Background and Aim: Extended-spectrum β-lactamase (ESBL) is an enzyme produced by the family of Enterobacteriaceae, especially Escherichia coli and Klebsiella pneumoniae, which can hydrolyze β-lactam antibiotics, such as penicillins, cephalosporins, cephamycin, and carbapenem. ESBL-producing bacteria are widely distributed from farms to slaughterhouses until food products originating from animals are available in the market, which plays an important role as a pathway for the exposure and transmission of ESBL-producing bacteria from food products of animal origin to humans. This study aimed to determine the antibacterial activity of Syzygium aromaticum (clove) and Cinnamomum verum (cinnamon) essential oils against strains resistant to ESBL-producing E. coli and K. pneumoniae isolates. Materials and Methods: The antibacterial activity of clove and cinnamon essential oils was tested against three strains of tested bacteria using the disk diffusion method. The minimum inhibitory concentration (MIC) of clove and cinnamon essential oils was determined using the broth microdilution method. The minimum bactericidal concentration (MBC) was determined using the MIC. Morphological changes on each tested bacteria were observed through scanning electron microscopy (SEM). Results: Both essential oils exhibited inhibitory effects toward all test organisms, indicated by inhibition zones around the disk. The MIC values of clove essential oil were 0.078% (v/v) for all tested bacteria, whereas the MICs of cinnamon essential oil ranged from 0.039% (v/v) to 0.156% (v/v) for all tested bacteria. MBC values of clove and cinnamon essential oils ranged from 0.078% (v/v) to 0.156% (v/v) for all tested bacteria. There were morphological changes in each tested bacterial cell that was observed through SEM. Each tested bacteria treated with clove and cinnamon essential oils showed shrinkage and cells lysis. Conclusion: It was concluded that clove and cinnamon essential oils have emerged as effective antibacterial agents by showing high antibacterial activity against ESBL-producing E. coli and K. pneumoniae isolates, as evidenced by the inhibition zone diameter and MIC value.

To clarify the epidemiological relationship between cattle and human infections of Shiga toxin-producing Escherichia coli (STEC), we studied the duration and magnitude of the excretion of STEC O157 and STEC O26 with rectal faeces from naturally infected cattle at a breeding farm in the Tohoku area of Japan, using microbiological methods. The prevalence of STEC O157 was 3·5% (11/324), whereas that of STEC O26 was 7·9% (14/178). Faecal shedding of STEC O157 persisted for <1 week to 10 weeks, whereas STEC O26 persisted from <1 week to 110000 c.f.u. for STEC O157 and from 3 to 2400 c.f.u. for STEC O26. All isolates of both STEC serotypes contained the stx1 or stx2 genes. Pulsed-field electrophoretic analysis of both STEC serotypes identified predominantly STEC O157 type III and STEC O26 type I in isolates, suggesting that a single STEC strain may be mutated in the intestinal tract of calves. These results indicate that STEC O157 is secreted for longer periods and in higher numbers than STEC O26 from healthy calves with natural infections, suggesting that STEC O157 may have more opportunities than STEC O26 to induce human disease.

Background and Aim: The feasibility assessment of food products on the market becomes one of the milestones of food safety. The quality of food safety of animal origin especially pork need to get attention and more real action from the parties related and concerned. Since pork is also a source of transmission for the contagion of foodborne disease so that the study of the existence of several agents in the pork and its products become the benchmark of safety level. This study aimed to isolate, identify, and detect the Shiga toxin 2a (stx2a) gene from Escherichia coli O157:H7 in pork, pig feces, and clean water in the Jagalan slaughterhouse. Materials and Methods: A total of 70 samples consisting of 32 pork samples, 32 pig fecal samples, and 6 clean water samples were used to isolate and identify E. coli O157:H7 and the stx2a gene. Isolation and identification of E. coli O157:H7 were performed using culture on eosin methylene blue agar and Sorbitol-MacConkey agar media and confirmed molecularly with polymerase chain reaction to amplify the target genes rfbE (317 bp) and fliC (381 bp). The isolates, which were identified as E. coli O157:H7, were investigated for the stx2a gene (553 bp). Results: The results of this study show that of the total collected samples, E. coli O157:H7 was 28.6% in Jagalan slaughterhouse and consisted of 25% of pork samples, 31.25% of pig fecal samples, and 33.3% of clean water samples. The isolates that were identified to be E. coli O157:H7 mostly contained the stx2a gene, which was equal to 75%, and consisted of seven isolates from pork samples, seven isolates from fecal samples, and one isolate from clean water samples. Conclusion: E. coli O157:H7 was found in 28.6% of pork, pig feces, and clean water in Jagalan slaughterhouse and 75% of identified E. coli O157:H7 contained the stx2a gene.

Shiga-like toxin (Stx) is an important factor in the pathogenesis of Escherichia coli O157:H7 infection and is responsible for some severe complications. Stx2 is usually associated with hemolytic uremic syndrome in humans. Its expression is regulated by elements located upstream of the stx2 gene, including stx2-promoter sequence, ribosome binding site, and the antiterminator q gene. The present study aimed to find the correlation between regulatory elements and the expression level of Stx2 in two local isolates of E. coli O157:H7. Two local E. coli O157:H7 strains SM-25(1) and KL-48(2), originating from human and cattle feces, respectively, and an E. coli reference strain, ATCC 43894, were investigated. The complete stx2 gene covering the sequences of promoter, ribosome binding site, and open reading frame and q gene of each strain was analyzed. The magnitude of Stx2 production was detected with a reverse passive latex agglutination method and Stx mediated cellular damage was determined with the Vero cell assay. A comparison of the complete stx2 gene contained stx2-promoter, ribosome binding site, and q genes of two local strains KL-48(2) and SM25(1), and the E. coli ATCC 43894 showed that the amino acid sequences were identical. Both local isolates were Stx negative in the reverse passive latex agglutination test and nontoxic in the Vero cell assay. The expression level of Shiga-like toxin of the two local isolates of E. coli O157:H7 did not only depend on the regulatory elements of the stx2 gene.

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