Juan P. Maestre

Bacterial community analysis of a gas-phase biotrickling filter for biogas mimics desulfurization through the rRNA approach.

The bacterial composition of a lab-scale biotrickling filter (BTF) treating high loads of H(2)S was investigated by the rRNA approach. Two 16S rRNA gene clone libraries were established 42 and 189 d after reactor startup, while fluorescent in-situ hybridization (FISH) with DNA probes was performed throughout 260d of reactor operation. Diversity, community structure and metamorphosis were studied from reactor startup to fully-established pseudo-steady state operation at near neutral pH and at an inlet H(2)S concentration of 2000 ppmv (load of 55.6g H(2)S m(-3)h(-1)). In addition, FISH was used for assessing the spatial distribution of sulfur-oxidizing bacteria (SOB) along the length of the reactor under pseudo-steady state operation. A major shift in the diversity of the community was observed with the operating time, from a well-diverse community at startup to pseudo-steady state operation with a majority of retrieved sequences affiliated to SOB of the sulfur cycle including Thiothrix spp., Thiobacillus spp., and Sulfurimonas denitrificans. Although aerobic species were predominant along the BTF, a vertical stratification was encountered, in which facultative anaerobes had a major relative abundance in the inlet part of the BTF, where the sulfide to oxygen ratio was higher. The observed changes were related to the trophic properties of the community, the DO concentration, the accumulation of elemental sulfur and the operation at neutral pH.

Long-term ammonia removal in a coconut fiber-packed biofilter: Analysis of N fractionation and reactor performance under steady-state and transient conditions

A comprehensive study of long-term ammonia removal in a biofilter packed with coconut fiber is presented under both steady-state and transient conditions. Low and high ammonia loads were applied to the reactor by varying the inlet ammonia concentration from 90 to 260 ppm(v) and gas contact times ranging from 20 to 36 s. Gas samples and leachate measurements were periodically analyzed and used for characterizing biofilter performance in terms of removal efficiency (RE) and elimination capacity (EC). Also, N fractions in the leachate were quantified to both identify the experimental rates of nitritation and nitratation and to determine the N leachate distribution. Results showed stratification in the biofilter activity and, thus, most of the NH(3) removal was performed in the lower part of the reactor. An average EC of 0.5 kg N-NH(3)m(-3)d(-1) was obtained for the whole reactor with a maximum local average EC of 1.7 kg N-NH(3)m(-3)d(-1). Leachate analyses showed that a ratio of 1:1 of ammonium and nitrate ions in the leachate was obtained throughout steady-state operation at low ammonia loads with similar values for nitritation and nitratation rates. Low nitratation rates during high ammonia load periods occurred because large amounts of ammonium and nitrite accumulated in the packed bed, thus causing inhibition episodes on nitrite-oxidizing bacteria due to free ammonia accumulation. Mass balances showed that 50% of the ammonia fed to the reactor was oxidized to either nitrite or nitrate and the rest was recovered as ammonium indicating that sorption processes play a fundamental role in the treatment of ammonia by biofiltration.

Characterization of the bacterial community in a biotrickling filter treating high loads of H2S by molecular biology tools

The diversity and spatial distribution of bacteria in a lab-scale biotrickling filter treating high loads of hydrogen sulfide (H(2)S) were investigated. Diversity and community structure were studied by terminal-restriction fragment length polymorphism (T-RFLP). A 16S rRNA gene clone library was established. Near Full-length 16S rRNA gene sequences were obtained, and clones were clustered into 24 operational taxonomic units (OTUs). Nearly 74% and 26% of the clones were affiliated with the phyla Proteobacteria and Bacteroidetes, respectively. Beta-, epsilon- and gamma-proteobacteria accounted for 15, 9 and 48%, respectively. Around 45% of the sequences retrieved were affiliated to bacteria of the sulfur cycle including Thiothrix spp., Thiobacillus spp. and Sulfurimonas denitrificans. Sequences related to Thiothrix lacustris accounted for a 38%. Rarefaction curve demonstrated that clone library constructed can be sufficient to describe the vast majority of the bacterial diversity of this reactor operating under strict conditions (2,000 ppm(v) of H(2)S). A spatial distribution of bacteria was found along the length of the reactor by means of the T-RFLP technique. Although aerobic species were predominant along the reactor, facultative anaerobes had a major relative abundance in the inlet part of the reactor, where the sulfide to oxygen ratio is higher.

Exploring the microbiome of the built environment: A primer on four biological methods available to building professionals

Building professionals are increasingly being called upon to conduct indoor microbial investigations as they remediate moisture-damaged buildings and design new, healthy, sustainable buildings. Characterizing the indoor microbial community present in the built environment is challenging and complicated by the vast array of biological methods available to building professionals. Furthermore, the particular biological technique employed to study an indoor environment can have a significant impact on the results obtained. This study evaluates the advantages and disadvantages of four biological methods suitable for indoor microbial investigations: culturing, quantitative polymerase chain reaction (qPCR), Sanger sequencing, and pyrosequencing. The results obtained from a study of four buildings are used to evaluate the merits of each bioanalytical approach. In each of the four study sites, the microbial-laden dust recovered on HVAC filters was used to provide a passive, long-term sample of the indoor air. Culturing of the microorganisms recovered from the dust was the least expensive method tested but provided a limited characterization of the microbial community present. qPCR provided the most specific information about the presence and quantity of target microorganisms but this method requires a priori knowledge of the species of interest and specifically designed primers that may not enumerate unanticipated species. Sanger sequencing provided microbial identification at the species level but lacked coverage to fully describe the microbial community present. Pyrosequencing provided in-depth sequence coverage of the microbial community present (to the genus level) but the vast dataset generated required increased computational analysis and data storage. Nevertheless, pyrosequencing when coupled with qPCR for target species quantification represents a viable approach that should become more accessible to building professionals as user-friendly software for analyzing sequencing results becomes available and more commercial laboratories offer these services.

Phthalates and organophosphates in settled dust and HVAC filter dust of U.S. low-income homes: Association with season, building characteristics, and childhood asthma

Phthalates and organophosphates are ubiquitous indoor semi-volatile organic contaminants (SVOCs) that have been widely used as plasticizers and flame retardants in consumer products. Although many studies have assessed their levels in house dust, only a few used dust samples captured by filters of building heating, ventilation, and air conditioning (HVAC) systems. HVAC filters collect particles from large volumes of air over a long period of time (potentially known) and thus provide a spatially and temporally integrated concentration. This study measured concentrations of phthalates and organophosphates in HVAC filter dust and settled floor dust collected from low-income homes in Texas, United States, in both the summer and winter seasons. The most frequently detected compounds were benzyl butyl phthalate (BBzP), di-(2-ethylhexyl) phthalate (DEHP), di-n-octyl phthalate (DnOP), tris (1-chloro-2-propyl) phosphate (TCIPP), triphenyl phosphate (TPHP), and tris (1,3-dichloroisopropyl) phosphate (TDCIPP). The median level of TCIPP in settled dust was 3- to 180-times higher than levels reported in other studies of residential homes. Significantly higher concentrations were observed in HVAC filter dust as compared to settled dust for most of the frequently detected compounds in both seasons, except for several phthalates in the winter. SVOC concentrations in settled dust in winter were generally higher than in summer, while different seasonality patterns were found for HVAC filter dust. Settled dust samples from homes with vinyl flooring contained significantly higher levels of BBzP and DEHP as compared to homes with other types of floor material. The concentration of DEHP and TDCIPP in settled dust also significantly associated with the presence of carpet in homes. Cleaning activities to remove dust from furniture actually increased the levels of certain compounds in HVAC filter dust, while frequent vacuuming of carpet helped to decrease the concentrations of some compounds in settled dust. Additionally, the size and age of a given house also correlated with the levels of some pollutants in dust. A statistically significant association between DEHP concentration in HVAC filter dust in summer and the severity of asthma in children was observed. These results suggest that HVAC filter dust represents a useful sampling medium to monitor indoor SVOC concentrations with high sensitivity; in contrast, when using settled dust, in addition to consideration of seasonal influences, it is critical to know the sampling location because the type and level of SVOCs may be related to local materials used there.