Handling leachate from glass cullet stockpiles.

Mixed glass cullet (crushed recycled glass containers) is stockpiled uncovered before use as roadway construction aggregate or daily cover in landfills. Rainwater that leaches through the stockpiles dissolves and suspends contaminants such as those from food residuals and paper labels. The objective of this study was to determine leachate quantity and quality from cullet stockpiles as a basis for development of Best Management Practices (BMPs). Four 35-tonne field stockpiles were set up for leachate analysis and to determine the effects of mechanical turning treatment on the leachate. Field-collected leachate and laboratory-generated washwater of cullet (water:cullet=3:1 by weight) were both analyzed for basic wastewater parameters, which showed pollutant levels comparable to or higher than those of untreated domestic wastewater or urban stormwater. While organic contamination decreased substantially (e.g., washwater BOD>95% reduction), TKN and total-phosphorus levels in leachate ranged between 11.6-154mgL(-1) and 1.6-12.0mgL(-1), respectively, and remained comparable to levels found in untreated domestic wastewater after four months. Turning enhanced the degradation of the organic constituents inside the stockpiles, which was confirmed by elevated temperatures. Based on this study, leachate from glass cullet stockpiles should not be released to surface water. For leachate from long-term cullet stockpiles, release to groundwater should be only done after treatment to reduce nitrogen levels.

Evaluation of autotrophic denitrification, heterotrophic nitrification, and PAOs in full scale simultaneous biological nutrient removal sysyems.

Anoxic and aerobic batch reactor assays using a two-by-two factorial design were developed to determine the potential activity of autotrophic denitrification and heterotrophic nitrification in biological nutrient removal (BNR) activated sludge systems, especially those accomplishing simultaneous BNR (SBNR). Results from the application of these assays to three full scale closed loop bioreactors previously documented to be accomplishing SBNR demonstrated that these activities were minimal in comparison with the conventionally recognized activities of heterotrophic denitrification and autotrophic nitrification. Activity within the mixed liquor consistent with current theories for phosphorus accumulating organisms (PAOs) was also observed. Along with other observations, this suggests the presence of PAOs in the facilities studied.

Biological removal of gaseous ammonia in biofilters: space travel and earth-based applications.

Gaseous NH3 removal was studied in laboratory-scale biofilters (14-L reactor volume) containing perlite inoculated with a nitrifying enrichment culture. These biofilters received 6 L/min of airflow with inlet NH3 concentrations of 20 or 50 ppm, and removed more than 99.99% of the NH3 for the period of operation (101, 102 days). Comparison between an active reactor and an autoclaved control indicated that NH3 removal resulted from nitrification directly, as well as from enhanced absorption resulting from acidity produced by nitrification. Spatial distribution studies (20 ppm only) after 8 days of operation showed that nearly 95% of the NH3 could be accounted for in the lower 25% of the biofilter matrix, proximate to the port of entry. Periodic analysis of the biofilter material (20 and 50 ppm) showed accumulation of the nitrification product NO3- early in the operation, but later both NO2- and NO3- accumulated. Additionally, the N-mass balance accountability dropped from near 100% early in the experiments to approximately 95 and 75% for the 20- and 50-ppm biofilters, respectively. A partial contributing factor to this drop in mass balance accountability was the production of NO and N2O, which were detected in the biofilter exhaust.

Composting on Mars or the Moon: II. Temperature feedback control with top-wise introduction of waste material and air.

Whereas Earth-based composting reactors that effectively control the process are batch operations with bottom-to-top airflow, in extraterrestrial application both the fresh waste and the air need to be introduced from above. Stabilized compost and used air would exit below. This materials flow pattern permits the addition of waste whenever generated, obviating the need for multiple reactors, and the incorporation of a commode in the lid. Top loading in turn dictates top-down aeration, so that the most actively decomposing material (greatest need for heat removal and O2 replenishment) is first encountered. This novel material and aeration pattern was tested in conjunction with temperature feedback process control. Reactor characteristics were: working, volume, 0.15 m3; charge, 2 kg dry biomass per day (comparable to a 3-4 person self-sufficient bioregenerative habitat); retention time, 7 days. Judging from temperature profile, O2 level, air usage, pressure head loss, moisture, and odor, the system was effectively controlled over a 35-day period. Dry matter disappearance averaged 25% (10-42%). The compost product was substantially, though not completely, stabilized. This demonstrates the compatibility of top-wise introduction of waste and air with temperature feedback process control.

Composting on Mars or the Moon: I. Comparative evaluation of process design alternatives.

As a candidate technology for treating solid wastes and recovering resources in bioregenerative Advanced Life Support, composting potentially offers such advantages as compactness, low mass, near ambient reactor temperatures and pressures, reliability, flexibility, simplicity, and forgiveness of operational error or neglect. Importantly, the interactions among the physical, chemical, and biological factors that govern composting system behavior are well understood. This article comparatively evaluates five Generic Systems that describe the basic alternatives to composting facility design and control. These are: 1) passive aeration; 2) passive aeration abetted by mechanical agitation; 3) forced aeration--O2 feedback control; 4) forced aeration--temperature feedback control; 5) forced aeration--integrated O2 and temperature feedback control. Each of the five has a distinctive pattern of behavior and process performance characteristics. Only Systems 4 and 5 are judged to be viable candidates for ALS on alien worlds, though which is better suited in this application is yet to be determined.

Development of field application vectors for bioremediation of soils contaminated with polychlorinated biphenyls.

Field application vectors (FAVs), which are a combination of a selective substrate, a host, and a cloning vector, have been developed for the purpose of expressing foreign genes in nonsterile, competitive environments in which the gene products provide no advantage to the host. Such gene products are exemplified by the enzymes for the cometabolism of polychlorinated biphenyls (PCBs) through the biphenyl degradation pathway. Attempts to use highly competent PCB-cometabolizing strains in the environment in the absence of biphenyl have not been successful, while the addition of biphenyl is limited by its human toxicity and low water solubility. Broad-substrate-specificity PCB-degradative genes (bphABC) were cloned from a naturally occurring isolate. Pseudomonas sp. strain ENV307, into broad-host-range plasmid pRK293. The resulting PCB-degrading plasmids were transferred to the FAV host Pseudomonas paucimobilis 1IGP4, which utilizes the nontoxic, water-soluble, nonionic surfactant Igepal CO-720 as a selective growth substrate. Plasmid stability in the recombinant strains was determined in the absence of antibiotic selection. PCB-degrading activity was determined by resting cell assays. Treatment of contaminated soil (10, 100, or 1,000 ppm of Aroclor 1242) by surfactant amendment (1.0% [wt/wt]Igepal CO-720 in wet soil) and inoculation with recombinant isolates of strain 1IGP4 (approximately 4 x 10(6) cells per g of soil) resulted in degradation of many of the individual PCB congeners in the absence of biphenyl.(ABSTRACT TRUNCATED AT 250 WORDS)

Development and use of field application vectors to express nonadaptive foreign genes in competitive environments.

Many potential applications of genetically engineered microorganisms in environmental and agricultural biotechnology involve introducing genetic capabilities into nonsterile competitive environments in which they provide no advantage to the host. Field application vectors have been designed for the purpose of creating a temporary niche for the host in such environments. This technique involves the addition to the target environment of a selective substrate readily utilizable by the host microorganism but unavailable to most indigenous species. Thirteen nonionic and anionic detergents, representing a wide range of structural complexities and molecular weights, were screened as potential selective substrates. Competition experiments in soil, using Warburg respirometry, indicated that isolates from six different detergent enrichment cultures were more active on their corresponding detergents than the indigenous microorganisms. Detergents of intermediate structural complexities and molecular weights were most effective for use as selective substrates. A field application vector that utilizes 1.0% Igepal CO-720 (detergent) as the selective substrate and Pseudomonas paucimobilis 1IGP4 as the host was tested for its ability to increase the presence of nonadaptive tetracycline resistance marker genes in soil. In soil amended with the selective substrate, strain 1IGP4 plate counts increased by three orders of magnitude and tetracycline-resistant transformant (pRK293) counts increased from 1.8 x 10/g of soil to 4.3 x 10/g in 2 days. Inoculation in the absence of substrate amendment or amendment with a nonselective substrate did not result in growth of strain 1IGP4. These results demonstrate the effectiveness of field application vectors for increasing the concentration of nonadaptive genes in competitive environments.

Effect of temperature on bacterial species diversity in thermophilic solid-waste composting.

Continuously thermophilic composting was examined with a 4.5-liter reactor placed in an incubator maintained at representative temperatures. Feed was a mixture of dried table scraps and shredded newspaper wetted to 55% moisture. One run at 49 degrees C (run A) employed a 1:4 feed-to-compost ratio, while the other runs used a 10:1 ratio and were incubated at 50, 55, 60, or 65 degrees C. Due to self-heating, internal temperatures of the composting mass were 0 to 7 degrees C hotter than the incubator. Two full-scale composting plants (at Altoona, Pa., and Leicester, England) were also examined. Plate counts per gram (dry weight) on Trypticase soy broth (BBL Microbiology Systems) with 2% agar ranged from 0.7 X 10(9) to 5.3 X 10(9) for laboratory composting and 0.02 X 10(9) to 7.4 X 10(9) for field composting. Fifteen taxa were isolated, including 10 of genus Bacillus, which dominated all samples except that from run A. Species diversity decreased markedly in laboratory composting at 60 degrees C and above, but was similar for the three runs incubated at 49, 50, and 55 degrees C. The maximum desirable composting temperature based on species diversity is thus 60 degrees C, the same as that previously recommended based on measures of the rate of decomposition.

Identification of thermophilic bacteria in solid-waste composting.

The thermophilic microbiota of solid-waste composting, with major emphasis on Bacillus spp., was examined with Trypticase soy broth (BBL Microbiology Systems) with 2% agar as the initial plating medium. Five 4.5-liter laboratory units at 49 to 69 degrees C were fed a mixture of dried table scraps and shredded newspaper. The composting plants treating refuse at Altoona, Pa., and refuse-sludge at Leicester, England, were also sampled. Of 652 randomly picked colonies, 87% were identified as Bacillus spp. Other isolates included two genera of unidentified nonsporeforming bacteria (one of gram-negative small rods and the other of gram-variable coccobacilli), the actinomycetes Streptomyces spp. and Thermoactinomyces sp., and the fungus Aspergillus fumigatus. Among the Bacillus isolates, the following, in order of decreasing frequency, were observed: B. circulans complex, B. stearothermophilus, B. coagulans types A and B, B. licheniformis, B. brevis, B. sphaericus, Bacillus spp. types i and ii, and B. subtilis. About 15% of the Bacillus isolates could be assigned to species only by allowing for greater variability in one or more characteristics than has been reported by other authors for their strains. In particular, growth at higher temperatures than previously reported was found for strains of several species. A small number of Bacillus isolates (less than 2%) could not be assigned to any recognized species.

Concentrations of nitrifying bacteria in sewages, effluents, and a receiving stream and resistance of these organisms to chlorination.

Estimates of NH4+-and NO2-oxidizers in samples from four activated sludge plants treating mainly domestic sewage were obtained using a most-probable-number (MPN) technique. Ranges of concentrations per milliliter of each, respectively, were 1,010 to 3,880 and 79 to 145 in settled sewages, 32 to 7,420 and 2 to 1,010 in secondary effluents, and less than 0.1 to 622 and 0.1 to 70 in chlorinated secondary effluents. The results of this field study indicated that nitrifiers were more resistant to chlorination than fecal streptococci, which were also enumerated. In laboratory studies the survivals of these bacterial groups in secondary effluents were determined after exposure to chlorine residuals of up to 2 mg/liter for 0 to 60 min. The nitrifiers proved considerably more resistant than fecal streptococci, with NO2-oxidizers showing greater resistance than NH4+-oxidizers. Below the outfall of one of the plants that discharges heavily chlorinated unnitrified effluent, NH4+-oxidizers amounted to approximately 200 X 10(5) per g of slime scraped from stream-bed rocks. Upstream of the outfall this was approximatley 3 X 10(5)/G.

Length of incubation for enumerating nitrifying bacteria present in various environments.

The effect of incubation time on most-probable-number estimates of autotrophic nitrifying bacteria was investigated by using waters, rooted aquatic plants, sediments, and slimes as inoculum sources. Maximum most probable numbers of the NH4+-oxidizing group were attained in 20 to 55 days (median, 25). Estimates of NO2-oxidizers were highest at termination (103 to 113) days.