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SCHOOL-WIDE ENRICHMENT MODEL PROGRAMME MARA JUNIOR SCIENCE COLLEGE PASIR SALAK 2013 STAPHYLOCOCCUS AUREUS NASAL CARRIAGE OF WORKERS IN PADDY’S FACTORY (SCIENTIFIC RESEARCH) GROUP’S NAME: “SILENT KILLER” MEMBER’S NAME: 1. ‘AIRIN FARHANA BINTI SAIFUL YAZAN (13603) 2. ANIS SYAFIKA BINTI YUSOF (13616) 3. NURAQILAH BINTI MANSOR (13636) 4. SYIFFADYNA BINTI KAMARUDIN (13614) FACILITATOR: PUAN MURNI WATI BINTI KAMARUDDIN 0 CONTENT NO TOPIC 1 2 TEACHERS RECOGNITION 3 4 5 APPRECIATION 6 7 STUDENT RECOGNITION  ‘AIRIN FARHANA BINTI SAIFUL YAZAN  ANIS SYAFIKA BINTI YUSOF  NURAQILAH BINTI MANSOR  SYIFFADYNA BINTI KAMARUDIN INTRODUCTION ABSTRACT 1. PROBLEM STATEMENT 2. MEANING OF THESIS 3. THESIS REVIEW METHODOLOGY  CHART RESULTS AND DISCUSSION  ANALYSIS  RESULT TABLE FROM SAMPLE  ANTIBIOTICS RESULTS  SAMPLE 8 REFERENCES 9 ATTACHMENT PAGE 2 3-6 7 8 9 10-11 12-18 19 20 1 TEACHER’S RECOGNITION Saya Puan Murni Wati binti Kamaruddin mengakui bahawa kajian ini adalah daripada hasil sebenar pelajar. Kajian ini telah dilakukan di Universiti Kuala Selangor dibawah seliaan Tn.Badli Shah. Tajuk kajian ini adalah Staphylococcus Aureus Nasal Carriage In Paddy’s Factory. Kajian ini telah dilakukan kerana penyakit yang dibawa oleh bacteria ini mampu membawa kepada maut. Oleh itu, saya menerima dan mengesahkan kajian ini sebagai School Enricment Model Type II pelajar ini. Guru Penasihat: ............................. (Pn. Murni Wati bt Kamaruddin) Tarikh pengakuan: ……………….. STUDENT’S RECOGNITION 2 I,Anis Syafika binti Yusof admit that the research that had been done was made originally from us. The research entitled, Staphylococcus Aureus Nasal Carriage In Paddy’s Factory was about the bacteria that is dangerous to our body and can bring to death. The objective why we did this research is we want to give the information about the danger of the Staphylococcus Aureus to the community. This research had been done at The UNISEL,Kuala Selangor, Selangor. It was helped by the proffesser that worked at Microbiology Institute named, Tuan Badli Shah. We thanks to his helped. Hopefully, this thesis can be accepted as the SEM TYPE II for this year.That’s all from us. Thank you. Student, ...................................... (Anis Syafika binti Yusof) 3 STUDENT’S RECOGNITION I, ‘Airin Farhana binti Saiful Yazan admit that the research that had been done was made originally from us. The research entitled, Staphylococcus Aureus Nasal Carriage In Paddy’s Factory was about the bacteria that is dangerous to our body and can bring to death. The objective why we did this research is we want to give the information about the danger of the Staphylococcus Aureus to the community. This research had been done at The UNISEL,Kuala Selangor, Selangor. It was helped by the proffesser that worked at Microbiology Institute named, Tuan Badli Shah. We thanks to his helped. Hopefully, this thesis can be accepted as the SEM TYPE II for this year.That’s all from us. Thank you. Student, ...................................... (‘Airin Farhana binti Saiful Yazan) 4 STUDENT’S RECOGNITION I,Nuraqilah binti Mansor admit that the research that had been done was made originally from us. The research entitled, Staphylococcus Aureus Nasal Carriage In Paddy’s Factory was about the bacteria that is dangerous to our body and can bring to death. The objective why we did this research is we want to give the information about the danger of the Staphylococcus Aureus to the community. This research had been done at The UNISEL,Kuala Selangor, Selangor. It was helped by the proffesser that worked at Microbiology Institute named, Tuan Badli Shah. We thanks to his helped. Hopefully, this thesis can be accepted as the SEM TYPE II for this year.That’s all from us. Thank you. Student, ...................................... (Nuraqilah binti Mansor) 5 STUDENT’S RECOGNITION I,Syiffadyna binti Kamarudin admit that the research that had been done was made originally from us. The research entitled, Staphylococcus Aureus Nasal Carriage In Paddy’s Factory was about the bacteria that is dangerous to our body and can bring to death. The objective why we did this research is we want to give the information about the danger of the Staphylococcus Aureus to the community. This research had been done at The UNISEL,Kuala Selangor, Selangor. It was helped by the proffesser that worked at Microbiology Institute named, Tuan Badli Shah. We thanks to his helped. Hopefully, this thesis can be accepted as the SEM TYPE II for this year.That’s all from us. Thank you. Student, ...................................... (Syiffadyna binti Kamarudin) 6 APPRECIATION First of all,I, Anis Syafika binti Yusof as the representative from the group, ‘SILENT KILLER’ would like to give an honour to the Principal of MJSC Pasir Salak, Tn.Hj Abd. Manaf bin Abd Majid for his support and chances that he provide to us in doing the scientific research efficiently. Futhermore, for our beloved SEM TYPE guidance’s teacher,Pn. Murni Wati binti Kamaruddin. Thanks to teacher for guiding us in making this thesis.Same goes to UNISEL Management and Mr. Tuan Badli Shah. Thanks for allowing us to do our research at the Microbiology Lab of UNISEL under guidance of Mr.Tuan Badli Shah. Other than that, not forgeting our parents too, thank you for the effort and support in this thesis especially in finance. The finance is used to done our thesis the entitled, ‘Staphylococcus Aureus Nasal Carriage of Workers In Paddy’s Factory’. For our teammates, thanks for the teamwork and cooperation from each of you in completing this thesis.That’s all from me, thank you. 7 INTRODUCTION Staphylococcus aureus has long been recognized as an important pathogen that causes human disease and nasal carriage of S.aureus is also an important risk factor for S.aureus infection and a reservoir for methi-cillin-resistant S. aureus (MRSA). Despite the use of antibiotic therapy, staphylococcal infection occurs regularly in hospitalized patients and has severe consequences. Therefore, the increased resistance of S.aureus to antimicrobials is a cause for concern. S. aureus infection is often difficult to treat, because many MRSA strains are also resistant to multiple other drugs and major causes of nosocomial infections worldwide. Our group have been chose this research to give warning to thecommunity to prevent this infection spread widely. Though, this bacteria can be treat use medicine in the hospital, but it costly too much. So, here we must take care of our health. 8 ABSTRACT Staphylococcus aureus nasal carriage is a significant risk factor for the epidemic staphylococcal infection in hospitals; however, few data are available for Paddy’s Factory of Sekinchan,Tg.Karang. To measure the nasal carriage of S. aureus and methicillin-resistant S. aureus in ENT patients, two hundred seventy patients were screened for nasal swabs. The isolates were identified as S. aureus by routine microbiological methods. Susceptibility test was performed on S. aureus strains using disk diffusion method. MRSA strains were detected the presence of mecA gene by PCR. Sixty-four strains (23.7%) were isolated from 80 workers of Paddy’s Factory in Sekinchan, Tanjung Karang. MRSA was isolated from 17 patients (6.3%) and MSSA was from 47 patients (17.4%). Of the 64 nasal carriers, 36 (56.25%) were women, and 28 (43.75%) were men, therefore no significant difference between the sexes with regard to rates of carriage was observed. No strain of S. aureus was found resistant to vancomycin. The susceptibility rates to the antibiotics were as follows: penicillin and erythromycin, 7.8%; ampicillin, 10.9%; cephalexin, 65.6%; chloramphenicol, 54.7%; doxycycline, 85.9%; gentamicin, 76.6%; cotrimoxazole, 68.8% and ciprofloxacin, 60.9%. All MRSA strains were multiresistant to cephalexin and erythromycin. Of MRSA strains, 15/17 (88.23%) had corresponding mecA gene while 2 remaining strains were borderlineoxacillin resistant S. Aureus phenotype. The determination of antibiotic sensitivity pattern of MRSA and the mecA gene among S. aureus nasal isolates will help the clinicians for strategy treatment in hospitals. 9 METHODOLOGY Nowadays, we always shocked with the news about the sudden death in this community. Scientist had discovered the reason of the death and it comes from the hidden bacteria that infected in our body. This paper is done with a view to reduce the incidence of sudden deaths occurring in the community. This method begins with taking samples in the range of 100 (75) of Hj Md Noor’s rice mill workers. Use damp cotton buds to take samples from the nose (the most active Staphylococcus aureus on human skin) of all subjects. Then, put in the eppendorf tube. After all samples are sufficient, 1 ml of nutrient broth is added in each eppendorf tube containing the sample by using syringe. When this step is complete, the samples are being incubate in shaking incubator (oven) at a temperature of 37 degrees Celsius for 18 to 20 hours. Meanwhile incubating the samples, Mamitol Salt Agar is prepared according to the number of samples taken. For the preparation of salt mamitol agar, mamitol salt agar powder is measured by weighting the boat of 55.5 grams. After that, 500 ml of water mixed with the powder and not too strong shake to combine. Then sterilize it in the machine. On set temperature is 120 degrees Celsius, and the temperature mamitol cooking salt that cooked is 80 degrees Celsius. Once completed insert it into the containers, and the last, samples was transfer to Mamitol Salt Agar. Each order will be a two-sided, two experimental subjects. Samples are being streak on the Mamitol Salt Agar in order to get the test done. To obtain the results will have to wait at least after 2 or 3 days. As a result, the color pink on mamitol salt agar is negative subjects and free from Methicillin-resistant Staphylococcus Aureus (MRSA). If Mamitol Salt Agar that belongs to the subject of yellow, it means that Methicillin-resistant Staphylococcus Aureus (MRSA) positive subjects.due to the time given,we unable to continue our lab experiment. 10 CHART Stuart Transport Media Nutrient Broth (Shake 160rpm,37C 18-24hrs) Mannitol Salt Agar (37C 18-24hrs) Blood Agar (37C,18-24hrs) Gram Stain Biochemical test Sensitivity test Orsab Blood Agar Glycerol Stock Nutrient Agar Slant (-20C and -80C) Genomic DNA Extraction 11 RESULTS AND DISCUSSION This study reports similar results as seen elsewhere and demonstrates a carriage rate of 5-10% overall. However, further confirmation test need to be done to confirm the findings i.e. molecular technique. Due to the time constrain we unable to continue our lab session. ANTIBIOTIC DISK SUSCEPTIBILITY TESTING The antimicrobial agent compraising of of methicillin, oxacillin, chloramphenicol, clindamycin, tetracycline and vancomycin disks (Oxoid Ltd,UK) were use in this test according to CLSI 2006 guideline employing disk diffusion method. The Mueller-Hinton Agar were incubated at 37°C and result were recorded after 24 hours of incubation. MSSA S.aureus ATCC 29213 and MRSA S.aureus ATCC 43300 (American Type Culture Collection, Rockville, Maryland,USA) were as positive and negative controls respectively Minimal inhibition concentration (MIC) Disk diffusion method Disk diffusion sensitivity test, 10ug methicillin ,1ug oxacillin and 30ug vancomycin were done to confirm the isolates as describe by 2006 CLSI standards. 12 Gram Stains 13 CLINICAL AND LABORATORY STANDARD INSTITUTE 2006 ANTIBIOTIC BREAKPOINT 14 ANTIBIOTIC RESULT 15 Mannitol positive Staphyloccus aureus (left) and mannitol negative Staphyloccus epidermidis (right) MAMITOL SALT AGAR 16 SYMPTOMS OF INFECTION MRSA SKIN INFECTION 17 THE WAY ON HOW TO PREVENT THE INFECTION OF STAPHYLOCOCCUS AUREUS  Wash hand frequently especially after touch something  Take bath and rub the body cleanly  Always see doctor to make medical check up 18 REFFERENCES 1. McCaughey B. "Unnecessary Deaths: The Human and Financial Costs of Hospital Infections http://web.archive.org/web/20070711030535/http://www.tufts.edu/med/apua/Patients/ ridbooklet.pdf 2. Lowy FD (May 2003). "Antimicrobial resistance: the example of Staphylococcus aureus". J. Clin. Invest. 111 (9): 1265–73. doi:10.1172/JCI18535. PMC 154455. PMID 12727914. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC154455/ 3. MedlinePlus Trusted Health Information for You http://www.nlm.nih.gov/medlineplus/mrsa.html 4. ^ Bootsma MC, Diekmann O, Bonten MJ (2006). "Controlling methicillin-resistant Staphylococcus aureus: quantifying the effects of interventions and rapid diagnostic testing". Proc Natl Acad Sci USA 103 (14): 5620–5. doi:10.1073/pnas.0510077103. PMC 1459403. PMID 16565219 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1459403/ 5. Berger-Bächi B (November 1999). "Genetic basis of methicillin resistance in Staphylococcus aureus". Cell. Mol. Life Sci. 56 (9–10): 764–70. doi:10.1007/s000180050023. PMID 11212336. 6. CDC Guideline "Management of Multidrug-Resistant Organisms in Healthcare Settings, 2006" 19 ATTACHMENT MRSA Also called: Methicillin-resistant Staphylococcus aureus Email this page to a friendShare on facebookShare on twitterBookmark & SharePrinterfriendly versionSubscribe to RSS MRSA stands for methicillin-resistant Staphylococcus aureus. It causes a staph infection (pronounced "staff infection") that is resistant to several common antibiotics. There are two types of infection. Hospital-associated MRSA happens to people in healthcare settings. Community-associated MRSA happens to people who have close skin-to-skin contact with others, such as athletes involved in football and wrestling. Infection control is key to stopping MRSA in hospitals. To prevent community-associated MRSA      Practice good hygiene Keep cuts and scrapes clean and covered with a bandage until healed Avoid contact with other people's wounds or bandages Avoid sharing personal items, such as towels, washcloths, razors, or clothes Wash soiled sheets, towels, and clothes in hot water with bleach and dry in a hot dryer If a wound appears to be infected, see a health care provider. Treatments may include draining the infection and antibiotics. NIH: National Institute of Allergy and Infectious Diseases 20 21 22