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In the present work, biosynthesis of silver nanoparticles (AgNPs) using isolate of Shigella sp. AS8 culture supernatant as a reducing agent has been demonstrated. Synthesis of AgNPs was completed within 180 min of incubation at 35 °C under bright light condition. The biosynthesized nanoparticles were characterized by UV-vis spectroscopy, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), Dynamic light scattering (DLS), Transmission electron microscopy (TEM), Field emission scanning electron microscopy (FE-SEM) equipped with energy dispersive X-ray (EDX) and Atomic force microscopy (AFM). The formation of brown color reaction product with strong UV-vis spectrum absorption maxima at 411 nm due to surface plasmon resonance (SPR) indicated the synthesis of AgNPs. FTIR spectrum confirmed association of supernatant protein molecules with synthesized nanoparticles. DLS, TEM, FE-SEM and AFM showed biosynthesized nanoparticles were spherical in shape with an average size of 20 nm. EDX data analysis reveals presence of metallic silver. While, XRD analysis revealed that synthesized particles were pure and crystalline in nature. Further, AgNPs were evaluated as an antibacterial agent against extended spectrum β-lactamase (ESBL) positive water-borne pathogens. The results of present study suggest that biosynthesized AgNPs can be used to combat ESBL producing multidrug resistant bacteria. Copyright © 2016 VBRI Press

Present study reports a facile synthesis of surface-active antimicrobial hyperbranched polyurethane coatings using oleo-ethers of boric acid (BA) as branching and biocidal moiety (BHPU). The antimicrobial branching center was synthesized via polycondensation reaction of BA and vegetable oil-based diol. The structural characterization of synthesized BHPU and its linear counterpart was investigated using Fourier-transform infrared (FTIR) and nuclear magnetic resonance (1H, 13C, and 11B NMR) spectroscopy techniques. The cured coatings were examined by physico-mechanical, thermogravimetric (TG) analysis and differential scanning calorimetry (DSC). The antimicrobial behavior of these polymers against Gram-positive and Gram-negative bacteria was carried out by well diffusion technique. The appearance of zone of inhibition (ZOI) in case of BHPU confirmed its antimicrobial activity, which arisen due to the presence of cationic moiety in its structure. These investigations showed that the utilization of oleo-ethers of BA as branching agent in synthesis of BHPU coatings induced prominent effect on its physico-mechanical, thermal, and biocidal properties. In addition, soil burial study for 210 days was conducted on BHPU film to confirm its contact-killing mechanism against soil-borne bacteria. These results suggest the potential scope of BHPU in various applications such as long-term antimicrobial surface-active coatings for medical devices, packaging industry, paints, etc. Keywords: Boric acid, Hyperbranched, Polyurethane, Antimicrobial coating, Sustainable polymers

Colistin, a polycationic antimicrobial peptide, is one of the last-resort antibiotics for treating infections caused by carbapenem-resistant Gram-negative bacteria. The antibacterial activity of colistin occurs through electrostatic interaction between the polycationic peptide group of colistin and the negatively charged phosphate groups of lipid A membrane. This study investigated the interaction of colistin with the outer membrane and surface constituents of resistant and susceptible strains of

Antibiotic resistance conceded as a global concern is a phenomenon that emerged from the bacterial response to the extensive utilization of antimicrobials. The expansion of resistance determinants through horizontal transfer is linked with mobile genetic elements (MGEs) like transposons, insertion sequences, and integrons. Heavy metals also create consequential health hazards. Metal resistance gene in alliance with antibiotic resistance genes (ARGs) and MGEs is assisting bacteria to attain exalted quantity of resistance.The present work was carried out to study ARGs blaCTX-M, AmpC, qnrS, MGEs like ISecp1, TN3, TN21, and Int I by performing PCR and sequencing from Wular and Dal lakes of Kashmir; India. The genetic environment analysis of blaCTX-M-15 was carried out using PCR amplification, and sequencing approach followed by in-silico docking and mutational studies. Co-occurrence of ARGs and HMRGs was determined. Plasmid typing was done using PCR-based replicon typing (PBRT) and conjugation assay was also performed.Out of 201 isolates attained from 16 locations, 33 were ESBLs producers. 30 ESBL displaying isolates were perceived positive for CTX-M gene, followed by AmpC (17), qnrS (13), ISecp1 (15), TN3 (11), TN21 (11), Int I (18), and SulI (14). The genetic environment of blaCTX-M-15 was observed as (ISEcp1-blaCTX-M-15-orf477), classical promoter-10 TACAAT and -35 TTGAA was found at the 3' region. The 3D structure of CTX-M-15 and ISEcp1 was generated and CTX-M-15-ISEcp1 (R299L) docking and mutation showed a reduction in hydrogen bonds. Co-occurrence of antibiotics and HMRGs (mer, sil, and ars) was found in 18, 14, and 8 isolates. PBRT analysis showed the presence of Inc. groups- B/O, F, I1, HI1, FIA, HI2, N, FIB, L/M. Molecular analysis of transconjugants showed the successful transfer of ARGs, MGEs, and HMRGs in the E. coli J53 AZThis study highlights the occurrence of ESBL producing bacteria in the aquatic environment of Kashmir India that can serve as a reservoir of ARGs. It also discussed the molecular mechanisms of MGEs which can help in containing the spread of antibiotic resistance.

Colistin regained global interest as a consequence of the rising prevalence of multidrug-resistant Gram-negative Enterobacteriaceae. In parallel, colistin-resistant bacteria emerged in response to the unregulated use of this antibiotic. However, some Gram-negative species are intrinsically resistant to colistin activity, such as Neisseria meningitides, Burkholderia species, and Proteus mirabilis. Most identified colistin resistance usually involves modulation of lipid A that decreases or removes early charge-based interaction with colistin through up-regulation of multistep capsular polysaccharide expression. The membrane modifications occur by the addition of cationic phosphoethanolamine (pEtN) or 4-amino-l-arabinose on lipid A that results in decrease in the negative charge on the bacterial surface. Therefore, electrostatic interaction between polycationic colistin and lipopolysaccharide (LPS) is halted. It has been reported that these modifications on the bacterial surface occur due to overexpression of chromosomally mediated two-component system genes (PmrAB and PhoPQ) and mutation in lipid A biosynthesis genes that result in loss of the ability to produce lipid A and consequently LPS chain, thereafter recently identified variants of plasmid-borne genes (mcr-1 to mcr-10). It was hypothesized that mcr genes derived from intrinsically resistant environmental bacteria that carried chromosomal pmrC gene, a part of the pmrCAB operon, code three proteins viz. pEtN response regulator PmrA, sensor kinase protein PmrAB, and phosphotransferase PmrC. These plasmid-borne mcr genes become a serious concern as they assist in the dissemination of colistin resistance to other pathogenic bacteria. This review presents the progress of multiple strategies of colistin resistance mechanisms in bacteria, mainly focusing on surface changes of the outer membrane LPS structure and other resistance genetic determinants. New handier and versatile methods have been discussed for rapid detection of colistin resistance determinants and the latest approaches to revert colistin resistance that include the use of new drugs, drug combinations and inhibitors. Indeed, more investigations are required to identify the exact role of different colistin resistance determinants that will aid in developing new less toxic and potent drugs to treat bacterial infections. Therefore, colistin resistance should be considered a severe medical issue requiring multisectoral research with proper surveillance and suitable monitoring systems to report the dissemination rate of these resistant genes.

In the present study, we have investigated prevalence and diversity of ESBL genes among Klebsiella isolates obtained from highly polluted stretch of river Yamuna, India. Phenotypic screenings of 116 Klebsiella isolates revealed ~30% were positive for ESBL production. Antibiotic profiling showed multidrug resistance phenotype among 90% isolates. Prevalence of blaTEM, blaSHV and blaCTX-M genes were found to be 57, 54 and 48% respectively. Furthermore, we identified eight variants of blaSHV (SHV-1, SHV-11, SHV-27, SHV-28, SHV-38, SHV-61, SHV-144, SHV-148), three each of blaTEM (TEM-1, TEM-116, TEM-206) and blaCTX-M (CTX-M-15, CTX-M-55, CTX-M-188) among Klebsiella spp. Co-occurrence of blaTEM, blaSHV and blaCTX-M (any two or all three) was observed among 45% Klebsiella isolates. Occurrence of blaCTX-M-188 and blaTEM-206 in environmental isolates of K. pneumoniae has not been reported earlier. Identification of blaTEM-206, blaSHV-27 and blaSHV-144 from Klebsiella spp. and blaTEM-116 from K. quasipneumoniae and K. variicola is the first report from India.

In this study, we investigated the prevalence of ESBLs producing bacteria from highly polluted stretch of river Yamuna and co-occurrence of ESBLs and silver resistance gene among tested isolates. Water samples were collected from ten different polluted sites from Delhi stretch of river Yamuna, India. Out of 399 non-duplicate isolates screened, 121 (~ 30%) were found to be ESBLs producers. ESBLs positive isolates showed high level of resistance for β-lactam and non-β-lactam antibiotics, viz. ampicillin, cephalosporins, ciprofloxacin and polymyxin-B. A significantly high level of resistance (26.4%) was observed for last resort antibiotic colistin. 69% of ESBLs positive isolates exhibited multidrug resistance phenotype and high MAR index of different sampling sites ranging from 0.21 to 0.68. Molecular characterization revealed that blaCTX-M was the most prevalent type of ESBL (60.3%) followed by blaTEM (47.9%) and blaSHV (15.7%). We further observed co-occurrence of silver resistance determinants silE, silP and silS in 49.3%, 53.4% and 36.9% isolates harboring blaCTX-M genes. Conjugation transfer assay confirmed the successful transfer of plasmid encoding ESBLs and silver resistance determinants together. To the best of our knowledge, this is first report of co-occurrence of CTX-M and silver resistance gene from aquatic environment. Moreover, this study reports occurrence of ESBL genes in Bacillus safensis and Brachymonas chironomi not reported earlier. Co-occurrence of sil and blaCTX-M gene strengthens the hypothesis of heavy metal exert selection pressure for the emergence of antibiotic resistance.

Increased use of colistin, a last resort drug due to failure of carbapenems, has possibly contributed in development and spread of resistance to colistin among Enterobacteriaceae. The colistin belongs to the family of polymyxins, cationic polypeptides, with broad-spectrum activity against Gram-negative bacteria. In this study, we obtained 253 non-duplicate bacterial isolates from sewage water in Delhi and phenotypically screened for colistin resistance. Of the 47 positive isolates, the colistin resistance gene mcr-1 was detected among 5 isolates. Based on 16S ribosomal RNA-based identification, bacterial isolates were found to be Escherichia coli, Aeromonas veronii, and Aeromonas dhakensis. Extended spectrum β-lactamases (ESBL)-resistant determinants CTX-M and TEM were detected in all five mcr-1 positive isolates. On the basis of literature survey, this is the first report of mcr-1 gene from Aeromonas veronii and Aeromonas dhakensis worldwide. Furthermore, mcr-1 gene has not been reported earlier from sewage water in India. Antibiotic susceptibility test of all five isolates against 9 different classes of drugs revealed multidrug-resistant phenotype with high minimum inhibitory concentration values. In vitro transconjugation studies showed successful transfer of mcr-1 and other ESBL-resistant determinants. The occurrence of colistin resistance phenotype conferred by plasmid-based mcr-1 gene in the environment and an ever-increasing list of bacterial isolates is a cause of concern. A comprehensive survey of different water bodies and epidemiological studies are required to assess the risk of dissemination of resistance determinants.

In the present study, we have investigated prevalence and diversity of ESBL genes among

History of mankind is regarded as struggle against infectious diseases. Rather than observing the withering away of bacterial diseases, antibiotic resistance has emerged as a serious global health concern. Medium of antibiotic resistance in bacteria varies greatly and comprises of target protection, target substitution, antibiotic detoxification and block of intracellular antibiotic accumulation. Further aggravation to prevailing situation arose on observing bacteria gradually becoming resistant to different classes of antibiotics through acquisition of resistance genes from same and different genera of bacteria. Attributing bacteria with feature of better adaptability, dispersal of antibiotic resistance genes to minimize effects of antibiotics by various means including horizontal gene transfer (conjugation, transformation, and transduction), Mobile genetic elements (plasmids, transposons, insertion sequences, integrons, and integrative-conjugative elements) and bacterial toxin-antitoxin system led to speedy bloom of antibiotic resistance amongst bacteria. Proficiency of bacteria to obtain resistance genes generated an unpleasant situation; a grave, but a lot unacknowledged, feature of resistance gene transfer.