Nitrite and nitrate as electron acceptors for biological sulphide oxidation.

Autotrophic denitrification with sulphide using nitrate (R1) and nitrite (R2) as electron acceptor was investigated at bench scale. Different solids retention times (SRT) (5 and 20 d) have been tested in R1 while R2 was operated at SRT=13 d. The results indicated that the process allows complete sulphide removal to be achieved in all tested conditions. Tested sulphide loads were estimated from the H2S produced in a pilot-scale anaerobic digester treating vegetable tannery primary sludge; nitrogen loads originated from the nitrification of the supernatant. Average nitrogen removal efficiencies higher than 80% were observed in all the tested conditions once steady state was reached. A maximum specific nitrate removal rate equal to 0.35 g N-NO3- g VSS(-1) d(-1) was reached in R1. Due to sulphide limitation, incomplete denitrification was observed and nitrite and thiosulphate tend to accumulate especially in the presence of variable environmental conditions in both R1 and R2. Lower SRT caused higher NO2accumulated/NO3reduced ratios (0.22 and 0.24, with SRT of 5 d and 20 d, respectively) using nitrate as electron acceptor in steady-state condition. Temperature decrease caused sudden NO2accumulated/NO3reduced ratio increase in R1 and NO2- removal decrease in R2.

Continuous hydrogen and methane production in a two-stage cheese whey fermentation system.

The feasibility of integrating biological hydrogen and methane production in a two-stage process using mixed cultures and cheese whey powder (CWP) as substrate was studied. The effect of operational parameters such as hydraulic retention time (HRT) and organic loading rate (OLR) on the volumetric hydrogen (VHPR) and methane (VMPR) production rates was assessed. The highest VHPR was 28 L H2/L/d, obtained during stable operation in a CSTR at HRT and OLR of 6 h and 142 g lactose/ L/d, respectively. Moreover, hydrogen (13 L/L/d) was produced even at HRT as low as 3.5 h and OLR of 163 g lactose/L/d, nonetheless, the reactor operation was not stable. Regarding methane production in an UASB reactor, the acidified effluent from the hydrogen-producing bioreactor was efficiently treated obtaining COD removals above 90% at OLR and HRT of 20 g COD/L/d and 6 h, respectively. The two-stage process for continuous production of hydrogen and methane recovered over 70% of the energy present in the substrate. This study demonstrated that hydrogen production can be efficiently coupled to methane production in a two-stage system and that CWP is an adequate substrate for energy production.

Effects of transient temperature increases on odor production from thermophilic anaerobic digestion.

The City of Los Angeles, Bureau of Sanitation, has implemented thermophilic anaerobic sludge digestion at the Hyperion and Terminal Island Treatment Plants (HTP and TITP). A two-stage continuous-batch process was established at HTP, while a single-stage sequencing batch process was established at TITP. This was to evaluate compliance with the Class A pathogen reduction requirements of U.S. EPA 40 CFR Part 503. A rapid increase of the digester temperature at TITP from 57.5 to 65.5 degrees C caused an increase of the volatile fatty acid to alkalinity ratio, a decline in digester performance, and an elevated production of methyl mercaptan and hydrogen sulfide. A rapid increase of the digester temperature at HTP from 54 to 58 degrees C caused an elevated production of methyl mercaptan, but the effect on the volatile fatty acid to alkalinity ratio and digester performance was insignificant. It is likely that these effects observed at TITP and HTP were transient responses to rapid changes in temperature.

Toxicity of di-(2-ethylhexyl) phthalate on the anaerobic digestion of wastewater sludge.

Previous studies on the microbial degradation of individual phthalic acid esters (PAEs) have demonstrated that the compounds with short ester hydrocarbon chains are easily biodegraded and mineralized, but PAEs with long ester chains are less susceptible to degradation and some of them are considered recalcitrant. Moreover, they inhibit methanogenesis. However, studies have not been made on the effect of feeding a combination of recalcitrant and biodegradable PAEs into anaerobic digesters treating wastewater sludge. The present study was conducted with wastewater sludge from the Los Angeles Bureau of Sanitation's Hyperion Treatment Plant. Di (2-ethylhexyl) phthalate (DEHP), the most common persistent PAE found in wastewater, and di-n-butyl phthalate (DBP), a common PAE with short ester chains, were sorbed into the sludge fed to a bench-scale digester for a period of 12 weeks. DEHP degradation was always poor, and accumulation of DEHP was correlated with inhibition of the microbial degradation of DBP and with process instability of the test digester. Inhibition of the DBP removal was completely reversed after DEHP addition was discontinued, but biogas production never recovered to the level observed in a control digester. Other process parameters of digester performance were not affected by DEHP accumulation. These results are similar to the toxic effects of long chain fatty acids on sludge digestion, suggesting that DEHP or its degradation products affect all the microbial populations in the anaerobic bioreactor. Our results imply that high levels of DEHP or other recalcitrant PAEs in wastewater sludge are likely to compromise methanogenesis and removal of biodegradable PAEs in sludge digesters.

Identification of anaerobic nonsporeforming gram-positive bacilli by biochemical tests and gas-liquid chromatography.

There are many methods to identify anaerobic nonsporeforming bacilli: histological, bacteriological (biochemical test, microsystem API 20 A), serological, cell wall composition analysis, molecular methods and gas-liquid chromatography (GLC). A comparison between biochemical tests and gas-liquid chromatography was made in this study for the identification of this group of microorganisms. GLC conditions were established with the aid of reference strains. These conditions were then applied to ten strains which were previously identified by biochemical tests. Strains were grown in PYG broth and fermentation end products were analyzed, volatile and non volatile fatty acids. Their qualitative determination was made by comparing the retention time of known standards and the chromatographic pattern of reference strains. In addition, a semiquantitative analysis was made. The results of identification by biochemical tests were: five strains belonged to Actinomyces genus; three were Propionibacterium acnes; one Propionibacterium granulosum and one P. propionicum. By the GLC only seven strains were identified: four were Actinomyces and three P. acnes. Only six strains showed identification correlation by both biochemical tests and GLC. GLC is a presumptive identification method that can be used along with other complementary tests for a definitive identification at genus level.

Bacterial artificial chromosome (BAC) library as a tool for physical mapping of the archaeon Methanosarcina thermophila TM-1.

We have used a variety of methods to characterize the genome of the archaeon Methanosarcina thermophila TM-1. Pulsed-field gel analysis indicates a genome size of 2.8 Mb. We have constructed a bacterial artificial chromosome (BAC) library of M. thermophila and have used it to generate physical maps for this organism. The library is made up of 384 clones with an average insert size of 58 kb representing 8.0 genome equivalents. The utility of the library for low-resolution physical mapping was shown by identifying NotI linking clones and using these to order the NotI macrorestriction fragments of M. thermophila into a 2.8 Mb map. Hybridization of nine single copy genes and a 16S rRNA sequence to these macrorestriction fragments forms the basis for the first genetic map in this organism. High-resolution physical maps, consisting of overlapping clones, have been created using HindIII fingerprints of BAC clones. In this way, we identified a minimal path of five clones that span a 270 kb NotI fragment. The ease of manipulating BAC clones makes the BAC system an excellent choice for the construction of low-resolution and high-resolution physical and genetic maps of archaeal genomes. It also provides a substrate for future genome-sequencing efforts.