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o Importance of limited use
o Measures for controlling antibiotic resistance
I wish to record my deep sense of gratitude and indebtedness to Almighty God for
supporting me in every walk of life and making me capable seeing dreams and ultimately
strive hard to achieve it.
I would like to take this opportunity to thank all the people in my life who have helped me
in completing third project and providing me with their abled support.
I owe my gratitude to the faculty and staff members of Hamdard Public School .Our
honorable Additional Secretary, Mr. Samar Hamid, and Madam Principal, Mrs. Zakia
Majid Siddiqui, who have always been a source of inspiration for us.
I owe special sense of gratitude to our Biotechnology teacher, Mr. Khalid Anwer, for his
learned suggestions and encouraging attitude. I am equally thankful to our Laboratory
Assistant, Mr. Anwer for providing me necessary aids during this project.
The internet services have been an immensely important part of my project in providing
all the necessary details, articles, information and pictorial representations made use of
in this project.
My sincere thanks are to my friends and classmates Sonam, Anam, Varsha and Shafaque
for their valuable suggestions, constant support and encouragement to do my best.
I would be failing in my duty if I do not acknowledge the indebted support of my family
members. I thank my beloved brother who lend a helping hand in completing this project
as well as my parents who have always stood by me in every right decision.
This project has only been able to be completed due to the trust, well wishes and
unending support of all the people associated with me.
Broadly defined, antibiotics include a chemically heterogeneous group of small organic
Molecules of microbial origin that, at low concentrations, are deleterious to the growth or
metabolic activities of other microorganisms. That soil is rich in microorganisms capable
of antibiotic synthesis is well accepted, but the frequency with which synthesis occurs at
ecologically significant levels in nature has been much less clear. Over the past decade,
however, genetic and molecular techniques, coupled with sensitive and bioanalytical
assays and equipment, have been applied to demonstrate conclusively that
microorganisms synthesize a variety of antibiotics, even under field conditions, in the
rhizosphere (that portion of the soil enriched in carbon and energy resources released by
plant roots). These antibiotics can contribute to microbial competitiveness and the
suppression of plant root pathogens, and the bacteria that produce them are therefore of
considerable interest as a practical means of plant disease control. More generally, the
techniques used to understand the role of antibiotics in the rhizosphere are applicable to
other habitats where mechanisms of microbial antagonism or the production of bioactive
metabolites are of interest.
When used together, the bioanalytical and molecular approaches are complementary,
allowing the detection and quantification of metabolites produced in situ as well as an
evaluation of their activity, and hence their ecological significance. The direct detection of
a metabolite provides irrefutable evidence that the genetic and physiological potentials
for its synthesis have been met, and the amounts recovered are in part a function of the
net rates of synthesis and turnover under particular experimental circumstances. Direct
measurements are most informative when the identity and physical properties of the
metabolite are known so that procedures for extraction can be optimized, and are of value
in assessing the relative amounts present over a range of conditions or in monitoring
persistence and dissemination in the environment. Even when the structure is known,
sensitivity of detection is likely to be the single most limiting factor to the direct analysis
of metabolites produced in situ. The comparatively large sample sizes from which
metabolites are extracted generally preclude direct analyses of substances produced by
localized populations in spatially restricted sites in soil or on plant surfaces. Molecular
approaches offer highly sensitive but indirect alternatives to the direct analysis of
bioactive metabolites produced in situ. These techniques detect either the potential for
synthesis as inferred from the presence or expression of biosynthetic genes, or an activity
attributable to the presence of the metabolite itself. For example, introduced and
indigenous antibiotic-producing strains can be detected and enumerated by using probes
and primers based on unique DNA sequences within genes specific for antibiotic
biosynthesis. Such sequences also have been applied to access novel biosynthetic genes
directly from soil without the need for culturing. Reporter gene systems (described
elsewhere in this volume) can be used to monitor the transcription of antibiotic
biosynthesis genes expressed in situ. When the impact of metabolites on other organisms
is of primary interest, as when antibiotic-producing agents are introduced for purposes of
biological control, bioremediation, or bio fertilization, antibiotic-nonproducing mutant
derivatives are indispensable in distinguishing between effects due specifically to the
antibiotic and those attributable to other activities of the introduced agents. This project
reviews factors known to affect the production, activity and detection of antibiotics in situ,
discusses methods for extraction and quantification from soil and plant materials, and
describes approaches to detecting biosynthetic genes, their expression, and the effects
of synthesis in soil habitats. Research until now has focused mainly on the activities of a
few bacterial genera producing compounds of known structure, but the techniques that
have been developed may be applicable to diverse taxa producing structurally undefined
bioactive metabolites as well.
From the data presented here it would appear widespread use of antimicrobials in both
inpatient and outpatient settings has been associated with the emergence of antibiotic
resistant microorganisms. Bacterial strains that have been susceptible to all antimicrobial
agents for decades have now developed resistance not only to those classic therapies,
but to newer agents as well. Other organisms have developed resistance to new
antimicrobials almost as soon as the drugs have been marketed, if not earlier. Organisms
that are resistant to several different groups of antimicrobials have become more
prevalent in recent years. Antibiotic resistance has been a concern in the medical
community since the 1950s, when an increase was documented in colonization and
infection rates of penicillin-resistant Staphylococcus aureus in hospitalized patients. In
March 1991, the Alabama Department of Public Health (ADPH) distributed a document,
“Position Paper on the Control of Methicillin-resistant Staphylococcus aureus in Hospitals
and Long-term Care Facilities”, to the acute and long-term care facilities in Alabama. As
there continues to be an evolution of knowledge concerning antibiotic resistance, this
document has been written to update the 1991 document and to incorporate the present
standards of care. As the need arises, it will be revised.
A rapid increase in the occurrence of vancomycin-resistant enterococci (VRE) reported
in United States hospitals has generated concern comparable to that observed when the
methicillin-resistant Staphylococcus aureus (MRSA) problem was first recognized. In
addition, the first clinical strain of vancomycin-resistant staphylococcus aureus (VRSA)
has recently been documented in the United States. In September, 1995, the Centers for
Disease Control and Prevention (CDC) published the document “Recommendations for
Preventing the Spread of Vancomycin Resistance, Recommendations of the Hospital
Infection Control Practices Advisory Committee (HICPAC)”. While this valuable CDC
document addresses prevention and control issues in the acute care setting, it does not
provide guidance for management of vancomycin resistance in other healthcare settings.
The purpose of this document is to update and expand the previous ADPH position paper
on MRSA and to include infection control recommendations concerning VRE resistance
in Alabama healthcare facilities/settings. Since today’s trend is toward shorter hospital
stays, more outpatient surgery, outpatient IV therapy, and a shift to home therapy, and
because a colonization/infection may not be resolved when the patient is discharged or
transferred, we have included recommendations for nontraditional healthcare settings in
an effort to prevent and control antibiotic-resistant organism transmission.
Preventing and controlling the spread of all potential antibiotic-resistant bacteria in a
variety of healthcare settings will require a coordinated, conscious effort from all
individuals participating in the healthcare delivery system. Elements of this effort will
require: (1) instituting a system for surveillance of clinical antimicrobial susceptibility
summary reports by location and risk, (2) prudent use of antibiotics, (3) early detection
and prompt reporting of MRSA, VRE, and other epidemiologically important antibiotic
resistant organisms by clinical microbiology laboratories, (4) immediate implementation
of Standard Precautions and other appropriate infection control measures with specific
emphasis on hand hygiene (to include monitoring of healthcare worker adherence) to
prevent further spread, (5) implementing protocols for removal of invasive devices when
they are no longer needed, (6) educating healthcare staff, as well as the general public,
regarding the problems of antibiotic resistance, (7) meticulous communication between
healthcare facilities/settings, and (8) incorporating the concept of infection control and
prevention of transmission of infections into the healthcare facility’s/setting’s safety
Since the discovery of penicillin in 1929 by Alexander Fleming the importance of
antibiotics as chemotherapeutic agent has been increasing year after year. More than
800 antibiotics are known though only few of them have a therapeutic importance. The
study of the biosynthetic pathway of many antibiotics have served as a way to design new
pathways and products.
Penicillin production can be studied as an example for the antibiotic world because it was
the first antibiotic produced on a large scale and also because today it is the one most
commonly used inthe treatment of human infectious diseasearound the world. In addition
many of the techniques used for the industrial production of penicillin have served as a
model for the industrial production of other antibiotics or secondary metabolites.
Traditionally increase in the production of antibiotics has been obtained using classical
improvement methods which have given good results and the use of these methods have
allowed researchers to improve the strains and the production processes. However over
the last few years molecular biology techniques have been implemented in order to
increase final antibiotic production and also to obtain products that are not naturally
After approximately 20 years of gene manipulation there is still one question open. Can
the DNA Recombinant technology improve natural evolution or simply make it go faster?
Since industry still uses classical mutation and screening methods to select for better
producing strains, molecular biology can probably serve as an additional tool to improve
strains which must be combined with the classical improvement techniques to get the
The emergence and spread of resistance to antibiotics among common pathogenic
bacteria is an important healthcare concern. Today, the magnitude of the problem has
become so great that it is threatening to reverse the scientific progress made so far.
Several factors contribute to the problem including misuse on the part of physicians and
patients, widespread use of antibiotics with high resistance potential, use of antibiotics as
animal growth promoters and in household products. Antibiotic resistance, especially the
development of bacteria resistant to multiple drugs, is a rapidly growing global problem.
Diverse factors, including patient’s expectation, over-the-counter availability of antibiotics,
rampant use of antibiotics with high resistance potential and use of antibiotics as growth
promoters in animals contribute to the problem. Effective strategies to control antibiotic
• Increasing public awareness through public education campaigns
• Promoting prudent use of antibiotics by health professionals
• Restricting the use of antibiotics with high resistance potential
• Checking over-the-counter sale of antibiotic
• Discouraging the use of antibiotics in animal husbandry and household products
• Formulating evidence-based guidelines for the treatment of common infections
• Surveillance at local and national level to assess the magnitude of the problem and
effectiveness of interventions.
Interventions may have limited effect as long as other regions of the world continue to
misuse antibiotics, select for resistant bacteria and spread them. Therefore, we must all
join hands in this fight to return to the world of susceptible microbes.
1. Asaka, O., and M. Shoda. 1996. Biocontrol of Rhizoctonia solani damping-off of tomato
with Bacillus subtilis RB14. Appl. Environ. Microbiol. 62:4081-4085.
2. Blum, U., S. B. Weed, and B. R. Dalton. 1987. Influence of various soil factors on the
effects of ferulic acid on leaf expansion of cucumber seedlings. Plant Soil 98:111-130.
3. Blum, U., A. D. Worsham, L. D. King, and T. M. Gerig. 1994. Use of water and EDTA
extractions to estimate available (free and reversibly bound) phenolic acids in Cecil soils.
J. Chem. Ecol. 20:341-359.
4. Bonsall, R. F., D. M. Weller, and L. S. Thomashow. 1997. Quantification of 2,4-
diacetylphloroglucinol produced by fluorescent Pseudomonas spp. in vitro and in the
rhizosphere of wheat. Appl. Environ. Microbiol. 63:951-955.
5. Burkhead, K. D., D. A. Schisler, and P. J. Slininger. 1994. Pyrrolnitrin production by
biological control agent Pseudomonas cepacia B37w in culture and in colonized wounds
of potatoes. Appl. Environ. Microbiol. 60:2031-2039.
6. Carroll, H., Y. Moënne-Loccoz, D. N. Dowling, and F. O'Gara. 1995. Mutational
disruption of the biosynthesis genes coding for the antifungal metabolite 2,4-
diacetylphloroglucinol does not influence the ecological fitness of Pseudomonas
fluorescens F113 in the rhizosphere of sugarbeets. Appl. Environ. Microbiol. 61:3002-
7. Chin-a-Woeng, T. F. C., G. V. Bloemberg, A. J. van der Bij, K. M. G. M. van der Drift,
J. Schripsema, B. Kroon, R. J. Scheffer, C. Keel, P. A. H. M. Bakker, J.-V. Tichy, F. J. de
Bruijn, J. E. Thomas-Oates, and B. J. J. Lugtenberg. 1998. Biocontrol by phenazine-1-
Carboxamide-producing Pseudomonas chlororaphis PCL 1391 of tomato root rot caused
by Fusarium oxysporum f. sp. radicis-lycopersici. Mol. Plant-Microbe Interact. 11:1069-
8. Chiou, C. T. 1989. Theoretical considerations of the partition uptake of nonionic organic
compounds by soil organic matter, pp. 1-29 In B. L. Sawhney and K. Brown (ed.),
Reactions and Movement of Organic Chemicals in Soils. SSSA Special Publication 22.
Soil Science Society of America, Madison, WI.
9. Corbell, N., and J. E. Loper. 1995. A global regulator of secondary metabolite
production in Pseudomonas fluorescens Pf-5. J. Bacteriol. 177:6230-6236.
10. Dalton, B. R., U. Blum, and S. B. Weed. 1983. Allelopathic substances in ecosystems:
effectiveness of sterile soil components in altering recovery of ferulic acid. J. Chem. Ecol.
11. Dalton, B. R., U. Blum, and S. B. Weed. 1989. Differential sorption of exogenously
applied ferulic, p-coumaric, p-hydroxybenzoic, and vanillic acids in soil. Soil Sci. Soc. Am.
J. 53:757- 762.
12. Dalton, B. R., S. B. Weed, and U. Blum. 1987. Plant phenolic acids in soils: a
comparison of extraction procedures. Soil Sci. Soc. Am. J. 51:1515-1521.
13. di Pietro, A., M. Gut-Rella, J. P. Pachlatko, and F. J. Schwinn. 1992. Role of antibiotics
produced by Chaetomium globosum in biocontrol of Pythium ultimum, a causal agent of
damping-off. Phytopathology 82:131-135.
14. Duffy, B. K., and G. Défago. 1997. Zinc improves biocontrol of Fusarium crown and
root rot of tomato by Pseudomonas fluorescens and represses the production of pathogen
metabolites inhibitory to bacterial antibiotic synthesis. Phytopathology 87:1250-1257.
15. Fried, B., and Sherma, J. 1982. Thin-Layer Chromatography: Techniques and
Applications. Marcel Dekker, New York.
16. Georgakopoulos, D. G., M. Hendson, N. J. Panopoulos, and M. N. Schroth. 1994.
Analysis of expression of a phenazine biosynthesis locus of Pseudomonas aureofaciens
PGS12 on seeds with a mutant carrying a phenazine biosynthetic locus-ice nucleation
reporter gene fusion. Appl. Environ. Microbiol. 60:4573-4579.
17. Hasset, J. J., and W. L. Banwart. 1989. The sorption of nonpolar organics by soils
and sediments,, pp. 31-44. In B. L. Sawhney and K. Brown (ed.), Reactions and
Movement of Organic Chemicals in Soils. SSSA Special Publication 22. Soil Science
Society of America, Madison, WI.
18. Homans, A. L., and A. Fuchs. 1970. Direct bioautography on thin-layer
chromatograms as a method for detecting fungitoxic substances. J. Chromatogr. 51:327-
19. Huang, P. M., T. S. C. Want, M. K. Wang, M. H. Wu, and N. W. Hsu. 1977. Retention
of phenolic acids by noncrystalline hydroxy-aluminum and -iron compounds and clay
minerals of soils. Soil Sci. 123:213-219.
20. Huddleston, A. S., N. Cresswell, M. C. P. Neves, J. E. Beringer, S. Baumberg, D. I.
Thomas, and E. M. H. Wellington. 1997. Molecular detection of streptomycin-producing
Streptomycetes in Brazilian soils. Appl. Environ. Microbiol. 63:1288-1297.
21. Joyner, D. C., and S. E. Lindow. 2000. Heterogeneity of iron bioavailability on plants
assessed with a whole-cell GFP-based bacterial biosensor. Microbiology 146:2435-2445.
22. Kaminsky, R., and W. H. Muller. 1987. The extraction of soil phytotoxins using a
neutral EDTA solution. Soil Sci. 124:205-210.
23. Keel, C., U. Schnider, M. Maurhofer, C. Voisard, J. Laville, U. Burger, P. Wirthner, D.
Haas, and G. Défago. 1992. Suppression of root diseases by Pseudomonas fluorescens
CHA0: importance of the bacterial secondary metabolite 2,4-diacetylphloroglucinol. Mol.
Plant-Microbe Interact. 5:4-13.
24. Kelley, W. T., D. L. Coffey, and T. C. Mueller. 1994. Liquid chromatographic
determination of phenolic acids in soil. J. AOAC International 4:805-809.
25. Kempf, H.-J., P. H. Bauer, and M. N. Schroth. 1993. Herbicolin A associated with
crown and roots of wheat after seed treatment with Erwinia herbicola B247.
Phytopathology 83:213- 216.
26. Kempf, H.-J., S. Sinterhauf, M. Müller, and P. Pachlatko. 1994. Production of two
antibiotics by a biocontrol bacterium in the spermosphere of barley and in the rhizosphere
of cotton, p. 114-116. In M. H. Ryder, P. M. Stephens, and G. D. Bowen (ed.), Improving
Plant Productivity with Rhizobacteria. CSIRO Division of Soils, Adelaide, Australia.
27. Kraus, J., and J. E. Loper. 1992. Lack of evidence for a role of antifungal metabolite
production by Pseudomonas fluorescens Pf-5 in biological control of Pythium damping-
off of cucumber. Phytopathology 82:264-271.
28. Kraus, J. and J. E. Loper. 1995. Characterization of a genomic region required for
production of the antibiotic pyoluteorin by the biological control agent Pseudomonas
fluorescens Pf-5. Appl. Environ. Microbiol. 61:849-854.
29. Lebuhn, M., and A. Hartmann. 1993. Method for the determination of indole-3-acetic
acid and related compounds of L-tryptophan catabloism in soils. J. Chromatog. 629:255-
30. Lehmann, R. G., H. H. Cheng, and J. B. Harsh. 1987. Oxidation of phenolic acids by
soil, iron and manganese oxides. Soil Sci. Soc. Am. J. 51:352-356.
31. Lumsden, R. D., J. C. Locke, S. T. Adkins, J. F. Walter, and C. J. Rideout. 1992.
Isolation and localization of the antibiotic gliotoxin produced by Gliocladium virens from
alginate prill in soil and soilless media. Phytopathology 82:230-235.
32. Maurhofer M., C. Keel, D. Haas, and G. Défago. 1995. Influence of plant species on
disease suppression by Pseudomonas fluorescens strain CHA0 with enhanced antibiotic
production. Plant Pathol. 44:40-50.
33. Mavrodi, O. V., B. B. McSpadden Gardener, D. V. Mavrodi, R. F. Bonsall, D. M.
Weller, and L. S. Thomashow. 2001. Genetic diversity of phlD from 2,4
diacetylphloroglucinolproducing fluorescent Pseudomonas spp. Phytopathology 91:35-
34. Mazzola, M., R. J. Cook, L. S. Thomashow, D. M. Weller, and L. S. Pierson. 1992.
Contribution of phenazine antibiotic biosynthesis to the ecological competence of
fluorescent pseudomonads in soil habitats. Appl. Environ. Microbiol. 58:2616-2624.
35. McSpadden Gardener, B. B., D. V. Mavrodi, L. S. Thomashow, and D. M. Weller.
2001. A rapid PCR-based assay characterizing rhizosphere populations of 2,4-DAPG-
producing bacteria. Phytopathology 91:44-54.
36. Metsä-Ketalä, M., V. Salo, L. Halo, A. Hautala, J. Hakala, P. Mäntsälä, and K.
Ylihonko. 1999. An efficient approach for screening minimal PKS genes from
Streptomyces. FEMS Microbiol. Lett. 180:1-6.
37. Nakayama, T., Y. Homma, Y. Hashidoko, J. Mizutani, and S. Tahara. 1999. Possible
role of xanthobaccins produced by Stenotrophomonas sp. strain SB-K88 in suppression
of sugar beet damping-off disease. Appl. Environ. Microbiol. 65:4334-4339.
38. Picard, C., F. di Cello, M. Ventura, R. Fani, and A. Gluckert. 2000. Frequency and
biodiversity of 2,4-diacetylphloroglucinol-producing bacteria isolated from the maize
rhizosphere at different stages of plant growth. Appl. Environ. Microbiol. 66:948-955.
39. Pierson, E. A., D. W. Wood, J. A. Cannon, F. M. Blachere, and L. S. Pierson III. 1998.
Interpopulation signaling via N-acyl-homoserine lactones among bacteria in the wheat
rhizosphere. Mol. Plant-Microbe Interact. 11:1078-1084.
40. Pierson III, L. S., and L. S. Thomashow. 1993. Cloning and heterologous expression
of the phenazine biosynthetic locus from Pseudomonas aureofaciens 30-84. Mol. Plant-
Microbe Interact. 5:330-339.
41. Raaijmakers, J. M., R. F. Bonsall, and D. M. Weller. 1999. Effect of population density
of Pseudomonas fluorescens on production of 2,4-diacetylphloroglucinol in the
rhizosphere of wheat. Phytopathology 89:470-475.
42. Raaijmakers, J. M., D. M. Weller, and L. S. Thomashow. 1997. Frequency of
antibioticproducing Pseudomonas spp. in natural environments. Appl. Environ. Microbiol.
43. Schnider-Keel, U., A. Seematter, M. Maurhofer, C. Blumer, B. Duffy, C. Gigot-
Bonnefoy, C. Reimmann, R. Notz, G. Défago, D. Haas, and C. Keel. 2000. Autoinduction
of 2,4- diacetylphloroglucinol biosynthesis in the biocontrol agent Pseudomonas
fluorescens CHA0 and repression by the bacterial metabolites salicylate and pyoluteorin.
J. Bacteriol. 182: 1215-1225.
44. Seow, K.-T., G. Meurer, M. Gerlitz, E. Wendt-Pienkowski, C. R. Hutchinson, and J.
Davies. 1997. A study of iterative type II polyketide synthases, using bacterial genes
cloned from soil DNA: a means to access and use genes from uncultured
microorganisms. J. Bacteriol. 179: 7360-7368.
45. Shanahan, P., A. Borro, F. O'Gara, and J. D. Glennon. 1992. Isolation, trace
enrichment and liquid chromatographic analysis of diacetylphloroglucinol in culture and
soil samples using UV and amperometric detection. J. Chromatogr. 606:171-177.
46. Shanahan, P., D. J. O'Sullivan, P. Simpson, J. D. Glennon, and F. O'Gara. 1992.
Isolation of 2,4-diacetylphloroglucinol from a fluorescent pseudomonad and investigation
of physiological parameters influencing its production. Appl. Environ. Microbiol. 58:353-
47. Stohl, E. A., S. F. Brady, J. Clardy, and J. Handelsman. 1999. ZmaR, a novel and
widespresd antibiotic resistance determinant that acetylates zwittermycin A. Appl.
Environ. Microbiol. 181:5455-5460.
NAME-: AQSA FATIMA
FATHERS NAME-: RAFEEQUDDIN
MOTHERS NAME-: SHABEENA RAFEEQ
ADDRESS-: RZ-605, 4TH FLOOR, STREET 21, TUGLAKABAD EXTN, NEW DELHI.
MOBILE NUMBER-: 9891393915, 9718482523