The Effect of Mercury Exposure to Escherichia Coli Bacteria Resistant to Mercury and Escerichia Coli Esbl in Vitro

Background: The level of pollution in Indonesia is still very high, consist of water pollution, air pollution and soil pollution. Mercury is one of the heavy metals that pollutes the waters of the sea, while Escherichia coli is exposed to mercury will try to defend itself by doing mercury detoxification so that it can live in an environment that contains mercury. Escherichia coli that tries to defend itself from mercury exposure in the environment will experience a change in its genes into mercury resistant Escherichia coli. In plasmids or transposons, it might also stimulate the formation of resistance genes for some antibiotics, include producing the ESBL enzyme, so that it can convert non ESBL Escherichia coli into ESBL Escherichia coli. Objective: This study aims to prove that the repeated exposure of mercury will change non ESBL-mercury sensitive Escherichia coli into ESBL- mercury resistant Escherichia coli. Method: This was an experimental study with 27 non-ESBL Escherichia coli isolates as identified from Phoenix. Non-ESBL Escherichia coli clinical isolates were tested by giving exposure to HgCl2 with concentrations of 0.02 ppm, 0.10 ppm, 0.20 ppm for 1-14 days until mercury resistant Escherichia coli was formed, and then ESBL screening was tested by giving Cefotaxime exposure to them. Results: On the first day of mercury exposure, there were 9 isolates of 0.02 ppm HgCl2 resistant Escherichia coli, 9 isolates of 0.10 ppm HgCl2 resistant Escherichia coli, 9 isolates of 0.20 ppm HgCl2 resistant Escherichia coli. Furthermore, this Escherichia coli isolate was exposed to Cefotaxim as ESBL screening. The final results of post-exposure HgCl2 0.02 ppm was obtained 3 (33.3%) isolates were still sensitive to Cefotaxime and 6 (66.7%) isolates that were resistant to Cefotaxime. The final results of post-exposure HgCl2 0.10 ppm was obtained all 9 (100%) isolates that were resistant to Cefotaxime. The final results of post-exposure HgCl2 0.20 ppm obtained 2 (22.2%) isolates were still sensitive to Cefotaxime and 7 (77.8%) isolate were resistant to Cefotaxime. Conclusion: Escherichia coli in urine had the phenotive change into mercury resistant Escherichia coli. Mercury exposure of 0.02 ppm, 0.10 ppm, 0.20 ppm for 1 day in vitro on isolates of non ESBL-mercury resistant Escherichia coli caused changes in 22 isolates of Escherichia coli in urine

Pengaruh pH dan Suhu terhadap Aktivitas Pereduksi Merkuri Bakteri Resisten Merkuri Tinggi Bacillus cereus yang Diisolasi dari Urin Pasien dengan Amalgam Gigi

Mercury is a very toxic compound to humans, therefore, a method to overcome its presence in the environment is required. Detoxification of mercury can be done by using mercury resistant bacteria. Mercury-resistant bacteria Bacillus cereus isolate FUA have been obtained from the urine of patients with dental mercury amalgam. This study was aimed to determine the mercury detoxification activity of Bacillus cereus isolate FUA at varying pH medium and incubation temperature. The study was carried out by growing Bacillus cereus isolate FUA on oblique media, then were planted in the growth media of LB broth containing mercury compounds of 10 ppm HgCl2 with varying pHs of 5, 7, and 9 and incubation temperatures of 15, 25 and 35oC. The amount of bacterial growth was analyzed by using spectrophotometer and mercury levels were analyzed by using CV-AAS method. The results showed that the growth and mercury reducing activity of Bacillus cereus isolate FUA were optimum at pH 7 and incubation temperature of 35oC. In conclusion, the growth of Bacillus cereus isolate FUA and its mercury reducing activity were optimum at pH 7 and temperature of 35oC. It is expected that the results of this study can be the basis for further research on the process of mercury detoxificationKeywords: Bacillus cereus, urine, mercury resistance, pH, temperature Abstrak: Merkuri adalah senyawa yang sangat beracun bagi manusia sehingga diperlukan metode untuk mengatasi keberadaannya di lingkungan. Detoksifikasi merkuri dapat dilakukan dengan menggunakan bakteri resisten merkuri. Bakteri yang resisten merkuri Bacillus cereus isolat FUA telah diperoleh dari urin pasien dengan amalgam gigi. Penelitian ini bertujuan untuk mengetahui aktivitas detoksifikasi merkuri Bacillus cereus isolat FUA pada berbagai variasi pH medium dan suhu inkubasi. Bakteri Bacillus cereus isolat FUA ditumbuhkan pada media miring, kemudian ditanam pada media pertumbuhan bakteri LB broth yang mengandung senyawa merkuri 10 ppm HgCl2 dengan berbagai pH 5, 7 dan 9 dan suhu inkubasi 15, 25 dan 35oC. Jumlah pertumbuhan bakteri dianalisis menggunakan spektrofotometer dan kadar merkuri dianalisis menggunakan metode CV-AAS. Hasil penelitian menunjukkan bahwa aktivitas pertumbuhan dan aktivitas pereduksi merkuri Bacillus cereus isolat FUA optimum pada lingkungan pertumbuhan dengan pH 7 dan suhu inkubasi 35oC. Simpulan penelitian ini ialah aktivitas pertumbuhan dan pereduksi merkuri Bacillus cereus isolat FUA yang optimum pada pH 7 dan suhu 35oC. Diharapkan hasil penelitian ini dapat menjadi dasar untuk penelitian lebih lanjut tentang proses detoksifikasi merkuri.Kata kunci: Bacillus cereus, urin, resistensi merkuri, pH, suhu

Mercury contamination imposes structural shift on the microbial community of an agricultural soil

Background The purpose of this study is to use shotgun next-generation sequencing to unravel the microbial community structure of an agricultural soil, decipher the effects of mercury contamination on the structure of the microbial community and the soil physicochemistry and heavy metals content. Results The soil physicochemistry after mercury contamination revealed a shift in soil pH from neutral (6.99 ± 0.001) to acidic (5.96 ± 0.25), a decline in moisture content to < 4 %, and a significant decrease in the concentrations of all the macronutrients and the total organic matter. Significant decrease in all the heavy metals detected in the agricultural soil was also observed in mercury inundated SL3 microcosm. Structural analysis of the metagenomes of SL1 (agricultural soil) and SL3 (mercury-contaminated agricultural soil) using Illumina shotgun next-generation sequencing revealed the loss due to mercury contamination of 54.75 % of the microbial community consisting of an archaeal domain, 11 phyla, 12 classes, 24 orders, 36 families, 59 genera, and 86 species. The dominant phylum, class, genus, and species in SL1 metagenome are Proteobacteria, Bacilli, Staphylococcus, and Sphingobacterium sp. 21; while in SL3 metagenome, Proteobacteria, Alphaproteobacteria, Singulisphaera, and Singulisphaera acidiphila were preponderant. Mercury contamination resulted in a massive upscale in the population of members of the phylum Planctomycetes and the genera Singulisphaera, Brevundimonas, Sanguibacter, Exiguobacterium, Desulfobacca, and Proteus in SL3 metagenome while it causes massive decline in the population of genera Staphylococcus and Brachybacterium. Conclusions This study revealed that mercury contamination of the agricultural soil imposed selective pressure on the members of the microbial community, which negatively impact on their population, alter soil physicochemistry, and enriched sizable numbers of members of the community that are well adapted to mercury stress. It also reveals members of microbial community hitherto not reported to be important in mercury detoxification process.

Genomic analysis for heavy metal resistance in S. maltophilia

Stenotrophomonas maltophilia is highly resistant to heavy metals, but the genetic knowledge of metal resistance in S. maltophilia is poorly understood. In this study, the genome of S.maltophilia Pho isolated from the contaminated soil near a metalwork factory was sequenced using PacBio RS II. Its genome is composed of a single chromosome with a GC content of 66.4% and 4434 protein-encoding genes. Comparative analysis revealed high syntney between S.maltophilia Pho and the model strain, S.maltophilia K279a. Then, the type and number of mechanisms for heavy metal uptake were analyzed firstly. Results revealed 7 unspecific ion transporter genes and 13 specific ion transporter genes, most of which were involved in iron transport. But the sulfate permeases belonging to the family of SulT/CysP that can uptake chromate and the high affinity ZnuABC/SitABCD were absent. Secondly, the putative genes controlling metal efflux were identified. Results showed that this bacterium encoded 5 CDFs, 1 copper exporting ATPase and 4 RND systems, including 2 CzcABC efflux pumps. Moreover, the putative metal transformation genes including arsenate and mercury detoxification genes were also identified. This study may provide useful information on the metal resistance mechanisms of S.maltophilia.