Evaluation of susceptibility testing methods for polymyxin

SummaryBackgroundThe widespread resistance in Gram-negative bacteria has necessitated evaluation of the use of older antimicrobials such as polymyxins. In the present study we evaluated the different susceptibility testing methods for polymyxins B and E against Gram-negative bacteria using the new Clinical and Laboratory Standards Institute (CLSI) guidelines.MethodsThe susceptibility of 281 multidrug-resistant (MDR) Gram-negative bacteria (GNB) to polymyxin B was evaluated, comparing broth microdilution (BMD; reference method), agar dilution, E-test, and disk diffusion. Disk diffusion testing of polymyxin B was also performed against 723 MDR GNB.ResultsTwenty-four of 281 (8.5%) isolates were found to be resistant to polymyxin B by the reference BMD method. The rates of very major errors for agar dilution and E-test (for polymyxin B) were 0.7% and 1%, respectively, and those for disk diffusion (for polymyxin B and polymyxin E) were 1% and 0.7%, respectively. For the 257 isolates found sensitive by reference BMD, the rates of major errors by agar dilution and E-test (for polymyxin B) were 2.4% and 0%, respectively, and those for disk diffusion (polymyxin B and polymyxin E) were 0% and 0.7%, respectively. Twenty-six (3.6%) of the 723 Gram-negative isolates were resistant to polymyxin B by disk diffusion.ConclusionThe E-test and agar dilution methods showed good concordance with BMD. The disk diffusion method can be useful for initial screening in diagnostic laboratories

Post-traumatic skin and soft tissue infection due to Aeromonas hydrophila

We report a case of posttraumatic skin and soft tissue infection in a patient who sustained laceration after being hit by a water tanker. Aeromonas hydrophila was isolated from pus and was identified to the species level by Vitek 2 and a battery of biochemical tests. The patient responded to thorough drainage, debridement of wound and 2 weeks of intravenous antibiotics. The patient was taken up for split skin grafting of the raw area. She was discharged with satisfactory graft uptake after 1 week without any further antibiotics advice. Follow-up after 3 weeks was satisfactory with healthy cover on the raw area and normal weight bearing on the left leg

Antimicrobial resistance in beta-haemolytic streptococci in India: A four-year study

Background & objectives: The incidence and severity of invasive and non-invasive infections demonstrate variability over time. The emerging resistance of Group A streptococci (GAS) to commonly used antibiotics is of grave concern. This study was conducted to assess the antimicrobial resistance of beta-haemolytic streptococci (βHS) in India and to ascertain the molecular mechanisms of resistance. Methods: All isolates of βHS from the Trauma Centre of All India Institute of Medical Sciences (AIIMS) (north India), and heavily populated area of old Delhi from 2010 to 2014 and Yashoda Hospital, Secunderabad (in south India, 2010-2012) and preserved isolates of βHS at AIIMS (2005-2009) were included. Phenotypic confirmation was done using conventional methods and the Vitek 2. Antibiotic sensitivity testing was done by disc diffusion and E-test. Detection of resistance genes, erm(A), erm(B), mef(A), tet(M) and tet(O), was done by polymerase chain reaction (PCR). Results: A total of 296 isolates of βHS (240 from north and 21 from south India) were included in the study. Of the 296 βHS, 220 (74%) were GAS, 52 (17.5%) were Group G streptococci and 11 (3.7%), 10 (3.3%) and three (1%) were Group B streptococci, Group C streptococci and Group F streptococci, respectively. A total of 102 (46%) and 174 (79%) isolates were resistant to tetracycline and erythromycin, respectively; a lower resistance to ciprofloxacin (21, 9.5%) was observed. A total of 42 (14%) and 30 (10%) isolates, respectively, were positive for tet(M) and erm(B) genes. Only 13 (5%) isolates were positive for mef(A). None of the isolates were positive for erm(A) and tet(O). There was discordance between the results of E-test and PCR for erythromycin and tetracycline. Interpretation & conclusions: A high level of resistance to erythromycin and tetracycline was seen in βHS in India. Discordance between genotypic and phenotypic results was reported. Absence of erm(A) and tet(O) with high prevalence of tet(M) and erm(B) was observed

Resistance pattern of mupirocin in methicillin-resistant Staphylococcus aureus in trauma patients and comparison between disc diffusion and E-test for better detection of resistance in low resource countries

Introduction: Mupirocin is an effective antibiotic for elimination of methicillin-resistant Staphylococcus aureus (MRSA) from nasal colonization and has been used to control outbreaks. Current reports show an increasing trend of resistance to this antibiotic. Objective: This study was conducted to analyze the resistance pattern of MRSA to mupirocin among the patients admitted following trauma to an apex trauma care center of India and to compare the efficacy between two methods of antimicrobial sensitivity testing. Materials and Methods: A total of 150 isolates of MRSA from various clinical samples of trauma patients over a period of 2 years were included in this study. These strains were confirmed for MRSA using VITEK ® 2 Compact and the Clinical Laboratory Standard Institute disc diffusion methods. The mupirocin susceptibility of the strains was tested by using E-test and 5 μg mupirocin disc in parallel each time, and the results were compared. Results: Clear zones of inhibition were observed in both tests. Though, good correlation was observed between the disc diffusion and E-tests in >98%, E-test showed a tendency to show lower minimum inhibitory concentration (MIC) in the remaining. These finding did not affect the final interpretation or outcomes. Of the total 150 strains, 138 (92%) showed sensitivity with the zone size in the range of 30-45 mm by 5 μg disc; rest (8%) showed sensitivity with the zone in the range of 18-30 mm by 5 μg disc, but 143 (95%) showed MIC ≤ 0.094 μg/ml and 8 (5%) gave MIC ≤ 0.75 μg/ml but ≥0.094 μg/ml by E-test. However, when both tests were compared, 5 (3.3%) showed zone size between 14 and 25 mm with ≤0.75 but >0.25 μg/ml MIC; 7 (5%) falling between 25 and 30 mm zone with MIC of ≤0.25 but >0.094 μg/ml and 138 (92%) showed zone >30 mm with MIC ≤0.094 but >0.064 μg/ml. Conclusions: All the MRSA isolates in our study were sensitive to mupirocin which is an encouraging finding. Though good screening for sensitivity can be done with 5 μg mupirocin disc, E-test provides a much clear and accurate results in clinical set-up. Hence, disc test can be used in resource poor countries and supplemented with E-test when needed

Post-traumatic skin and soft tissue infection due to Aeromonas hydrophila

We report a case of posttraumatic skin and soft tissue infection in a patient who sustained laceration after being hit by a water tanker. Aeromonas hydrophila was isolated from pus and was identified to the species level by Vitek 2 and a battery of biochemical tests. The patient responded to thorough drainage, debridement of wound and 2 weeks of intravenous antibiotics. The patient was taken up for split skin grafting of the raw area. She was discharged with satisfactory graft uptake after 1 week without any further antibiotics advice. Follow-up after 3 weeks was satisfactory with healthy cover on the raw area and normal weight bearing on the left leg

Antibiotic susceptibilities, streptococcal pyrogenic exotoxin gene profiles among clinical isolates of group C or G Streptococcus dysgalactiae subsp. equisimilis & of group G S. anginosus group at a tertiary care centre

Background & objectives: Group C and group G streptococci (together GCGS) are often regarded as commensal bacteria and their role in streptococcal disease burden is under-recognized. While reports of recovery of GCGS from normally sterile body sites are increasing, their resistance to macrolides, fluoroquinolone further warrants all invasive β haemolytic streptococci to be identified to the species level and accurately tested for antimicrobial susceptibility. This study was aimed to determine the prevalence, clinical profile, antimicrobial susceptibility and streptococcal pyrogenic exotoxin gene profile (speA, speB, speC, speF, smeZ, speI, speM, speG, speH and ssa) of GCGS obtained over a period of two years at a tertiary care centre from north India. Methods: The clinical samples were processed as per standard microbiological techniques. β-haemolytic streptococci (BHS) were characterized and grouped. Antimicrobial susceptibility of GCGS was performed using disk diffusion method. All GCGS were characterized for the presence of streptococcal pyrogenic exotoxins (spe) and spe genes were amplified by PCR method. Results: GCGS (23 GGS, 2GCS) comprised 16 per cent of β haemolytic streptococci (25/142 βHS, 16%) isolated over the study period. Of the 25 GCGS, 22 (88%) were recovered from pus, two (8%) from respiratory tract, whereas one isolate was recovered from blood of a fatal case of septicaemia. Of the total 23 GGS isolates, 18 (78%) were identified as Streptococcus dysgalactiae subsp equisimilis (SDSE, large-colony phenotype), five (21%) were Streptococcus anginosus group (SAG, small-colony phenotype). The two GCS were identified as SDSE. All GCGS isolates were susceptible to penicillin, vancomycin, and linezolid. Tetracycline resistance was noted in 50 per cent of SDSE isolates. The rates of macrolide and fluoroquinolone resistance in SDSE were low. Twelve of the 20 SDSE isolates were positive for one or more spe genes, with five of the SDSE isolates simultaneously carrying speA+ speB+ smeZ+ speF or speB+ smeZ+speF, speI+speM+speG+speH or, speI+spe M+speH or speA+ speB+ speC+ smeZ+ speF. One notable finding was the presence of spe B in four of the five isolates of the Streptococcus anginosus group. No isolate was positive for ssa. Interpretation & conclusions: Our study showed no association between GCGS isolates harbouring streptococcal pyrogenic exotoxins and disease severity. This might be attributed to the small sample size of spe-positive isolates