Pneumonia, and other infectious diseases are leading causes of death among
children younger than five years in low and middle -income countries and also
in India (Black et al. 2003 and
Bassani et al. 2010). However, little
is known about the causes of death in children after age five years. The Global
Burden of Disease and Risk Factors (GBD) estimates that in 2004 there were
approximately 69,000 deaths from pneumonia and 1000 deaths from diarrheal
diseases among children aged 5-14 years in India, (Lopez AD et al. 2006).
vast majority of deaths due to pneumonia and diarrhea occur in the poorest
regions–nearly 90 percent of them in sub-Saharan Africa and South Asia. About
half the world’s deaths due to pneumonia and diarrhea occur in just five mostly
poor and populous countries: India, Nigeria, the Democratic Republic of the
Congo, Pakistan, and Ethiopia (Liu et al.
among children under age 5 due to pneumonia and diarrhea are high in India. In
2005, 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.
and Klebsiella pneumonia are two of
the main causative agents of most of the
infectious diseases. These are common bacteria to produce Extended-spectrum
?-lactamases (ESBL) and plasmid-mediated quinolone resistance (PMQR), which of
the main factor that confers resistance to these bacteria against beta-lactam
and quinolone. And these resistance species are emerging clinical challenge for
1: Causes of deaths in children under 5 years in India, 2005
?-lactamases (ESBLs) have become widespread in a significant percentage of
Gram- negative bacteria, especially in Enterobacteriaceae throughout the world
(Bradford, 2001). Generally, the ESBLs are enzymes that arise by mutations in
genes 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).
ESBLs are encoded by blaTEM, blaSHV or blaCTX-M genes in Enterobacteriaceae and other Gram-negative
bacteria (Monstein et al., 2007). At
the same time, plasmid-mediated quinolone resistance (PMQR) is also a threat to
the veterinary clinical therapy and human public health. Fluoroquinolones are
among the most commonly prescribed antimicrobials because of their
broad-spectrum antimicrobial activity, and fluoroquinolone-resistant
gram-negative pathogens have emerged worldwide. Quinolone resistance is
traditionally mediated by the mutation of chromosomal genes encoding DNA gyrase
and/or topoisomerase IV or by the mutation of genes regulating the expression
of efflux pumps (Hooper, D. C. 1999, Hooper, D. C. 2000). To date, at least
three types of PMQR determinants, including qnr
families (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, qnr
genes have been commonly detected among isolates producing ESBLs (Robicsek et al., 2006) which raises the risk for
transfer of multidrug-resistant Escherichia
coli to humans.
was thought that quinolone resistance could be acquired only by chromosomal
mutations until plasmid-mediated resistance to quinolones was described in a
clinical isolate of Klebsiella pneumonia
in 1998 (Martinez-Martinez, L., A. Pascual, and G. A.
1998). Since then, four major groups of qnr
determinants, qnrA, qnrB, qnrC, and qnrS, have been
identified (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) genes
have been described— aac(6_)-Ib-cr, which encodes a variant aminoglycoside
acetyltransferase that modifies ciprofloxacin (Robicsek, A., J. Strahilevitz,
G. A. Jacoby, M. Macielag, D. Abbanat, K. Bush, and D. C. Hooper. 2006), and
qepA, which encodes an efflux pump belonging to the major facilitator subfamily
(Pe’richon, B., P. Courvalin, and M. Galimand. 2007). These PMQR determinants
are increasingly being identified worldwide in clinical isolates of
Enterobacteriaceae (Gay, K., A. Robicsek, J. Strahilevitz, C. H. Park, G.
Jacoby, T. J. Barrett, F. Medalla, T. M. Chiller, and D. C. Hooper. 2006) and
in clinical and environmental Aeromonas species isolates (Cattoir, V., L.
Poirel, C. Aubert, C. J. Soussy, and P. Nordmann. 2008). The global occurrence
of antibiotic resistance genes in bacteria found in aquatic environments is an
increasing concern. Microorganisms that carry genes encoding resistance to a
broad range of antibiotics have been found in hospital wastewater and excreted
animal product as well as in sewage, surface water, river water, groundwater
and drinking water (Zhang X et al.,
2009). Aquatic environments are described as natural reservoirs of antibiotic-resistant
bacteria (Ferreira et al., 2007).In
addition, the presence of antibiotics and their metabolites in sewage may
promote both the selection of resistant strains, and the horizontal transfer of
antibiotic resistance genes (Andersen SR et
al., 1993). All natural waters support and contain a variety of organisms,
both plants, and animals as the natural flora and fauna.
all 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 present
as glaciers and permanent snow and 0.3% is present as groundwater, stored in
the aquifer both as confined and unconfined and is perched aquifers. The
available freshwater content in terms of direct use is only thus limited to 1%.
India is a developing nation facing a serious problem of natural resource
scarcity, especially that of water in view of rapidly population growth and
economic development. In India, water bodies are increasingly getting contaminated
with sewage, domestic waste, industrial wastes; and due to sociocultural activities. The
bacteria causing cholera, typhoid fever and bacillary dysentery may be present
in sewage water or polluted water. Polluted water also contains several
pathogenic viruses, out of which the prominent one is the one causing
infections such as hepatitis (jaundice) and poliomyelitis, etc. One should
always keep a check on the presence of biological as well as chemical
pollutants present in water. Municipal water and food products are routinely
monitored for the presence of pathogens, but there are many challenges in doing
so. Even non-pathogenic bacteria, if present in large enough numbers can cause
deterioration of food products and water. The alternative for the testing of
many different types of pathogens is to test for those bacteria that indicate
the presence of pathogens. The most common source of pathogens in food and
water is fecal contamination, of either animal or human origin. Feces also
contain innumerable non-pathogenic bacteria, and the presence of these other
bacteria in food or water indicates that fecal contamination has occurred and
hence the presence of pathogens. Microbes are susceptible to antibiotics and
most of the diseases of bacterial origin are treated with these antibiotics.
But of late these pathogenic bacteria upon indiscriminate exposure to the
antibiotics have started developing resistance towards them and thus posing an
alarming scenario in the world of medical sciences. The presence of ESBL and
PMQR in bacteria are yet other elements to the development and acquisition of
antibiotic resistance among the pathogens. In this study, therefore I studied
and investigated the occurrence of genetic elements responsible for the ESBL
and PMQR in the fresh water bodies, and the river Yamuna in and around Delhi
region to assess and predict the danger posed by the ESBL and PMQR pathogens
particularly in E. coli and K pneumonia.
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 at
different environmental stress condition.
To investigate the occurrence of ESBL
and PMQR genes in above-selected isolates to reveal genetic variation among
ESBL and PMQR.