Diarrhea,Pneumonia, and other infectious diseases are leading causes of death amongchildren younger than five years in low and middle -income countries and alsoin India (Black et al. 2003 andBassani et al. 2010). However, littleis known about the causes of death in children after age five years. The GlobalBurden of Disease and Risk Factors (GBD) estimates that in 2004 there wereapproximately 69,000 deaths from pneumonia and 1000 deaths from diarrhealdiseases among children aged 5-14 years in India, (Lopez AD et al. 2006).
Thevast majority of deaths due to pneumonia and diarrhea occur in the poorestregions–nearly 90 percent of them in sub-Saharan Africa and South Asia. Abouthalf the world’s deaths due to pneumonia and diarrhea occur in just five mostlypoor and populous countries: India, Nigeria, the Democratic Republic of theCongo, Pakistan, and Ethiopia (Liu et al.2012).Deathsamong children under age 5 due to pneumonia and diarrhea are high in India. In2005, 2.
35 million deaths (Mortality Rate-85.8) were found. In North India,there is also high mortality rate. In 2005, 65.6 mortality rates were reported(Black et al. 2003 and Bassani et al.2010).
Escherichia coliand Klebsiella pneumonia are two ofthe main causative agents of most of theinfectious diseases. These are common bacteria to produce Extended-spectrum?-lactamases (ESBL) and plasmid-mediated quinolone resistance (PMQR), which ofthe main factor that confers resistance to these bacteria against beta-lactamand quinolone. And these resistance species are emerging clinical challenge forus. Fig.1: Causes of deaths in children under 5 years in India, 2005 Extended-spectrum?-lactamases (ESBLs) have become widespread in a significant percentage ofGram- negative bacteria, especially in Enterobacteriaceae throughout the world(Bradford, 2001). Generally, the ESBLs are enzymes that arise by mutations ingenes for common plasmid- mediated ?-lactamases, such as TEM-1, SHV-1, and OXA –10, or may be distantly related to a native enzyme, as in the case of CTX-M?-lactamases (Smet et al., 2009).
Usually,ESBLs are encoded by blaTEM, blaSHV or blaCTX-M genes in Enterobacteriaceae and other Gram-negativebacteria (Monstein et al., 2007). Atthe same time, plasmid-mediated quinolone resistance (PMQR) is also a threat tothe veterinary clinical therapy and human public health. Fluoroquinolones areamong the most commonly prescribed antimicrobials because of theirbroad-spectrum antimicrobial activity, and fluoroquinolone-resistantgram-negative pathogens have emerged worldwide. Quinolone resistance istraditionally mediated by the mutation of chromosomal genes encoding DNA gyraseand/or topoisomerase IV or by the mutation of genes regulating the expressionof efflux pumps (Hooper, D. C.
1999, Hooper, D. C. 2000).
To date, at leastthree types of PMQR determinants, including qnrfamilies (qnrA, qnrB, qnrS, qnrC and qnrD), aac (60)-Ib-cr and quinolone efflux pump (qepA and oqxAB)have been extensively reported (Strahilevitz et al., 2009). Moreover, qnrgenes have been commonly detected among isolates producing ESBLs (Robicsek et al., 2006) which raises the risk fortransfer of multidrug-resistant Escherichiacoli to humans.Itwas thought that quinolone resistance could be acquired only by chromosomalmutations until plasmid-mediated resistance to quinolones was described in aclinical isolate of Klebsiella pneumoniain 1998 (Martinez-Martinez, L., A.
Pascual, and G. A. Jacoby.1998). Since then, four major groups of qnrdeterminants, qnrA, qnrB, qnrC, and qnrS, have beenidentified (Jacoby, G., V.
Cattoir, D. Hooper, L. Mart?´nez-Mart?´nez, P.
Nordmann, A. Pascual, L. Poirel, and M. Wang.
2008, Wang, M., X. Xu, and S. Wu.2008), and two additional plasmid-mediated quinolone resistance (PMQR) geneshave been described— aac(6_)-Ib-cr, which encodes a variant aminoglycosideacetyltransferase that modifies ciprofloxacin (Robicsek, A., J. Strahilevitz,G. A.
Jacoby, M. Macielag, D. Abbanat, K. Bush, and D.
C. Hooper. 2006), andqepA, which encodes an efflux pump belonging to the major facilitator subfamily(Pe’richon, B., P. Courvalin, and M. Galimand. 2007).
These PMQR determinantsare increasingly being identified worldwide in clinical isolates ofEnterobacteriaceae (Gay, K., A. Robicsek, J. Strahilevitz, C.
H. Park, G.Jacoby, T. J.
Barrett, F. Medalla, T. M.
Chiller, and D. C. Hooper. 2006) andin clinical and environmental Aeromonas species isolates (Cattoir, V.
, L.Poirel, C. Aubert, C. J.
Soussy, and P. Nordmann. 2008).
The global occurrenceof antibiotic resistance genes in bacteria found in aquatic environments is anincreasing concern. Microorganisms that carry genes encoding resistance to abroad range of antibiotics have been found in hospital wastewater and excretedanimal product as well as in sewage, surface water, river water, groundwaterand drinking water (Zhang X et al.,2009). Aquatic environments are described as natural reservoirs of antibiotic-resistantbacteria (Ferreira et al., 2007).
Inaddition, the presence of antibiotics and their metabolites in sewage maypromote both the selection of resistant strains, and the horizontal transfer ofantibiotic resistance genes (Andersen SR etal., 1993). All natural waters support and contain a variety of organisms,both plants, and animals as the natural flora and fauna. Almostall the world’s water (97%) is located in the oceans, but as might be expected,the high concentration of salts renders the sea water virtually unusable. 1.74% of water is presentas glaciers and permanent snow and 0.
3% is present as groundwater, stored inthe aquifer both as confined and unconfined and is perched aquifers. Theavailable freshwater content in terms of direct use is only thus limited to 1%.India is a developing nation facing a serious problem of natural resourcescarcity, especially that of water in view of rapidly population growth andeconomic development. In India, water bodies are increasingly getting contaminatedwith sewage, domestic waste, industrial wastes; and due to sociocultural activities.
Thebacteria causing cholera, typhoid fever and bacillary dysentery may be presentin sewage water or polluted water. Polluted water also contains severalpathogenic viruses, out of which the prominent one is the one causinginfections such as hepatitis (jaundice) and poliomyelitis, etc. One shouldalways keep a check on the presence of biological as well as chemicalpollutants present in water. Municipal water and food products are routinelymonitored for the presence of pathogens, but there are many challenges in doingso. Even non-pathogenic bacteria, if present in large enough numbers can causedeterioration of food products and water. The alternative for the testing ofmany different types of pathogens is to test for those bacteria that indicatethe presence of pathogens. The most common source of pathogens in food andwater is fecal contamination, of either animal or human origin. Feces alsocontain innumerable non-pathogenic bacteria, and the presence of these otherbacteria in food or water indicates that fecal contamination has occurred andhence the presence of pathogens.
Microbes are susceptible to antibiotics andmost of the diseases of bacterial origin are treated with these antibiotics.But of late these pathogenic bacteria upon indiscriminate exposure to theantibiotics have started developing resistance towards them and thus posing analarming scenario in the world of medical sciences. The presence of ESBL andPMQR in bacteria are yet other elements to the development and acquisition ofantibiotic resistance among the pathogens. In this study, therefore I studiedand investigated the occurrence of genetic elements responsible for the ESBLand PMQR in the fresh water bodies, and the river Yamuna in and around Delhiregion to assess and predict the danger posed by the ESBL and PMQR pathogensparticularly in E. coli and K pneumonia.
Objectives: · Isolation and identification of E.coli and K. pneumonia from Yamuna water as well as sewage water in Delhi.· Antimicrobial susceptibility testing,and phenotypic detection of ESBL and PMQR.· To study the protein expression atdifferent environmental stress condition.
· To investigate the occurrence of ESBLand PMQR genes in above-selected isolates to reveal genetic variation amongESBL and PMQR.
