243

SUMMARY AND CONCLUSIONS

An experimental facility was built to test the feasibility of recovering nutrients and energy from swine manure. A source of industrial waste heat was simulated to enhance the biological processes involved. Algae and bacteria were used to convert nutrients into biomass and to generate biogas. Manure was removed from the animal quarters by a gutter flushing system and solids were separated from liquids by gravity settling. Solids were subjected to anaerobic digestion in a 14 m 3 digester to release plant nutrients, reduce the volume of the solids, stabilize the manure biologically to reduce odor problems, and recover energy in the form of methane gas. The liquid phase of the manure was used as a nutrient substrate for the growth of algae and bacteria in 12 outdoor basins with a combined surface area of 24 m 2 and a combined volume of 6,000 I. The waste heat needed to maintain above-ambient temperatures in the bioconversion units was simulated by heating water in heat exchangers with electrical resistance heaters. An accurate heat balance for digester and basins was obtained by using kWh meters. The experimental facility was designed so that the waste water could either be recycled for flushing or discharged into a neaby lagoon. In the latter case, only fresh water was used for flushing of the animal quarters. The experimental facility was used to study: 1. the functioning of the digester and the algal growth basins, 2. the degree of nutrient removal, 3. dry matter and protein yields, 4. production of methane gas, 5. the energy requirements of the digester and the outdoor basins, 6. the quality of the waste water after removal of the biomass, 7. the biological stability of the conversion units, 8. the practical and economical feasibility of the nutrient recovery system, 9. the development of design criteria for a full-scale operation, and 10. the formulation of other bioconversion processes which might improve the recovery of nutrients as well as the quality and quantity of end products from swine manure.

Growth of Algae
In early laboratory tests, the high temperature strain 211/8K of Ch/orella vulgaris was found to grow better in swine waste than Scenedesmus obliquus Sc D 3 WT, Scenedesmus quadricauda, Selenastrum capricornutum, Spirulina major, and Botyrococcus, and was therefore selected for use in the field experiments.

244 Experiments were designed to test the ability of Chlorella vulgaris 211/8K to establish itself as the predominant algal species and to find the combinations of culture depth, retention time, and temperature which produce the highest cell concentrations under prevailing climatic conditions. It was determined that the fresh manure must be diluted before the liquid phase is suitable for algal growth. In terms of inorganic nitrogen the manure must be diluted to contain 150 to 250 mg of elemental nitrogen in the ammonium form per liter. This is achieved by a 50-fold dilution of the volume of manure excreted by the animals, including liquids and solids. When the manure is not sufficiently diluted, the high concentration of organic matter promotes bacterial growth over algal growth.

Light Requirements It was found that Chlorella vulgaris 211/8K requires a monthly average of the daily rate of solar radiation in excess of 2,000 kcal/m 2 day and a photoperiod of at least 11 h, in order to maintain a sufficiently high concentration of cells for harvesting at retention times of 8 days or less. Growth continues at less than 2,000 kcal/m 2 day but at a reduced rate, so that adjustments in the retention time and/or culture depth are necessary to prevent depletion of the algae and eventual predominance of bacterial growth. Adjustments in retention time and/or culture depth are also necessary whenever a prolonged period of cloudiness occurs during which the rate of solar radiation remains below 2,000 kcal/m 2 day, although the monthly average of the daily rate may still be above 2,000 kcal/m 2 day.

Culture Depth The highest concentrations of algae were found at the culture depth of 10 cm. Increasing the culture depth to 20 cm reduced the concentration of algae by 50 percent. However, the total biomass, including algae and bacteria, was nearly the same at either depth.

Temperature The use of waste heat to maintain a culture temperature of at least 25 C was essential in achieving high concentrations of algae, except during the summer months when the monthly average of the daily maximum air temperature was 25 C and the monthly average of the daily minimum air temperature was not less than 10 C. The statement assumes continuous mixing of the algae and equilibrium of temperature between the culture medium and air. Maximum concentrations of Chlorella were obtained at 30 C or 35 C. However, the contribution made by the algae to the total biomass, including bacteria, was higher at the temperature of 35 C than at 30 C. This was attributed to the production of anti-bacterial substances by Chlorella at 35 C but not at 30 C.

245 Retention Time The concentration of algal cells was found to increase with retention time. Maximum concentrations were obtained at retention times of 6 to 8 days, indicating a generation time of Ch/ore//a of 72 to 96 h under field conditions.

Concentrations of Algal Cells

Concentrations of cells of Ch/ore//a greater than 10 7/ml could be maintained in the properly diluted waste water at retention times of 3 to 8 days, temperatures of 25 to 35 C, and culture depths of 7.5 to 20 cm when the monthly average of the daily rate of solar radiation was at least 2,000 kcal/m 2 day. Experiments conducted during the months of October through December showed that a culture temperature of 35 C, a shallow culture depth of 10 cm, and a retention time of 8 days do not compensate for the loss in algal growth due to the short photoperiod of less than 11 h/day and a monthly average of the daily rate of solar radiation of less than 2,000 kcal/m2 day. Under these conditions, a retention time of 8 days exceeds the generation time of Ch/ore//a vulgaris 211/8K, so that the harvesting process depletes the algal population and bacterial growth dominates. When the cell density of Ch/ore//a could be maintained above 10 7/ml, the culture fluid had a deep green color which was used as visual evidence of algal growth. The typical odor of swine manure was absent. The photosynthetic activity of the algae caused daily changes in the pH and the dissolved oxygen content of the swine waste substrate. The magnitude of these changes was a function of the intensity of solar radiation, culture depth, and temperature. In general, the photosynthetic activity of the algae, as measured by the DO content of the culture, increased soon after sunrise and reached a maximum value sometime between 13:00 and 16:00 h. The DO concentration frequently rose above 20 mg/I and the pH rose as high as 9 in the 20 cm deep cultures and above 10 in the 7.5 cm deep basins. Nighttime respiratory activity of algae and bacteria increased the concentration of dissolved CO2 and reduced the pH to about 8 by early morning. During the same time the DO decreased to concentrations between 4 and 6 mg/I. Periods of prolonged cloudiness reduced the photosynthetic activity of the algae so that the pH fluctuated only between 7.8 and 8.5 and the DO content between 4 and 8 mg/I. Temperatures of less than 25 C also reduced photosynthetically induced changes in pH and DO concentrations. Cultures with cell densities of less than 10 6/ml showed no visual evidence of algal growth. The medium had the appearance of activated sewage and the odor was that typical of swine manure. The pH in these cultures remained at 7.6 and the DO concentrations at less than 1 mg/I. This condition was favored at radiation intensities of less than 2,000 kcal/m 2 day in combination with short retention times of 2 to 3 days and culture depths greater than 10 cm. It was concluded that the most effective way of maintaining a high density of Ch/ore//a cells during prolonged periods of low intensity of solar radiation was to lower the culture depth to 10 cm or less, to stop harvesting, and to stop the addition of fresh substrate to the culture.

246 Contribution of Algal Cells to Total Biomass Efforts were made to distinguish between the contribution made by algal cells and the contribution made by bacterial cells to the total biomass. Algal cell counts were made using a hemocytometer and Coulter counter. Concentrations of algal dry matter were estimated as the product of mean cell volume and algal cell counts, taking into consideration a moisture content of 75 percent w/w and a specific gravity of the algae of 1.1 g/cm 3 . The mean cell volume was determined by a MCV-computer attached to the Coulter counter. It was assumed that the difference between the concentrations of total dry matter obtained by the Millipore filter technique and the estimated concentrations of algal dry matter represented the contribution made by the bacterial cells to the total biomass. In sufficiently diluted waste containing 150 to 250 mg NH4-N/I, the highest concentration of algal dry matter, namely 0.70 g/I, was obtained in 10 cm deep basins when the retention time was 6 days, the temperature of the culture was 35 C, and the mean of the intensity of solar radiation was 3,000 kcal/m 2 day. Essentially all of the biomass consisted of algae under these conditions. In contrast, the concentration of algal dry matter was 0.36 g/I in the 20 cm deep basins and the algae were estimated to have contributed about 50 percent to the concentration of total biomass of 0.74 g/I. The total biomass harvested per day from either 10 cm or 20 cm deep basins would thus be the same in a large scale operation. But the product recovered from the 10 cm deep basins would be algae whereas the product recovered from the 20 cm deep basins would consist of an equal mixture of algae and bacteria. Given a constant volume of waste which must be treated each day, the recovery of an exclusively algal product from 10 cm deep basins would require twice as much surface or land area than the recovery of a mixture of algae and bacteria from 20 cm deep basins. At either depth, the concentration of 1,600 mg COD/I of waste water was reduced by 80 percent, the concentration of 900 mg BOD/I by 90 percent, and 30 percent of the inorganic nitrogen in the waste was recovered as organic nitrogen in the biomass. When the retention time was 4 days instead of 6 days, the concentration of algal dry matter was reduced from 0.70 g/I to 0.45 g/I in the 10 cm deep basins and the algae contributed between 40 and 70 percent of the dry matter to the total biomass. The concentration of algal dry matter was reduced from 0.36 g/I to 0.22 g/I in the 20 cm deep basins and the algae contributed between 20 and 40 percent of the dry matter to the total biomass.

Yields of Algal Biomass Maximum yields of 10 to 12 g/m 2 day of algal dry matter were obtained in 10, 15, and 20 cm deep basins when the retention time was 4 to 6 days, the temperature of the cultures 30 to 35 C, and the mean rate of solar radiation 3,000 kcal/m 2 day. Under these conditions, Chlorella vulgaris 211/8K converted 1 to 2 percent of the total radiation into chemical energy. Although the yields in g/m 2 were similar in the 10, 15, and 20 cm deep basins at both the 4-day and the 6-day retention times, operation of the basins at a depth of 10 cm and a retention time of 6 days would be preferred, because the concentration of algal dry matter and the

247 percentage of contribution by the algae to the total biomass were higher for this combination of parameters than for the other combinations of culture depth and retention time. When the quantity of algal dry matter recovered from the waste was related to the live weight of the animals which produced the waste, maximum yields of 2 to 3 g/kg pig per day were obtained in 7.5 and 10 cm deep basins operated at 6-day and 8-day retention times.

Recovery of Nitrogen The nitrogen balance in the outdoor basins showed that 21 to 47 percent of the ammonium nitrogen available to the algae was converted into harvested biomass. The biomass contained 6 to 11 percent nitrogen on a dry matter basis. The low percentage of conversion of ammonium nitrogen into organic nitrogen was undoubtedly due in part to the need of the algae to make continuous physiological adjustments in response to the daily variations in the intensity of the solar radiation and the pH and DO content of the medium. High concentration of oxygen and pH values above 8 favor photorespiration, a process which reduces biomass production by inhibiting protein synthesis. High concentrations of OH- ions also cause the precipitation of micro- and macronutrients such as iron, magnesium, manganese, and calcium which are essential for photosynthesis and protein synthesis. It is therefore essential to control the pH, preferably in the range between 7.5 and 7.9, to improve the recovery of nitrogen from swine waste. This range of pH enhances the activity of the enzymes responsible for the fixation of CO2, in particular ribulose-1,5-diphosphate carboxylase. Control of the pH is essential for another reason: 50 to 75 percent of the ammonium nitrogen in the waste could not be accounted for in the biomass and in the effluent of the basins. This loss in N was in part attributed to volatilization as NH3 at pH values above 8, induced by the photosynthetic activity of the algae. The remainder of the N not accounted for settled to the bottom of the basins as sludge. The bottom sediment or sludge is considered a desirable source of nitrogen and other nutrients when algae are grown in dilute waste such as domestic sewage. In contrast, swine manure is a rich source of nitrogen so that the layer of sludge no longer serves a useful purpose. In fact, the sludge was found to reduce the efficiency of heat transfer from the heat exchanger pipes placed near the bottom of the basins to the culture medium and, more importantly, it served as an ideal environment for the proliferation of algal predators such as Vorticella spp. Furthermore, the sludge not only represented a loss of harvestable biomass but also an inefficient use of energy because its organic nitrogen had to be recycled again, first into inorganic nitrogen by bacterial ammonification and then back into organic nitrogen by algal and bacterial assimilation. Quality of Treated Waste Water The water quality of the liquid swine waste was measured in terms of chemical and biological oxygen demands. Rapid removal of COD and BOD occurred at retention times of 2 to 4 days. Thereafter, further increases in the retention time had only a small effect on the reduction of COD and BOD.

248 The COD of the waste pumped into the basins ranged from 700 to 4,800 mg/I and the BOD ranged from 340 to 2,300 mg/I. Algal-bacterial growth reduced the COD by 80 percent, leaving a COD of 140 to 960 mg/I in the effluent after removal of the biomass by centrifugation. The BOD was reduced by 90 percent, leaving 34 to 230 mg/I in the effluent. These levels of COD and BOD are acceptable in a system where the water is recycled for flushing. The residual COD and BOD is too high to permit discharge of the centrifuged waste water into public waters.

Stability of Algal Cultures Continuous mixing, at least during daylight hours, was found to be essential for maintaining a predominant and competitive Chlorella population. Chlorella vulgaris 211/8K was able to dominate the cultures to the practical exclusion of other algal species. Only small numbers of Euglena, Ankistrodesmus, Oscrnatoria, and Scenedesmus spp. as well as diatoms, in particular Nitzchia sp., were observed. However, an ever present threat to the dominance of the non-motile Chlorella were cells of Chlamydomonas spp. in both the motile and palmaloid stages. It was not established why the number of Chlamydomonas did not increase as long as mixing continued. The paddle wheels turned at 5 rpm, imparting the culture fluid a speed of about 15 cm/sec. When the mixing action was stopped, Chlamydomonas cells usually covered the entire basin surface in a thick layer of scum within two to three days, thus excluding most of the light from the Chlorella population below. A return to mixing would simply move the entire layer along the surface of the cultures. Attempts to remove the Chlamydomonas by skimming were found to be ineffective. The layer of cells broke up easily and the cells dispersed into the culture medium only to form another layer a few hours later. Because of the reduced availability of light and perhaps the production of antagonistic substances by the Chlamydomonas, the Chlorella population declined rapidly. The opportunistic invasion of Chlamydomonas occurred most often during the months of May and June. The organism is a common inhabitant of soil. Its prevalence during these two months may have been related to the plowing and tilling activities in the farmlands adjacent to the experimental facility. Invasion by predatory rotifers was not observed, but protozoa occurred in large numbers. Those in the circulating culture fluid consisted mostly of ciliates and flagellates which were non-predatory to the algae. In the accumulating bottom sediment, however, Vorticella, Amoeba, and Paramecium spp. were found to feed on Chlorella. Of these three algal predators, Vorticella was found to be most numerous and voracious when in the actively reproducing stage of its life cycle. It was not established why the number of Vorticella did not increase sufficiently to threaten the survival of the Chlorella population. Another threat to the stability of the algal population was the apparently random invasion of a fungus. During July, 1976, when all 12 basins were operated under identical conditions, five basins became infested with a fungus which formed star-shaped particles ranging in size from 5 to 20 mm in diameter. These particles tended to form floating patches of aggregated clumps. The infection disappeared on its own within 10 days after its onset.

249 A complete loss of the Chlorella populations was experiences at all culture depths when the basins were operated at a retention time of 6 days and received rates of solar radiation in excess of 4,000 kcal/m 2 day. Autoflocculation was avoided at a retention time of 4 days. The spontaneous clumping and settling of the algal cells was attributed to the precipitation of essential nutrient elements and changes in cell surface charges at pH values above 8.5, induced by the photosynthetic activity of the algae. The loss of Chlorella vulgaris 211/8K by autoflocculation can be prevented by controlling the pH of the culture fluid. Control should be exercised between pH 7.5 and 7.9 by CO2 injection to keep micro- and macronutrient elements available to the algae in soluble form and to maximize production of the enzyme ribulose-1,5-disphosphate carboxylase in the carbon reduction cycle for maximum photosynthesis. Although total quantities of essential elements such as iron, magnesium, manganese, calcium, and sulfur were found to be adequate in the swine manure for algal growth, the availability of these elements to the algae in a metabolically suitable form may actually be less than indicated, so that the supplemental addition of a mixture of minerals may be advisable. The loss of cultural stability due to the invasion of predators, fungi, and undesirable species of algae remains unpredictable until research determines which factors enable Chlorella, or any other chosen alga, to maintain predominance. Continuous mixing and control of pH are but two such factors. A third factor may be the recycling of a portion of the harvested algae to increase the concentration of Chlorella in the basins.

Harvesting of Algae The algal-bacterial biomass in the basin effluents was concentrated to a paste with a solids content of 15 to 20 percent on a dry matter basis. The centrifuge which was selected for harvesting the biomass is commonly used in industry to separate crystals and other heavy particles from solutions. It was acquired for the experimental facility because of its low cost of $6,800 relative to a cost of $13,000 to $15,000 for the smallest centrifuges available on the market for concentrating single cell protein. The centrifuge was adequate for harvesting the algae for feeding trials but it was found impractical for large scale harvesting. Its disadvantages include a fairly large motor of 2.2 kW, a low centrifugal force of 2,000 x G, and the necessity to stop operations periodically to scrape the algal paste from the centrifuge bowl. At a flow rate of 5 1/m in, the centrifuge removed about 75 percent of the algal cells while most of the bacterial cells passed through because of the low centrifugal force. At a flow rate of 3 l/min, approximately 85 percent of the algae were recovered. Given the highest concentration of algal dry matter produced in the outdoor basins, namely 0.70 g/I, and a cost of 50/kWh of electricity, the cost of concentrating the algae was $0.69 to $1.02 per kg. If a centrifuge specifically designed to harvest single cell protein had been available, the cost of concentrating the algae would have been $0.07 per kg algae or $0.13 per kg of crude protein. This compares with a cost of $0.50 to $0.70 per kg of soybean meal protein, F.O.B. Portland, Oregon, 1977. To be competitive with soybean meal as a protein supplement in livestock rations, the cost of producing and processing the algae, in addition to the cost of harvesting should therefore not exceed $0.40 to $0.60/kg.

250 The most efficient and reliable methods of harvesting micro-algae, but also the most expensive ones, are centrifugation and chemical flocculation. As compared to centrifugation, chemical flocculation has the disadvantage of contaminating the algal product with the coagulating chemical, usually alum. This makes the use of the algae as a feed supplement questionable because of possible toxicity effects as well as reduced digestibility and palatability. The least expensive but also the least efficient methods of harvesting algae are sedimentation, autoflocculation, and filtration through microstrainers. The efficiency of microstrainers could be improved significantly by growing filamentous algae instead of single-celled or colonial algae such as Chlorella and Scenedesmus and by increasing the concentration of the filamentous algae. Treatment ponds with continuous flow-through of waste water usually contain mixed populations of algae. At short retention times, single-celled or colonial type green algae predominate over slower growing, filamentous blue-green algae such as Oscillatoria. By lengthening the retention time and recycling a portion of the harvested algae to the pond it may be feasible to establish the slow growing filamentous algae as the predominant species over the fast growing unicellular algae. The selection of filamentous algae for the purpose of decreasing the cost of harvesting has several disadvantages, however. Filamentous algae are most often blue-green algae, many of which are known to produce potent toxins lethal to both man and animal. Strict operational controls as well as product quality controls must therefore be maintained to ensure the safe use of the final product as a protein supplement in livestock rations. Furthermore, the long retention time needed to assure the predominance of filamentous algae, as compared to the short retention time needed to maintain the predominance of unicellular algae such as Chlorella, will require a correspondingly larger surface area of the algae pond in order to treat the same volume of waste water each day. Thus trade-offs have to be considered in evaluating the cost of harvesting algae. Microstraining filamentous blue-green algae is less costly than centrifuging unicellular green algae such as Chlorella and Scenedesmus, but some blue-green algae are known to produce deadly toxins whereas green algae are not known to produce toxins and, given a fixed volume of waste which needs to be treated each day, the filamentous algae require more surface/land area for growth than the unicellular algae because of the difference in growth rates. Growing Chlorella or some other green algae and harvesting them by centrifugation may therefore be more cost effective for a large scale operation than growing filamentous algae and concentrating them by microstrainers.

Nutritional Value of Algae

The harvested biomass was freeze-dried and its nutritional value determined in feeding trials with Long-Evans rats. It was found that the raw algae had a low protein efficiency ratio (PER) of 0.84 which improved to 131 by steam autoclaving for 30 min at 120 C. The digestibility of the raw algae was 59 percent which improved to 68 percent after autoclaving. In comparison, the digestibility of soybean meal is 85 percent. Although the PER was low, when used as a protein supplement to corn, the algae gave favorable growth and feed efficiency responses. At a dietary level of 18 percent, the algal protein performed as well as fish and soybean meal at a dietary level of 13 percent.

251 Amino acid supplementation of a corn diet containing autoclaved algae, indicated that lysine was the first-limiting amino acid while methionine was present in adequate quantities. A corn-algae diet supplemented with 0.3 percent lysine gave a growth rate of 5.5 g/day, exceeding a growth rate of 5 g/day obtained with a corn-soybean meal diet at the same protein level. Because the lysine content of the raw algae was high, namely 4.83 percent on a dry matter basis, further study is needed to identify the reasons for its low availability. Lysine inactivation during heat processing and lysine "tie-up" as constituent of the indigestible cell wall are two possibilities to consider. Development of processing methods to increase protein digestibility and the availability of lysine are needed before the algae can be used successfully as a replacement for soybean meal in swine rations.

Bacterial Growth
Batch cultures of mixed populations of bacteria indigenous to the liquid phase of swine manure reached maximum concentrations in 6 h, at which time the cells could be harvested. Temperatures in the range between 15 and 35 C did not influence maximum concentrations of bacterial dry matter but determined which species of bacteria were present. Maximum concentrations of bacterial dry matter increased linearly as a function of the concentration of ammonium N in the range from 100 to 800 mg/I. The crude protein content was quite variable and often below 50 percent of the dry matter. This was attributed to the occurrence of autolysis. Maximum concentrations of bacteria in batch cultures are obtained near the stationary phase of growth when many species of bacteria undergo autolysis. The recovery of nutrients from swine waste by bacteria would be more efficient in a continuous culture system which maintains the bacterial populations at an exponential rate of growth. Autolysis would be largely avoided and a crude protein content of 50 to 75 percent of the dry matter could be expected. Yields of 0.38 g cells/g COD and yields of 2.2 g cells/kg pig per day appear feasible. The continuous culture of bacteria is probably the simplest biological method of recovering nutrients from swine waste. Many of the different genera and species of bacteria indigenous to the swine waste tolerate or can adapt to a wide range of pH, temperature, and nutrient concentrations. The need to control particular environmental and nutritional conditions is therefore reduced to a minimum and the system can be operated throughout the year. Automated systems for the continuous culture of bacteria are commercially available. However, given the genera and species of bacteria present in swine waste, it is conceivable that toxic substances and organisms which are potentially pathogenic to both man and animal can be produced. The harvested biomass may therefore have to be processed to eliminate the potential problems of toxicity and pathogenicity before the bacteria can be used as a protein supplement in livestock feed. This aspect of nutrient recovery by bacteria from swine waste needs support for further research.

252 Anaerobic Digestion Operational Characteristics The settled manure solids from 50 pigs were loaded into an underground, concrete digester with a volume of 14 m 3 at a rate of 0.96 kg VS/m 3 day (0.06 lb VS/ft3 day). The digester content was continuously mixed and maintained at a temperature of 37 C. Its pH stabilized at 7.2. The hydraulic residence time was 47 days. The digester influent had a COD/VS ratio of 1.5 and a VS/TS ratio of 0.8. Gas Production The digestion process removed 55 percent of the total solids from the swine manure. The destruction of the volatile solids averaged 56 percent, and the COD was reduced by 41 percent. These values are in general agreement with values cited in the literature for small laboratory digesters ranging in size from a few liters to several hundred liters. The daily gas production averaged 8.4 m 3 (300 ft3 ), containing 68 percent CH4 and 32 percent CO2. Based on the destruction of volatile solids, the production of biogas corresponded to 1.06 m 3/kg VS removed (17.2 ft3/Ib VS). This value is consistent with values obtained for laboratory sized digesters. Energy Requirements The energy requirements for heating the digester ranged from 13,000 kcal/day during the summer to 64,000 kcal/day during the winter. These values are greatly exaggerated because of the unusual arrangement of placing two concrete tanks side by side into the ground and connecting them to make a single functional unit, and because the top surfaces of the tanks were used as work areas and left exposed to the air without insulation.

Potential for Improving Yields of Biogas

The production of biogas may be enhanced by the following methods: 1. The loading rate may be increased from 0.96 kg VS/m 3 day to 3.8 kg VS/m3 day while decreasing the hydraulic residence time from 47 days to 10 to 17 days. 2. The production of biogas with a high fuel value is favored when the C/N ratio is near 30. Swine manure has a C/N ratio of approximately 10. By adding properly pretreated cellulosic wastes to the digester, such as grass and cereal straws as well as other crop residues, the C/N ratio of the swine manure may be raised to 30 and the yield of CH 4 by 0.44 m 3/kg of cellulose. Pure cellulose is readily digested under anaerobic conditions. However, in its natural state it is chemically bound to hemicellulose and lignin in a complex structure which is largely inaccessible to the extracellular enzymes of the bacteria in the digester. Pretreatment of the cellulosic waste is therefore necessary.

253 The practical feasibility of pretreating cellulosic wastes and adding them to manure to improve the yield of biogas remains to be determined. 3. The growth of yeast or microfungi in the liquid phase of diluted manure would generate considerable quantities of heat which could be diverted to the anaerobic digester. By coupling the heat producing aerobic fermentation of yeast or microfungi in the liquid phase of the manure with the heat requiring anaerobic digestion of the manure solids, the energy requirements normally expected for cooling of the yeast fermenter can be eliminated while at the same time the energy requirements of the digester for heating can be reduced, if not eliminated also. Most of the biogas could then be used for purposes other than heating the digester. 4. At present, the anaerobic digestion of organic wastes is practiced most often in a single vessel. However, the digestion process itself is mediated by two distinct populations of bacteria which differ from each other in growth characteristics and sensitivity to environmental stress. Separating the acidogenic and methanogenic populations of bacteria and permitting them to grow in separate reactors under environmental and nutritional conditions most suitable for each group, may optimize the anaerobic digestion of organic matter. Phase separation may be expected to provide substantial benefits, including greater removal of volatile solids and therefore higher yields of biogas as well as reduced digester volume and capital cost requirements.

Problems
An important element in the development of the anaerobic digester for farm use is the acceptance of the technology by the farmer. The probability of acceptance is increased if the digester system returns a profit, which may not necessarily be a monetary one, if the digester system is easy to understand, install, and operate, and if adequate service is available. The quantity and quality of manure available for digestion from a livestock operation or farming enterprise are likely to vary during the year. The energy needs also vary from season to season. An important research need is therefore the development of systems of waste management which aim at the full utilization of organic wastes from all sources, including manure and crop residues, and find a continuous use for the gas.

Energy Balance of Basins

Solar radiation, atmospheric radiation, back radiation, convection, evaporation, precipitation, and the addition of cold water to replace losses due to harvesting and evaporation contributed to the energy balance of the basins. Equations for each component of heat transfer were developed and programmed into a computer to predict the net energy requirements of outdoor basins under varying environmental conditions. Energy requirements measured for outdoor basins at 30 C ranged from 27.8 mcal/cm2sec in February to an average of 10.6 mcal/cm 2 sec in August. Energy requirements for cultures maintained at 15 C ranged from 9.8 mcal/cm 2 sec in February to 4.8 mcal/cm 2 sec in April.

254 The computer model predicted the energy requirements within 10 percent of the measured values. The model indicated that 73 to 115 percent of the net energy loss was due to evaporation and convection for the 30 C cultures. The relative contribution made by radiation to the total loss of energy varied from a net energy gain of 20.3 percent in July to a net energy loss of 17 percent in January. An average of 5.6 percent of the loss in energy was due to the addition of make-up water.

Methods of Nutrient and Energy Recovery The necessity to dilute the swine waste with large volumes of water in order to make the liquid phase of the manure suitable for algal growth, and the potentially high cost of harvesting and processing the algae prompted further considerations of the biological recovery of nutrients and energy from swine manure. To reduce the large volumes of dilution water, it is proposed to pretreat the liquid waste by first using it as a substrate for the growth of selected heterotrophs such as yeasts and microfungi. As an alternative to harvesting the algae by centrifugation and to avoid the potentially high cost of processing the algae to improve their low digestibility and the low availability of lysine, it is proposed to use a polyculture of fish, thereby gaining a high quality protein which is easy to harvest and already accepted as a supplement in livestock feed. An effort was made to bring the several possible bioconversion processes together into functional management systems, which have in common that the solids are treated by anaerobic digestion to recover biogas, to solubilize nutrients, and to reduce the solid waste by 40 to 60 percent. Most also conserve water by recycling the waste water to flush the animal quarters and dilute the manure. The harvested protein is recycled as a supplement in livestock feed. The proposed bioconversion methods include: OPTION A: Methane and Fertilizer The manure is digested to produce gas and a biologically stabilized waste for use as a fertilizer. OPTION B: Methane and Algae The manure is digested to produce gas and to solubilize plant nutrients. The nutrients in the liquid phase of the digested manure are recovered by algae. OPTION C: Methane, Yeast, Microfungi, and Algae The manure solids are digested to produce gas. Yeast, microfungi, and algae are cultivated in the liquid phase of the manure. OPTION D: Methane and Fish The manure is digested to produce gas. The digester effluent is used as a source of nutrients for fish.

255
OPTION E: Methane and Bacteria

The manure is digested to produce gas from the solids. The liquid portion of the manure and the supernatant liquid from the digester are treated in aeration basins, operated to maximize the production of bacteria. Architectural perspectives and plan views were developed together with schematic diagrams showing the flow of energy and materials through each system based on the feed and energy needs and the waste discharge of 100 pigs. The quantities of biogas and protein which can be recovered from swine waste are limited by the availability of carbon and nitrogen. Algae, yeast, or microfungi could recover about 5 percent of the digestible energy present in the feed or replace 30 to 60 percent of the soybean meal protein. About 90 percent of the combustible energy present in the manure could be recovered. However, yields of each bioconversion process can be increased as desired by supplying the bioconversion units with additional sources of carbon and nitrogen. Technical difficulties need to be overcome, principally the conversion of cellulosic wastes to reducing sugars, the stability of the algal cultures under field conditions, the efficient transfer of oxygen to growing cells of yeast, bacteria, or microfungi, and improvement of the anaerobic digestion process by phase separation. No new technologies need be developed.

257 REFERENCES

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