Genomic potential of anaerobic methane- and ammonium
Anaerobic wastewater treatment offers several advantages; However, waste from anaerobic digester still contains high levels of dissolved ammonium and methane that need to be removed before the waste can be discharged into surface water. Elimination of simultaneous anaerobic methane and ammonium by denitrification (N-Damo) methanotrophs in combination with the anaerobic ammonium-oxidizing (Anammox) bacteria could be a potential solution to this challenge. After surveying the molecules of waste brewery waste water treating plants, which indicates the presence of both N-Damo and bacteria Anammox, we started an anaerobic bioreactor with a continuous supply of methane, ammonia and nitrite to enrich this anaerobic microorganisms.
After 14 months of operation, stable enrichment cultures containing two types of ‘Candidatus Methylomirabilis oxyfera’ bacteria and two strains of ‘Ca. Brocadia’-like bacteria Anammox achieved. In this community, the bacteria Anammox converted 80% of the ammonium nitrite, while ‘Ca. Methylomirabilis’ contribute 20% of the consumption of nitrite. Analysis of 16S rRNA metagenomic reads and fluorescence in situ hybridization (FISH) correlate well and show that, after 14 months, ‘Ca. Methylomirabilis’ and Anammox bacteria is about 30 and 20% of the total microbial community. In addition, the majority (10%) of society consists of Planctomycetes Phycisphaera related.
Assembly and binning of metagenomic sequence resulting in a high-quality draft genome of two ‘Ca. Methylomirabilis species’ contains a marker gene pmoCAB, xoxF, and NIR and allegedly NO dismutase gene. The Anammox draft genome is closely related to ‘Ca. Brocadia Fulgida ‘including hzsABC marker gene, hao, and HDH. Whole-batch anaerobic reactor and measuring the activity of methane, ammonia, nitrite, and nitrate revealed the average anaerobic methane oxidation rate of 0.12 mmol h-1 L-1 and ammonium oxidation rate of 0.5 mmol h-1 L-1.
Together, these studies illustrate the draft genome enrichment and anaerobic methanotrophs of brewery wastewater treatment, where this organism together with Anammox bacteria can contribute significantly to the elimination of methane and ammonium in a more sustainable way. KEY POINTS: • An enrichment culture containing N-Damo and bacteria Anammox obtained. • Simultaneous consumption of ammonia, nitrite, and methane under anoxic conditions. • In-depth analysis of metagenomic biodiversity inoculum and cultural enrichment.
Diversity, enrichment, and genomic potential of anaerobic methane- and ammonium-oxidizing microorganisms from a brewery wastewater treatment plant
Description: Small Intestine tissue lysate was prepared by homogenization in modified RIPA buffer (150 mM sodium chloride, 50 mM Tris-HCl, pH 7.4, 1 mM ethylenediaminetetraacetic acid, 1 mM phenylmethylsulfonyl fluoride, 1% Triton X-100, 1% sodium deoxycholic acid, 0.1% sodium dodecylsulfate, 5 μg/ml of aprotinin, 5 μg/ml of leupeptin. Tissue and cell debris was removed by centrifugation. Protein concentration was determined with Bio-Rad protein assay. The product was boiled for 5 min in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 50 mM DTT.
Description: Small intestine tissue lysate was prepared by homogenization in modified RIPA buffer (150 mM sodium chloride, 50 mM Tris-HCl, pH 7.4, 1 mM ethylenediaminetetraacetic acid, 1 mM phenylmethylsulfonyl fluoride, 1% Triton X-100, 1% sodium deoxycholic acid, 0.1% sodium dodecylsulfate, 5 μg/ml of aprotinin, 5 μg/ml of leupeptin. Tissue and cell debris was removed by centrifugation. Protein concentration was determined with Bio-Rad protein assay. The product was boiled for 5 min in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 50 mM DTT.
Description: Small intestine cancer, metastatic lymph nodes and normal tissue high density tissue microarray, 69 cases/208 core, with stage and grade data
Tissue cDNA, First Strand, Mouse Adult Normal, Small Intestine, BioGenomics
Description: Small Intestine tumor tissue lysate was prepared by homogenization in lysis buffer (10 mM HEPES pH7.9, 1.5 mM MgCl2, 10 mM KCl, 1 mM ethylenediaminetetraacetic acid, 10% glycerol, 1% NP-40, and a cocktail of protease inhibitors). Tissue and cell debris was removed by centrifugation. The product was boiled for 5 min in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 50 mM DTT.
copper nanowires as nanofertilizers to plant alfalfa: Understanding the interaction of nano-bio systems of microbial genomics, plant molecular response and spectroscopic studies
The latest applications of nano copper (Cu) in agrosystem compounds have demonstrated the potential to improve the performance of plant physiological and agronomic parameters. We grew alfalfa (Medicago sativa) in potting mix changed with bulk, nano, and Cu ionic compound at 80 and 280 mg Cu / kg; then, we evaluate the performance of the plant at the physiological and molecular level.
Plants treated with the precipitation / Cu nano served better agronomic response. P and S content is reduced in bulk and ion Cu-exposed plants, compared with controls (p ≤ 0.05). All forms of Cu increase the content of Fe and Zn in the Fe in the roots and leaves, compared to controls (p ≤ 0.05).
Leaf-superoxide dismutase expression was plus ~ 27-fold and Rubisco mRNA was not affected in bulk / nano Cu-treated plants, compared with controls (p ≤ 0.05). Bulk / nano Cu increased relative abundance of microorganisms involved in uptake. These results indicate that the nano Cu improve the physiology of alfalfa and could be considered as a potential nanofertilizers.
Description: Bovine kidney tissue lysate was prepared by homogenization using a proprietary technique. The tissue was frozen in liquid nitrogen immediately after excision and then stored at -70°C. The bovine kidney tissue total protein is provided in a buffer including HEPES (pH 7.9), MgCl2, KCl, EDTA, Sucrose, Glycerol, Sodium deoxycholate, NP-40, and a cocktail of protease inhibitors. For quality control purposes, the kidney tissue pattern on SDS-PAGE gel is shown to be consistent for each lot by visualization with coomassie blue staining. The kidney tissue is then Western analyzed by either GAPDH or β-actin antibody, and the expression level is consistent with each lot.
Description: Guinea Pig kidney tissue lysate was prepared by homogenization using a proprietary technique. The tissue was frozen in liquid nitrogen immediately after excision and then stored at -70°C. The guinea pig kidney tissue total protein is provided in a buffer including HEPES (pH 7.9), MgCl2, KCl, EDTA, Sucrose, Glycerol, Sodium deoxycholate, NP-40, and a cocktail of protease inhibitors. For quality control purposes, the kidney tissue pattern on SDS-PAGE gel is shown to be consistent for each lot by visualization with coomassie blue staining. The kidney tissue is then Western analyzed by either GAPDH or β-actin antibody, and the expression level is consistent with each lot.
Description: Monkey (Cynomolgus) kidney tissue lysate was prepared by homogenization using a proprietary technique. The tissue was frozen in liquid nitrogen immediately after excision and then stored at -70°C. The monkey (Cynomolgus) kidney tissue total protein is provided in a buffer including HEPES (pH 7.9), MgCl2, KCl, EDTA, Sucrose, Glycerol, Sodium deoxycholate, NP-40, and a cocktail of protease inhibitors. For quality control purposes, the kidney tissue pattern on SDS-PAGE gel is shown to be consistent for each lot by visualization with coomassie blue staining. The kidney tissue is then Western analyzed by either GAPDH or β-actin antibody, and the expression level is consistent with each lot.
