STUDY OF DRUG RESISTANCE USING ANTIBIOTICS

CERTIFICATE OF AUTHENTICITE This is to certify that “ swati Chhachhia” a

student of class XII B has successfully completed investigatory project under the

guidance of Mrs. Manju Phatak .

Signature External (subject teacher) Examiner

ACKNOWLEDGEMENT

I feel proud to present my investigatory project in Biology on the “STUDY OF DRUG

RESISTANCE IN BACTERIA USING ANTIBIOTICS” . This project would not have been feasible without the proper rigorous guidance of biology teacher Mrs. Manju Phatak who guided me throughout this project in every possible way . An

investigatory project involves various difficult lab experiments, which have to obtain the observations and conclude the reports on a meaningful note. Thereby, I would like to thanks Mrs. Manju Phatak for

guiding me on a sustematic basis and ensuring that in completed all my research with ease. Rigorous hard work has to put in this project to ensure that it proves to be the best. I hope that it proves to be the best. I hope that this project will to be proves to be a breeding ground for the next generation of students and will

guide them in ever possible way .

INTRODUCTION

An antibacterial is an agent that inhibits bacterial growth or kills bacteria. The term is often used synonymously with the term antibiotic(s). Today, however, with increased knowledge of the causative agents of various compounds, including anti-fungal and other compounds. Antibacterials must be distinguished from disinfectants(sanitizing agents), which are less-selective substance used to destroy microorganisms.The term antibiotic was first used in 1942 by Selman Waksman and his collaborators in

the journal articles to describe any subatance produced by a microorganisms that

is antagonistic to the growth of other microorganisms in high dilution. This

defination excluded substances that kill bacteria but are not produced by a

microorganisms (such as gastric juices and hydrogen peroxide). It also excluded

synthetic anti bacterial compounds such as the sulfonamides. Many antibacterial

compounds are relatively small molecules with a molecular weight of less than 2000

atomic mass units.

CLASSES Antibacterial antibiotics are commonly classified based on their mechanism of

action, chemical structure, or spectrun of activity . Most targer bacterial functions or growth processes. Those that target the

bacterial cell wall (penicillins and cephalosporins) or the cell membrane

(polymyxins), or interfere with essential bacterial enzymes (rifamycins,

lipiarmycins, quinolones, and sulfonamides) have bactericidal activities. Those that target protein synthesis (macrolides,

lincosamides and tetracyclines) are usually bacteriostatic (with the exception of

bactericidal aminoglycosides)

MEDIAL USES TREATMENT

•Bacterial infection•Protozoan infection e.g.Metronidazole is effective against several parasitics•Immunomodulation e.g.Tetracycline is effective against periodontal inflammation, &

Dapsone which is effective against autoimmu- -ne diseases oral mucous membrane. PREVENTION OF INFECTION

•Surgical wounds•Dental antibiotic prophylaxis•Conditions of neutropenia e.g. Cancer-related

SIDE EFFECTSAntibacterial are screened for any negativeeffects on humans or other mammals beforeapproval foe clinical use, and are usually

considered safe and most are well-tolerated. However, some antibacterials

have been associated with a rangeof adverse effects. Side-effects range from mild to very serious depending on the antibiotics

used, the microbial organisms targeted, and the individual patient. Safety profiles of

newer drugs are often not as well-established as for those that have a long history of use. Adverse effects range from

fever and nausea to major allergic reactions, including photodermatitis and anaphylaxis. Common side-effects include diarrhea, resulting from disruption of the

species composition in the intestinal flora, resulting, for example, in

overgrowth of pathogenic bacteria,such as Clostridium difficle. Antibacterials can

also affect the vaginal flora, and may lead to overgrowth of yeast species of the genus

Candida in the vulvo-vaginal area. Additional side-effects can result

frominteraction with other drugs, such as elevated risk of tendon damage from

administration of a quinolone antibiotic wit a systemic corticosteroi

Antibacterial-resistant strains and species, sometimes referred to as “superbugs”, now contribute to the

emergence of diseases that were for a while well-controlled. For example, emergent bacterial strains causing

tuberculosis(TB) that are resistant to previously effective antibacterial treatments pose many therapeutic

challenges. Every year, nearly half a million new cases of multidrug-resistant

tuberculosis(MDR-TB) are estimated to occur worldwide. For example, NDM-1 is a newly identified enzyme conveying bacterial

resistance to a broad range of beta- lactam antibacterials. The United Kingdom's Health Protection Agency has stated that “most

isolates with NDM-1 enzyme are resistant to all standard intravenous antibiotics for

treatment of severe infection”.

RESISTANT PATHOGEN ACINETOBACTERAcinetobacter is a group of bacteria commonly found in soil and water . While there are many types or “species” of Acinetobacter and all can cause human disease, Acinetobacter baumannii accounts for about 80% of reported infections.Outbreak of Acinetobacter infections typically occur in intensive care units and healthcare settings housing very ill patients. Acinetobacter infection rarely occur outside of healthcare setting.

ANTHARXAntharx is a serious disease caused by Bacillus anthracis, a bacterium that forms spores. Antharx most commonly occours in wild and domestic mammalian species, but it can also occour in humans when they are exposed to infected animals or to tissue form infected animals or when anthrax spores are used as a bioterrorist weapon. Some starins of B. Anthracis may be naturally resistant to certain antibiotics and not other. In addition, there may be biologically mutant strains that are engineered to be resistant to various antibiotics. CAMPYLOBACTERCampylobacter is estimated to cause over 1.3million infections and 76 death in the United States each year. Many of these infections are foodborne. Campylobacter infections can result in long-term consequences, such as arthritis or a type of paralysis called Guillain-Barre syndrome.

GROUP B STREPTOCOCCUS

Group B Streptococcus (group B strep) is a type of bacteria that causes illness in newborn babies, the elderly, and the adults with other illness , such as diabetes or liver diseases . Group B strep has shown confirmed resistance to certain antibiotics

DO'S AND DON'TS OF

ANTIBIOTICS•DO take the antibiotic exactly as indicated

on the container.•DO finish the prescription even if you

being to feel better.•DON'T demand antibiotics when your health care provider says they are not needed.

•DON'T skip doses.•DON'T share the prescription with anyone

else.•DON'T save the prescription for later time.

MATERIAL REQUIRED•Sterilised petridishes

•Sterilised culture tube with media•Transfer loops

•Forcep•Flask•Beaker•Burner

•Penicillin•Aureomycin

•Hay•Alcohol

•Agar•Starch

•Distilled water etc.

EXPERIMENTAL PROCEDURE•To 200ml distilled water in a flask, add 8gms of agar powder and 2gms of starch. Put a few pieces of dry hay into the medium. Cover flask with an inverted beaker. Boil the medium for 5min and then cool the medium to room temperature. Place the flask in warm place. Within 2-3days, formation of scum of cloudy suspension appear on the medium indicating the growth of Bacillus subtilis.•Take culture tubes with agar medium and heat the test tubes in warm water to melt agar. Cool each test tube so that you can hold it in your hand and the agar remains liquid. Remove the cotton plug and pass the mouth of the test tube through the burner flame twice. Flame the transfer loop after dipping it in alcohol and let it cool. Pick up a loop full of bacterial culture from the flask and transfer it to the warm agar in the culture tube. Flame the loop and the mouth of the culture tube and replace the cotton plug. Roll the culture tube of warm agar between plams to mix the bacteria well with agar. Transfer of bacteria should be done as quickly as possible.

•Take sterilised petridishes. Remove the cotton plug and flame the mouth of the culture tube. Lift the cover of the petridish at an angle of 45degree and quickly pour the medium of the culture tube into the bottom half of the dish. Remove the culture tube and replace the cover of the petridish. Move the covered petridish along the table top in a figure eight pattern to distribute the medium evenly. Allow the agar to cool. Prepare two petridishes and mark them A and B.•Prepare the penicillina nd aeromycin solution by dissolving the powdered drugs in distilled water. Cut discs of filter paper of 1cm diameter. Soak a disc in each of the penicillin and aeromycin solution. Dip the forceps in alcohol and pass the forceps tip quickly over the burner flame. Use the sterilised forceps to put penicillin and aeromycin soaked discs at two distant sites of petridish A. Consider the petridish B as control. Keep both the petridishes undisturbed in warm place or in an incubator to allow the bacteria to grow . Observe the petridish for several days.

OBSERVATIONThe area around the antibiotic discs in the petridish will be clear. In other areas, colonies of bacteria will be observed. Examine the clear area in each petridish for few more days. A few very small colonies may appear in the areas. These are the colonies of resistant strains of the bacteria. CONCLUSIONSAntibiotic drugs kill most of bacetria strains, hence the areaa appear clear. However, a few strains which were resistant in the bacterial population survived and produced colonies later.This proves the resistant starin to antibiotics were present in the bacterial population.

RESISTANCESEM depiciting methicillin – resistance Staphylococcus aureus bacteria.The emergence of resistance of bacteria to antibacterial drugs is a common phenomenon. Emergence of resistance often reflects evolutionary processes that take place during antibacterial drug therapy. The antibacteria; treatment may select for bacteria; strains with physiologically or genetically enhanced capacity to survive high doses of antibacterials. Under certain conditions, it may result in preferential growth of susceptible bacteria is inhibited by the drug. For example, antibacterials selection for strain having previously acquired antibacterial-resistance genes was demonstrated in 1943 by the Luria-Delbruck experiment. Antibacterials such as penicillin and erythromycin, which used to have high efficacy against many bacterial species and strains,have become less effective, because of increased resistance of many bacterial strains.