BACKGROUND: We developed a new disposable anaerobic culture system, namely, the Quick anaero-system, for easy culturing of obligate anaerobes. METHODS: Our system consists of 3 components: 1) new disposable anaerobic gas pack, 2) disposable culture-envelope and sealer, and 3) reusable stainless plate rack with mesh containing 10 g of palladium catalyst pellets. To evaluate the efficiency of our system, we used 12 anaerobic bacteria. We prepared 2 sets of ten-fold serial dilutions of the 12 anaerobes, and inoculated these samples on Luria-Bertani (LB) broth and LB blood agar plate (LB-BAP) (BD Diagnostic Systems, USA). Each set was incubated in the Quick anaero-system (DAS Tech, Korea) and BBL GasPak jar with BD GasPak EZ Anaerobe Container System (BD Diagnostic Systems) at 35-37degrees C for 48 hr. The minimal inoculum size showing visible growth of 12 anaerobes when incubated in both the systems was compared. RESULTS: The minimal inoculum size showing visible growth for 2 out of the 12 anaerobes in the LB broth and 9 out of the 12 anaerobes on LB-BAP was lower for the Quick anaero-system than in the BD GasPak EZ Anaerobe Container System. The mean time (+/-SD) required to achieve absolute anaerobic conditions of the Quick anaero-system was 17 min and 56 sec (+/-3 min and 25 sec). CONCLUSIONS: The Quick anaero-system is a simple and effective method of culturing obligate anaerobes, and its performance is superior to that of the BD GasPak EZ Anaerobe Container System.
Bacteria, Anaerobic/*growth &development ; Bacteriological Techniques/instrumentation/methods ; Culture Media/chemistry ; Gases/chemistry ; Palladium/chemistry ; Reagent Kits, Diagnostic

Anaerobic ammonium oxidation (Anammox) process is a high-rate nitrogen removal technology that has been applied in sludge dewatering effluents treatment with nitrogen removal rate as high as 9.5 kg/(m x d). However, due to the slow growth rate of the autotrophic Anammox bacteria and the susceptivity to environmental conditions, the start-up of Anammox process is very long; the operation is unstable; and the nitrogen removal from organic-containing and/or toxicant-containing ammonium-rich wastewaters using Anammox process becomes difficult. Thus, the application of this high-rate process is significantly limited. In this paper, a newly-developed Anammox process with sequential biocatalyst (Anammox biomass) addition was established based on the procedure in fermentation engineering. We introduced the Anammox process with sequential biocatalyst addition on start-up, stable operation and the treatment of organic-containing and toxicant-containing ammonium-rich wastewaters. Results show that supplementing high-activity Anammox biomass into reactors will increase the amount of as well as the ratio of Anammox bacteria. Thus, the innovative Anammox process with sequential biocatalyst addition not only accelerates the start-up course, but also enhances the stability of Anammox process. Furthermore, it overcomes the drawbacks of wastewaters containing high organic content and toxic substances. Therefore, the application of Anammox process may be further enlarged.
Bacteria, Anaerobic ; enzymology ; growth & development ; metabolism ; Biocatalysis ; Biomass ; Bioreactors ; microbiology ; Enzymes ; chemistry ; Nitrogen ; isolation & purification ; metabolism ; Oxidation-Reduction ; Quaternary Ammonium Compounds ; isolation & purification ; metabolism ; Waste Disposal, Fluid ; methods

The recalcitrance of lignocellulosic biomass makes its hydrolysis by cellulases less effective, and the consolidated bioprocessing (CBP) strategy that combines enzyme production, cellulose hydrolysis and fermentation, particularly the synergetic role of different microbes in attacking cellulose component could be a solution. In this article, a facultative anaerobe microbial consortium named H was isolated, which exhibited high stability even after 30 subcultures, with pH ranging from 6 to 9. Within three days, 0.5 g filter paper immerged in 100 mL PCS buffer was completely degraded, and 1.54 g/L ethanol was produced, correspondingly. Further analysis on the component of the microbe consortium was carried out though 16S rDNA and DGGE, and Clostridium thermosuccinogene, Clostridium straminisolvens and Clostridium isatidis that can directly convert cellulose to ethanol were identified, indicating that Clostridium spp. played important role in cellulose degradation through the synergistic coordination of different species, and the characterization of the consortium will benefit the analysis of the underlying mechanisms as well as the optimization of the CBP process for more efficient cellulose degradation and ethanol production.
Bacteria, Anaerobic ; metabolism ; Cellulase ; metabolism ; Cellulose ; metabolism ; Clostridium ; classification ; growth & development ; metabolism ; Clostridium thermocellum ; growth & development ; metabolism ; Culture Techniques ; methods ; Ethanol ; metabolism ; Fermentation ; Hydrolysis ; Industrial Microbiology ; methods ; Microbial Consortia ; physiology ; Microbial Interactions