Microbiological and kinetic aspects of a biofilter for the removal of toluene from waste gases

Microbiological and kinetic aspects of a biofilter inoculated with a consortium of five bacteria and two yeast adapted to remove toluene vapors were investigated. Initially the toluene sorption isotherm on peat and the effect of different environmental conditions on the toluene consumption rates of this consortium were measured. The fast start-up of the biofilter and the decay in the elimination capacity (EC) were reproduced using microcosm assays with toluene successive additions. Nutrient limitation and a large degree of heterogeneity were also detected. EC values, extrapolated from microcosms, were higher than biofilter EC when it was operating close to 100% efficiency but tended to relate better as the biofilter EC diminished. In studies on the microbial evolution in the biofilter, an increase in the cell count and variation in the ecology of the consortium were noted. Bacterial counts up to 10 x 10(11) cfu/gdry peat were found in 88 days, which corresponds to about a 10(4) increase from inoculum. Observations with SEM showed a nonuniform biofilm development on the support and the presence of an extracellular material. The results obtained in this work demonstrated that activity measurement in microcosms concomitant to the biofilter operation could be an important tool for understanding, predicting and improving the biofiltration performance. Copyright 1999 John Wiley & Sons, Inc.     

Design of a biofilter packed with crab shell and operation of the biofilter fed with leaf mold solution as a nutrient

After measuring toluene adsorption (15.7 mg-toluene/g-material), water holding capacity (18.5%), organic content (53.8%), specific surface area (18.1 m²/g-material), and microbial attachment, crab shells were chosen as the main packing material for a biofilter design. The crab shells, cheap and abundant in the Gangneung area, also have relatively rigid structure, low density, and ability to neutralize acids generated during mineralization of toluene. Since towel scraps have water holding capacity as high as 301.2%, 10% of the total packing was supplemented with them to compensate for low water holding capacity of the crab shells. The biofilter fed with defined chemical medium under 0.8∼1.3 mg/L of inlet toluene concentration and 18 seconds of residence time showed satisfactory removal efficiency of over 97% and 72.8 g/h·m³ of removal capacity. For the purpose of deceasing operation costs, leaf mold solution was tried as an alternative nutrient instead of a defined chemical medium. The removal efficiency and removal capacity were 85% and 56.3 g/h·m³, respectively, using the same inlet toluene concentration and residence time. This research shows the possibility of recycling crab shell waste as packing material for biofilter. In addition, leaf mold was able to serve as an alternative nutrient, which remarkably decreased the operating cost of the biofilter.     

Start-up and the effect of gaseous ammonia additions on a biofilter for the elimination of toluene vapors

Biotechnological techniques, including biofilters and biotrickling filters are increasingly used to treat air polluted with VOCs (Volatile Organic Compounds). In this work, the start-up, the effect of the gaseous ammonia addition on the toluene removal rate, and the problems of the heat accumulation on the performance of a laboratory scale biofilter were studied. The packing material was sterilized peat enriched with a mineral medium and inoculated with an adapted consortium (two yeast and five bacteria). Start-up showed a short adaptation period and an increased toluene elimination capacity (EC) up to a maximum of 190 g/m3/h. This was related to increased CO2 outlet concentration and temperature gradients between the packed bed and the inlet (Tm-Tin). These events were associated with the growth of the microbial population. The biofilter EC decreased thereafter, to attain a steady state of 8 g/m3/h. At this point, gaseous ammonia was added. EC increased up to 80 g/m3/h, with simultaneous increases on the CO2 concentration and (Tm-Tin). Two weeks after the ammonia addition, the new steady state was 30 g/m3/h. In a second ammonia addition, the maximum EC attained was 40 g/m3/h, and the biofilter was in steady state at 25 g/m3/h. Carbon, heat, and water balances were made through 88 d of biofilter operation. Emitted CO2 was about 44.5% of the theoretical value relative to the total toluene oxidation, but accumulated carbon was found as biomass, easily biodegradable material, and carbonates. Heat and water balances showed strong variations depending on EC. For 88 d the total metabolic heat was -181.2 x 10(3) Kcal/m3, and water evaporation was found to be 56.5 kg/m3. Evidence of nitrogen limitation, drying, and heterogeneities were found in this study.     

Toluene biofiltration by the fungus Scedosporium apiospermum TB1

The performance of biofilters inoculated with the fungus Scedosporium apiospermum was evaluated. This fungus was isolated from a biofilter which operated with toluene for more than 6 months. The experiments were performed in a 2.9 L reactor packed with vermiculite or with vermiculite-granular activated carbon as packing material. The initial moisture content of the support and the inlet concentration of toluene were 70% and 6 g/m3, respectively. As the pressure drop increased from 5-40 mm H2O a strong initial growth was observed. Stable operation was maintained for 20 days with a moisture content of 55% and a biomass of 33 mg biomass/g dry support. These conditions were achieved with intermittent addition of culture medium, which permitted a stable elimination capacity (EC) of 100 g/m3(reactor)h without clogging. Pressure drop across the bed and CO2 production were related to toluene elimination. Measurement of toluene, at different levels of the biofilter, showed that the system attained higher local EC (200 g/m3(r)h) at the reactor outlet. These conditions were related to local humidity conditions. When the mineral medium was added periodically before the EC decreases, EC of approximately 258 g/m3(r)h were maintained with removal efficiencies of 98%. Under these conditions the average moisture content was 60% and 41 mg biomass/g dry support was produced. No sporulation was observed. Evaluation of bacterial content and activities showed that the toluene elimination was only due to S. apiospermum catabolism.     

Improving hexane removal by enhancing fungal development in a microbial consortium biofilter

The removal of hydrophobic pollutants in biofilters is often limited by gas liquid mass transfer to the biotic aqueous phase where biodegradation occurs. It has been proposed that the use of fungi may improve their removal efficiency. To confirm this, the uptake of hexane vapors was investigated in 2.6-L perlite-packed biofilters, inoculated with a mixed culture containing bacteria and fungi, which were operated under neutral or acid conditions. For a hexane inlet load of around 140 g.m-3.h-1, elimination capacities (EC) of 60 and 100 g.m-3.h-1 were respectively reached with the neutral and acid systems. Increasing the inlet hexane load showed that the maximum EC obtained in the acid biofilter (150 g.m-3.h-1) was twice greater than in the neutral filter. The addition of bacterial inhibitors had no significant effect on EC in the acid system. The biomass in the acid biofilter was 187 mg.g-1 (dry perlite) without an important pressure drop (26.5 mm of water.m-1reactor). The greater efficiency obtained with the acid biofilter can be related to the hydrophobic aerial hyphae which are in direct contact with the gas and can absorb the hydrophobic compounds faster than the flat bacterial biofilms. Two fungi were isolated from the acid biofilter and were identified as Cladosporium and Fusarium spp. Hexane EC of 40 g.m-3.h-1 for Cladosporium sp. and 50 g.m-3.h-1 for Fusarium sp. were obtained in short time experiments in small biofilters (0.230 L). A biomass content around 30 mg.g-1 (dry perlite) showed the potential for hexane biofiltration of the strains.     

Fungal/bacterial interactions during the biodegradation of TEX hydrocarbons (toluene, ethylbenzene and p-xylene) in gas biofilters operated under xerophilic conditions

The treatment of air contaminated with toluene, ethylbenzene, and p-xylene was assayed in three laboratory-scale biofilters, each consisting of two modules connected in series, packed with a pelletized organic fertilizer and inoculated with a toluene-degrading liquid enrichment culture. Biofilters were operated in parallel for 185?days in which the volumetric organic loading rate was progressively increased. The operation regime was subjected to drying out, so that packing humidity generally remained below 40%. Significant process failure occurred with ethylbenzene and p-xylene, but the toluene biofilter comparatively sustained a significant elimination capacity. Microbial community characterization by quantitative PCR and denaturing gradient gel electrophoresis showed substantial fungal enrichment in the toluene biofilter. Ribotypes identical to the well-known toluene-degrading black yeast Exophiala oligosperma (Chaetotyriales) were found among the dominant species. The microbial community structure was similar in the biofilters loaded with toluene and ethylbenzene but with p-xylene was quite specific and encompassed other chaetothyrialean fungi. Several species of Actinomycetales were found in the packing while the inoculum was dominated by representatives of the Burkholderiales and Xanthomonadales. One single fungal ribotype homologous to Acremonium kiliense was detected in the inoculum. The implications of xerophilic biofilter operation on process biosafety and efficiency are discussed.     

Thermophilic biofiltration of benzene and toluene

In the current studies, we characterized the degradation of a hot mixture of benzene and toluene (BT) gases by a thermophilic biofilter using polyurethane as packing material and high-temperature compost as a microbial source. We also examined the effect of supplementing the biofilter with yeast extract (YE). We found that YE substantially enhanced microbial activity in the thermophilic biofilter. The degrading activity of the biofilter supplied with YE was stable during long-term operation (approximately 100 d) without accumulating excess biomass. The maximum elimination capacity (1,650 g x m(-3) h(-1)) in the biofilter supplemented with YE was 3.5 times higher than that in the biofilter without YE (470 g g x m(-3) h(-1)). At similar retention times, the capacity to eliminate BT for the YE-supplemented biofilter was higher than for previously reported mesophilic biofilters. Thus, thermophilic biofiltration can be used to degrade hydrophobic compounds such as a BT mixture. Finally, 16S rDNA polymerase chain reaction-DGGE (PCR-DGGE) fingerprinting revealed that the thermophilic bacteria in the biofilter included Rubrobacter sp. and Mycobacterium sp.     

Application of biological activated carbon as a low pH biofilter medium for gas mixture treatment

Packing material is a crucial component of a bioreactor as it is the microbial population's habitat. This study assessed potential improvements to current biofiltration processes by investigating use of a novel support medium. Biological activated carbon (BAC) with microorganisms growing on granular activated carbon can produce a novel medium in which both adsorption and biodegradation contribute to pollutants removal. Investigation of carbon characteristics demonstrated that BAC was an ideal packing medium for biofiltration. The application of the novel packing medium for gas mixture treatment was evaluated in a low pH biofilter. Results demonstrated that BAC biofilter obtained high removal efficiency for both H(2)S and toluene. The removal mechanisms of BAC were investigated after the biofilter operation and it demonstrated that the performance of the BAC system was mainly controlled by the additive contributions of two removal mechanisms - adsorption and biodegradation. This study also indicated the potential for simultaneous treatment of hydrogen sulfide and toluene at low pH condition.     

Characterization of a biofilter treating toluene contaminated air

The removal of toluene from an experimental gas-stream was studied in an industrial biofilter filled with poplar wood bark. Toluene degradation, approximately 85% through the operating period, resulted in low levels of toluene in the off-gas effluent. For a toluene load of 6.7 g m^-3 h^-1 the elimination capacity of the biofilter was found to be 6.0 g m^-3 h^-1. Toluene removal was due to biodegradative activity of microorganisms in the filter bed; the most probable number counts of toluene degraders increased from 2.4×10² to 6.4×10⁷ MPN/g dry packing material in about seven months of air-toluene supply. The degradative capacity of a Burkholderia (Pseudomonas) cepacia strain, isolated from the biofilter material, as an example of the effectiveness of microbial toluence removal was tested in batch culture. The microorganism degraded completely 250 ppm of toluence supplied as sole carbon source in 24 hours. The high performance demonstrated for a long period and the mechanical and physico-chemical stability of the biofilter favour its use in industrial full-scale off-gas control.     

Biotreatment of a gas-phase volatile mixture from fibreglass and composite manufacturing industries

Acetone, toluene and styrene (ATS) are representative air pollutants emanating during the production process in fibreglass and composite manufacturing industries. In this study, the performance of a steady-state biofilter inoculated with the fungus Sporothrix variecibatus was tested at different empty bed residence times (EBRTs), and at different inlet concentrations of ATS, corresponding to total pollutant loading rates ranging from 30 to 490 gm(-3)hour(-1). Styrene was somewhat better removed (47-100%) in the biofilter than acetone (34-100%) and toluene (42-100%), with maximum elimination capacities (EC(max)) of 108, 72 and 144 gm(-3)hour(-1), for ATS, respectively. Besides, it was observed that, although increasing the concentration of ATS decreased their removal, the presence of toluene also decreased the EC(max) of both acetone and toluene in the ternary mixture. During transient operations, the biofilter was subjected to intermittent shutdown and re-start operations where the gas-phase pollutant flow was stopped for either 5 or 16d. It was observed that, for longer shutdown periods (16d), the biofilter required nearly 8-10d to reach similar removal patterns to those observed before the shutdown phase. Batch biodegradation tests were conducted, using Sporothrix-like microorganisms present in the leachate of the biofilter, with a mixture of ATS as the sole carbon source. Complete removal of ATS was observed within the test period of 168 hours. Styrene was degraded faster, with a specific substrate utilization rate of 0.9 mg styrenemg biomass(-1)hour(-1), followed by toluene (0.6) and acetone (0.44). The effectiveness of the biofilter to reach high total EC (321.3 gm(-3)hour(-1)), and withstand transient operations shows the robustness of this fungal-bioreactor and its suitability to handle emissions from a fibreglass and composite manufacturing industry.     

Microbial response and elimination capacity in biofilters subjected to high toluene loadings

Elimination capacity (EC) is frequently used as a performance and design criterion for vapor-phase biofilters without further verification of the microbial quantity and activity. This study was conducted to investigate how biofilters respond to high pollutant loadings and ultimately how this affects the EC of the biofilter. Two identical laboratory-scale biofilters were maintained at an initial toluene loading rate of 46 g m⁻³ h⁻¹ for a period of 24 days. After the initial biofilm development stage, the loading rates were increased to 91 g m⁻³ h⁻¹ and 137 g m⁻³ h⁻¹, respectively. Following a short period of pseudo-steady state, toluene removal efficiencies rapidly declined in both biofilters, with a concurrent decline in both critical and maximum ECs. The decline was mainly due to deterioration in the biodegradation activity of the biofilm and a decline in the toluene-degrading bacterial population within the biofilm phase. The findings imply that high toluene loadings accelerated the deterioration in overall performance due to a rapid accumulation of inactive biomass. As a result, care must be used when relying on EC values for biofilter design and operational purposes, since the values do not appropriately reflect the temporal changes in biodegradation activity and active biomass quantities that can occur in biofilters subjected to high inlet loadings.     

Fungal removal of gaseous hexane in biofilters packed with poly(ethylene carbonate) pine sawdust or peat composites

The removal of volatile organic compounds (VOC) in biofilters packed with organic filter beds, such as peat moss (PM) and pine sawdust (PS), frequently presents drawbacks associated to the collapse of internal structures affecting the long-term operation. Poly(ethylene ether carbonate) (PEEC) groups grafted to these organic carriers cross linked with 4,4'-methylenebis(phenylisocyanate) (MDI) permitted fiber aggregation into specific shapes and with excellent hexane sorption performance. Modified peat moss (IPM) showed very favorable characteristics for rapid microbial development. Water-holding capacity in addition to hexane adsorption almost equal to the dry samples was obtained. Pilot scale hexane biofiltration experiments were performed with the composites after inoculation with the filamentous fungus Fusarium solani. During the operation of the biofilter under non-aseptic conditions, the addition of bacterial antibiotics did not have a relevant effect on hexane removal, confirming the role of fungi in the uptake of hexane and that bacterial growth was intrinsically limited by an adequate performance of the composites. IPM biofilter had a start-up period of 8-13 days with concurrent CO(2) production of approximately 90 g m(-3) h(-1) at day 11. The final pressure drop after 70 days of operation was 5.3 mmH(2)O m(-1) reactor. For modified pine sawdust (IPS) packed biofilter, 5 days were required to develop an EC of about 100 g m(-3) h(-1) with an inlet hexane load of approximately 190 g m(-3) h(-1). Under similar conditions, 12-17 days were required to observe a significant start-up in the reference perlite biofilter to reach gradually an EC of approximately 100 g m(-3) h(-1) at day 32. Under typical biofiltration conditions, the physical-chemical properties of the modified supports maintained a minimum water activity (a(w)) of 0.925 and a pH between 4 and 5.5, which allowed the preferential fungal development and limited bacterial growth.     

Removal of a high load of ammonia by a marine bacterium,Vibrio alginolyticus in biofilter

A newly isolated heterotrophic marine bacterium,Vibrio alginolyticus, was used to remove a high load of ammonia gas under non-sterile condition. The cells were inoculated onto an inorganic packing material in a fixed-bed reactor (biofilter), and a high bad of ammonia, in the range of ammonia gas concentration of 170 ppm to 880 ppm, was introduced continuously. Sucrose solution and 3% NaCl was supplied intermittently to supplement the carbon source and water to the biofilter. The average percentage of gas removed exceeded 85% for 107-day operation. The maximum removal capacity and the complete removal capacity were 19 g-N kg⁻¹ dry packing material day⁻¹ and 16 g-N kg⁻¹ dry packing material day⁻¹, respectively, which were about three times greater than those obtained in nitrifying sludge inoculated onto the same packing material. On day 82, the enhanced pressure drop was restored to the normal one by NaOH treatment, and efficient removal characteristics were later observed. During this operation, the non-sterile condition had no significantly adverse effect on the removability of ammonia byV. alginolyticus.     

Removal of benzene and toluene in polyurethane biofilter immobilized with Rhodococcus sp. EH831 under transient loading

The performance of a polyurethane (PU) biofilter inoculated with Rhodococcus sp. EH831 was evaluated under different transient loading conditions, such as shutdown, intermittent and fluctuating loading. A mixture of benzene and toluene vapors was employed as model pollutants. When the biofilter was restarted after a 2 week-shutdown, during which neither clean air nor water was supplied, the benzene and toluene removal capacities were rapidly restored after a re-adaptation period of only 1 day. A comparison of the removal capacity under continuous and intermittent loading revealed that constant and periodic loading (8 h on/16 h off per day) and a 2 day-shutdown did not significantly influence the biofilter performance, although the removals of benzene and toluene were relatively unstable and lower under intermittent loading during the initial week. The result of quantitative real-time PCR showed that Rhodococcus sp. EH831 could be maintained during transient loading periods (10¹⁰–10¹¹ CFU/g-dry PU) irrespective of the different operating conditions.     

Styrene degradation by <Emphasis Type="Italic">Pseudomonas</Emphasis> sp. SR-5 in biofilters with organic and inorganic packing materials

Pseudomonas sp. SR-5 was isolated as a styrene-degrading bacterium. In liquid culture containing 1% (v/v) styrene, more than 90% styrene was degraded in 53 h and the doubling time of SR-5 was 2 h. The removal of styrene gas was investigated in biofilters for 31 days using an organic packing material of peat and an inorganic packing material of ceramic inoculated with SR-5. The maximum-styrene-elimination capacities for peat and ceramic packing materials were 236 and 81 g m^–3 h^–1, respectively. The percentage of styrene converted to low molecular weight compounds including CO₂ in the peat and ceramic biofilters during a 10-day operation were estimated to be 90.4 and 36.7%, respectively. As the pressure drop in the peat bioflter at the end of experiment was significantly higher than that in ceramic biofilter, a biofilter using a mixture of peat and ceramic was tested. We determined that the maximum elimination capacity was 170 g m^–3 h^–1 and the production of low molecular weight compounds was 95% at a low pressure drop for this mixed packing material filter.     

Evaluation of full-scale biofilter with rockwool mixture treating ammonia gas from livestock manure composting

NH3 removal by a full-scale biofilter with rockwool packing materials was studied by measuring the gases and potential nitrification and denitrification activities of those materials in order to improve the biofiltration technology used in livestock farms. The rockwool biofilter was a durable and effective system for removing NH3, which was varied with the turning of manure composts. Furthermore, NH3 could be treated in the absence of an extra increase in two greenhouse gases, N2O and CH4. Potential nitrification and denitrification activities of the packing materials were estimated to be 8.2-12.2 mg N, and 1.42-4.69 mg N/100 g dry samples per day, respectively. The results suggested that potential nitrification and denitrification activities would increase within the biofilter where substrates, NH3 or NO3(-), have accumulated as a result of its operation. However, since percolate water contained high concentrations of NH4(+) and NO3(-), further improvement is required by reducing nitrogenous compounds within both the biofilter and percolate water.     

Correlation of biological activity and reactor performance in biofiltration of toluene with the fungus Paecilomyces variotii CBS115145

A biofiltration system inoculated with the mold Paecilomyces variotii CBS115145 showed a toluene elimination capacity (EC) of around 250 g/m3 of biofilter/h, which was higher than the values usually reported for bacteria. P. variotii assimilated m- and p-cresols but not the o isomer. Initial toluene hydroxylation occurred both on the methyl group and through the p-cresol pathway. These results were corroborated by detecting benzyl alcohol, benzaldehyde, and p-cresol as volatile intermediates. In liquid cultures with toluene as a substrate, the activity of toluene oxygenase (TO) was 5.6 nmol of O2/min/mg of biomass, and that of benzyl alcohol dehydrogenase was 16.2 nmol of NADH/min/mg of protein. Toluene biodegradation determined from the TO activity in the biofilter depended on the biomass distribution and the substrate concentration. The specific enzymatic activity decreased from 6.3 to 1.9 nmol of O2/min/mg of biomass along the reactor. Good agreement was found between the EC calculated from the TO activity and the EC measured on the biofilter. The results were confirmed by short-time biofiltration experiments. Average EC measured in different biofiltration experiments and EC calculated from the TO activity showed a linear relation, suggesting that in the biofilters, EC was limited by biological reaction. As the enzymatic activities of P. variotii were similar to those reported for bacteria, the high performance of the fungal biofilters can possibly be explained by the increased transfer of the hydrophobic compounds, including oxygen, from the gas phase to the mycelia, overcoming the transfer problems associated with the flat bacterial biofilms.     

Removal characteristics of high load ammonia gas by a biofilter seeded with a marine bacterium, Vibrio alginolyticus

When the cells of the newly isolated marine bacterium, Vibrio alginolyticus, were inoculated on to an inorganic packing material in biofilter, and a load of ammonia of 2.4–22.5 g-N kg^–1dry packing material was introduced continuously under non-sterile conditions, the average amount of NH₃removed exceeded 85% over 61-d operation. The maximum removal capacity and the complete removal capacity were 22.8 g-N kg^–1dry packing material dand 18.6 g-N kg^–1dry packing material d, respectively, which were about four times larger than those obtained in autotrophic nitrifying sludge inoculated on the same packing material.     

Biofiltration for the removal and ‘detoxification’ of a complex mixture of volatile organic compounds

A study was performed to determine the effectiveness of using biofiltration for the removal of a complex mixture of volatile organic compounds (VOCs) air-stripped from petroleum hydrocarbons. A biofilter was constructed which contained 264 cm³ of packing material (Celite^? R-635). The unit was inoculated with a mixed culture containing a hydrocarbon-degrading Pseudomonas sp and an Alcaligenes sp. Several of the major compounds in the VOC mixture were monitored individually, along with the total VOCs, using gas chromatography. The average influent concentration of the VOC mixture was 320 ppmv and the average total VOC removal rate was over 56%, with the average removal rate of the monitored individual compounds ranging from 49–90%. After 30 days of operation the average overall removal rate was 69% and the removal of the major compounds averaged 92%. The toxicity and mutagenicity of the air stream was monitored using the Microtox and Ames assays, respectively. These data show marked decreases in toxicity and mutagenicity of the air stream as a result of the biofiltration treatment. The biofiltration system, therefore, was not only effective in removing VOCs from the air stream over an extended time-period, but was also effective in greatly reducing the toxicity and mutagenicity associated with the remaining VOCs. Received 03 July 1997/ Accepted in revised form 25 November 1997     

Treatment of MEK and toluene mixtures in biofilters: effect of operating strategy an performance during transient loading

In recent years, biofiltration has been increasingly applied as an air pollution control technology to minimize or eliminate emissions of volatile organic compounds from industrial sources and environmental remediation activities. Although the ability of this technology to maintain high removal efficiency during relatively steady loading conditions has been well established for many waste streams, relatively little research has focused on development of operating strategies that could improve treatment performance during transient loading conditions typical of industrial operations. In the research described herein, two operating strategies were evaluated over a period of 295 days in biofilters treating a model waste gas stream containing a two-component mixture of methyl ethyl ketone (MEK) and toluene. One biofilter was operated as a sequencing batch biofilter (SBB), and the other was operated as a conventional continuous-flow biofilter (CFB). During "normal" steady loading conditions, the model waste stream contained MEK concentrations ranging from 80 to 89 ppmv and toluene concentrations ranging from 28 to 30 ppmv. Both biofilter operating strategies resulted in stable long-term performance with greater than 99% contaminant removal during these normal loading conditions. On a regular basis, the influent MEK and toluene concentrations were temporarily increased to five times the normal influent concentration for the duration of 1 h to test performance during transient "shock loading" conditions. Biofilter performance during the model shock loading conditions demonstrate that SBB operating strategies can result in superior treatment in two important areas: (1) overall mass of contaminants removed and (2) minimum instantaneous removal efficiency.