• Microbiology and Biotechnology Letters
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• v.51 no.3
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• pp.268-279
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• 2023

The pharmaceutical industry in Bangladesh produces a diverse range of antibiotics for human and animal use, however, waste disposal management is inadequate. This results in substantial quantities of antibiotics being discharged into water bodies, which provide suitable environment for the growth of antibiotic-resistant bacteria, capable of spreading resistance genes. This study intended for exploring the bacterial antibiotic resistance profile in adjoining aquatic environmental sources of pharmaceutical manufacturing facilities in Bangladesh. Seven surface water samples were collected from the vicinity of two pharmaceutical industries located in the Savar area and 51 Escherichia coli isolates were identified using both phenotypic and genotypic methods. Antibiotic susceptibility tests revealed the highest percentage of resistance against ampicillin, azithromycin, and nalidixic acid (100%) and the lowest resistance against meropenem (1.96%) out of sixteen different antibiotics tested. 100% of the study E. coli isolates were observed with Multidrug resistance phenotypes, with the Multiple Antibiotic Resistance (MAR) value ranging from 0.6-1.0. Furthermore, 69% of the isolates were Extended Spectrum Beta-Lactamases (ESBL) positive as per the Double Disk Diffusion Synergy Test (DDST). ESBL resistance genes bla_TEM, bla_CTX-M-13, bla_CTX-M-15, and bla_SHV were detected in 70.6% (n = 36), 60.8% (n = 32), 54.9% (n = 28), and 1.96% (n = 1) of the isolates, respectively, by Polymerase Chain Reaction (PCR). Additionally, 15.68% (n = 8) of the isolates were positive for E. coli specific virulence genes in PCR. These findings suggest that pharmaceutical wastewater, if not properly treated, could be a formidable source of antibiotic resistance spread in the surrounding aquatic environment. Therefore, continued surveillance for drug resistance among bacterial populations around drug manufacturing facilities in Bangladesh is necessary, along with proper waste disposal management.

• Journal of Microbiology and Biotechnology
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• v.19 no.9
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• pp.1065-1069
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• 2009

Of 143 clinical isolates of Klebsiella pneumoniae collected from Korean non-tertiary hospitals, 24 (16.8%) showed an extended-spectrum ${\beta}$-lactamase-positive phenotype. PCR and sequence analysis revealed the presence of TEM-116 (n=13), CTX-M-3 (n=5), CTX-M-14 (n=2), CTX-M-15 (n=3), and SHV-12 (n=16). Each of the 24 isolates encoded more than one ${\beta}$-lactamase, and seven isolates (29%) harbored two different SHV-type ${\beta}$-lactamase genes ($bla_{SHV-11}$ and $bla_{SHV-12}$) bounded by insertion sequence IS26 in a single transferable plasmid.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1993.04a
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• pp.34-35
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• 1993

The Development of new asymmetric induction methods useful for syntheses of biologically active natural products and drugs, using C4-chiral 1,3-th-iazolidine-2-thiones, has been a recent focus of interest. 1-8) The present account describes the significance of particular heterocycles in the synthetic development of new 1${\beta}$-substituted carbapenems. A fungal metabolite, (＋)-thienamycin (1) has attracted one's attention as a hopeful candidate for new-generation antibiotic drugs because of its strong antimicrobial activities and wide antimicrobial spectra due to the extensive inhibition against various ${\beta}$-lactamases. However, it has been serious problems toward a practically useful drug that (＋)-thienamycin is fairly labile in the solution and can be metabolized by renal dehydropept- idase-I (DHP-I). Recently, a Merck Sharp ＆ Dohme research group exploited a non-natural ${\beta}$-lactam, imipenem (2) which has been appeared in the drug market as the first carbapenem-type antibiotic drug. 9) However 2 must be used with a DHP-I inhibitor, cilastatin sodium (3).9) Thus, a 1,${\beta}$-methyl- carbapenem derivative 4 has been disclosed by the same group. 10) It seems to be more hopeful candidate as a new-generation antibiotic because it can directly resist against metabolism by the renal DHP-1 without an enzyme inhibitor 3. 10)

• Microbiology and Biotechnology Letters
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• v.51 no.4
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• pp.517-525
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• 2023

The emergence and spread of multidrug-resistant Pseudomonas aeruginosa (MRPA) have become a serious problem worldwide. The involvement of metallo-β-lactamases (MBLs) in inducing carbapenem resistance is particularly acute. However, unlike other members of the Enterobacteriaceae genus, new clones of P. aeruginosa are constantly emerging and rapidly replacing previously prevalent dominant clones. Therefore, this study aimed to perform antimicrobial resistance gene analysis, integron gene cassette analysis using DNA sequencing, and plasmid transfer analysis by conjugation to investigate the antimicrobial resistance dynamics of 18 P. aeruginosa strains isolated from various medical samples at a general hospital in Busan from September 2017 to September 2019. All 18 strains showed extensively drug-resistant (XDR) phenotype and were resistant to most antibiotics, except colistin (100%) but were susceptible to aztreonam (22.2%) and ceftazidime (16.6%). Approximately 66.7% of the strains had Class 1 integrons showing various antimicrobial resistances. Notably, IMP-6 ST235 (66.7%), VIM-2 ST357 (16.7%), and IMP-1 ST446(16.7%) were identified. The identification of IMP-1-producing ST446, previously unreported in Korea, is noteworthy considering the emergence and prevalence of another MRPA high-risk clone.

• Biomedical Science Letters
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• v.25 no.3
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• pp.275-281
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• 2019

This study was performed to characterize the chromosomal metallo-${\beta}$-lactamases (MBLs) of Chryseobacterium indologenes isolated from Korea and to propose a clustering method of IND MBLs based on their amino acid similarities. Chromosomal MBL genes were amplified by PCR from 31 clinical isolates of E. indologenes. Nucleotide sequencing was performed by the dideoxy chain termination method using these PCR products. Antimicrobial susceptibilities were determined by the agar dilution method. PCR experiments showed that all 31 E. indologenes isolates contained all $bla_{IND}$ genes. DNA sequence analysis revealed that E. indologenes isolates possessed ten types of $bla_{IND}$ gene, including seven novel variants ($bla_{IND-8}$ to $bla_{IND-14}$). The most common combination of MBL was IND-2 (n = 18). Minimum inhibitory concentrations of imipenem and meropenem for the isolates harboring novel IND MBLs were ${\geq}16{\mu}g/mL$. IND MBLs were grouped in three clusters, based on amino acid similarities.

• Korean Journal of Microbiology
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• v.46 no.1
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• pp.38-45
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• 2010

The study performed to identify the type of ESBL against strains which are producing extendedspectrum ${\beta}$-lactamases and isolated from sewage in Suyeong sewage disposal plant, Busan Environmental Corporation. By the standard activated sludge method, Suyeong sewage disposal plant purify living and lavatory sewage gathering from the northeast Busan and the facility purify total 550,000 tons of living sewage disposal a day. 14 strains were isolated by double disk synergy test and the third generation cepha-antibiotics test. Indole, methyl-red, Voges-Proskauer, Simmon's citrate, decarboxylasedihydrolase and sugar-fermentation tests identified as Klebsiella pneumoniae (n=4) and Escherichia coli (n=10). Plasmid-mediated transmission test against isolated 14 strains proved 11 strains transmitted resistance to recipient E. coli J53 (sodium $azide^R$, $ceftazidime^S$). 9 strains of conjugant were expressed ESBL genes transferred from parental strain but 2 conjugants did not expressed. The type of ESBL from each strain was determined by isoelectric focusing points, DNA and amino acids sequencing. The results indicated that the types of ESBL transmitted to recipient E. coli J53 were TEM-1, the parental TEM type and SHV-12 type.

• Korean Journal of Microbiology
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• v.42 no.2
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• pp.125-130
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• 2006

The emergence of extended spectrum ${\beta}$-lactamase producing bacteria is causing very serious problems in Korea. Although there have been many reports about these bacteria Isolated from patients and clinical specimens, there is no report of extended spectrum ${\beta}$-lactamase producing organisms isolated from natural environment in Korea. This study was conducted to investigate the biological characteristics and extended spectrum ${\beta}$-lactamase types of eighteen strains of extended spectrum ${\beta}$-lactamase producing Klebsiella pneumoniae and Escherichia coli isolated from a slaughterhouse in Pusan in Korea during 2002 to 2004. Extended spectrum ${\beta}$-lactamases were identified by double-disk synergy test, conjugation, isoelectric focusing values and gene sequencing. Eight strains of Klebsiella pneumoniae and two strains of Eschericha coli were isolated kom pigs and transferred extended spectrum ${\beta}$-lactamase genes to recipient Escherichia coli J53 (sodium azide resistant and ceftazidime senstive) strain by conjugation. The conjugants of extended spectrum ${\beta}$-lactamase genes were alignments and translated to amino acids by BCM and NCBI blast. Eight conjugants of Klebsiella pneumonae were typed TEM-52, and two strains of Escherichia coli, SHV-12, but CMY-1 type were not detected in this study.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.44 no.6
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• pp.597-604
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• 2011

Using 108 strains of Vibrio parahaemolyticus isolated from seawater, we investigated ampicillin-resistance profiles and the genetic characterization of ${\beta}$-lactamase (VPA0477). All of the strains studied, except one strain, were resistant to ampicillin. However, the strain that was susceptible to ampicillin had the same ${\beta}$-lactamase gene as the ampicillin-resistant strains. We compared ${\beta}$-lactamase promoter region sequences among five strains, including both ampicillin-resistant and -susceptible strains. In the susceptible strain, a nucleotide at position -19 in the methionine initiation codon for ${\beta}$-lactamase was not present in the ampicillin-resistant strains. The genes in the region containing the gene VPA0477 were present in all of the tested strains, and LA-PCR analysis showed that the distance between VPA0474 and VPA0479 in all of the V. parahaemolyticus samples was precisely 5.7 kb. In V. parahaemolyticus ${\beta}$-lactamase, four important structural features that are conserved in Class A ${\beta}$-lactamases were present in the deduced amino acid sequences. Taken together, our study demonstrates that V. parahaemolyticus ${\beta}$-lactamase is included in the Class A ${\beta}$-lactamase group, and some nucleotides within the promoter region are of particular importance for ${\beta}$-lactamase activity.

• Archives of Pharmacal Research
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• v.24 no.6
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• pp.590-596
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• 2001

A new extended-spectrum ${\beta}-lactamase$ with an isoelectric point (pl) of 6.2 was detected in Klebsiella pneumoniae Fl 61 that was isolated from a patient with infection. This strain was highly resistant to the third or fourth generation cephalosporins such as cceftazidime ceftriaxone, cefoperzaone, and cefpirome. Analysis of this strain by the double disk diffusion test showed synergies between amoxicillin-clavulanate (AMX-CA) and cefotaxime, and AMX-CA and aztreonam, which suggested that this strain produced a extended-spectrum ${\beta}-lactamase$ (ESBL). Cenetic analysis revealed that the resistance was due to the presence of a 9.4-kb plasmic, designated as pkpl 61, encoding for new ${\beta}-lactamase$ gene (bla). Sequence analysis showed that a new bla gene of pkpl 61 differed from $bla_{TEM-1}$ by three mutations leading to the following amino acid substitutions: $Val_{84}{\rightarrow}lie,{\;}Ala_{184}{\rightarrow}Val,{\;}and{\;}Gly_{238}{\rightarrow}Ser$. These mutations have not been reported previously in the TIM type ${\beta}-lactamases$ produced by clinical strains. The novel ${\beta}-lactamase$ was overexpressed in E. coli and purified by ion exchange chromatography on Q-Sepharose and CM-Sepharose, and then further purified by gel filtration on Sehadex G-200. The catalytic activity of th8 purified ${\beta}-lactamase$ was confirmed by the nitrocefin disk.

• Microbiology and Biotechnology Letters
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• v.34 no.4
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• pp.342-351
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• 2006

The aim of this study was to survey susceptibilities of Escherichia coli and Klebsiella pneumoniae isolates against cefotaxime and to determine the prevalences of CTX-M type extended-spectrum $\beta$-lactamases (ESBLs) producing E. coli and K. pneumoniae in Korea. During the period of February to July, 2005, 153 E. coli and 52 K. neumoniae isolates were collected from 2 hospitals in Busan. Antimicrobial susceptibilities to cefotaxime were tested by the disk diffusion method. ESBL production of E. coli and K. pneumoniae was determined by the double disk synergy test. MICs of $\beta$-lactam antibiotics were determined by the agar dilution method. Blac$_{CTX-M}$ genes of the organism were detected by PCR. Among 153 isolates of E. coli and 52 isolates of K. neumoniae, 27 (17.6%) and 25 (48.0%) were intermediate or resistant to cefotaxime, respectively. Twenty-three (15.0%) isolates out of 153 E. coli and 13 (25.0%) out of 52 K. neumoniae isolates showed positive results for ESBL by the double disk synergy test. Twenty isolates out of 23 ESBL producing E. coli and 12 out of 13 ESBL producing K. neumoniae isolates harbored biacTx-M gene,11 of ESBL producing E. coli and 12 of ESBL producing K. neuinoniae isolates harbored bla$_{CTX-M}$ gene, 11 of the ESBL producing E. coli and 2 of ESBL producing K. neumoniae isolates harbored bla$_{TEM}$ gene, and 1 of the ESBL producing E. coli and 12 of ESBL producing K. neumoniae isolates harbored bla$_{SHV}$ gene. E. coli and K. neumoniae isolates producing CTX-M-type ESBLs were not uncommon in Korea. It is thought that continuous survey are necessary for inspecting the spread and novel variants of CTX-M-type ESBL genes. Further me]'e investigation and research on ESBL producing strains are needed in order to prevent the spread of resistant bacteria.