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Introduction: Enterobacteriaceae such as Escherichia coli and Klebsiella pneumoniae are the most prominent bacterial species resistant to almost all commonly used antibiotics. Carbapenem is one of the last resort drugs for treating such emerging multidrug-resistant bacteria. This study aimed to detect carbapenem-resistant blaNDM-1 gene in ESBL producing E. coli and K. pneumoniae isolates., Methodology: A total of 190 E. coli and 350 K. pneumoniae isolates were screened for extended spectrumβ-lactamase (ESBL), carbapenemase and metallo β-lactamase (MBL) production via double-disk synergy test (DDST), modified Hodge test and combined-disk diffusion method. The blaNDM-1 gene was detected by PCR and confirmed via Sanger sequencing method., Results: Of the 540 isolates tested, 71.8% were found to be multidrug-resistant. Overall rate of ESBL-positive isolates were 57.89% E. coli and 31.42% K. pneumoniae. Among ESBL positive isolates, 49.09% E. coli and 40% K. pneumoniae were positive for carbapenemase production whereas MBL production was detected in 29% E. coli and 22% K. pneumoniae isolates. In MBL positive isolates, (37%) E. coli and (40%) K. pneumoniae isolates harboured blaNDM-1 gene. The pair-wise DNA was aligned with the NDM-1 sequence from GenBank. The alignment score was 243 and the blast nucleotide sequencing results showed 97% sequence similarity with the sequences in GenBank for the blaNDM-1 gene., Conclusions: The blaNDM-1 gene was found to be the most prevalent in urine samples. There is a dire need to conduct screening tests in hospitals and communities to find out the exact prevalence of the blaNDM-1 spread in our population., Competing Interests: No Conflict of Interest is declared, (Copyright (c) 2021 Sohni Naeem, Hazrat Bilal, Hafsah Muhammad, Muhammad Asif Khan, Fareeha Hameed, Sher Bahadur, Tayyab Ur Rehman.)

Background and Aims: To examine antimicrobial resistance of commonly isolated pathogens in elderly hospitalized patients., Methods: Data regarding all clinically significant isolates from blood and urine cultures of patients admitted to a multilevel geriatric hospital during March 2015 to April 2016 were collected. Antimicrobial susceptibility testing was performed according to Clinical and Laboratory Standard Institute guidelines., Results: Escherichia coli, Proteus mirabilis, and Klebsiella pneumoniae were the most common isolates, with proportions of extended spectrum beta-lactamase positivity of 60, 40, and 61% respectively. Adjusted logistic regression models indicated that resistance of Escherichia coli to ceftriaxone [odds ratio (OR) 2.8, 95% confidence interval (CI) 1.5-5.1], ceftazidime (OR 2.8, 95% CI 1.5-5.1), ciprofloxacin (OR 2.2, 95% CI 1.2-4.0), amoxicillin/clavulanic acid (OR 2.3, 95% CI 1.2-4.3), and trimethoprim/sulfamethoxazole (OR 2.4, 95% CI 1.4-4.3) was significantly higher in skilled nursing wards than in acute geriatric wards. Resistance of Proteus mirabilis to ceftriaxone (OR 3.1, 95% CI 1.5-6.4) and Klebsiella pneumoniae to ciprofloxacin (OR 3.2, 95% CI 1.3-7.9) was significantly higher in skilled nursing wards than in acute wards., Conclusions and Discussion: Antimicrobial resistance was found to be high in a multilevel geriatric hospital, especially in skilled nursing wards. These findings call for rethinking of the empirical antimicrobial therapy and of the efforts for prevention of nosocomial infection.

Between July 2011 and August 2011, the New Delhi metallo-beta-lactamase 1 (NDM-1) gene was detected in Klebsiella pneumoniae and Escherichia coli isolates obtained from six patients hospitalised in four healthcare facilities in northern Italy. The patient who had been hospitalised in New Delhi, India, from February to May 2011 and subsequently in the Bologna area, Italy, from May to July 2011, may have been the source of the outbreak. Our findings suggest ongoing spread of this carbapenem-resistance gene in Italy and highlight the need for intensive surveillance.

We prospectively studied an accelerated phenotypic method by incorporating the double disk synergy test in the standard Kirby-Bauer disk diffusion susceptibility testing, to evaluate a protocol for the rapid detection of extended-spectrum beta-lactamases (ESBL) in urinary isolates of Escherichia coli (E. coli) and Klebsiella pneumoniae (K. pneumoniae). All ESBL-positive isolates were confirmed by the standard Clinical Laboratory Standards Institute (CLSI) confirmatory disk diffusion method. Between November 2004 and December 2005, a total of 6988 urine specimens were analyzed of which, 776 (11%) showed significant growth. They included E. coli in 577 cases (74%) and K. pneumoniae in 199 (25.6%). Of these, 63 E. coli (8%) and 15 K. pneumoniae (7.5%) were positive for ESBL by the accelerated and CLSI methods. Compared to the standard CLSI method, the accelerated method reduced the ESBL detection time from two days to one day. We conclude that the accelerated ESBL detection technique used by us in this study is a reliable and rapid method for detecting ESBL in urinary isolates of E. coli and K. pneumoniae.