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Wayan Tunas Artama
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IDENTIFIKASI ESCHERICHIA COLI O157:H7 SERTA DETEKSI GEN SHIGA LIKE TOXIN 1 DAN 2 ASAL FESES HEWAN, DAGING, DAN FESES MANUSIA Suardana, I Wayan; Tunas Artama, Wayan; Asmara, Widya; Setiadi Daryono, Budi
Jurnal Veteriner Vol 11, No 4 (2010)
Publisher : Jurnal Veteriner

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Abstract

Escherichia coli O157:H7 with the ability to produce shiga-like toxin was isolated from beef, cattle, chicken, and human feces. Due to its importance to human health, it is necessary to identify the genes encoding the production of shiga-like toxin, stx1 and stx2 respectively to further understand the pathogenesis. Isolation of E. coli was done on Eosin Methylene Blue Agar (EMBA), followed by identification on Sorbitol MacConkey Agar (SMAC), latex agglutination test, and H7 antiserum test, respectivelly. The existence of genes stx1 and stx2 in E. coli O157:H7 was confirmed molecularly using PCR method with specific primers LP 30/31 and LP 43/44, Stx2 (F)/Stx2 (R) respectively. Escherichia coli O157:H7 was isolated from 22 out of 344 samples (6,4%). Some isolates showed gene stx1 and stx2 was detected in two isolates as indicated by a 384 bp band (stx1 gene), 584 bp and 1588 bp bands (stx2 gene) respectivelly. The results indicated that local isolates E. coli O157:H7 are potential as a zoonoses agent.
Studi Epidemiologi Agen Zoonosis Escherichia coli O157:H7 melalui Analisis Random Amplification of Polymorphic DNA (RAPD) Suardana, I Wayan; Tunas Artama, Wayan; Asmara, Widya; Setiadi Daryono, Budi
Jurnal Veteriner Vol 12, No 2 (2011)
Publisher : Jurnal Veteriner

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Epidemiological studies of zoonotic agent Escherichia coli O157:H7 have been analyzed pheneticallyand or phylogenetically. In a phenetic classification, micoorganisms are arranged into groups (phena) onthe basis of high overall similarity using both phenotypic and genotypic methods without judgementaspect of its ancestry or evolutionary. Due to its importance to epidemiological aspect, the study of geneticvariation of isolates origin from some sources need to be conducted in order to trace the routes of infection.A total of 20 samples obtained from some sources i.e clinically human feces, non-clinically human feces,cattle feces, chicken feces, and beef feces were used in this study. The study was started by confirming allof the isolates using O157 latex agglutination test and H7 antiserum test, followed by genomic DNAanalysis by random amplification of polymorphic DNA /RAPD methods. RAPD results were analyzed using a simple matching coeficient (Ssm) and alogorhythm unweighted pair group method using arithmeticaverages (UPGMA) programe. Results showed there were range of genetic DNA from local isolates (75.1–99,6%) which was almost similar to ATCC 43894 control isolate. The highest similarity (99.6%) to ATCC43894 control was showed by SM-7(1) isolate obtained from cattle fecal and KL-68(1), isolate obtainedfrom clinically human fecal. In addition, KL-52(7) obtained from clinically human fecal had high similarity(99.6%) to MK-35 isolate obtained from chicken fecal. On the other hand, DS-21(4) and DS-16(2) isolatesthat were obtained from beef had high similarity (84.9%) to other isolates including ATCC 43894 controlisolate. The highest similarity of E. coli O157:H7 isolates that were obtained from cattle feces, beef, andchicken feces to human feces isolate indicated that there were both cattle and chicken were potentialreservoirs of the zoonotic agen which can be transmitted to human.