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Elucidation of Microbial Diversity in Wastewater Treatment System Through Molecular Tools: Molecular Tools, Techniques, and Applications in Wastewater Treatment
Published in Maulin P. Shah, Wastewater Treatment, 2022
Nikhil Nair, Sanchita Patwardhan, Nilesh S. Wagh, Jaya Lakkakula, Nomvano Mketo
In another study 16S rDNA-based DGGE was applied for ammonia-oxidizing beta (subclass Proteobacteria [β-AOM]). Cross-feeding experiments were carried out for a period of 4 months to obtain the information on the inhibition of the processed water, and a two-stage laboratory plant was set up to analyze the processed water. To determine the nitrification capacity of activated sludge samples, autotrophic oxygen uptake rates (OURN max) were calculated. DNA was extracted from Nitrosomonas europaea ATCC 19718 and Escherichia coli type strain DSM 30083 to be used as positive and negative controls during PCR and DGGE. PCR was performed using triplicate reactions of 10–1, 10–2, and 10–3, from respective DNA templates. DGGE analysis was performed with the recovered amplicons containing the attached GC-clamp (the primer PRBA 338f bears a 5′ attached GC-rich sequence) for rapid screening of the DNA sequence. Results from DGGE were definite and useful for the understanding of other experiments and assisted in finding solutions to nitrification problems in wastewater treatment [8].
Interconnection between PHA and Stress Robustness of Bacteria
Published in Martin Koller, The Handbook of Polyhydroxyalkanoates, 2020
Stanislav Obruca, Petr Sedlacek, Iva Pernicova, Adriana Kovalcik, Ivana Novackova, Eva Slaninova, Ivana Marova
Nonetheless, the presence of PHA also protects cells against severe consequences of osmotic fluctuations as was observed, for instance, in the experiments performed by Zhao et al. who compared the survival rate of Aeromonas hydrophila and its PHA synthase knock-out mutant. When exposed to various stress factors including osmotic challenge, the wild-type strain capable of PHA synthesis demonstrated higher resistance to osmotic pressure [26]. Comparable results were obtained by Kadouri et al. who compared the wild-type strain of Azospirillum brasilense and its PHA synthase deletion mutants [61] and intracellular PHA depolymerase deletion mutants [62]. Similarly, transgenic Escherichia coli capable of PHA synthesis was more robust against osmotic challenge than the wild-type strain [63].
Japanese Approach to Validation
Published in James Agalloco, Phil DeSantis, Anthony Grilli, Anthony Pavell, Handbook of Validation in Pharmaceutical Processes, 2021
Satoshi Sugimoto, Mitsuo Mori, Kiyoshi Mochizuki, Keisuke Nishikawa, Takuji Ikeda, Yusuke Matsuda, Hiroaki Nakamura, Yasuhito Ikematsu
To validate a protocol/procedure, it is required to demonstrate that the detection target is a suitable index/indicator for bacterial number or quantity. It is also important to state whether any special precautions are necessary in applying the protocol/procedure. When using a type strain, the result of validation should be equivalent to or better than that of the conventional method. However, because the detection principles of new methods are usually different from that of conventional methods, the correlation between them is not always required. For detection of environmental bacteria, it is important that the physiological state of the type strain should be maintained as close as possible to that of environmental bacteria, in order to obtain reliable results.”
Isolation and identification of an osmotolerant Bacillus amyloliquefaciens strain T4 for 2, 3-butanediol production with tobacco waste
Published in Preparative Biochemistry & Biotechnology, 2022
Ju Han, Fan Wang, Zhihao Li, Lijuan Liu, Ge Zhang, Guoqiang Chen, Jing Liu, Haibo Zhang
A single colony of strain T4 was inoculated in 5 mL LB broth and cultured at 37 °C for 12 h and subcultured into 50 mL of the three different media (LB culture, high glucose concentration medium and TWE medium) at the same concentration, respectively. The experiment was carried out in 250 mL conical flask at 37 °C for 24 h. During this process, the optical density (OD) that determined the concentration of biomass was tested at 600 nm in an ultraviolet spectrophotometer (Cary 50 UV-Vis, Varian) for every 2–3 hours. Escherichia coli BL21 (E. coli BL21) and a type strain Bacillus subtilis 168 (B. subtilis 168) were used as control.