Screening and Potential of the Incidence of Resistance Transfer Among the Multidrug and Heavy Metal Resistant Gram-Negative Isolates from Hospital Effluents of Northern India

摘要

Background: Hospital wastewater has a high amount of both organic and inorganic matter,as well as high densities of living organisms, including pathogenic, and environmental bacteria. It hasbeen suggested that genes encoding resistance to an antibiotic can be located together with heavymetals resistance genes on either the same genetic structure (plasmid) or different genetic structureswithin the same bacterial strain. Resistance transfer is mainly attributed to conjugation since many antimicrobialresistance genes are situated on mobile elements, such as plasmids and conjugative transposons,whereas renovation and transduction are usually more limited. Our study confirmed the flowof resistance genes between indigenous and foreign organisms and indicated the possibility of resistancetransfer from environmental reservoirs to pathogenic strains, which should be underlined inthe future. The recent patents on drug resistance (US20030130169, WO/2001/060387,WO/2016/151092) and gene transfer (JP2003189855, JP2010094090), helped in this study.Methods: Water samples were collected from three different sites of hospital wastewater. Isolation ofGram-negative bacteria from hospital wastewater samples was done using the standard microbial procedure.The heavy metal resistance was determined by the minimum inhibitory concentration (MIC)against the test bacterial strain by spot plate method. The antibiotic resistance was determined by astandard disc diffusion technique. The bacterial resistance transfer studies were determined betweendonor and recipient strain in nutrient as well as wastewater. The antibiogram and MIC of the donorsand transconjugants were studied by above-described methods.Results: A high number of Gram-Negative Bacterial Isolates (GNB) exhibited antibiotic and metal resistancetransfer into E. coli K-12 and similar GNB isolates in nutrient broth as compared towastewater. The microbial conjugation experiments showed that a high percentage of multi-resistantGNB (75% and 66%) was able to transfer their single or multidrug resistance patterns to E. coli K-12among antibiotic while 58%, 66% of the multiresistant isolates were able to transfer their single ormulti-metal resistance patterns to E. coli K-12 among metal in nutrient medium and wastewater, respectively.In the present conjugation study, 97.5% and 70% of the total tested GNB isolates wereable to transfer an antibiotic-resistant marker to recipient GNB in both the medium (nutrient mediumand wastewater), whereas 92.5% and72.5% of the isolates were able to transfer metal resistant markerto recipient GNB in nutrient medium and wastewater from all the site tested. The higher (6.8x10-1 and5.9x10-1) frequency of transfer was observed among antibiotic and metal while the lower frequency oftransfer was (7.0x10-3 and 2.0x10-3) exhibited against antibiotic and metal in both the medium fromthe entire site tested, respectively.Conclusion: We can recommend that the hospital water is heavily polluted with several types of antibiotics,toxic metals as well as the potentially hazardous bacterial flora because of their capacity to resistone or the other well known antibiotic and chemotherapeutic agents. These studies provide evidencethat a wide variety of clinically important antibiotic and metal resistance genes is mobile withinaquatic bacterial communities one step ahead of the above, we can envisage the alarming situationprevailing in our system and surrounding in the light of transmissible nature of R-plasmids.