The population of buffaloes in Indonesia was 1.37 million in 2012, representing an increase of 5.5 % over its population the previous year. Buffaloes have been in Indonesia for such a long time, they have become a part of the lives of the majority of the Indonesian society. Research has been conducted to know the bacteria diversity in domestic buffalo meat and bowels from traditional markets in Pandeglang, Banten, in order to ascertain their safety based on their initial contamination and also to study the sensitivity of several of the bacteria to irradiation and antibiotics. The total bacterial was assessed by total plate count method as index of quality. The buffalo meat and bowel samples were taken from livers, intestines, lymph, lungs and tripe. Results showed that the contaminating bacteria were aerobic bacteria, coliform bacteria including Escherichia coli (E. coli), and Staphylococcus spp. in buffalo meat and bowel. The numbers of aerobic bacteria were in the 1.7×10 5 -2.3×10 6 CFU/g range, while the total coliform bacteria were in the 2.0×10 3 -6.8×10 4 CFU/g range. The total number of E. coli was in the 2.0×10 3 -6.0×10 4 CFU/g range, and Staphylococcus spp. was in the 2.0×10 4 -2.7×105 CFU/g range. No Salmonella was detected in any of the samples observed. The total coliform bacteria, E. coli, and Staphylococcus spp. in all buffalo meat and bowel samples exceeded the maximum numbers of microbes permitted by the Indonesian National Standard (SNI). The maximum of total coliform, E. coli, and Staphylococcus spp. permitted by SNI are 1.0×10 2, 1.0×10 and 1.0×10 2 CFU/g, respectively. The D 10 values of S. aureus were in the 0.13 -0.23 kGy range, while for E. coli they were in the 0.07 -0.13 kGy range. The isolate of S. aureus from the lungs was the most resistant to cefoxitin, tetracycline, and amoxicillin antibiotics. The isolate of E. coli from buffalo bowels were almost sensitive to cefoxitin, tetracycline, and amoxicillin antibiotics.
Biofilm is an aggregate of consortium bacteria that adhere to each other on a surface. It is usually protected by the exopolysaccharide layer. Various invasive medical procedures, such as catheterization, endotracheal tube installation, and contact lens utilization, are vulnerable to biofilm infection. The National Institute of Health (NIH) estimates 65% of all microbial infections are caused by biofilm. Periplasmic α-amylase (MalS) is an enzyme that hydrolyzes α-1, 4-glicosidic bond in glycogen, starch, and others related polysaccharides in periplasmic space. Another protein called hemolysin-α (HlyA) is a secretion signal protein on C terminal of particular peptide in gram negative bacteria. We proposed a novel recombinant plasmid expressing α-amylase and hemolysin-α fusion in pSB1C3 which is cloned into E.coli to enable α-amylase excretion to extracellular for degrading biofilm polysaccharides content, as in starch agar.Microtiter assay was performed to analyze the reduction percentage of biofilm by adding recombinant E.coli into media. This system is more effective in degrading biofilm from gram positive bacteria i.e.: Bacillus substilis (30.21%) and Staphylococcus aureus (24.20%), and less effective degrading biofilm of gram negative i.e.: Vibrio cholera (5.30%), Pseudomonas aeruginosa (8.50%), Klebsiella pneumonia (6.75%) and E. coli (-0.6%). Gram positive bacteria have a thick layer of peptidoglycan, causing the enzyme to work more effectively in degrading polysaccharides.
AbstrakEkspresi dan Penelitian Fungsi Protein Fusi α-Amilase dan Hemolisin-α sebagai suatu Penerapan dalam Penurunan Polisakarida Biofilm. Biofilm adalah sekumpulan bakteria yang saling melekat satu sama lain pada suatu permukaan. Biofilm ini biasanya dilindungi oleh lapisan eksopolisakarida. Berbagai prosedur medis yang pro-aktif, seperti kateterisasi, instalasi alat bantuan pernafasan, dan penggunaan lensa kontak mata, rentan terhadap infeksi biofilm. NIH (National Institute of Health -Institusi Kesehatan Nasional) memperkirakan 65% dari semua infeksi mikroba disebabkan oleh biofilm. Enzim α-amilase periplasma (MalS) merupakan suatu enzim yang menghidrolisis α-1, ikatan 4-glikosidik dalam glikogen, zat tepung, dan lainnya terkait polisakarida pada ruang periplasma. Protein lainnya yang disebut hemolisin-α (HlyA) merupakan protein sinyal sekresi pada terminal C dari peptida tertentu dalam bakteria gram-negatif. C merupakan protein sinyal sekresi pada terminal C dari peptida tertentu dalam bakteria gram-negatif. Kami mengusulkan suatu plasmid rekombinan baru mengekspresikan fusi α-amilase and hemolisin-α dalam pSB1C3 yang diklon menjadi E. coli untuk memungkinkan ekskresi α-amilase ke luar sel tubuh (ekstraselular) untuk menurunkan isi polisakarida biofilm, seperti dalam agar zat tepung. Tes dengan tabung kecil dilakukan untuk menganalisis persentase pengurangan biofilm dengan menambahkan E. coli rekombinan ke dalam media. Sistem ini lebih efektif dalam menurunkan tingkat biofilm dari bakteria gram-positif, seperti Bacillus subst...
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