where as nitrofurantoin resistance was also considerably low (17.7%)6 The other antibiotics like piperacillin/tazobactum, cefaperazone/sulbactum and Amikacin have shown significant activity against the E.coli isolates. The high end antibiotics like colistin, Carbapenems (IPM, MRP) have shown the greatest activity with almost all the isolates being susceptible to them. Though Tigecycline had also shown absolute sensitivity in vitro, it is not a drug of choice for UTIs as the urinary levels are low. Klebsiella has also shown almost similar resistance pattern as that of E.coli. In contrast to Enterobacteriaceae members Pseudomonas is not susceptible to routine antibiotics like Amoxyclav, ceftriaxone, Co-trimoxazole, Tetracyclines, Nitrofurantoin, etc. Among Pencillins, only ureido-pencillins (Piperacillin, Ticarcillin), among Cephalosporins only ceftazidime, Cefaperazone & Cefepime have got the anti- pseudomonal activity which can be used for the treatment. Polymyxins, Carbapenems & Aminoglycosides are having highest anti- pseudomonal activity. Pseudomonas was absolutely sensitive to colistin & polymyxin B followed by Aminoglycosides. Among the Fluoroquinolones, Ciprofloxacin has shown the greatest activity against Pseudomonas with only 1/4th of the isolates being resistant.

Among the Gram positive organisms, methicillin resistance was observed in 44% of the Staphyloccocal isolates. Levofloxacin has shown better activity compared to other Fluoroquinolones. Almost all the isolates were sensitive to Vancomycin & Linezolid.

Similar findings were observed in Prakash et al study7 where Escherichia coli was the most common isolate among the uropathogens (42.58%) followed by Klebsiella pneumoniae (18.71%)  Pseudomonas aeruginosa (12.90%), Staphylococcus aureus (9.68%). Carbapenems have shown very good activity for majority of the gram negative isolates in their study.

Staphylococcus aureus was the most common etiological agent isolated from wound infections (Pus & Wound swabs) followed by E.coli, Klebsiella & P seudomonas. No resistance was observed for Vancomycin & Linezolid among Staphylococcus aureus. Resistance pattern of Gram negative isolates were almost similar to that of Urinary isolates. Similar findings were observed in different studies by various researches.8,9

Among Respiratory specimens (ET secretions, BAL, Sputum etc.,) Klebsiella was the most common organism followed by Pseudomonas, Acinetobacter, Streptococcus pneumoniae and others. Klebsiella and Pseudomonas are the common isolates of Sputum whereas almost all the isolates of Acinetobacter were from ET secretions. Klebsiella was highly resistant to amoxyclav and third generation cephalosporins. Carbapenems remained as the drugs of choice in majority of the gram negative respiratory isolates as the resistance was considerably lower to these group of drugs with the exception being acinetobacter where more than 50% of the isolates being resistant to carbapenems.

Among the blood cultures E.coli was the most common gram negative organism isolated followed by Klebsiella. Though coagulase negative staphylococci were the most common gram positive cocci isolated from the blood cutures, the significance of these organisms is questionable in majority of the cases as they are the most common contaminants encountered in the blood culture specimens. Pseudomonas and S.aureus were also isolated in significant number of cases. For the enterobacteriacea isolated from blood cultures, carbapenems were highly effective followed by cefaperazone sulbactum/piperacillin tazobactum and amikacin. 36% of the S.aureus isolates from the blood culture were MRSA. Similar findings were observed in many other studies from India.10,11 In Gohel et al study E.coli and Klebsiella were the most common gram negative isolates from blood culture and staphylococcus was the most common gram positive organism. Carbapenems were highly active against majority of the gram negative isolates where as for staphylococci vancomycin and linezolid were very effective.11

Overall Gram Negative isolates were highly sensitive to Polymyxins, Tigecycline, Carbapenems, Aminoglycosides, Piperacillin-Tazobactum, Cefaperazone-sulbactum, Fosfomycin, Nitrofurantoin (for Urine Only). Resistance to other Beta-lacta ms including Cephalosporins, amoxyclav and Fluoroquinolones, Co-trimoxazole was high. Gram Positive isolates were highly sensitive to Vancomycin & Linezolid

Conclusion

Periodic Antimicrobial resistance surveillance is one of the important factors for improvising the standards of the health care. Understanding the local resistance patterns, systematic analysis and application of the surveillance data will help the clinicians in appropriate management of the patients & also prevents the emergence of drug resistance among the clinical isolates.

Source of Funding

None.

Conflict of Interest

None.

References

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A Dixit N Kumar S Kumar V Trigun Antimicrobial resistance: Progress in the decade since emergence of New Delhi metallo--lactamase in IndiaIndian J Community Med20194448

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K Walia J Madhumathi B Veeraraghavan A Chakrabarti A Kapil P Ray Establishing Antimicrobial Resistance Surveillance & Research Network in India: Journey so farIndian J Med Res201914916479

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CLSI, “Performance standards for antimicrobial susceptibility testing,” Twentieth informational supplement, Clinical and Laboratory Standards Institute Doc. M100ED28-2018.201810028

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R Laxminarayan A Duse C Wattal A K Zaidi H F Wertheim N Sumpradit Antibiotic resistance-the need for global solutions. Lancet Infect Dis201313105798

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S B Levy B Marshall Antibacterial resistance worldwide: causes, challenges and responsesNature Med20041012122129

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Ahmed Sardar Comparative Evaluation of Fosfomycin Activity with other Antimicrobial Agents Against E.coli Isolate. J Clin Diagn Res20171122629

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D Prakash R S Saxena Distribution and antimicrobial susceptibility pattern of bacterial pathogens causing urinary tract infection in Urban Community of Meerut City India. ISRN Microbiol2013201313

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H B Mantravadi M R Chinthaparthi V Shravani Aerobic isolates in pus and their antibiotic sensitivity pattern: a study conducted in a teaching hospital in Andhra PradeshInt J Med Sci Public Health2015410761079

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9.Jamatia Bacteriological Profile and Antimicrobial Resistance Patterns isolates in pus samples at Agartala Government Medical CollegeAsian J Pharm Clin Res201710113

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S Gupta B Kashyap Bacteriological profile and antibiogram of blood culture isolates from a tertiary care hospital of North IndiaTrop J Med Res20161994103

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K Gohel A Jojera S Soni S Gang R Sabnis M Desai Bacteriological profile and drug resistance patterns of blood culture isolates in a tertiary care nephrourology teaching instituteBiomed Res Int2014153747153747



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