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Evaluation and Characterization of Organic Waste Products as Nutritional Sources of Bahiagrass

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EVALUATION AND CHARACTERIZATION OF ORGANIC WASTE PRODUCTS AS NUTRITIONAL SOURCES FOR BAHIAGRASS By CAROLINE BORGES REIS HAMILTON A THESIS PRESENTED TO THE GRADUATE SCHOOL OF THE UNIVERSITY OF FLOR IDA IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE UNIVERSITY OF FLORIDA 2006

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Copyright 2006 by Caroline Borges Reis Hamilton

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iii ACKNOWLEDGMENTS I would like to express my si ncere thanks and appreciati on to Dr. Jerry B. Sartain, the chairman of my supervisory committ ee, for his guidance, patience and support throughout my entire degree program. I could no t have succeeded if was not for his trust and belief in an unknown student. I also woul d like to extend my appreciation to my committee members: Dr. Jack E. Rechcigl, Dr. Martin B. Adjei and Dr. Craig D. Stanley. Sincere thanks and appreciation go to Ed Hopwood Jr., Nahid Varshovi and Martin Sandquist for all their guidance and help in all the lab and glasshouse work. Special thanks go to my husband, Kevin Hamilton, for his invaluable support, friendship and encouragement during the past ye ars. Also thanks go to my parents, Jose Gaspar Reis and Maria Alice Borges Reis, my brother and sisters, and my grandmother for all their love and support through my whole life Without their help this thesis would not be a reality today.

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iv TABLE OF CONTENTS page ACKNOWLEDGMENTS.................................................................................................iii LIST OF TABLES.............................................................................................................vi LIST OF FIGURES.........................................................................................................viii ABSTRACT....................................................................................................................... ix CHAPTER 1 INTRODUCTION........................................................................................................1 2 LITERATURE REVIEW.............................................................................................3 Manures and Biosolids in General................................................................................3 Biosolids...................................................................................................................... .5 Manure......................................................................................................................... .8 Bahiagrass ( Paspalum notatum )...................................................................................9 Organic Matter and Microbial Activity......................................................................10 Nitrogen Mineralization..............................................................................................13 Phosphorus Mineralization.........................................................................................16 Leaching of Phosphorus.............................................................................................18 Phosphorus Runoff.....................................................................................................19 Manure Management..................................................................................................21 Micronutrients.............................................................................................................23 Heavy Metal Additions...............................................................................................24 Environmental Aspects...............................................................................................28 3 MATERIALS AND METHODS...............................................................................30 Study 1: Chemically Characterize the Ma terials Relative to Their Nutritional Value and Potential Toxic Environmental Impact.................................................30 Study 2: Determination of the Minera lization Rates of the Materials........................32 Study 3: Glass House Study.......................................................................................33

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v 4 RESULTS AND DISCUSSION.................................................................................35 Study 1: Chemical Characterization of Organic Materials Relative to Their Nutritional Value and Potential Toxic Environmental Impact..............................35 Study 2: Determination of the Minera lization Rates of Study Materials...................41 CO2 Evolution.....................................................................................................44 Study 3: Glass House Study 1st Application............................................................49 Bahiagrass Dry Matter Production......................................................................49 Bahiagrass Root Dry Matter Production.............................................................49 Visual Quality Rating..........................................................................................50 Bahiagrass Chemi cal Composition......................................................................52 Leaching Evaluation............................................................................................56 Study 4: Glass House Study 2nd Application............................................................61 Bahiagrass Dry Matter Production......................................................................61 Bahiagrass Root Dry Matter Production.............................................................62 Bahiagrass Quality Rating...................................................................................63 Bahiagrass Chemi cal Composition......................................................................65 Leaching Evaluation............................................................................................69 5 CONCLUSIONS........................................................................................................74 LITERATURE CITED......................................................................................................76 BIOGRAPHICAL SKETCH.............................................................................................86

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vi LIST OF TABLES Table page 2-1 Chemical composition of the two biosolids...............................................................6 2-2 Inorganic and organic P soluble in water in different amendments.........................19 4-1 Heavy metal concentrations of bi osolids in relation to USEPA ceiling concentration limits for land appli cation (on a dry weight basis)............................40 4-2 Percentage of N mineralized in 256 days in relation to biosolids type and the resulting soil pH and EC..........................................................................................43 4-3 Analysis of variance (ANOVA) table us ed for the determination of statistical differences in the analysis for effects of biosolids and rate source on total CO2 evolved.....................................................................................................................48 4-4 Bahiagrass stolon dry ma tter production after 112 days..........................................50 4-5 Visual rating during Fall 2004 compari ng the biosolids at different rates...............51 4-6 Analysis of variance (ANOVA) table us ed for the determination of statistical differences in the analysis for effects of biosolids and rate source on total dry matter accumulation.................................................................................................52 4-7 Total N uptake by bahiagrass from biosolids in first 112 days................................54 4-8 Quantity of P applied by each material and percentage of P uptake by bahiagrass.55 4-9 Total NH4-N and NO3-N content in leachates from organic materials....................57 4-10 Concentration of NO3-N and NH4 –N in leachate as affected by the incubation period........................................................................................................................5 8 4-11 Leachate pH values from 7 to 112 days after treatment application........................60 4-12 Leachate EC values from 7 to 112 days after treatment application........................60 4-13 Bahiagrass root dry matter production 112 days after reapplication of the materials...................................................................................................................62

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vii 4-14 Visual rating during Spring 2005 compari ng N sources at different rates after biosolid reapplication...............................................................................................63 4-15 Total N uptake by bahiagrass from bioso lids in 112 days afte r reapplication of the materials.............................................................................................................66 4-16 Quantity of P applied by each material and percentage of P uptake by bahiagrass after reapplication of the materials...........................................................................68 4-17 Total NH4-N and NO3 -N content in leachates from the different organic materials after reapplication.....................................................................................70 4-18 Concentration of NO3-N and NH4 –N in leachate as affected by biosolid reapplication.............................................................................................................70 4-19 pH measurements in the leachate from 7 to 112 days after reapplication of the materials...................................................................................................................72 4-20 EC measurements in the leachate from 7 to 112 days after reapplication of the materials...................................................................................................................73

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viii LIST OF FIGURES Figure page 4-1 Data represent total nitrogen, phos phorus and potassium. A. Total Kjeldahl Nitrogen, B. Phosphorus and C. Potassium concentration of biosolids (on a dry weight basis).............................................................................................................37 4-2 Data represent total calcium, magne sium, iron and manganese. (A) Ca, (B) Mg, (C) Fe and (D) Mn concentrations of biosolids (on a dry weight basis)..................38 4-3 Data represent CO2 evolved from biosolids at weeks 1(A), 2(B), 8 (C), 16 (D), 24(E) and 33(F), respectively, at 90 Kg N ha-1. Data represent means and standard error of three replicates. Means with the same letter are not different at P>0.05......................................................................................................................46 4-4 Data represent CO2 evolved from biosolids at weeks 1(A), 2(B), 8 (C), 16 (D), 24(E) and 33(F), respectively, at 180 Kg N ha-1. Data represent means and standard error of three replicates. Means with the same letter are not different at P> 0.05.....................................................................................................................47 4-5 Data represent total CO2 evolved from different biosolids at 90 Kg N ha-1 (A) and 180 Kg N ha-1 (B). Data represent means and standard error of three replicates. Means with the same le tter are not different at P>0.05..........................48 4-6 The effect of nitrogen rate on cumu lative forage dry matter accumulation over 112 days....................................................................................................................51 4-7 The effect of nitrogen source and ra te on cumulative bahiagrass tissue dry matter accumulated over 112 days......................................................................................64

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ix Abstract of Thesis Presen ted to the Graduate School of the University of Florida in Partial Fulfillment of the Requirements for the Degree of Master of Science EVALUATION AND CHARACTERIZATION OF ORGANIC WASTE PRODUCTS AS NUTRITIONAL SOURCES FOR BAHIAGRASS By Caroline Borges Reis Hamilton May 2006 Chair: Jerry B. Sartain Co-chair: Jack E. Rechcigl Major Department: Soil and Water Science Biosolids can be used as nutrient sources in agricultural and horticultural areas. The objectives were to chemically characterize bios olid materials relative to their nutritional value and potential toxic environmental impac t; determine the mineralization rates of the materials relative to their ability to supply nut ritional fertility and compare the biosolids in a glasshouse setting relative to thei r potential for inducin g growth response of bahiagrass in a typical flatwoods soil. The mate rials used in this study were limed slurry, limed cake, black kow, black hen, disney compost, milorganite, n-viro and baltimore pellets. All materials were char acterized for N, P, K, Ca, Mg, Fe, Mn and heavy metals (Cd, Zn, Cu, Pb, Mo and As) using four replic ations. Milorganite contained the highest N content followed by Black hen and Baltimore pe llets. Limed slurry contained more than 2% N and P; limed cake, limed slurry and Nviro had adequate amounts of Ca while all the sources had adequate amounts of Mg. Milo rganite and baltimore pellets contained the

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x highest levels of Fe (39500 and 39000 ppm, re spectively) while ba ltimore pellets and black hen had the largest am ount of Mn (850 and 550, respec tively). All the biosolids tested contained EPA acceptable heavy metals concentrations. For the mineralization study, the materials (90 kg N ha-1 and 180 kg N ha-1) were mixed with an uncoated white sand (1710g) and a surface la yer of Arredondo fine sand (90 g) and placed into incubation lysimeters. A CO2 trap of 1N NaOH was placed in the head space of the lysimeters to estimate microbial decomposition rate. Limed slurry had the highest percent mineralization at both N rates, followed by Milorganite and Baltimore pellets. Based on CO2 evolved, the system was biologically active. Biosolids appear to have satisfactory nutritional values and mineralization rates to support plant growth. For the glasshouse study, bahiagrass was established by sodding in tubs using a typical flatwood soil from Pine Acres area for a growth period of 112 days. Materials were surface applied following establishment at rates of 0Kg ha-1, 90Kg ha-1 (1.74g N) and 180Kg ha-1 (3.48g N) on a dry weight basis. Amm onium sulfate was used as a mineral N source. Parameters estimated were bahiagrass root and ti ssue dry matter producti on, bahiagrass tissue chemical composition (N and P) and leachat e composition (pH, EC, N and P analysis). Biosolids induced bahiagrass forage producti on similar to AS and superior to the untreated control. Tissue N and P increased wi th increasing rates of application; limed slurry and N-viro induced N upt ake similar to AS. None of the biosolids contributed as much N to the leachate as AS. No detectab le P was found in leachates through out the study, including after reapplica tion. Therefore, biosolids used in this study have the potential for use in agriculture as a supplemental source of fertilizer and soil conditioner not representing a potential toxic environmental impact.

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1 CHAPTER 1 INTRODUCTION Manures have been used as fertilizer for centuries, but crop fertilization with manure has received renewed attention in re cent years as concern for water pollution potential from excess manure application has increased. Most people commonly view biosolids as waste materials, but they can also be used as nutrient sources in agriculture and horticultural areas .With proper treatm ent and processing, biosolids have the potential to be used as fertilizers. Biosolids are generated when solid s accumulate during domestic sewage processing. Wastewater residua ls are produced wherever th e population is concentrated enough to require a centralized domestic wastew ater treatment facility. These treatment plants continuously generate se wage sludge that must be di sposed of by one of several means. Sewage sludge becomes biosolids wh en it undergoes pathogen control treatment to meet federal and state sewage sludge regulatory requirements, followed by land application to beneficially r ecycle it. During the past decade s, some disposal practices have been restricted. Landfill space is scarce sittings of incineration facilities are difficult and certain types of surface wa ters can be adversely impacted by excess nutrient load and contaminants from treated effluents. Because of the wide range of environmental and societal problems associated with m unicipal wastewater and sludge disposal, municipalities have a need to find new ways to dispose of or, make better use of these materials. For coastal cities in particular regulations requiring elimination of ocean disposal of sludge have pr ecipitated the need for othe r management alternatives.

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2 Fortunately, these materials have the potenti al for use in agriculture. The simultaneous increase in the demand for water, coupled with tighter regulation over the safety of sludge, widens an opportunity to use treated effluent for irrigati on, and to use treated sludge as a supplemental source of fertilizer and soil conditioner. In the quest for more appropriate utiliza tion of the biosolids on agricultural lands, the following studies were conduc ted with the following objectives: 1. Chemically characterize several biosolid s materials relative to their nutritional value and potential toxic environmental impact. 2. Determine the mineralization rates of the mate rials relative to their ability to supply required nutritional fertility and/or their potential for supplying excess loading of identified hazardous heavy metals. 3. Compare the materials in a glasshouse set ting relative to their potential for inducing growth response of bahiagrass grow n in a typical flatwoods soil.

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3 CHAPTER 2 LITERATURE REVIEW Manures and Biosolids in General The most widely used organic amendments are sewage sludge and compost, which contain high organic matter, N and P, maki ng them suitable for agricultural purposes (Usman et al., 2004). Biosolids are widely used as a soil conditi oner and inexpensive source of nutrients within agriculture (Webber et al., 1996; Petersen et al., 2003). Biosolids provides short-term input of plan t available nutrients and stimulation of microbial activity, and it cont ributes to long-term maintena nce of nutrient and organic matter pools (Petersen et al., 2003). Land applic ation of the residual biosolids (sludge) from municipal wastewater treatment has in creased in recent decades to become a common disposal method in North America and Europe (National Research Council 2002). Where biosolids are used on agricultural cropland or forestland, the economical benefits of improving soil structure and addi ng nutrients for plant uptake can partially offset, the additional processing and transportation costs necessary to implement such a program (Bastian, 1986; Kelty et al., 2004). The main alternative disposal methods of landfilling and incineration have no such offse tting benefits. A third widely used disposal method ocean dumping was banned in the US in 1992 (National Research Council 2002). Biosolids application has been used more commonly in agriculture and wasteland reclamation than in forestry (Kelty et al., 2004). Manure is an important “commodity” that can enhance plant growth and reduce the necessity for application of mineral fertilizer. Manure contai ns a range of compounds that

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4 have rapid or intermediate N mineralizat ion characteristics, or that are strong immobilizers of N. Manures vary compositionally dependi ng upon source (Serna and Pomares, 1991; Van Kessel et al., 2000) and in vitro incubation experiments indicate that the organic C and N are typically less mineralizable in cattle (Bos taurus) manure than in swine (Sus scrofa) and poultry (primarily chicken, Gallus gallus domesticus) manures (Serna and Pomares, 1991; Saviozzi et al., 1993; Van Kessel et al., 2000). The dairy industry is under in creasing pressure to more efficiently use and recycle nutrients and to minimize detrimental losses to the environment. One of the critical nutrient issues involves land application manure. Over-application of nutrients can lead to contamination of surface or ground water whil e under-application results in reduced crop yield. Soils in some regions of the United St ates are replete in P and manure application is P-based in these areas, howev er, application rates have more typically been dictated by N requirements. Information required to es timate the appropriate rate of manure application includes: crop nutrient requireme nts, soil nutrient supply, and manure nutrient supply (Van Kessel et al., 2002). Manure N is an important resource that c ould be used more efficiently in crop production. However, manure contains a wi de variety of N compounds of varying complexity that makes estimation of plant N availability difficult (Van Kessel et al., 2000). Nitrogen mineralization is a relatively sl ow microbial process th at is affected by factors such as amendment composition, so il type, temperature, pH, aeration, and moisture (Mikkelsen et al. 1995; Van Kessel et al., 2002). Organic N availability is estimated to be 35%, 12% and 2% of the initia l organic N in the firs t through to the third year after dairy manure applic ation (Van Kessel et al., 2002 ). Although fixed values are

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5 generally used, estimates of mineralized organic N from cow manure in the 1st year of application are highly variab le, and range from 0% to 50% (Serna and Pomares 1991; Kirchmann and Lundvall 1993; Paul and B eauchamp 1994; Van Ke ssel et al., 2002). According to Van Kessel et al. (2002) wh en manure-amended soils were incubated for 56 days, all of the NH4 +-N was depleted; NO3 -N concentrations increased during this time period whereas NO2 -N did not accumulate in these samples and concentrations were always negligible (<1% of total inor ganic N). Overall, net mineralization ranged from –29.2% to 54.9%. But if the average mineralizability of 12.8% was used to determine crop-available N for a manure application, and the N from manure was actually immobilized, the crop w ould have insufficient N. Alte rnatively, if the true net mineralization was the other extreme, inorga nic N would be added in excess and this could have detrimental environmental and ec onomical implications. Accurate estimates of manure N composition and avai lability are an essential pa rt of an efficient nutrient utilization plan. Biosolids One main characteristic of biosolids is th at they are very variable in composition and do not form a group of fertilizers with a co nsistent nutrient content. As an example, we have the chemical composition of two diffe rent types of biosolids in Table 2-1. The fertilizer value of biosolids can be signi ficant, but varies considerably depending on origin and processing prior to application (Smith et al., 1998; Petersen et al., 2003).The pools of dissolved nutrients in biosolids are t ypically small, and plant uptake must await mineralization of organic constituents or, fo r P, dissolution of preci pitates. This makes information about the extent and temporal dynamics of nutrient release an important aspect of waste characterizat ion (Petersen et al., 2003).

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6 Phosphorus content ranges ge nerally from 34 to 57 g P2O5 Kg-1 dry matter (Sommellier et al., 1996; Quilbe et al., 2005), but this content, as well as speciation (mineral/organic form), depends on original effluent composition and types of treatment (raw, digested or composted sludges). Surface runoff is the major cause of phosphorus loss from cultivated fields, and this process was shown to be closely linked to sediment transport (Sharpley et al., 1992; Quilbe et al ., 2005). Nitrogen in bios olids is mainly in organic forms, while mineral forms are gene rally in low concentration and are mainly represented by NH4 +-N. It was shown that, in biosolids, this element can be volatilized or rapidly mobilized by runoff and leaching (Gangb azo et al., 1995; Quilbe et al., 2005). In contrast, nitrate transfers occu r only after a long-term nitrific ation process in soil (Garau et al., 1986; Serna and Pomare s, 1992; Quilbe et al., 2005). Table 2-1: Chemical composition of the two biosolids Parameter Anaerobically Digested Limed Sludge Dry matter (%) 60.4 50.5 pH 7.6 12.4 Organic matter (g kg-1 dry matter) 328 172 Organic Carbon (g C kg-1 dry matter) 164 87 C/N ratio 11.1 8.9 Total Nitrogen (g N kg-1 dry matter) 15 19 Ammonium N (g N-NH4 kg-1 dry matter) 2.4 0.2 Total Phosphorus (g P2O5 kg-1 dry matter) 78 22 Quilbe et al., 2005 Pellets are a more highly processed materi al that requires greater initial production costs but has important advantages over liqui d or dewatered forms. Most importantly, the heating process used to produce pellets s ubstantially reduces microbial populations, including pathogenic species, wh ich changes the biosolid materi al from class B to class A in US Environmental Protection Agency (EPA ) regulations (Nationa l Research Council

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7 2002). A consequence for land application is th at precaution must be taken to limit access to land for up to 1 year where class-B bi osolids have been spread, whereas no such requirement exists for applying class-A bi osolids (Kelty et al., 2004). In addition, transportation costs are lower for pellets th an for liquid sludge, because the moisture content is reduced by heat treatment to 10% or less. For use of liquid sludges (90%-95% water), the moisture content of the material is often reduced for transportation and then rewatered at the site of app lication, but the dewatered form still has a moisture content of 70%-85% (Tchobanoglous and Burton 1991; Ke lty et al., 2004). Cake (dewatered) sludges lose a substantial proporti on of their inorganic soluble NH4-N in the liquid phase during the dewatering process. Therefore, th e type of sludge, and hence the form in which the N is present in the soil, will aff ect the N mineralization dynamics of the sludge amended soil (Smith et al., 1998; Smith et al ., 2004). This is important from both the estimation of N requirement for cr op nutrition and the potential for NO3-N leaching (Smith et al., 2004). Since biosolid production is increasing, more and more a ttention is being given to anaerobic digestion as a Wa ste Activated Sludge (WAS) st abilization process. This interest is due to the capac ity of anaerobic digestion to reduce the amount of organic solids and also to the formation of biogas wh ich is a renewable energy source (Valo et al., 2004). In anaerobic digestion of WAS, hydrolysis is considered to be the limiting step. Indeed, after aerobic treatment in a wastewater treatment plant, the main part of the sludge’s organic matter is enclosed in bact erial flocs which redu ce its availability to anaerobic microorganisms (Valo et al., 2004).

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8 Manure Application of animal manures to agricultural fields is a widely used method of increasing soil organic matter and fertility (K haleel et al., 1981; Loecke et al., 2004). Most solid livestock manures can be applie d directly to crop fi elds or piled for composting. Composting manure can reduce fi eld application cost s by increasing bulk density and reducing volume. Co mposting can also increase application uniformity due to a reduction in particle size, and decrease am ounts of viable weed seeds (Wiese et al., 1998; Loecke et al., 2004) and phytotoxic substa nces (Tiquia and Tam, 1998; Loecke et al., 2004) contained in manure or manure bedding mixtures. However, with composting, there are potentially greater production and environmental costs associated with extra handling and possible lo sses of nutrients. Nitrogen losses during composting of anim al manures have ranged from 200 to 700g kg-1 of total N (Martins and Lewes, 1992; Ra o Bhamidimarri and Pandey ,1996; Eghball et al., 1997; Tiquia et al., 2002; Lo ecke et al., 2004). Lower N-use efficiency of compost is typically attributed to increased humification of the compost relative to its feedstock fresh manure (Loecke et al., 2004). Sewage sl udge stabilization through composting is a successful way to reduce the negative effect s of the unstable organic matter in the soil (Sanchez-Monedero et al., 2004; Bernal et al ., 1998a), and increase the agricultural value of the sludge as a consequen ce of the organic matter humifi cation during the process. Furthermore, the reduction in size of the wa stes during stabilization reduces the costs of transportation (Sanchez-Monedero et al., 2004). Composting manure is a useful method of producing a stabilized product that can be stored or spre ad with little odor or flybreeding potential. The other advantages of composting are that it kills pathogens and weed seeds, and improves the handling char acteristics of manu re by reducing manure

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9 volume and weight. However, composting has so me disadvantages that include nutrient and C loss during composting, the cost of land, equipment, and labor required for composting, and odor associated with composting (Eghball, 2002). Bahiagrass ( Paspalum notatum ) Bahiagrass ( Paspalum notatum Flugge.) was introduced fr om Brazil in 1914. It was first used as a pasture grass on the sandy soils of the southeastern United States (Trenholm et al., 2003). Bahiagrass ( Paspalum notatum Flugge.), a warm-season perennial, is grown throughout Florida and in th e Coastal Plain and Gulf Coast regions of the southern United States. Bahiagrass is adapted to climatic conditions throughout Florida and can be grown on upland well-drai ned sands as well as the moist, poorlydrained flatwoods soils of peni nsular Florida. In Florida, bahiagrass is used on more land area than any other single pasture species, covering an estimated 2.5 million acres. Most of this acreage is used for grazing with so me hay, sod, and seed harvested from pastures (Chambliss, 2000). Bahiagrass is relatively easy and inexpens ive to establish. It produces moderately well on soils with very low fertility, yet has good response to fertilization (Chambliss, 2000). Bahiagrass is very deeply rooted, but to lerant of flooding. It is able to maintain sod even under conditions of extremely low soil fertility and to continuously cycle nutrients while building up a store of nut rients and carbon reserves. Bahiagrass is commonly grown for a number of years prior to incorporation. During this time, it is able to accumulate a large amount of dry matter and N. Incorporation of bahiagrass sod improves soil tilth and slightly increases so il organic matter (Chambliss, 2000). It is likely that the presence of growing sod increa ses the total microbial biomass of the soil. This microbial biomass may present an a dditional source of organic N available for

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10 mineralization. Concentration of N in bahiagrass aboveground herbage varies substantially according to fertilization pr actices and generally ranges between15-25 g kg-1 (Johnson, 2004). Much of the total biomass of bahiagrass is composed of non-herbage components such as rhizomes, and roots. Th e N concentration of these parts is much lower than it is in the forage. The N c oncentration of the residue also depends on fertilization regime. Nitrogen is a limiting growth factor for many of the fora ge species grown on sludge amended soils. Much work has been undertaken to determine the nut rient benefits of sewage sludge derived N (Epstein et al., 1978; Parker and Sommers 1983; Coker et al., 1987; Smith et al., 1998; Hallett et al., 1999; Smit h et al., 2004). In particular, Smith et al. (1998) categorized different types of sludge according to whether they showed high NO3N production potential (liquid digested and lagooned liquid undigested sl udge), initial immobilization of native soil N followed by limited NO3-N production (dewatered undigested sludge), or high resist ance to mineralization and NO3-N production (air dried digested sludge) (Smith et al., 2004). The c oncentration of N will de termine both the rate and total quantity of N released. Organic Matter and Microbial Activity Franco-Hernandez et al., 2003 obs erved that the application of biosolids to soil will increase its organic matter. An increase in organic matter increases infiltration of water, increases CEC, improves soil structure and prevents erosion. Approximately 28% of organic C of the biosolids mineralized within 42 days if no priming effect was considered (Kuzyakov et al., 2000; Franco-Hernandez et al., 2003). Franco-Hernandez et al. (2003) observed that production of CO2 increased when biosolids were added to soil, and there was no difference in the production of CO2 between the different types of biosolids added

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11 to the soil. Application of biosolids treated with Ca(OH)2 and manure treatment, reduced the production of CO2 compared to the other types of biosolids treatments (FrancoHernandez et al., 2003). In order to determ ine microbial activity, Sanchez-Monedero et al. (2004) utilized the specific respiration activity, which represents the amount of CO2 mineralized per unit of biomass carbon per day, which was calculated by dividing the daily amount of CO2 evolved by the amount of biomass carbon, and expressed as mg CO2 per gram of biomass per day. This paramete r has been successfully used to detect disturbance or stress of the soil microbial biomass due to extern al inputs of organic matter or the presence of toxic substances as heavy metals that were usually reflected as an increase of specific respirati on (Sanchez-Monedero et al., 2004). Pascual et al. (1997) demonstrated that the addition of urban organic wastes (municipal solid wastes, sewage sludge and co mpost) to the soil increases the values of biomass carbon, basal respiration, biomass C/to tal organic C ratio, and metabolic quotient (qCO2), indicating the activati on of soil microorganisms (Usman et al., 2004). The organic carbon mineralization during the expe riment was recorded as cumulative CO2evolution (g kg-1 soil). The addition of sewage sludge and compost led to an increase in organic carbon mineralization compared to unamended soil (Usman et al., 2004). The cumulative amount of C mineralized increased w ith increasing applica tion rate of sewage sludge and compost. Usman et al., 2004 obs erved that the highest organic carbon mineralization was found for sewage sludge. Hi gh loss of organic C by mineralization in sewage sludge amended soil could have been due to the high microbial activity as a consequence of the high concentration of di ssolved organic C introduced with sewage sludge. Dissolved organic C is the most im portant source of energy for microorganisms

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12 (Usman et al., 2004). Whereas the evaluation of the size and activity of soil microbial biomass could give useful information about deviation of soil environment from its equilibrium, a combination of these two pa rameters as in the case of the specific respiration (qCO2), that expresses the amounts of CO2-C produced per unit biomass and time, could be a more sensitive indicator of environmental changes, both permanent (stress) and temporary (disturbance). This pa rameter has been successf ully used to detect disturbance or stress of the soil microbial biomass due to extern al inputs of organic matter (Anderson and Donsch, 1990; Sanchez-M onedero et al., 2004) or the presence of toxic substances as heavy metals (Brookes and McGrat h, 1984 and Sanchez-Monedero et al., 2004) that were usually refl ected as an increase of qCO2. Franco-Hernandez et al. (2003) observed that the higher pH in the soil at the onset of the incubation when biosolids treated with Ca(OH)2 was added might have affected microbial activity directly or indi rectly through the formation of NH3. It might be worthwhile to follow the salinity and sodicity in soil when biosolids are applied more than once. High sodicity and salinity are know n to inhibit plant growth and affect soil processes (Franco-Hernandez et al., 2003). A pplication of biosolids treated with Ca(OH)2 might have the additional effect of increas ing pH in acid soils but it might inhibit mineralization in more alkaline so ils (Franco-Hernandez et al., 2003). Microbial biomass is a dynamic component of soil organic matter and therefore of particular interests as a source of, or competito r for, plant nutrients (Petersen et al., 2003). Smith (1998) concluded that the N flow th rough the microbial biomass would in many cases be able to supply all th e N required for crop growth. Petersen et al. (2003) observed that microbes catalyze mineralization processes and therefore presumably multiplied

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13 during sludge degradation. Add ition of a fresh sewage sludge mixture caused the highest soil qCO2 increase, from 27 to 210 mgCO2 g-1 day –1, after 3 days of incubation (Sanchez-Monedero et al., 2004). After the initial increase, th e specific respiration tended towards the values of the control, indicating the pattern of the soil to recover its initial equilibrium status (Sanchez-Monedero et al., 2004). Sanchez-Monedero et al. (2004) observed that the highest qCO2 obtained for the soils ame nded with the less stabilized sewage sludge mixtures implied that th e newly developed soil microbial biomass mineralized a larger amount of the added or ganic C, per unit biomass C than the soil amended with mature compost. In fact, th e carbon mineralization during incubation of soil amended with the fresh mixture was 20% of the added C, whereas the C mineralization did not exceed 5% when the mature compost was added (SanchezMonedero et al., 2004). Nitrogen Mineralization In 1994, the U.S. broiler industry produced 7 billion broilers and generated about 10 million Mg of litter (Georgia Agricultural Statistics Se rvice, 1995), which is a mixture of chicken excreta and bedding material. A ssuming an average concentration of 30g N Kg-1, this litter contained 300,000 Mg of N, a valuable resource for fertilizing crops (Gordillo et al., 1997). In a study invol ving manures, sewage sludge, and soil amendments, Douglas and Magdoff (1991) found a good relationship (r2=0.82) between the fraction of organic N mineralized and N released by Walkely-Black acid-dichromate digestion. According to Gordillo et al. 1997, ni trogen mineralization from broiler litter started immediately after the litter was inco rporated into the soil, with inorganic N accumulating very rapidly thereafter and on av erage, approximately 50% of the total N released was mineralized during the first 24 hou rs. In addition, other easily mineralizable

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14 N compounds, such as urea and creatina, are pr esent in fresh chicken manure (Gordillo et al., 1997) and may also be pr esent in broiler litter. Composted dairy manures are a major source of N in organic fa rming where the use of manufactured chemicals is prohibited (H adas et al., 1994). Th e composting process reduces the viability of weed seeds in manures, and suppressive effects of composts on soil pathogens have also been found; thereby, the need for pesticides for the following crop is reduced. The rate of net N mineralization of compos ts in the soil must be known to optimize their use, minimize hazards of NO3 contamination, and predict the amount of additional N needed during the growth period for optimal crop production (Hadas et al., 1994). Castellanos and Pratt (1981) showed that composted manures released considerably less available N than fresh ma nures. Hadas and Portnoy (1994) showed that differences in the composition of the compos ts demonstrated the general problem that composts are not a defined product, even if they are all produced from cattle manure. Yard waste compost materials that are we ll cured during production, or that have had additional curing time in the soil, appear to be able to release N at amounts and rates similar to moderately fertile reference soils Compost amendment levels of 500 kg total N ha-1 appears to provide similar cumulative amoun ts of N release as the granitic subsoils under these experimental conditions. A pplication of about 1000 kg total N ha-1 is estimated to approach the cumulative N releas e amounts of the reference granitic topsoil. The rates of long-term N release also appro ach those of the sel ected reference soil materials, depending on compost type. Initial periods of immobilization appear to occur for uncured or fibrous yard waste compost materials. Use of such materials could potentially reduce initial plant establishm ent and growth, although such materials may

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15 function well when used as surface mulch a pplications that decompose more slowly (Claassen et al., 2004). Net nitrogen mineralization rates from co -composted materials are much higher than from yard waste composts initially, although the long-term N release rates were more similar. Sizable initial releases of available N may occur from co-composts, which may potentially allow leaching losses to wate rsheds or may facilitate weedy invasion (Claassen et al., 2004). Approximate ly 1 to 7% of the total N contained in the yard waste composts was released during the incubati on period, suggesting that the yard waste composts have the potential to continue to mineralize N fo r many years afte r application to field soils. In contrast, the co-compost materials released about 27%, providing much more short term available N, but less potenti al for continued N rel ease (Claassen et al., 2004). Immature composts may be high in ammonium, which is phytotoxic to plants. Low availability of N and other macronutrien ts limits growth, if the feedstock for composting is carbonaceous or otherwise lo w in nutrients (Barker, 2001). FrancoHernandez et al., 2003 observed that N mineralization was in creased following amendment with limed biosolids. In the past, nitrogen availability has been assessed on the basis of either anaerobic or aerobic soil incubation (Petersen et al., 2003). Nitrogen release by short-term anaerobic incubation was largely of microbial origin (Petersen et al., 2003). However, net N mineralization in laboratory systems wit hout plants may not give a quantitative estimate of plant N availability in the fiel d, where crop roots compete with microbial immobilization processes (Petersen et al., 2003) and, probably, field observations provide a more realistic picture of nutrient availa bility. The mineralization of the nitrogen

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16 contained in organic residuals depends on a va riety of factors, in cluding the type and form of the residuals. Aerobically digested sludge had higher rates of N mineralization than anaerobically digested sludge (19-50% versus 16-41%) after 16 weeks of incubation at 30 C (Adgebidi et al., 2003). Composted sewage sludge exhibits a lowe r N mineralization rate compared to noncomposted sludge. Tyson and Cabrera observed organic mineralization rates of 0.4-5.8% and 25.4-39.8% for composted and non-compos ted poultry litter, respectively, after 8 weeks of incubation at 25 C (Adgebidi et al., 2003). Nitr ogen immobilization, attributed to relatively wide C/N ratios, which occurs when the nitrogen conten t of the sludge is not adequate enough to meet the demands of the soil microbial community, has also been reported for composts (Adgebidi et al., 2003). As the decomposition of the sludge occurs, carbon evolved as carbon dioxide (CO2), and the C/N ratio is gradually diminished, enhancing nitrogen availability. The physical and chemical conditions prevailing in a decomposition medium also affect its minera lization rates in poultry litter (Adgebidi et al., 2003). Phosphorus Mineralization Most Florida forages are grown in sandy, infertile soils, and many mineral deficiencies observed in grazing species can be related to the characteristics of soils of the region (Tiffany et al., 2001).Municipal biosolid s (sludge) have been used as fertilizer for some time often due to their high phosphorus (P) content, which promotes growth of agricultural crops (Tiffany et al., 2001). Mineral uptake by plan ts is affected by biosolid load rate and chemistry, soil type and pH and plant genetics. (Tiffany et al., 2001). Today, dried, heat treated, anaerobically di gested, pathogen-free, biosolids are widely

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17 marketed. These biosolids contain varying amounts of mineral elements which could prove beneficial, if they improve soils and forage nutritive value without creating toxicity. (Tiffany et al., 2001). Approximately 1.85 x 108 Kg of P enters the environment in the form of animal manure per year in the USA (Wodzinski and Ullah, 1996; Parham et al., 2002). Sound waste management in ecosystems or waters heds demands prudent consideration of P inputs (Pierzynski et al., 1994) Unfortunately biosolids a nd manure-borne P reactions have not received nearly the study that wast e-bound N has. We do know that (as with N) calculations based on total waste-P concentrations to determine allowable P loads to a soil can be grossly over-conservative, as al l the (total) P may not be bioavailable or soluble enough to leach. Phosphorus in biosolids and manures ex ists in a variety of forms (e.g. McCoy et al., 1986; Pierzyns ki et al., 1994) resulting in reported bioavalia bilities (to plants) ranging from 10% to 100% of that of soluble fe rtilizer P (e.g. de Haan, 1980; Gestring and Jarrel, 1982; Si kora et al., 1983; MacCoy et al ., 1986). Most agricultural best management practices for animal wa stes are now based on providing sufficient nitrogen (N), in a timely manner, to meet crop N requirements at a realistic yield. Documented problems with nitrate-N (NO3-N) contamination of ground waters in areas with high animal densities have been the driving force behind this approach. However, a number of emerging environmental issues, such as the eutrophication of surface waters by phosphorus (P) in runoff; the fate of trace el ements, antibiotics, pesticides, and growth hormones in wastes; and the effects of pathoge ns in wastes on human and animal health have forced us to re-evaluate the N-based ma nagement of animal wastes (Sims and Wolf,

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18 1994). Franco-Hernandez et al., 2003 observed that phosphorus mineralization occurred but the application of biosolids did not increase rate of mineralization. Inorganic phosphate (Pi) partakes in many abiotic processes, such as sorption to soil particles and precipitation. Phosphorus is also an essential nutrient and as such is involved in biological reactions in the soil (Petersen et al ., 2003). Biotic and abiotic processes occur simultaneously and interactively, and both as pects should be considered in discussions of soil P dynamics (Petersen et al., 2003). The concentration of inorganic phosphate in the soil solution is low and crops to a large extent draw upon P associated with the solid phase, which complicates the assessment of P availability to the crop (Petersen et al., 2003). Petersen et al. (2003), observed that inorganic N accounted for only a small fraction of total N in both sludges (anaerobically di gested sludge and activate sludge), and the crop N uptake obs erved thus implies a significant net N mineralization. The continued application of fertilizers and manures in many areas has resulted in the buildup of soil P concentrations above those required for optimum plant growth, because of these elevated concentrations the po tential for P loss is increased. This loss is a function of, but not exclusively, topogr aphy, soil type, soil test phosphorus (STP) concentration, and soil hydrol ogy (McDowell and Sharpley, 2 001). Both the properties of the biosolids (P forms, solubility) and th e soil being amended (P fixing capacity) determine the potential environm ental hazard of such accumulated P (Anjos et al., 2000). Leaching of Phosphorus Areas with intensive livestock farming ofte n have soils enriched with P due to the addition or disposal of manure (Withers et al., 2001). In areas of Europe and North America, P-rich soils are common (Fixe n, 1998; Skinner and Todd, 1998). Until about 15

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19 years ago, there was a common misconception within the literature that P loss via subsurface pathways was negligible as P is st rongly sorbed to soil and thus, only surface pathways were of concern (Sharpley and Me nzel, 1987). However in free-draining soils and those where drainage is enhanced, P can be lost in environmentally significant quantities (Heckrath et al., 1995; Hesketh a nd Brookes, 2000). A comparison of inorganic and organic P soluble in wate r in different amendments is showed in table 2-2: Table 2-2: Inorganic and or ganic P soluble in water in different amendments Fraction From Dairy manure P (%) Dairy compost P (%) Poultry manure P (%) Poultry compost P (%) Superphosphate P (%) Water Inorganic 51 15 26 21 100 Organic 12 1 8 1 ---Total Inorganic 63 92 84 87 100 Organic 25 5 14 11 ---(Derived from Macdowell, R.W. and A.N. Sharpley. 2003) MacDowell and Sharpley (2003) noted that for high-flow lysimeters, more P was leached from dairy compost and poultry manure amended soils compared with mineral superphosphate and dairy manur e. In both cropped and grassl and situations, it has been shown that increasing soil P is associated w ith higher concentrations of P in drainage water than in low P soils (Sharpley et al., 1996; Sims et al., 1998; Sh arpley et al., 2000). Measures to reduce the potential for P enri chment to runoff include manure treatment, composting, peletizing, and transport to areas with a nutrient defic it (Sharpley et al., 1998; Sharpley et al., 2000). Phosphorus Runoff Phosphorus loss in runoff can cause eutrophi cation of streams and lakes resulting in surface water quality problems. An important component in managing P losses to the environment is understanding crop P uptake fr om various nutrient sources. Plant P

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20 availability may differ from one organic P source to another as a result of different origins (e.g. municipal waste, dairy, swine, poultry) or variations in management (e.g., storage time or temperature, chemical tr eatments, manure separation, inclusion of bedding) (Ebeling et al., 2003). Knowing th e differences in P availability among P sources will guide application rate decisions so that crop P requirements are satisfied while minimizing soil P accumulation and runoff losses (Ebeling et al., 2003). Beneficial use of biosolids as a soil amendment is based on their potential to positively alter soil properties such as plant nutrient availability, water holding capacity, tilth (physical conditions of so il related to tillage, seed bed and rooting media), and cation exchange capacity (related to enhanced soil organic matter status) (Camberato et al.,1997). However, due to their high nutrient content and other chemicals such as trace elements, sewage sludges may create potential health and environmental problems that might question their use in agriculture. In particular, mobilization of P and N in runoff after sludge spreading is likely to contribu te to eutrophication of downstream surface waters (Quilbe et al., 2005). The wastewater treatment process involves a primary settling step to separate heavy organic solids from sewage. A secondary biol ogical process uses cycling anaerobic and aerobic conditions to remove P from plant effluent. The solids from primary settling and the waste biomass from secondary treatment ar e digested anaerobically to decompose and stabilize the solids. The dige sted solids are thickened by us e of a gravity belt thickener and organic polymer addition prior to being re cycled to agricultural land (Ebeling et al., 2003).

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21 Data from a greenhouse and field stu dy conducted by Goss and Stewart (1979) indicated that feedlot manure produced hi gher crop utilization efficiency than superphosphate fertilizer. They suggested that the microbial activity in the soil increased manure P availability, reduced luxury P cons umption by plants, and extended the time period of adequate P availabili ty levels (Ebeling et al., 2003). Ebeling et al.(2003) studies indicated that P sources with relatively high soluble P or bioavailable P concentrations (CaHPO4 and biosolids) provided hi gher levels of P for plant uptake in poorly buffered systems such as sand media. Manure Management Manure management recommendations must account for site vulnerability to surface runoff, macropore leaching, and erosion, as well as soil P content and saturation, because not all soils and fields have the sa me potential to transfer P by surface runoff and leaching to surface waters. As a result, thres hold soil P levels should be indexed against P transport potential with lower values for P s ource areas than for area s not contributing to water export (Sharpley et al., 2000). When applied to the soil, inorganic P (M CP and DCP) and ammoniated phosphate remain localized around the point of applic ation unless cultiva tion or other mixing processes occur (James et al., 1996).Kissel et al. (1985) described the chemistry of P in calcareous soils as a continuum of ad sorption, formation of amorphous calcium phosphate compounds, and gradual formation of crystalline phospha te compounds (James et al., 1996). In soils exposed to manure ove r extended periods, P enrichment occurs below the surface 20-cm layer (James et al., 1996). Dalal (1977) concluded that the accumulation of organic P in soils was primar ily a result of microbial activity and the organic P may become available to plants by mineralization (James et al., 1996). James et

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22 al.(1996) concluded that longtime manure disposal on land increased the concentration of extractable inorganic P in subsoil layers as deep as 210cm and that manuring increased extractable organic P concentrations markedly in the 0 to 30 cm layer but this organic P dissipated to background levels within 2 to 3 years after manuring ceased. According to Parham et al (2002) animal manure-P is rela tively more mobile but less available for plants than inorganic fertiliz er-P. Long-term application of cattle manure did not result in excessive accumulation of P in the surf ace 0-30cm soils, but promoted microbial activities and P cycling in soil. James et al. (1996) observed that or ganic P and inorganic P from heavy manuring do not threaten soil or groundwater quality under deep calcareous soils if manure application is limited by acceptable N-loading rates. Wastewater treatment and sludge processing methods marked ly influence biosolid s P mobility (Elliott et al., 2002). Lu and O’Connor (1999) observed that sludge materials when treated with Fe and Al to remove P from the corresponding liquid wastes, there by containing total Fe and Al of 10% or more may increase soil P so rption when applied to the land, especially when the sludge is applied to soils with lo w native P adsorption capacity such as Myakka sand soils. Lu and O’Connor (2001) observed th at increases in P sorption were correlated with increases in oxalate-extractab le Fe and Al contents of amended soils. Although temporary, the increased retention of P effected by biosolid s applications can have important implications. Phosphorus in bi osolids containing (or tailored to contain) abundant Fe and/or Al can be expected to be have as a slowly available P source, and to be less subject to excessive leaching losses than completely soluble sources. This can be very important in many areas of Florida domin ated by soils that sorb P poorly and allow extensive P leaching.

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23 Soils common to most of the USA contain sufficient P-sorptive capacity (Al and Fe oxides) to prevent P leaching and to mask i nherent differences in the P solubilities of biosolids materials. Recent Florida legislation require s P-based biosolids application rates in watersheds associated with P-se nsitive water bodies (Sec. 373.4595 FL Statutes). Compared with N-based nutrient management, a P-based approach dictates lower waste application rates (Elliott et al., 2002). Eghball (1999) found that application of beef cattle feedlot manure or compost increased the soil surface (0-15cm) pH while N application as NH4NO3 reduced the pH (from 6.4 to 5.6). The increase in soil pH with manure and compost application was attributed to a beef cattle diet that cont ains approximately 15g CaCO3 kg-1. Application of manure with an increased N: P ratio at rates based on crop N requirements would more closely match crop up take of P and reduce the potential for soil P accumulation (Sharpley et al., 2000). Controls over agricultural P inputs are required to prevent soil P build-up, causing increased background losses; better management of fresh P inputs is required to avoid in cidental losses of P that o ccur when storm events follow applications of manures to th e soil surface and more precis e land management is required on naturally dispersive soils in runoff producing areas (Sharp ley et al., 2000). Micronutrients Moral et al. (2002) studies i ndicated that sewage sludge ap plications can be useful to increasing the available micronutrients (Fe, Mn, Cu and Zn) concentrations in calcareous soils. They suggested that metal ava ilability in the calcareous soils was closely related to texture due to th eir influence in the organic matter dynamics. The dynamics of the metal fraction are dependent upon the stab ilization of the organic matter (composted vs non-composted), rate of amen dment applied (non lin ear increments with rate), and also

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24 the texture of the soils (loamy vs sandy soils) (Moral et al., 2002). In addition, Kuo (1990) observed that sewage sludge mineralization incr eased the soluble organic compounds, especially low molecular weight or ganic acids in soils, that could mobilize not only the exogenous micronutrien ts from the sewage sludge but also the non-available metal fraction in the soil. Heavy Metal Additions Unlike sewage sludge application, where land application is limited based on allowable metal loadings by EPA 503 re gulations (U.S. EPA, 1993), regulations governing livestock and poultry manure by-produc ts are generally governed at the state level, based on total N and/or P loading (B olan et al., 2004). The U.S. Environmental Protection Agency (EPA) Part 503 rule regulates nine metals in sewage sludge that is to be land applied. These include As, Cd, Cu, H g, Mo, Ni, Pb, Se, and Zn (U.S. EPA, 1993). U.S. EPA Part 503 risk assessment indicates th at other metals do not pose potential health or environmental risks at land a pplication sites at typical appl ication rates (Bolan et al., 2004). It has often been observed that metals added through organic amendments, such as sewage sludge and manure by-products, accu mulate in the surface layer, indicating a strong retention by the amended soil (Adriano, 2001; Bolan et al., 2004). Distribution of metals in sewage sl udge and manure by-products and their subsequent redistribution in soils amended with these materials have generally been investigated by selective seque ntial chemical extraction tech niques (Bolan et al., 2004). The principal metal forms in manure and manur e-treated soils are soluble, exchangeable, adsorbed, organic-bound, oxide-bound, and precipitated (Bolan et al., 2004). Concentrations of heavy meta ls recovered from dry biosolids were lower than 100 mgKg1 except for zinc (Zn) and chromium (Cr) on materials treated with lime. (Franco-

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25 Hernandez et al., 2003). Applicat ions rates to soil should be limited so that heavy metals do not accumulate and end up into the food ch ain or groundwater, that pathogens can not infect people or that concentrations of toxic compounds become dangerous (FrancoHernandez et al., 2003). Whereas the risk asso ciated with the presence of heavy metals and persistent organic pollutants can only be controlled by limiting the amounts of sludge applied to the soil, the amount of pathoge ns can be reduced by sludge stabilization treatment prior to addition to the soil (Sanchez-Monedero et al., 2004). Sanchez-Monedero et al. (2004) observed th at the bioavailable DTPA-extractable Cu in the soil amended with any of the orga nic mixtures (38% sewa ge sludge + 62% of cotton waste in four composting periods: initial, during the thermophilic phase – after 21 days of composting, the end of the active phase – after 49 da ys of composting and mature compost – after 109 days of composting) wa s on average 14% lower in the soil amended than the control soil, indicating a positiv e effect of the organic matter on the immobilization of this heavy metal in the so il. The opposite effect was shown for Zn, as all the soils amended with the organic mixt ures showed higher c oncentration of the available metal (on average about 39% higher than the control soil). They also observed that the amount of heavy metals in both soil and organic materials were under the maximum levels accepted by the current legisl ation. The increase in total metal content following a single sludge addition was negligible (<1 mg kg-1) for Cu, Ni, Cd, Cr and Pb). In the case of Zn, amendment of soil w ith the mature compost caused an increase of total content of 2.9 mg kg-1. Organic wastes such as sewage sludge and compost increase the input of carbon and nutrients to the soil. However, sewa ge sludge-applied heavy metals, and organic

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26 pollutants adversely affect soil biochemical properties (Usman et al., 2004). It is evident, that the heavy metals introduced with sewage sludge or compost cause accumulation of organic matter and decrease the turnover rate of organic matter, presumably because of inhibitory effects on the microbial biomass (Usman et al., 2004). Moreno et al, (1999) found that the addition of cadmium-contaminat ed sewage sludge compost to the soil decreased microbial biomass C and stimulated the metabolic activity of the microbial biomass (Usman et al., 2004).The addition of sewage sludge to soils could affect the potential availability of heavy metals (U sman et al., 2004). Mineralization of sludge organic matter may release heavy metals into more soluble forms that may harm sensitive crops and microbes (Usman et al., 2004). The ava ilability of heavy metals in the sewage sludge and soils treated with sewage sludge depends on many factors such as the properties and amount of heavy metals, the partitioning of he avy metals between solution and solid phase, and soil characteristics (Usman et al., 2004). It is widely known that the bioavailability of heavy metals in soil is strongly influenced by the amount and the quality of organic matter that can react w ith the heavy metals, forming complexes and chelates of varying stability (Usman et al., 2004). Soil organic matter is quite effective in retaining heavy metals. Heavy metal-organic associations can occur both in soil solution and at the solid surfac es of either native soil constituents or any added material (e.g., Biosolids) (Usman et al., 2004). In a hea vy metal-polluted soil, Kiikila et al. (2002) studied the effect of biosolids as organi c immobilizing agents and observed that the exchangeable Cu concentration decreased (U sman et al., 2004). On the other hand, the heavy metal could be complexed by dissolved organic matter (DOC), which enhances leaching in the field (Usman et al., 2004).

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27 Numerous laboratory and fields studies have demonstrated that most trace elements (including common industrial metal elements) are relatively immobile in soils and once added will remain in the layer of incorporati on. Therefore, problems associated with trace elements entering ground water beneath land where sludge has been applied is unlikely (Yingming and Corey, 1993). Results of field studies demonstrated that crops grown on sludge-amended soils benefited from the plant nutrients in sewage sludge, but would not accumulate As, Cr, Cu, Pb, Hg, Ni, or Zn in amounts sufficient to be harmful to consumers. Depending upon the soil conditions, however, there was a potential, for Cd, molybdenum (Mo), and Se to be taken up by cr ops in amounts harmful to humans (Cd) and animals (Cd, Mo, and Se) who consumed the affected harvest (Logan and Chaney, 1983). Cadmium is one of the most controvers ial of the contaminant elements. Unlike Cr, Cu, Zn or Se it plays no known role in plan t or animal metabolis m (Zhang et al., 2000). Neither total nor available heavy metal con centrations were increased in the soil by sewage sludge compost application, in fact the composting process reduces the heavy metal availability in the raw material, po ssibly due to adsorpti on on or complexing by humic substances (Korboulews ky et al., 2002). Available B is strongly associated with organic matter in soil. Highly organic materi als like compost may have a large portion of the B they contain in the available and pot entially toxic forms (Zhang et al., 2000). Copper sulfate (CuSO4) is a potentially beneficial feed additive in swine ( Sus scrofu domesticus ) and poultry ( Gallus gallus domesticus ) production, but while supplementing copper (Cu) levels in swine pr oduction, concern has been expressed over elevated Cu concentrations (up to 1000 mg Cu kg-1) in the manure resulting from this practice (Miller et al., 1986) Studies using sequential me tal extraction techniques on

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28 sewage sludges and Cu in manures have s uggested sulfides, carbonates, and organic complexes of Cu as major chemical forms (Miller et al., 1986). Calibration of sequential extractants against standard Cu minerals allowe d Stover et al. (1976) to infer that sulfides may be an important Cu form in sewage sludge (Miller et al., 1986). Miller et al. (1986) concluded that Cu in swine manure is prim arily bound to organics contrary to earlier findings, and that sludges also contains predominately alkali-soluble Cu, although differences in origin and composition of th e waste have been shown to be important. Miller et al. (1986) further c oncluded that drying of manure prior to chemical extraction substantially increases th e solubility of Cu. Environmental Aspects Continuous heavy manuring may pose envi ronmental problems, such as odor, pollution of ground and surface water due to leaching and run-off of organics and nutrients, and soil accumulation of heavy meta ls (Hsu et al., 2001). The composting of organic wastes is mainly a biological decomposition pro cess of organic materials, resulting in a net loss of total organic ma tter (OM) and a concen tration of inorganic constituents (Hsu et al., 2001).The main pr oducts of the composting process are fully mineralized materials such as CO2, H2O, mineral ions, stabilized OM (mostly humic substances), and ash. Composting reduces the volume and weight of the raw material resulting in a stable product that can be applied to soil as a valuable fe rtilizer and soil amendment (Hsu et al., 2001). In cases where th e potential toxic metal concentrations of compost are high, the leachability of metals associated with compost is of concern. Studies have shown that the chemical form of an element is more important than the total concentration in determining its leachabil ity into ground water (H ung Hsu et al., 2001). According to Hung Hsu et al. (2001), during composting, the major portions of Cu, Mn,

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29 and Zn were in the organically-bound, solid particulate and or ganically complexed fractions, respectively. Metal dist ributions in different chemical fractions were generally independent of composting age and, thus, independent of respective total metal concentrations in the composts. Jackson et al. (2003) obser ved that trace elements Ni, Cu, and As were found to be readily soluble in poultry litter and that mo re than 70% of Ni and Cu was in the cationic form, or bound in re latively labile complexes that dissociated to the cationic species, therefore trace metals from poultry litter are e xpected to be readily sorbed by soil mineral phases on land app lication of poultry litt er. It is becoming increasingly important for producers to clos ely monitor manure nutrient application and adjust fertilizer application based on manure inputs (Van Kessel et al., 2000).

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30 CHAPTER 3 MATERIALS AND METHODS This research consisted of three separate studies conducted at the University of Florida, Gainesville, Florida. The firs t study was conducted on an array of sludge materials to determine their nutritional value and potential toxic environmental impact in the spring of 2004 from February through Ap ril. The second study was conducted in the laboratory on the same sludge materials to determine the mineralization rates in the spring of 2004 from April through December. The third study was conducted in a glasshouse on bahiagrass established from sod during the fall of 2004 from September through January 2005. A second growth respon se evaluation was conducted during the spring of 2005 from March through July to doc ument the nutrients build up in the soil and to assess their nutrient availability to bahiagrass. Study 1: Chemically Characterize the Mate rials Relative to Their Nutritional Value and Potential Toxic Environmental Impact. This portion of the project the organic ma terials were chemically characterized for the following elements: N, P, K, Ca, Mg, Fe, Mn and heavy metals (Cd, Zn, Cu, Pb, Mo and As), using four replications for each material. The materials evaluated were: Limed Slurry (Anaerobically digeste d, lime stabilized slurry bios olid (Class B)); Limed Cake (Lime-stabilized cake bioso lids (Class B)); Black Kow (Composted cow manure (Class A)); Black Hen ( Composted Poultry manur e (Class A)); Disney Compost (Composted biosolid) (Class A)); Milorgan ite (Anaerobically digested sewage sludge (Class A));

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31 Tarpon Springs N-Viro (Aerobically dige sted (Class A)); Baltimore pellets (Anaerobically digested sludge (Class A)). Materials were prepared for analysis for Ca, Mg, K, Fe, Mn and P using the ashing method described by Hanlon et al. (1994). Five grams of the dried material were placed in 50ml pyrex beakers. Samples were ashed in a furnace at 550C for 5 hours. Samples were then moistened using a small amount of deionized water and 3.0 M Nitric acid. They were then placed on a hot plate and dried. Samples were placed back in the furnace and heated at 450C for an additional 10 minut es. Samples were allowed to cool and 1.0 ml of 5.0 M HCl was added. The sample was scraped from the beaker using an angleedged rubber policemen, decanted into a 50 ml volumetric flask, and diluted to 50 ml using deionized water. Calcium, Mg, K, Fe and Mn were determined using a Varian Atomic Absorption Spectrophotometer (Varian Analytical Inst ruments, Sugar Land, TX). Phosphorus was determined using the Molybdate Blue method (Hanlon et al., 1994). Nitrogen was determined by TKN, Total Kjeldahl Nitroge n, method described by Horneck and Miller (1998). A two grams tissue sub-sample was place d in a 50 ml digestion tube to which 1.5 g of digested catalyst (Kel –Pak 1.0g K2SO4, 0.3g CuSO4) and 3mL of 35N H2SO4 were added. The mixture was mixed using vortex sh aker and heated on a Tecator Digestion System 40-1016 Digestor (Tecator AB, Hoga nas, Sweden) at 370C for 3 hours. Once the solution cooled, it diluted to 50 ml usi ng deionized water. Total N was determined using Alpkem RFA-300 auto analyzer. For h eavy metals the materi als were digested according to Jackson (1958) as presented by Fiskell (1965). One half gram of the materials was transferred to a digestion tube 25 ml of water and 2ml of concentrated

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32 HNO3 was added and evaporated to dryness on a hot plate. Ten ml of 30% hydrogen peroxide was added and the solution was ev aporated to near dryness at 90C. This treatment was repeated until no effervesces occurred on addition of H2O2. Three drops of concentrated H2SO4 and 10 ml of HF were added. Sa mples were placed on the block digester (200C) and the solution evaporated to dryness. Finally, 15 ml of concentrated HNO3, 2 ml of concentrated H2SO4, and 5 ml of HCLO4 were added and samples were continuously heated until strong fumes of SO3 were produced. Samples were allowed to cool, 25 ml of water was added, the sides of th e containers were rins ed and policed with a polyethylene rod to complete the extraction of the metals. Solution was transferred to a 50 ml volumetric flask and diluted to volum e (Page et al., 1982). Copper and Zn were determined by atomic absorption and Cd, Pb, Mo and As were determined by ICP (Page et al., 1982). Study 2: Determination of the Min eralization Rates of the Materials The mineralization rates of the materials was determined in order to evaluate their potential for supplying nutrients over time. In cubation lysimeters (30 cm section of 7.5cm diameter PVC tubing fitted with fiberglass mat across the lower e nd, held in place by a PVC cap) were used to determine the mineralization rates. The materials were applied at rate equivalent to 90 kg N ha-1 and 180 kg N ha-1 respectively, on a dry weight basis. Uncoated white sand (1710 g) and a surface layer (0 to 5cm depth) of Arredondo fine sand (90 g) we re mixed with the materials and placed in the incubation lysimeters. The mixture was br ought to 10% moisture (approximately 80% water holding capacity). A CO2 trap of 1N NaOH was placed in the head space of the incubation lysimeter to estimate mi crobial decomposition rate. The CO2 trap was replaced and titrated with 1N HCl weekly to estimate CO2 evolution. After 7, 14, 28, 56,

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33 84, 112, 140, 168, 208 and 263 days each lysimeter was leached with 0.02 N citric acid and ammoniacal-N, nitrate-N and P were dete rmined on the leachate. Lysimeters were maintained at near optimum moisture, but to induce leaching an additional pore volume of excess water (ca. 500 ml) was a dded every leachate day for a total of 10 leachates. These leachates were designed to represent the natural removal of the mineralized N. The leached volume was r ecorded and an aliquot was taken for NH4 NO3-N and P analysis (Hor neck and Miller, 1998). Study 3: Glass House Study This study was set up to compare the biosolid s in a glasshouse setting relative to their potential for inducing grow th response of bahiagrass in a typical flatwoods soil. The study was conducted at the University of Fl orida’s Turfgrass Envirotron, Gainesville, Florida. Bahiagrass (Paspalum notatum) wa s established by sodding during the fall of 2004 in a glass house in tubs (0.19m2) using a typical flatwood soil from Pine acres area (Candler fine sand – sandy, siliceous, uncoa ted, hyperthermic Typic Quartzipsamments) for a growth period of 112 days. Materials (L imed cake, Limed slurry, Disney compost, Baltimore pellets and N-viro) were surface applied following bahiagrass establishment at rates of 0 kg N ha-1, 90 kg N ha-1 (1.74 g N) and 180 kg N ha-1 (3.48 g N) on a dry weight basis. A standard inorganic N source, Ammonium sulfate, was used for comparison. The parameters estimated were: bahiagrass root dry matter production, bahiagrass quality rating, bahiagrass chemical composition a nd leaching evaluation (pH, EC, N and P analysis). The experiment was arrang ed in RCB design with 4 replicates. Glasshouse conditions were maintained rela tively constant by means of computer integration of overhead fans, an exhaust fan, a wall length humidifier, and automatic roof window. Leachates were collected after 7, 14, 28, 56, 84 and 112 days. Clipping for dry

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34 matter production were taken as needed for a total of 7 collections. Stolon samples were collected at termination and washed free of soil. Tissue and stolon samples were dried at 70oC for 48 hours. Tissue samples were ground to 2 mm mesh and ashed (Hanlon et al., 1994c). Samples were analyzed for P by spectrophotometry using the molybdenumreduced molybdophosphoric blue color method, in a sulfuric acid syst em (Hanlon et al., 1994a). Leachates were collected at 7, 14, 28, 56, 84 and 112 days. The leachate volume was recorded and an aliquot was taken for pH, EC, P, NH4 and NO3 analysis (Hanlon et al., 1994). A second growth response evaluati on was conducted during the spring of 2005 from March through July to document the nutrient s build up in the soil and to assess their nutrient availability to bahiagrass. The same t ubs with the established stand of bahiagrass were employed. Statistical analysis for all st udies was performed using SAS for analysis of variances (SAS Institute, 1987). Duncan’s Multiple Range Test (DMRT) was used to identify differences among means.

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35 CHAPTER 4 RESULTS AND DISCUSSION Study 1: Chemical Characterization of Organic Materials Relative to Their Nutritional Value and Potential Toxic Environmental Impact. Most biosolids contain 40 g kg-1 to 70 g kg-1 of N, 1 to 20 g kg-1 P2O5 and very little K 0.001 to 10 g kg-1. The stabilization process used to treat wastewat er alters the nutrient composition of the resulting bios olids (Muchovej et al., 2001). In this study Milorganite contained the highest N content (59 g kg-1) followed by Black hen and Baltimore pellets with 31.5 g kg-1. Limed slurry contained 23 g kg-1, Disney compost 16 g kg-1 and Limed cake 12.9 g kg-1. The materials N-viro a nd Black kow contained the lowest content of total kjeldahl nitrogen (TKN) with 1.8 and 3.5 g kg-1, respectively (Fig 4-1A.). Concentrations of 16 g Kg-1 N are usually sufficient to increase mineralization under aerobic conditions (Sartain, persona l communication). Nitr ogen availability depends largely on microbial activity which in turn depends on soil conditions (e.g., moisture and temperature) and the nature of the biosolid. Conditions favorable for conversion of N from organic to inorganic forms are roughly the same as those favorable for crop growth. For waste materials, it is generally estimated that 40 to 50% of the organic N will become available during the year following application (Kidder, G., 2001). Black hen contained the hi ghest P content (27.3 g kg-1) followed by Baltimore pellets (25.6 g kg-1), Milorganite and Limed slurry (22 g kg-1), Disney compost (11.7g kg1) and Limed cake (9.1g kg-1). Black kow and N-viro had the lowest P content, 3.7 and

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36 2.6g kg-1, respectively (Fig 4-1B.). Materials were generally very low in total K, ranging from 5.6 g kg-1 (Limed slurry) to 0.7 g kg-1 (Baltimore pellets). The only exception to this trend was Black hen which contained 46 g kg-1 of K (Fig 4-1C.). The materials Limed cake, N-viro and Limed slurry contained 2.4, 1.2 and 1.1 g kg-1 of Ca and 6.9, 6.5 and 6.6 g kg-1 of Mg, respectively, suggesting that th ese materials could be used as liming sources (Fig 4-2A. and 4-2B.). Waste lime is ge nerally fine and reacts rapidly with soil, giving quick increases in soil pH especially if cultivated. These materials can often be obtained at little or no cost to the farmer or rancher (Ki dder, G., 2001). Muchovej et al., 2001, suggested that lime stabilized bioso lids contained lower N, P, and metal concentrations, but higher Ca concentrations than digested biosolids, due to the large amount of lime added to the material. Milorganite (39.5 and 0.21 g kg-1), Baltimore pellets (39 and 0.9 g kg-1) and Disney compost (14.5 and 0.9 g kg-1) contained the highest concentrations of Fe and Mn, respectively (Fig 4-2C. and 4-2D.). Iron and Mn (hydrous oxides forms) adsorb metals in the biosolid and in amended soils, thus being able to reduce phytoavailability of many trace elemen ts (for example Cadmium). Nitrogen and P receive the most attention in management of organic wastes because of their importance in both plant nutrition and water contamination. However, all the other nutrient elements are also returned to the soil in organic waste applications and can c ontribute to improved crop performance (Kidder,G. 2001).Based on th eir N and P compositions, these biosolids can be used as low-grade N and P fertilizer and as a source of calcium, especially the lime-stabilized residuals.

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37 A. 015304560Limed slurry Limed cake Black kow Black hen Disney compost Baltimore pellets N-viro Milorganite Total Kjeldahl Nitrogen (g Kg-1) B. 0102030Limed slurry Limed cake Black kow Black hen Disney compost Baltimore pellets N-viro Milorganite Total Phosphorus (g Kg-1) Fig 4-1: Data represent to tal nitrogen, phosphorus and pot assium. A. Total Kjeldahl Nitrogen, B. Phosphorus and C. Potassium concentration of biosolids (on a dry weight basis).

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38 C. 01020304050Limed slurry Limed cake Black kow Black hen Disney compost Baltimore pellets N-viro Milorganite Total Potassium (g Kg-1) Fig 4-1: Continued A. 0.00.51.01.52.02.53.0Limed slurry Limed cake Black kow Black hen Disney compost Baltimore pellets N-viro Milorganite Total Calcium concentration (g kg-1) Fig 4-2: Data represent to tal calcium, magnesium, iron a nd manganese. (A) Ca, (B) Mg, (C) Fe and (D) Mn concentrations of biosolids (on a dry weight basis).

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39 B. 024681012Limed slurry Limed cake Black kow Black hen Disney compost Baltimore pellets N-viro Milorganite Total Magnesium concentration (g kg-1) C. 010203040Limed slurry Limed cake Black kow Black hen Disney compost Baltimore pellets N-viro Milorganite Total Iron concentration (g kg-1) Fig 4-2: Continued

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40 D. 0.00.10.20.30.40.50.60.70.80.91.0Limed slurry Limed cake Black kow Black hen Disney compost Baltimore pellets N-viro Milorganite Total Manganese concentration (g kg-1) Fig 4-2: Continued Table 4-1: Heavy metal concentrations of biosolids in relation to USEPA ceiling concentration limits for land application (on a dry weight basis). Biosolids Arsenic Cadmium Lead Copper Zinc Molybdenum -------------------------------------mg Kg-1---------------------------------Cumulative pollutant loading rates 41 39 300 1,500 2,800 NL Ceiling Concentration Limits 75 85 840 4,300 7,500 75 Limed slurry 3.98 0.72 6.79 317 330 8.14 Limed cake 6.97 0.51 15.30 197 198 5.93 Black kow 2.29 0.06 2.86 26 138 1.48 Black hen 15.60 0.24 1.55 672 641 3.54 Disney compost 22.50 0.71 15.60 291 296 15.30 Baltimore pellets 5.86 17.90 202 393 1010 8.94 N-viro 37.20 0.38 16.90 177 92 7.20 Milorganite 4.34 2.83 37.20 232 640 9.02 NL: No limit (EPA, 1994). According to EPA document 40 CFR Part 503 biosolids applicati on rates should be based on certain heavy metals ceiling concen tration limits. All materials in this study contained concentrations below the ceiling concentration limits and below the cumulative pollutant loading rates. These materials do not represent a potential toxic environmental

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41 risk (Table 4-1). Similar results were obs erved by Adjei et al.(2002). The use of these materials in the production of crops for human consumption when practiced in accordance with existing federal guidelines and re gulations, present negligible risk to the consumer, to crop production, and to the e nvironment (NRC 1996). Franco-Hernandez et al., 2003 reported concentrations of heavy me tals recovered from dry biosolids were lower than 0.1 g kg-1 except for zinc (Zn) and chromium (Cr) in materials treated with lime. Similar results were observed by Adje i et al. (2002) and Si gua et al.(2005), where concentrations of As, Cd, Cu, Pb, Mo, Ni a nd Zn of sewage sludge were far below the national USEPA health risk limits. Since the level of trace metals in the biosolid materials that were used in this study were below the USEPA risk limits, it is not expected that their levels will be harmful on soil and forage quality once they were applied for agricultural purposes (e.g. forage and pasture establishments). Study 2: Determination of the Mineralization Rates of Study Materials Nitrogen mineralization is a relatively slow microbial pr ocess that is affected by factors such as amendment composition, so il type, temperature, pH, aeration, and moisture (Van Kessel et al., 2002; Mikkelsen et al. 1995). The stabilization process used to treat wastewater alters th e nutrient composition of the resu lting biosolids. The rate of nutrient release, or mineralization, is also a ffected by the process. Mineralization values ranging from 41 to 50% of the organic N fr om aerobically-digested sewage sludge and 23 to 41% of the organic N from anaerobically -digested sludge have been reported during the initial crop season (Muchovej et al., 2001) The N release char acteristics of the materials in this study differed widely. Li med slurry had the highest percent of N mineralized when applied at 180 kg N ha-1 with 81% of the total N being mineralized (Table 4-2). Lindemann et al (1984) observed total minera lization rates ranging from

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42 56% to 72% for sewage sludge after 32 weeks of incubation. Mineralization rates of all materials increased when applied N was increased, except for limed cake. When applied at 180 Kg N ha-1, limed cake increased pH to 11.6 and EC to 2811 S cm-1, which resulted in reduced CO2 evolved implying that microbial activity was reduced (Fig 4-5). The lower rate of net N mineralization in th e first week, in the presence of compost, such as black kow, black hen and disney comp ost could be partly due to N assimilation by the growing microbial biomass, that was stimulated by both compost and soil organic C. Composted sewage sludge exhibits a lowe r N mineralization rate compared to noncomposted sludge (Adegbidi et al., 2003). Ba ltimore pellets had the second highest percent of N mineralized at 90 Kg N ha-1 (41%) while Milorganite had the second highest percent of N mineralized at 180 Kg N ha-1(58%). Based on the variable mineralization rates of all materials relative to rate applied there was no defi nite trend, each material has a specific pattern of N release based on its composition and type of treatment received (Table 4-2). Adegbidi et al (2003) observed that for se wage sludges, N mineralization rates of composts are specific to the material s utilized and to the experimental conditions and that on average non-composted organic resi duals had higher mineralization rates than composted organic residuals. Leachates were analyzed for water solubl e P and no detectable P was found in the leachates through all 263 days of incubation. Unlike the case with nitrogen, leaching of P to ground water has not traditionally been a major transport process. In, many soils, high P-sorbing oxide components keep leachate P levels well below eutrophication thresholds (0.01 to 0.05 mg L-1, Sims et al., 1998).

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43 It is very important to ob serve the pH values when bi osolids are being applied to the soil, because the soil pH also influences the rate of mineraliza tion. The pH values in the columns ranged from 6.2 to 11.6, with that of the untreated control soil around pH 6.6. Limed materials and manur e application (Black kow and Black hen) increased soil pH, while composted materials application, li ke Baltimore pellets, resulted in slightly lower soil pH values (Table 4-2). Table 4-2: Percentage of N mineralized in 256 days in relation to biosolids type and the resulting soil pH and EC. Biosolids Total N applied (Kg N ha-1) Total N Mineralized (mg) %N mineralized pH EC (S/cm) Limed cake 90 20 25 9.3 498 180 3 2 11.6 2811 Limed slurry 90 44 55 7.2 359 180 130 81 7.2 612 Black kow 90 5 6 7.2 198 180 11 7 7.2 311 Black hen 90 12 15 7.0 287 180 20 12 7.0 466 Disney compost 90 15 19 6.8 188 180 24 15 6.8 260 Baltimore pellets 90 31 39 6.2 208 180 65 41 6.2 336 N-viro 90 21 27 8.3 2409 180 30 19 8.7 3456 Milorganite 90 29 36 6.7 201 180 92 58 6.7 338 N-Viro application at 90 kg N ha-1 and 180 kg N ha-1increased the EC in the soils up to 2409 S cm-1 and 3456 S cm-1, respectively (Table 4-2) Usman et al. (2004) found that the addition of biosolids raised the soil salinity level more than the addition of compost and also that the salinity increased with increasing applica tion rate of organic wastes. Wong et al. (2001) found that the EC of soil increased due to the addition of biosolids.

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44 CO2 Evolution A CO2 trap of 1N NaOH was placed in the head space of the incubation lysimeter to estimate microbial decomposition rate. The CO2 trap was replaced and titrated with 1N HCl weekly to estimate CO2 evolution. The amount of CO2 evolved was used as an indicator that the materials were being mine ralized and microbial activity was occurring in the system. An interaction between bios olids and rate was observed at P>0.05 (Table 4-3). The intensive microbial activity during the fi rst week is shown by the rate of CO2 evolution that was enhanced in the presence of compos t (Fig 4-3A. and 4-4A.). Similar results were observed by Hadas et al., 1994. The effect of the added organic materials on CO2 evolution decreased rapidly with time, and in the first two weeks was greater in limed slurry and milorganite at 90 kg N ha-1 and 180 kg N ha-1, whereas the organic materials contributed very little to soil resp iration after 16 weeks. At 90 kg N ha-1 and 180 kg N ha1 there was a distinctive difference between CO2 evolved from biosolids at week 1 and week 2. At week 8 these difference was not not iceable anymore, as we observe in figure 4-3C. and 4-4C., there was no difference betw een biosolids. At week 16, we begin to observe differences between biosolids on ce more, however now the differences are related to the rapid decrease in CO2 evolved from some biosolid s, like limed slurry. By week 33, very little CO2 evolution was taking place (Fig. 4-3 and 4-4). We observed that the materials milorganite, limed slurry, limed cake and N-viro showed the same amount of CO2 evolved during the first week at 90 kg N ha-1 and 180 kg N ha-1 (Fig 4-3A and 44A). With milorg anite and limed slurry at 180 kg N ha-1 the quantity of CO2 being evolved was so high that it neutralized all of base solution, thus these data do not show the correct amount of CO2 evolved for those materials at 180 kg N ha-1 during the first

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45 week of the study. Therefore, if more base solution had been adde d to the lysimeters containing these treatments, we would proba bly had have a much higher amount of CO2 evolved from them, consequently th e difference between rates, 90 kg N ha-1 and 180 kg N ha-1, for those materials would be much higher in figures 4-3A and 4-4A. In the case of N-viro and Limed cake at 180 kg N ha-1, N-viro increased the EC values (3456 S cm1) while Limed cake produced a large increa se in pH (11.6), this high EC and pH probably inhibited microbial activity consequently the CO2 evolved at 180 kg N ha-1 was the same as at 90 kg N ha-1. The cumulative CO2 evolution represents the sum of measured weekly quantities of CO2 evolved. Overall, CO2 production increased as the rate of applied biosolid increased. Black kow produced the highe st amount of total CO2 evolved at 90 kg N ha-1 with 3646 mg of CO2 column-1, followed by Disney compost and Baltimore pellets, 3536 and 2947 mg CO2 column-1, respectively (Fig. 4-5). Black he n, N-viro and limed cake produced the lowest CO2 evolved with 2624, 2433 and 1978 mg CO2 column-1 respectively (Fig. 45). Because a true estimate was not obtain ed for Milorganite and Limed slurry, as explained above, they are not included in this discussion, even though those materials would probably have produced the highest CO2 evolved at 180 kg N ha-1 we can not make this assertion. Comparing all othe r biosolids when applied at 180 kg N ha-1, Black kow produced the highest amount of CO2 evolved (5802 mg CO2 column-1) followed by Disney compost with 5017 mg CO2 column-1 (Fig. 4-5). These findings are in agreement with those reported by Franco-Hernandez et al. (2003) who observe d that production of CO2 increased as the rate of biosolids added increased.

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46 A. Week 1 D. Week 16 d dc ba dc ba ba a bc 050100150200250300350400450500Limed Cake Limed Slurry Black Kow Black Hen Disney Compost Baltimore Pellets N-Viro MilorganiteBiosolidsCO2 evolved (mg wk-1) B. Week 2 E. Week 24 bc c a bc ba bac bac bc 050100150200250300350400450500Limed Cake Limed Slurry Black Kow Black Hen Disney Compost Baltimore Pellets N-Viro MilorganiteBiosolidsCO 2 evolved (mg wk-1) C. Week 8 F. Week 33 c cb a b b cb b a 050100150200250300350400450500Limed Cake Limed Slurry Black Kow Black Hen Disney Compost Baltimore Pellets N-Viro MilorganiteBiosolidsCO2 evolved (mg wk-1) Fig 4-3: Data represent CO2 evolved from biosolids at weeks 1(A), 2(B), 8 (C), 16 (D), 24(E) and 33(F), respectively, at 90 Kg N ha-1. Data represent means and standard error of three replicates. Means with the same letter are not different at P>0.05. f a e c d d f b 050100150200250300350400450500Limed Cake Limed Slurry Black Kow Black Hen Disney Compost Baltimore Pellets N-Viro MilorganiteBiosolidsCO2 evolved (mg wk-1) f a e d c dc f b 050100150200250300350400450500Limed Cake Limed Slurry Black Kow Black Hen Disney Compost Baltimore Pellets N-Viro MilorganiteBiosolidsCO2 evolved (mg wk-1) ba ba b ba ba ba b a 050100150200250300350400450500Limed Cake Limed Slurry Black Kow Black Hen Disney Compost Baltimore Pellets N-Viro MilorganiteBiosolidsCO2 evolved (mg wk-1)

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47 A. Week 1 D. Week 16 ba bc dc ba dc a d dc 050100150200250300350400450500Limed Cake Limed Slurry Black Kow Black Hen Disney Compost Baltimore Pellets N-Viro MilorganiteBiosolidsCO2 evolved (mg wk-1) B. Week 2 E. Week 24 d cb a cd b cbd cbd cd 050100150200250300350400450500Limed Cake Limed Slurry Black Kow Black Hen Disney Compost Baltimore Pellets N-Viro MilorganiteBiosolidsCO2 evolved (mg wk-1) C. Week 8 F. Week 33 ed e a ed cb cd ed b 050100150200250300350400450500Limed Cake Limed Slurry Black Kow Black Hen Disney Compost Baltimore Pellets N-Viro MilorganiteBiosolidsCO2 evolved (mg wk-1) Fig 4-4: Data represent CO2 evolved from biosolids at weeks 1(A), 2(B), 8 (C), 16 (D), 24(E) and 33(F), respectively, at 180 Kg N ha-1. Data represent means and standard error of three replicates. Mean s with the same letter are not different at P> 0.05. a d d a c b b a 050100150200250300350400450500Limed Cake Limed Slurry Black Kow Black Hen Disney Compost Baltimore Pellets N-Viro MilorganiteBiosolidsCO2 evolved (mg wk-1) f a e d b c f b 050100150200250300350400450500Limed Cake Limed Slurry Black Kow Black Hen Disney Compost Baltimore Pellets N-Viro MilorganiteBiosolidsCO2 evolved (mg wk-1) c ba bc bc ba c c a 050100150200250300350400450500Limed Cake Limed Slurry Black Kow Black Hen Disney Compost Baltimore Pellets N-Viro MilorganiteBiosolidsCO2 evolved (mg wk-1)

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48 c cb a a b cb 01000200030004000500060007000 Limed cake Black kow Black hen Disney compost Baltimore pellets N-viroBiosolidsTotal CO2 evolved (mg column-1) e a ed cb cd ed 01000200030004000500060007000 Limed cake Black kow Black hen Disney compost Baltimore pellets N-viroBiosolidsTotal CO2 evolved (mg column-1) Fig 4-5: Data represent total CO2 evolved from different biosolids at 90 Kg N ha-1 (A) and 180 Kg N ha-1 (B). Data represent means and standard error of three replicates. Means with the same letter are not different at P>0.05. Table 4-3: Analysis of variance (ANOVA) tabl e used for the determin ation of statistical differences in the analysis for effects of biosolids and rate source on total CO2 evolved. Source DF F value P0.05 > F Biosolid 6 122.69 <0.0001 Rate 1 137.99< 0.0001 Biosolid x rate 6 20.92<0.0001 Replication 2 Error 26 Total 41 A. 90Kg N ha-1 B. 180Kg N ha-1

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49 In summary, the amount of CO2 evolved was a good indicator that the materials were being mineralized, and according to the N mineralization rates observed in this study, biosolids have the ability to su pply nutritional fertility to plants. Study 3: Glass House Study 1st Application Bahiagrass Dry Matter Production All sources of N produced a hi gher level of dry matter than the unfertilized control. Dry matter production varied within biosolid and rates (90 kg N ha-1 and 180 kg N ha-1). Dry matter accumulation was gr eater with the 180 kg N ha-1 compared with the 90 kg N ha-1(Fig 4-6A. and 4-6B.). At 90 kg N ha-1 ammonium sulfate induced the highest dry matter accumulation (250.7 g m-2) followed by Limed slurry (239.3 g m-2) and N-viro (229 g m-2) (Fig 4-6A.). At 180 kg N ha-1 limed slurry induced highest dry matter accumulation (346.1 g m-2) followed by ammonium sulfate (323.8 g m-2) and N-Viro (318.1 g m-2) (Fig 4-6B.). Limed cake and disney compost induced the lowest dry matter accumulation at 90 kg N ha-1 and 180 kg N ha-1. Nitrogen is the most limiting nutrient for grass production on Florida’s spodic sandy soil s as evidenced by the lowest cumulative yield for the nonfertilized contro l compared with the N-fertili zed treatments (Adjei et al., 2002). Bahiagrass Root Dry Matter Production At the end of the experiment, a 45.6cm2 diameter x 15cm depth sample was taken from each tub to estimate differences in stolon growth between biosolids and applied rates (90 kg N ha-1 and 180 kg N ha-1). Stolon dry weight did not differ between biosolids or rates (90 kg N ha-1 and 180 kg N ha-1) after 112 days (Table 4-4). Ammonium sulfate stolon dry weight did not differ from biosolids. Generally, when biosol ids are applied as a sole N source at normal application rates the release of N is not su fficient to induce an

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50 optimum turf grass response relative to quali ty or growth. Biosolid s application in this study were based on normal application rates, therefore the N releas e after 112 days of application was probably not sufficient to i nduce bahiagrass stolon dry matter production to the point that differences be tween biosolids could be observed. Table 4-4: Bahiagrass stolon dry matter production after 112 days Biosolids Kg N ha-1 stolon weight ns -------------g -------------Limed cake 90 9.74 180 10.81 Limed slurry 90 10.62 180 11.73 Disney compost 90 11.58 180 10.03 Baltimore pellets 90 10.31 180 12.94 N-viro 90 10.98 180 10.16 Ammonium sulfate 90 12.57 180 13.05 control 0 10.34 ANOVA P-value CV 0.4 21 ns: not significant at 0.05. Visual Quality Rating Bahiagrass quality was rated visually on the basis of color, uniformity, and density on a 1 to 9 scale, where 1 was dead, 5.5 or above was acceptable, and 9 was superior quality. Bahiagrass quality was different from the control for all biosolids at 90 kg N ha-1 and 180 kg N ha-1 (Table 4-5). All biosolids produced acceptable quality turfgrass but none of the biosolids produced superior quality. Ammonium sulfate is an inorganic source of N used for a comparison with the bi osolids. Limed slurry and N-viro at 180 kg N ha-1 produced a turf quality comparable and s lightly higher than the one observed when ammonium sulfate was applied (Table 4-5). While stolon growth and visual quality were

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51 not affected, these data suggest that application rates (90 kg N ha-1 and 180 kg N ha-1) may have affected dry matter accumulation. Table 4-5: Visual rating duri ng Fall 2004 comparing the bios olids at different rates. Biosolids kg N ha-1 Visual Quality Ratings Scale: 1 to 9 Limed cake 90 7.0c* 180 7.4ba Limed slurry 90 7.1b 180 7.6a Disney compost 90 6.7c 180 7.4ba Baltimore pellets 90 6.7c 180 7.1b N-viro 90 7.2ba 180 7.4ba Ammonium sulfate 90 7.2ba 180 7.3ba control 0 5.1d ANOVA Pvalue CV <0.0001 3.7 Means with the same lette r are not different at 0.05. A. 90kg N ha-1 e b f d c a g0 50 100 150 200 250 300 Limed cakeLimed slurryDisney compost Baltimore pellets N-viroAmmonium sulfate ControlDry matter accumulation (g m-2) Fig 4-6: The effect of nitrogen rate on cu mulative forage dry matter accumulation over 112 days.

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52 B. 180 Kg N ha-1 g b c d f a e0 50 100 150 200 250 300 350 400 Limed cakeLimed slurryDisney compost Baltimore pellets N-viroAmmonium sulfate ControlDry matter accumulation (g m-2) *Means with same letter are not di fferent at 0.05 by Duncan test. Fig 4-6: Continued Table 4-6: Analysis of variance (ANOVA) tabl e used for the determin ation of statistical differences in the analysis for effects of biosolids and rate source on total dry matter accumulation. Source DF F value P0.05 > F Biosolid 6 17.10 <0.0001 Rate 1 33.96< 0.0001 Biosolid x rate 5 7.95<0.0001 Replication 3 Error 35 Total 50 Bahiagrass Chemical Composition Total N Uptake : Nitrogen uptake by the crop is im portant when biosolids are land applied because it is the means by which N is removed to prevent NO3-N leaching to the ground water. The uptake of N by bahiagrass (Table 4-7) increased with increasing biosolid application rate (90 kg N ha-1 vs 180 kg N ha-1), consequently, applying biosolid increased plant uptake of N compared to the control (Table 4-7).Limed slurry,

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53 ammonium sulfate and N-viro induced the highest N uptake at 90 kg N ha-1 and 180 kg N ha-1. N-Viro, at 90 kg N ha-1 and 180 kg N ha-1, response was probably related to the fact that this material is slow-release form of N, as showed in the previous study, allowing more time for bahiagrass uptake. These results are in agreement with the N mineralization data and CO2 evolution data showed at previously study, but when observing the CO2 evolution data, we see that N-viro (90 kg N ha-1 and 180 kg N ha-1) had one of the lowest CO2 evolved at week 1, probably b ecause the conditions, high pH (8.3 and 8.7) and EC (2409 S cm-1 and 3456 S cm-1), created in the soil when N-viro was applied inhibited microbial activity at firs t, however after week 8 N-viro showed an increase in CO2 evolution, suggesting an increase in microbial activity, increasing N mineralization and ultimately increasi ng N available for bahiagrass uptake. N-Recovery: Mineralization rates of approximate ly 40%, 15%, and 8% have also been estimated for waste-activated sludge, an aerobically-digested sludge, and composted sludge, respectively (Muchovej et al., 2001). At 90 kg N ha-1, the apparent nitrogen recovery was slightly higher when inorganic fertilizer, ammonium sulfate, was applied, (35%) compared with Limed slurry (33%) and N-viro (27%). At 180 kg N ha-1, limed slurry surpassed the inorgani c fertilizer inducing a 30% ni trogen recovery by bahiagrass followed by N-viro that induced a 24 % appare nt nitrogen recovery (Table 4-7). The apparent nitrogen recovery of biosolid N range d from 12 to 33%, which is lower than the 47 to 85% ANR of fertilizer N by forage grasses reported in previous studies (Lynch et al., 2004). A higher recovery of N in the gras s-soil system is de sirable and would not only increase N application efficiency, thereby reducing fertilizers co sts to producers, but would also reduce NO3 and NH4 + losses to the environment.

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54 Table 4-7: Total N uptake by bahiagrass from biosolids in first 112 days. Biosolids Kg N ha-1 Total N uptake % N recovered ---------g of N m-2-------Limed cake 90 3.80 de 21 180 5.20 b 19 Limed slurry 90 4.80 bcd 33 180 7.15 a 30 Disney compost 90 2.95 ef 12 180 5.10 b 18 Baltimore pellets 90 3.90 cde 22 180 5.10 b 18 N-viro 90 4.30 cd 27 180 6.20 a 24 Ammonium sulfate 90 4.95 bc 35 180 6.70 a 27 control 0 1.95 f ANOVA Pvalue CV biosolid x rate <0.001 14.6 Pr>F 0.12 Means followed by the same letter ar e not different at 0.05 by Duncan test. Total P Uptake: All applications were N based, so each material had a different quantity of P applied depending on concentrat ion of P in each material (Table 4-8). Phosphorus uptake is a product of dry matter accumulation and tissue P concentration. Phosphorus uptake increased with increasing rate of biosolids. All biosolids had an effect on bahiagrass P uptake (Table 4-8). Bahiagra ss P uptake in the treated tubs was greater than the untreated control (Table 4-8). Th is suggests that P in the biosolids became available for plant absorption within a relativ ely short period of time (112 days). At 90 Kg N ha-1, limed slurry induced the highest P uptake, differing from all treatments, followed by ammonium sulfate and N-viro (T able 4-8). Disney compost supplied the lowest quantity of P differing from all tr eatments at P< 0.05. These results are in agreement with the P findings in the biosolid s prior to application and based on the total amount of P applied. Ammonium sulfate does not contain P, however when applied a

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55 significant drop in pH is obser ved (from 6.0 to 5.0). It is suspected that this pH drop caused a release of P forms in the soil that were previously unavailable for uptake by the grass. Table 4-8: Quantity of P applied by each material and percentage of P uptake by bahiagrass Biosolids Kg N ha-1 Quantity of material applied on each tub Quantity of P applied by each material P uptake (*) Tissue P concentration % Recovery of applied P -----g-----g tub-1----g ---------g Kg-1--90 135 1.22 0.14 ef 3.80 11.5 Limed cake 180 270 2.45 0.20 abcd3.90 8.2 90 76 1.68 0.19 bcde4.20 11.3 Limed slurry 180 151 3.34 0.25 a 3.70 7.5 90 109 1.28 0.12 f 4.00 9.4 Disney compost 180 218 2.55 0.18 cde 3.90 7.1 90 55 1.40 0.16 def 3.90 11.4 Baltimore pellets 180 110 2.80 0.20 abcd3.90 7.1 90 967 2.54 0.17 de 3.90 6.7 N-viro 180 1933 5.08 0.23 abc 3.80 4.5 90 8 -0.18 cde 3.80 Ammonium sulfate 180 17 -0.24 ab 3.80 Control 0 --3.80 ANOVA Pvalue <0.0001 CV 20.98 biosolid x rate Pr>F 0.9229 (*)Means with the same letter are not different at 0.05 by Duncan test. In previous studies, biosolid fertil izer rates as low as 4.5 to 9 t ha-1 improved bahiagrass P concentrations to about 3 g kg-1 (Tiffany et al., 2001). Kincheloe et al. (1987) and Rechcigl et al. ( 1992) showed that adequate pr oduction of bahiagrass is achieved with a tissue P concentration of approximately 2.0 g kg-1. Increases in biosolids

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56 loading rate did not increase percentage P removal in bahiagrass over 112 days of biosolids application (Table 4-8). Although, as observed in table 4-8, the amount of P applied with each biosolid was different si nce the application was N based, the percent recovery calculation was based in each P load separately. Bahiagrass recovery of P was higher with limed cake, followed by baltimore pellets and limed slurry, all at 90 Kg N ha1. A low percentage of P was recovered in bahiagrass harvested over 112 days. Johnson et al., 2004, found that a low percentage of wast ewater or poultry litter sources of P was recovered in forage harv ested over four years. Leaching Evaluation Total N in Leachate: The leachate was analyzed for NH4 +, NO3 -, and P to assess N and P movement in soils amended with wastes. At 90 Kg N ha-1, limed slurry contributed the highest quantity of inorganic N to the leachate. In excess of 11.04% of the N applied as limed slurry was found in the leacha te, followed by 7.56% of the applied N for ammonium sulfate and 1.10% of the N applied as disney compost. At 180 Kg N ha-1, ammonium sulfate contributed the highest qua ntity of N to the leachate with 15.70% of the N applied, followed by limed slurry w ith 10.31% and Disney compost with 3.60% (Table 4-9). Ammonium sulf ate is commonly used in tu rf fertilization. Therefore, ammonium sulfate was used to establish a comparison be tween inorganic and organic sources of N. Besides limed slurry at 90 Kg N ha-1, none of the biosolids contributed as much N to the leachate as ammonium sulfate, suggesting these biosolids can be used to reduce the threat of environmental c ontamination (Table 4-9). Leachate NO3-N concentration was influenced by loading rate (90 Kg N ha-1 and 180 Kg N ha-1) and the length of time after the initia l application. As expected, NO3-N concentration increased with increasing level of applied biosolids and decreased with time after application

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57 (Table 4-9, 4-10). Leachate collected on day 84 did not contain detectable NO3-N (Table 4-10). The NH4-N data are presented in table 4-9 a nd 4-10. In general, the concentration of NH4-N was several fold lower than the NO3-N concentration. By day 56, there was no detectable levels of NH4-N in the leachate (T able 4-10). Most of the materials produced maximum levels of leached NH4 within 7 days (Table 4-10). As for NO3, microbial activity had to mineralize the materials to soluble forms, becoming available for plant uptake and runoff. According to Van Kessel et al. (2002) when manure-amended soils were incubated for 56 days, all of the NH4 +-N was depleted; NO3 -N concentrations increased during this time period. Nitrogen in sludge is mainly in organic forms, while mineral forms are generally in low concentration and are mainly represented by ammonium nitrogen. Total P in Leachate: No detectable P was found in the leachates through all the leachate collections, 7 to 112 days. Biosolid s and manure-borne P reactions have not been researched as much as waste bond N. We do know that (as with N) calculations based on total waste-P concentrations to determine allowable P loads to a soil can be grossly over-conservative, as all the (total) P may not be bioavailable or soluble enough to leach (Pierzynski et al., 1994). Soils comm on to most of the USA contain sufficient Psorptive capacity (Al and Fe oxides) to prevent significant P leaching and to mask inherent differences in the P solubilities of biosolids materials, th erefore leaching of P appears to be quite small for most bioso lids, even when applied to meet crop N requirements on sandy soils with limited Psorbing capacity (Ell iott et al., 2002). Table 4-9: Total NH4-N and NO3-N content in leachates from organic materials Biosolids Kg N ha-1 Total NH4 in leachate Total NO3 in leachate Total N in leachate (7 through 112 % of N applied leached

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58 days) --mg ---mg -------mg ------90 8.40 6.30 14.70 0.34 Limed cake 180 9.20 42.90 52.10 1.25 90 12.70 188.10 200.80 11.04 Limed slurry 180 64.80 302.60 367.40 10.31 90 5.00 22.80 27.80 1.10 Disney compost 180 37.40 96.10 133.50 3.60 90 4.50 4.35 8.85 0.01 Baltimore pellets 180 17.30 13.10 30.40 0.62 90 5.80 19.10 24.90 0.93 N-viro 180 6.87 117.40 124.27 3.32 90 29.70 110.60 140.30 7.56 Ammonium sulfate 180 173.80 380.80 554.60 15.70 Control 0 4.95 3.75 8.70 Table 4-10: Concentration of NO3-N and NH4 –N in leachate as affected by the incubation period Biosolids Kg N ha-1 7 days 14 days 28 days 56 days 84 days mg NO3 NH4NO3NH4NO3NH4NO3NH4 NO3 NH4 90 3 4 3 5 0 0 0 0 0 0 Limed cake 180 12 4 18 5 12 0 1 0 0 0 90 45 9 74 4 69 0 1 0 0 0 Limed slurry 180 46 43 157 22 97 0 3 0 0 0 90 11 3 9 2 3 0 0 0 0 0 Disney compost 180 25 9 47 29 23 0 2 0 0 0 90 2 2 2 2 1 0 0 0 0 0 Baltimore pellets 180 3 10 5 7 6 0 0 0 0 0 90 5 4 8 2 6 0 0 0 0 0 N-viro 180 12 2 27 4 66 1 12 0 0 0 90 32 20 62 8 17 2 0 0 0 0 Ammonium sulfate 180 64 127 162 35 148 12 7 0 0 0 Control 0 1 2 1 2 2 0 0 0 0 0 pH and EC in Leachate: The relationship between pH and EC and incubation period at different rates of biosolid applic ation is presented in table 4-11 and 4-12. The leachate pH ranged from 5.3 to 6.6, with the untreated control around 6.0. Limed materials increased the pH while ammonium sulfate reduced soil pH to about 5.3. The

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59 incubation period affected the pH of the leachat e, with or without bi osolid. Interestingly, the leachate pH increas e observed in the mineralization study for limed cake (9.3 at 90 Kg N ha-1 and 11.6 at 180 Kg N ha-1) and N-viro (8.3 at 90 Kg N ha-1 and 8.7 at 180 Kg N ha-1) was not observed in the glasshouse study. This difference is probably due to the interaction between the biosolids and the turf and to the biosolids to soil rates. In the incubation study the biosolids were applied based on a dry weight basis while in the glasshouse study the biosolids were surface a pplied on an area basis. On the other hand, in the incubation study the absence of the tu rf meant that all elements that were mineralizing from the biosolids were free, to be adsorbed by the soil and to change in the soil characteristics, like pH. When biosolids were appl ied in the glass house study, the same mineralization was occurring, but now, inte raction between the bi osolid and the turf was taking place, nutrients were also being absorbed by the turf, ions attached to the surface of root hairs, such as H+ ions, may exchange with t hose held on the soil surface, keeping the pH from increasing as much as they increased without the turf. Electrical conductivity (EC, an estimate of soluble salts) increased for soils receiving biosolids. N-Viro applied at 180 Kg N ha-1 produced the highest EC from 7 days through 112 days going from 1571 uS cm-1 to a peak of 3227 uS cm-1 at 56 days, from there decreasing to 1141 uS cm-1 at 112 days (Table 4-12). Electrical conductivity of the leachate from ammonium sulfate trea ted soils peaked at 14 days at 2270 uS cm-1. N-Viro at 90 Kg N ha-1 produced a peak EC at 56 days at 2072 uS cm-1. All the other treatments differ from the control, increasi ng the EC when compared to the control.

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60 Table 4-11: Leachate pH values from 7 to 112 days after treatment application. (*) Means with same letter are not different at 0.05 (**) Interactions effects be tween biosolids and rates ar e not significant; therefore comparisons were made across a ll biosolids at both rates. Table 4-12: Leachate EC values from 7 to 112 days after treatment application. BiosolidsKg N ha-17 days14 days28 days56 days84 days112 days ECECECECECEC Limed cake90414 f409d408d534b430b379b 180464 ef484d497d593e451b413b Limed slurry90571 ef771c721bc521b413b362b 180702 e975c847c546g409f370d Disney compost90585 ef593dc528cd649b410b377b 180712 e935c857c769c416e375c Baltimore pellets90437 ef 455d432d547b405b355b 180477 ef535d550d649d406g350f N-viro901267 c1712a2039a2072a1136a745a 1801571 b2161b2987a3227a1668a1141a A mmonium sulfat e 90993 d1036b751b642b399b326b 1801938 a2270a1649b934b449c360e Control0381 f405de434dd585bf417bd361be (*) Means with same letter are not different at 0.05 (** ) Rates 90 Kg N ha-1 and 180 Kg N ha-1 statistics analyzed separately from 14 to 112 days. Because the EC was measured from the soil leachate, these results indicate movement of salt from surface applied bios olid and inorganic fertilizer (ammonium sulfate) applications. Kinger y et al. (1994) found higher EC values in soil under fescue

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61 pastures with a long-term history of poultr y litter application compared to pastures receiving no poultry litter. Study 4: Glass House Study 2nd Application A second growth response evaluation was conducted during the spring of 2005 from March through July to document the nutrient s build up in the soil and to assess their nutrient availability to bahiagrass. Biosolids were reapplied to the same tubs with the established stand of bahiagrass was employed. Bahiagrass Dry Matter Production After reapplication of treatments the same experimental methodologies were employed through 112 days. All sources of N pr oduced a higher dry matter mass than the unfertilized control. Bahiagrass dry matter re sponse to N source varied within sources and rates (90 kg N ha-1 and 180 kg N ha-1). Dry matter accumulation was greater with the 180 kg N ha-1 compared with the 90 kg N ha-1(<0.0001). At 90 kg N ha-1 Limed slurry provided the highest dry matter accumulation (355 g m-2) followed by Limed cake (340 g m-2) and Ammonium sulfate (325 g m-2) (Fig 4-7A.). At 180 kg N ha-1 Limed slurry provided highest dry matter accumulation (575g m-2) followed by Ammonium sulfate (475 g m-2) and Baltimore pellets (468 g m-2) (Fig 4-7B.).The increased bahiagrass dry matter response to biosolids compared to th e unfertilized control is probably related to the nutrients, especially N (most limiting nutri ent), incorporated to the soil and its availability to the grass after mineraliza tion. Statistical comparisons were made to compare the bahiagrass dry matter producti on after first application and after reapplication. At 0.05 probability (two tailed test), the F value was 2.05; when comparing the MSE values of the first application with the reapplication the F value was 3.70, which

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62 was higher than 2.05, consequently the respons e variation for both st udies are different, therefore a comparison between both studies was not made. Bahiagrass Root Dry Matter Production A sample was taken from each tub at termination to evaluate the influence of biosolids on stolon growth. Stol on dry weight differed between N-sources and rates (90 kg N ha-1 and 180 kg N ha-1) 112 days after reapp lication (Table 4-13). Stolon growth of bahiagrass receiving baltimore pellets at 180 kg N ha-1 was greater than the stolon growth of the unfertilized control. This might be b ecause baltimore pellets has a high P content, as shown in study 1, therefore bahiagrass r eceiving baltimore pellets contained the highest tissue P concentration (T able 4-17) and P influence in root growth is evidenced as shown in Tisdale and Nelson, 1999. Stolon growth in response to all the other biosolids did not differ from the unfertilized control. Table 4-13: Bahiagrass root dry matter produc tion 112 days after r eapplication of the materials. Biosolids Kg N ha-1 root weight -----------g -----------Limed cake 90 9.95 ba* 180 8.68 b Limed slurry 90 9.55 b 180 8.78 b Disney compost 90 12.01 ba 180 9.61 b Baltimore pellets 90 12.00 ba 180 13.20 a Ammonium sulfate 90 9.70 ba 180 10.30 ba control 0 8.7 b ANOVA P-value CV 0.004 20 Means with the same lette r are not different at 0.05. Statistical comparisons were made to compare the bahiagrass root dry matter production after first applicati on and after reapplication. At 0.05 probability (two tailed

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63 test), the F value was 2.05; when comparing th e MSE values of the first application with the reapplication the F value was 1.04, whic h was lower than 2.05, therefore there was no difference between the root ha rvests (first and second appl ication), thus the parameter root weight was analyzed as a combined analysis. When analyzed combined, bahiagrass root growth receiving ba ltimore pellets at 180 kg N ha-1, remained greater when compared to the root growth of the unfer tilized control and th e other biosolids. Bahiagrass Quality Rating Bahiagrass quality was rated visually on the basis of color, uniformity, and density on a 1 to 9 scale, where 1 was dead, 5.5 or above was acceptable, and 9 was superior quality. Bahiagrass quality was different from the control for all treatments at 90 Kg N ha-1 and 180 Kg N ha-1 (Table 4-14). Table 4-14: Visual rating duri ng Spring 2005 comparing N sources at different rates after biosolid reapplication. Biosolids kg N ha-1 Visual rating means Scale: 1 to 10 Limed cake 90 7.20 f* 180 7.60 d Limed slurry 90 7.40 e 180 7.70 c Disney compost 90 7.00 h 180 7.80 a Baltimore pellets 90 7.10 g 180 7.60 d Ammonium sulfate 90 7.60 d 180 7.75 b control 0 5.40 i ANOVA Pvalue CV <0.0001 0 *Means with the same lette r are not different at 0.05. All the treatments exhibited acceptable qual ity after reapplication but none of the treatments exhibited superior quality. Disney compost and ammonium sulfate at rate 180 Kg N ha-1 produced a higher turf quality than th at of other treatments (Table 4-14).

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64 Forage production often requires significan t inputs of lime, N fertilizer, and less frequently of P and K fertilizers; all the Nsources used in this experiment to induce bahiagrass growth contained from 59 g kg-1 to 13 g kg-1 N, as shown in study 1. e c c d a b0.0 50.0 100.0 150.0 200.0 250.0 300.0 350.0 400.0 Limed cakeLimed slurryDisney compostBaltimore pellets Ammonium sulfate ControlDry matter accumulation (g m-2) A. 90kg N ha-1 f b c e a d0.0 100.0 200.0 300.0 400.0 500.0 600.0 700.0 Limed cakeLimed slurryDisney compostBaltimore pelletsAmmonium sulfate ControlDry matter accumulation (g m-2) B. 180 Kg N ha-1 *Means with same letter are not di fferent at 0.05 by Duncan test. Fig 4-7: The effect of nitrogen source and ra te on cumulative bahiagrass tissue dry matter accumulated over 112 days.

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65 Bahiagrass Chemical Composition Total N Uptake: Nitrogen applied to soils in mine ral fertilizers and biosolids, and also indigenous soil organic N, is subject to a complex series of in terrelated biochemical and physical processes which collectively form the nitrogen cycle. Nitrogen applied as nitrate may be subject to leaching; therefor e nitrogen uptake by the crop is important when biosolid land application is used becau se it is the means by which N is removed to prevent NO3-N leaching to the ground water. The uptake of N for bahiagrass forage after reapplication (Table 4-15) increased with in creasing biosolid app lication rate (90 kg N ha-1 and 180 kg N ha-1), Pr>F 0.0004, consequently, biosol id application increased plant uptake of N compared to the control (Table 4-15). Limed slurry and Limed cake supplied the highest level of N to the plant at 90 kg N ha-1 while Limed slurry and AS supplied the highest level of N at 180 kg N ha-1.Limed slurry response at 90 kg N ha-1 was probably related to the fact that its N was readily available at 90 kg N ha-1, as shown in the previous study limed slurry mineralized 55% of the N applied, while limed cake mineralized 25% of the N applied, being readily available for bahiag rass uptake. Both of those materials are limed materials, which resulted in a soil pH close to neutrality, which probably increased the N availability for plant uptake. Limed slurry at 180 kg N ha-1, had the highest percentage of N being mineralized in the incubation st udy (81%), therefore N was readily available for plant upta ke. Ammonium sulfate at 180 kg N ha-1, reduced the pH (4.2) in the system in the first 14 days. Statistical comparisons were made to compare the N uptake by bahiagrass after first app lication and after re application. At 0.05 probability (two tailed test), the F value was 2.05; when comparing the MSE values of the first application with the reapplicati on the F value was 2.10, which was higher than 2.05, consequently the response variation fo r both studies are di fferent, therefore a

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66 comparison between both studies was not made The apparent nitrogen recovery (ANR) of biosolid N by bahiagrass was calculated by difference as: ANR (%) = [(N uptake for biosolid – N uptake for unfertilized control)/ N reapplied] x 100.The recovery of biosolid N is recorded in table 4-15. Recovery of biosolid N ranged from 44.5 to 39% for Limed slurry N and 44.2 to 32.2% for Baltimore pellets was recovered in the harvested grass, which is related to the ANR of N-sources (47 to 85%) by forage grasses reported in previous studies (Lynch et al., 2004). This hi gh percentage N recove ry might be related to residual effect from previous applicati on. Muchovej and Rechcigl (1997) estimated a minimum recovery rate from biosolids by bahiagrass crop of 70% in 2 years with the remaining 30% becoming available with additional time. Table 4-15: Total N uptake by bahiagrass from biosolids in 112 days after reapplication of the materials. Biosolids Kg N ha-1Total N uptake % N recovered ---------g of N m-2-------90 5.00 c 35.0 Limed cake 180 7.35 b 31.0 90 5.35 c 39.0 Limed slurry 180 9.70 a 44.5 90 2.85 e 10.3 Disney compost 180 4.70 dc 15.8 90 4.75 dc 32.2 Baltimore pellets 180 7.70 b 44.2 90 4.55 dc 29.9 Ammonium sulfate 180 8.00 b 34.8 control 0 1.95 e ANOVA Biosolid x rate Pvalue CV Pr>F <0.0001 20.02 0.0837 Means followed by the same letter are not different by Duncan test. The recovery of biosolid N reported here shows a good relationship with the recovery estimated by Muchovej and Rech cigl (1997), however these results are

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67 estimated within 7 months after the firs t application and with in 3 months after reapplication of the materials. Total P Uptake: At 90 kg N ha-1application of limed slurry resulted in the highest P uptake by bahiagrass, differi ng from all N-sources, followe d by baltimore pellets and limed cake (Table 4-16). Application of disn ey compost resulted in the lowest P uptake by bahiagrass differing from all N-sources at P< 0.05. At 180 kg N ha-1, limed slurry was the material that induced th e highest P uptake by bahiagrass differing from all N-sources followed by baltimore pellets and limed cake. Disney compost induced the lowest P uptake by bahiagrass differing from all N-sour ces (Table 4-16). Ammonium sulfate is a material that has no P on its composition; howev er this material caused a drop in soil pH at first, going from 6.2 to 4.2. This pH drop mi ght have caused a release of P forms that before were unavailable for uptake by th e grass. Comparing tables 4-8 and 4-16, ammonium sulfate was the only N-source th at did not enhance the total P uptake by bahiagrass after reapplication, implicating that the increas e in P uptake by bahiagrass when AS was added was related to the pH. Ebeling et al., 2003, suggested that P sources with relatively high soluble P or bioavailable P concentrations (CaHPO4 and biosolids) provided high levels of P for plant uptake in poorly buffered systems such as sand media, which has little capacity of converting solubl e P to less available forms by reaction with or sorption by Fe, Al, Mg, or Ca compounds. All applications were N based, so each material had a different quantity of P applied depending on concentration of P in th e material (Table 4-16). Phosphorus uptake is a product of dry matter accumulation and tissue P concentration. Statistical comparisons were made to compare the bahi agrass P uptake after first application and

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68 after reapplication. At 0.05 pr obability (two tailed test), the F value was 2.05; when comparing the MSE values of the first applic ation with the reapplication the F value was 2.15, which was higher than 2.05, consequently the response variation for both studies are different, therefore a comparison between both studies was not made. Bahiagrass P uptake in the tubs that received biosolids application was greater than the untreated control (Table 4-16). This sugge sts that P in the biosolids became partly available for plant absorption within a relatively short period of time (112 days). Table 4-16: Quantity of P applied by each material and percentage of P uptake by bahiagrass after reapplication of the materials Biosolids Kg N ha-1 Quantity of material applied on each tub Quantity of P applied by each material P uptake (*) Tissue P concentration % Recovery of applied P -------g-------g tub-1-----g-------g Kg-1---90 135 1.22 0.25 de 4.00 20.5 Limed cake 180 270 2.45 0.37 bc 4.10 15.1 90 76 1.68 0.28 cde 4.20 16.7 Limed slurry 180 151 3.34 0.48 a 4.30 14.4 90 109 1.28 0.15 f 3.90 11.7 Disney compost 180 218 2.55 0.25 de 4.00 9.8 90 55 1.40 0.26 de 4.30 18.6 Baltimore pellets 180 110 2.80 0.43 ab 4.80 15.4 90 8 -0.19 ef 3.10 Ammonium sulfate 180 17 -0.30 cd 3.40 Control 0 --3.60 ANOVA P-value <0.0001 CV 20.37 Biosolids x rate Pr>F 0.38 (*)Means with the same lette r are not different at 0.05. Repeated use of biosolids may result in either increased soil P accumulation or P surface water runoff which could pose potenti al eutrophication problems for sensitive

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69 water bodies such as the Lake Okeechobee Ba sin (Adjei and Rechcigl, 2002; Reddy and Flaig, 1995; Walker and Havens, 1995). Leaching Evaluation Total N in Leachate: The leachate was analyzed for NH4 +, NO3 -, and P to assess N and P movement in soils after bi osolids reapplication. At 90 kg N ha-1, ammonium sulfate induced the highest total N content in l eachate at 1.26% of the N applied followed by limed slurry with 1.18% of N. At 180 kg N ha-1, ammonium sulfate produced the highest total N in the leachate with 4.19% of the N applied, followed by limed slurry with 1.01% and limed cake with 0.24%. Ammonium sulfate was the only inorganic fertilizer used in this study in order to establish a comparison between inorganic fertilizer and biosolids. None of the biosolids analyzed supplied as much N to the le achate as ammonium sulfate, suggesting that these biosolids do not represen t a hazardous to the wa ter table when used as fertilizer (Table 4-17). When dividing the MSE for first application with the MSE for reapplication the F value was 0.16, which is lower than 2.05, th erefore there was no difference between the total N in leachate (first and second app lication), thus, the parameter total N in leachate was combined for analyzes. Overall, there was a difference in total N in leachate when compared to the first application, Pr>F was 0.0011, which was lower than 0.05. The total N in leachate after rea pplication was lower than in the first application leachate. This could be e xplained by the bahiagrass N uptake after reapplication, which increased compared with the first application, so more N is being removed from the soil preventing NO3-N leaching to the ground water. The concentration of NO3-N in the leachate from the different N-so urces was influenced by loading rate (90 kg N ha-1 and 180 kg N ha-1) and the length of time after th e initial application. Nitrate-N concentration increased with increasing leve l of applied biosolid s and decreased with

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70 time after application (Table 4-17, 4-18). L eachate collected on day 28 did not contain detectable NO3-N (Table 4-18). Table 4-17: Total NH4-N and NO3 -N content in leachates from the different organic materials after reapplication Biosolids Kg N ha-1 Total NH4 in leachate Total NO3 in leachate Total N in leachate (7 through 112 days) % of N applied leached ---mg -----mg ---------mg ------90 6.75 0.00 6.75 0.39 Limed cake 180 5.25 3.00 8.25 0.24 90 14.75 5.75 20.5 1.18 Limed slurry 180 22.75 12.50 35.25 1.01 90 2.00 0.00 2.00 0.11 Disney compost 180 0.00 0.25 0.25 0.01 90 1.00 0.00 1.00 0.06 Baltimore pellets 180 0.00 0.00 0.00 0.00 90 22.00 0.00 22.00 1.26 Ammonium sulfate 180 138.00 8.00 146.00 4.19 Control 0 0.00 0.00 0.00 0.00 Table 4-18: Concentration of NO3-N and NH4 –N in leachate as affected by biosolid reapplication. Biosolids Kg N ha-1 7 days 14 days 28 days 56 days 84 days --------------------------------mg------------------------------------NO3 NH4 NO3NH4NO3 NH4NO3 NH4 NO3NH4 90 0 7 0 0 0 0 0 0 0 0 Limed cake 180 0 4 3 2 0 0 0 0 0 0 90 6 15 0 0 0 0 0 0 0 0 Limed slurry 180 10 20 3 3 0 0 0 0 0 0 90 0 2 0 0 0 0 0 0 0 0 Disney compost 180 0 0 0 0 0 0 0 0 0 0 90 0 1 0 0 0 0 0 0 0 0 Baltimore pellets 180 0 0 0 0 0 0 0 0 0 0 90 0 20 0 2 0 0 0 0 0 0 Ammonium sulfate 180 5 107 3 32 0 0 0 0 0 0 Control 0 0 0 0 0 0 0 0 0 0 0

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71 The NH4-N data are presented in table 4-17 a nd 4-18. In general, the concentration of NO3-N was several fold less than the NH4-N concentration. By day 28 all the materials had no detectable NH4 being leached (Table 4-19). All NH4 was readily available in soluble forms because most of the materials produced their leached NH4 peaks at 7 days (Table 4-18). As for NO3, microbial activity had to occur in order to mineralize the materials and break them into soluble forms, becoming available for plant uptake and runoff. Total P in Leachate: No detectable P was found in the leachates through all the leachate collections, 7 to 112 days. Unlike N, leaching of P has not traditionally been viewed as a major ground water problem. In many soils, abundant P-sorbing oxide components keep leachate P levels well bello w eutrophication threshol ds (Elliot at al., 2002). Peterson et al., (1994) concluded that ther e is no need to worry about P leaching to ground water because leaching wa s practically zero. Certainl y, areas with shallow ground water and course-textured soils of low P-sorbing capacity were not taken under consideration. Studies have reported that leach ing of biosolids P is minor or negligible (Elliot et al., 2002; Sui et al., 1999; Peters on et al., 1994). Then, for most locations, restricting biosolids application rates to the P needs of the crops would normally be unnecessary to minimize leaching concerns (Elliot et al., 2002). pH and EC: The relationship between pH and EC of the leachate and incubation period at different rates of biosolid after r eapplication is presente d in table 4-19 and 4-20. The pH values in the leachate ranged from 4.2 to 7.2, with the untreated control around 6.3. Limed materials increased the pH from 6.3 to 7.2 while ammoni um sulfate reduced soil pH from 6.3 to 4.2 at 7days. The incubati on period had a marked effect on the pH of

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72 the leachate, with or without biosolids. Similar to the first application of the biosolids, the pH increase observed in the incubation study wa s not observed here as well, most likely for the same reason stated in the previous study. Eghball (1999) found that application of beef cattle feedlot manure or compost increas ed the soil surface (0-15cm) pH while N application as NH4NO3 reduced the pH (from 6.4 to 5.6). Table 4-19: pH measurements in the leachate from 7 to 112 days after reapplication of the materials Biosolids (*) (**) Kg N ha-1 7 days 14 days 28 days 56 days 84 days 112 days -------------------------------pH----------------------------------90 6.5 a 6.5ab 6.0ab 6.8b 6.8 b 7.2a Limed cake 180 6.7 a 6.8a 6.9a 7.1a 7.2 a 7.2a 90 6.4 ab 6.3b 6.5cd 6.3cd 6.5 c 6.6cd Limed slurry 180 6.5 a 6.5ab 6.6bc 6.6bc 6.7 bc 6.9b 90 6.1 c 6.3b 6.4cd 6.3cd 6.5 c 6.5d Disney compost 180 6.1 c 6.2b 6.4cd 6.3cd 6.5 c 6.5d 90 6.0 c 6.2b 6.4cd 6.3cd 6.5 c 6.5d Baltimore pellets 180 6.0 c 6.2b 6.5cd 6.3cd 6.4 cd 6.6d 90 5.6 d 6.2b 6.3cd 6.3cd 6.6 bc 6.7c Ammonium sulfate 180 4.2 e 5.1c 5.7e 5.7e 6.1 d 6.2d Control 0 6.2 bc 6.3 b 6.4 cd 6.5 bcd6.6 bc 6.7 c (* ) Means with same letter are no t different at 0.05 by Duncan test. (**) Interactions effects be tween biosolids and rates ar e not significant; therefore comparisons were made across a ll biosolids at both rates. Electrical conductivity (EC, an estimate of soluble salts) levels increased for soils receiving biosolids. Leachate of soils rece iving ammonium sulfate reached a peak at 7 days with 910 uS cm-1 at 90 Kg N ha-1 and 1450 uS cm-1 at 180 Kg N ha-1. All other Nsources differed from the control, increasi ng the EC when compared to the control. Because the EC was measured from the soil l eachate, these results indicate movement of salt from surface applied biosolid and i norganic fertilizer (ammonium sulfate) applications, as shown by Eghball (2002).

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73 Table 4-20: EC measurements in the leachate from 7 to 112 days after reapplication of the materials Biosolids (*) (**) Kg N ha-1 7 days 14 days28 days56 days 84 days 112 days -----------------------------EC------------------------------Limed cake 90 574 cd 509 abc534 a 561 a 424 a 541a 180 675 bcd 579 ab 549 a 560 a 416 a 476 ab Limed slurry 90 589 cd 377 bc 418 b 452 bc 335 b 413 bc 180 768 bc 473 abc457 ba 494 b 368 ab 429 abc Disney 90 576 cd 439 abc464 ba 503 b 319 b 419 bc compost 180 837 b 488 abc455 ba 501 b 368 ab 445 abc Baltimore 90 490 d 374 bc 407 b 418 c 299 b 342 c pellets 180 554 cd 364 c 393 b 429 c 320 b 374 bc Ammonium 90 910 b 389 bc 387 b 420 c 303 b 359 bc sulfate 180 1450 a 629 a 433 b 486 b 326 b 377 bc Control 0 467 d 379 bc 410 b 422 c 315 b 359 bc (*) Means with same letter are not different at 0.05 by Duncan test. (**) Interactions effects be tween biosolids and rates ar e not significant; therefore comparisons were made across a ll biosolids at both rates.

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74 CHAPTER 5 CONCLUSIONS (i). Nutritional value and potent ial toxic environmental impact: All the materials contained sufficient N and P to s upport mineralization and nutrient release for bahiagrass growth. Limed slurry, N-viro and limed cake contained higher Ca concentration, due to the large amount of lim e added to these materials. Based on their N and P compositions, these biosolids can be used as low-grade N and P fertilizer and as a source of calcium, especially the lime-stabilized residuals. All the materials used in this study were of class A or cl ass B in terms of USEPA’s pathogens and pollutant concentration limit, not exceeding the ceiling a nd loading concentration limits, thus not representing a potential t oxic environmental impact. (ii) Mineralization: Biosolids N is an important res ource that could be used more efficiently in crop production. Percentage N mineralized from biosolids in 263 days of incubation ranged from 2 to 81% of their total N content. All biosolids increased mineralization rates when N a pplied was increased (180 kg N ha-1) except for limed cake, that when applied at 180 kg N ha-1 increased pH to 11.6 and EC to 2811 uS cm-1, which may have inhibited microbial activity in those columns thus reducing mineralization. Intensive microbial activity during the first week wa s shown by the rate of CO2 evolution that was enhanced in the presence of biosolids. Overall, CO2 production increased as the rate of applied biosolid increased. (iii) Biosolids potential for induc ing growth response of bahiagrass: The potential use of biosolids as an turf gra ss amendment is important to producers and

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75 environmentalists, since their use may improve forage nutritive value and at the same time be a means for biosolids di sposal. Most of the biosolids used in this study, specially the lime-stabilized materials, induced bahiagra ss forage production at a similar rate and to the same extent as the inorganic ammonium sulfate fertilizer. A ll N-sources gave better forage production when compared to the untr eated control. After r eapplication the large cumulative dry matter production obtained for N-sources indicate that biosolids can provide sufficient in season and residual plan t available N to support forage crops. Tissue N and P increased with increasing rate of bi osolids. Limed slurry and N-viro induced N uptake by bahiagrass to the same extent as the inorganic fertilizer. Besides limed slurry at 90 kg N ha-1, none of the biosolids contributed as much N to the leachate as ammonium sulfate, suggesting these bios olids can be used to reduce the threat of environmental contamination. As for P, no detectable P was found in the leachates throughout both study periods, including after reapplication. There is much to be learned about the use of biosolids on agricultural crops. Society will benefit from wise a pplication of this material an d thus from the recycling of a valuable source.

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76 LITERATURE CITED Adegbidi, H.G., and Briggs, R.D., 2003. Nitr ogen mineralization of sewage sludge and composted poultry manure applied to willow in a greenhouse experiment. Biomass and Bioenergy. 25: 665-673. Adjei, M.B., and Rechcigl, J.E., 2002. Bahiagrass production and nutritive value as affected by domestic wastewater residuals. Agron. J. 94: 1400-1410. Adriano, D.C., 2001. Trace elements in terrest rial environments. Springer-Verlag, New York, NY. Anderson, J. P. E., and Domsh, K H., 1990. Application of ecophysiological quotients (qCO2 and qD) on microbial biomasses from soil of different cropping histories. Soil Biology and Biochemistry. 22: 251-255. Anjos, J.T., Sarkar, D., and O’Connor G.A ., 2000. Extractable-P in biosolids-amended, soils: an incubation study. Revista de Estudos Ambientais, 2(2-3): 68-76. Barker, A.V., 2001. Evaluation of composts fo r growth of grass sod. Communications in Soil Science and Plant Analysis. 32 (11&12): 1841-1860. Barton, L., Shipper, L.A., Barkle, G.F., Mc Leod, M., Speir, T. W., Taylor, M. D., McGill, A.C., van Schaik, A.P., Fitzgerald, N.B., and Pandey, S. P., 2005. Land application of domestic effluent onto f our soil types: Plant uptake and nutrient leaching. J. Environ. Qual. 34: 635-643. Bastian, R. K., 1986. Overview on sludge utilization. In The forest alternative for treatment and utiliza tion of municipal and industrial wastes. Edited by D.W. Cole, C. L. Henry, and W. L. Nutter. Universi ty of Washington Press, Seattle. 7-25. Bernal, M. P., Sanchez-Monedero, M. A., Paredes, and C., Roig, A.,1998b. Carbon mineralization from organic wastes at different composting stages during their incubation with soil. Agriculture, Ecosystems and Environment 69: 175-189. Bolan, N.S., Adriano, D.C., and Mahimairaja, S., 2004. Distribution and bioavailability of trace elements in livestock and poultr y manure by-products. Critical Reviews in Environmental Science and Technology. 34: 291-338. Bole, J. B., and Bell, R. G., 1978. Land applic ation of municipal sewage waste water: Yield and chemical composition of fora ge crops. J. Environ. Qual. 7: 222-226.

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84 Tiffany, M.E., MacDowell, L.R., O’Connor, G.A., Martin, F.G., Wilkinson, N.S., Cardoso, E.C., Percival, S.S., and Rabian sky, P.A., 2000. Effects of pasture applied biosolids on performance and mineral status of grazing beef heifers. Journal of Animal Science. 78: 1331-1337. Tiquia, S. M., T. L. Richard, and M. S. Honeyman., 2002. Carbon, nutrient, and mass loss during composting. Nutr. Cy cling Agroecosyst. 62: 15-24. Tiquia, S. M., and F. Y. Tam., 1998. Elimin ation of phytotoxicity during co-composting of spent pig-manure sawdust litter an d pig sludge. Bioresour. Technol. 65: 43-49. Trenholm, L.E., Cisar, J.L., and Bryan Un ruh, J., 2003. Bahiagrass for Florida Lawns. Florida Cooperative Extension Service, Inst itute of Food and Agricultural Sciences, University of Florida ENH6. USEPA, 1993. Land application of sewage sludge: a guide for land-appliers on the requirements of the federal standards for the use or disposal of sewage sludge, 40 CFR Part 503. EPA-831-B-93-002b. Usman, A.R.A, Kuzyakov, Y., and Stahr, K., 2004. Dynamics of organic C mineralization and the mobile fraction of heavy metals in a calcareous soil incubated with organic wastes. Wate r, Air, and Soil Pollution. 158: 401-418. Valo, A., Carrere, H., and Delgenes, J.P., 2004. Thermal, chemical, and thermo-chemical pre-treatment of waste activated slud ge for anaerobic digestion. Journal of Chemical Technology and Biotechnology. 79: 1197-1203. Van Kessel, J.S., and Reeves, J.B., III, 2002. Nitrogen mineralization potential of dairy manure and its relationship to com position. Biol. Fert. Soils 36:118-123. Van Kessel, J.S., Reeves, J.B., III, and Meisinger, J.J., 2000. Nitrogen and carbon mineralization of potential manure com ponents. J. Environ. Qual. 29: 1669-1677. Webber, M. D., Rogers, H. R., Watts, C. D., Boxall, A. B.A., Davis, R. D., and Scoffin, R., 1996. Monitoring and prioritization of or ganic contaminants in sewage sludge using specific chemical analysis and pred ictive non-analytical methods. Sci. Tot. Environ. 185: 27-44. Wiese, A. F., Sweeten, J. M., Bean, B. W ., Salisbury, C. D., and Chenault, E. W., 1998. High temperature composting of cattle feed lot manure kills weed seed. Appl. Eng. Agric. 14: 377-380. Whithers, P.J.A., Clay, S.D., and Breeze, V.G., 2001. Phosphorus transfer in runoff following application of fertilizer, manure, and sewage sludge. J. Environ. Qual. 30: 180-188. Wodzinski, R. J., and Ullah, A. H. J., 1996. Phytase. Adv. Appl. Microbiol. 42: 263-302.

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85 Yingming, L., and Corey, R.B., 1993. Redistri bution of sludge-borne cadmium, copper and zinc in a cultivated pl ot. J. Environ. Qual. 22: 1-8. Zhang, M., Heaney, Solberg, D., E., and Heri quez, B., 2000. The effect of MSW compost on metal uptake and yield of wheat, barl ey and canola in less productive farming soils of Alberta. Compost Scienc e and Utilization, 8(3): 224-235.

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86 BIOGRAPHICAL SKETCH Caroline Reis was born in Uberlndia, Mina s Gerais, Brazil, in April of 1979. After graduating from high school, she attended Fede ral University of Uberlndia, where she graduated in 2003 with a bachelor’s degree in agronomy and where she was influenced by Dr. Korndorfer’s interest in soil science and fertilizer. Upon graduation, Caroline applied to the Un iversity of Florida where she met Dr. Jerry Sartain. She was accepted by Dr. Sartain into his program and will graduate with a M.S. degree in soil a nd water science in 2006.


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EVALUATION AND CHARACTERIZATION OF ORGANIC WASTE PRODUCTS
AS NUTRITIONAL SOURCES FOR BAHIAGRASS
















By

CAROLINE BORGES REIS HAMILTON


A THESIS PRESENTED TO THE GRADUATE SCHOOL
OF THE UNIVERSITY OF FLORIDA IN PARTIAL FULFILLMENT
OF THE REQUIREMENTS FOR THE DEGREE OF
MASTER OF SCIENCE

UNIVERSITY OF FLORIDA


2006

































Copyright 2006

by

Caroline Borges Reis Hamilton















ACKNOWLEDGMENTS

I would like to express my sincere thanks and appreciation to Dr. Jerry B. Sartain,

the chairman of my supervisory committee, for his guidance, patience and support

throughout my entire degree program. I could not have succeeded if was not for his trust

and belief in an unknown student. I also would like to extend my appreciation to my

committee members: Dr. Jack E. Rechcigl, Dr. Martin B. Adjei and Dr. Craig D. Stanley.

Sincere thanks and appreciation go to Ed Hopwood Jr., Nahid Varshovi and Martin

Sandquist for all their guidance and help in all the lab and glasshouse work.

Special thanks go to my husband, Kevin Hamilton, for his invaluable support,

friendship and encouragement during the past years. Also thanks go to my parents, Jose

Gaspar Reis and Maria Alice Borges Reis, my brother and sisters, and my grandmother

for all their love and support through my whole life. Without their help this thesis would

not be a reality today.
















TABLE OF CONTENTS

page

A C K N O W L E D G M E N T S ......... .................................................................................... iii

LIST OF TABLES ............. ..... ......................... .......... ............ vi

LIST OF FIGURES ............. .. ..... ...... ........ ....... .......................... viii

ABSTRACT ........ .............. ............. ...... ...................... ix

CHAPTER

1 INTRODUCTION ............... ................. ........... ................. ... .... 1

2 LITER A TU R E REV IEW ............................................................. ....................... 3

M anures and B iosolids in G eneral..................................................................... .... 3
B io so lid s ..................................................... ........................... 5
M anu re ............... ............................................................... 8
Bahiagrass (Paspalum notatum) ................................ ............... 9
O organic M matter and M icrobial A activity ........................................... .....................10
N nitrogen M ineralization............... .................................................... ............... 13
Phosphorus Mineralization ............ ..... ................................... 16
L teaching of Phosphorus ........................................... ....................................... 18
Phosphorus Runoff ................................................. ....... ............... 19
M anure M anagem ent ............................................................................ ............. 2 1
M icronu trients .............................................................................................. ..... 2 3
H heavy M etal A dditions......................................................................... .............24
Environm mental A aspects ............................................................. ............. ............... 28

3 M ATERIALS AND M ETHOD S ........................................ ......................... 30

Study 1: Chemically Characterize the Materials Relative to Their Nutritional
Value and Potential Toxic Environmental Impact...............................................30
Study 2: Determination of the Mineralization Rates of the Materials.....................32
Study 3: G lass H house Study ............................................... ............................ 33









4 RESULTS AND DISCU SSION ........................................... .......................... 35

Study 1: Chemical Characterization of Organic Materials Relative to Their
Nutritional Value and Potential Toxic Environmental Impact. ..........................35
Study 2: Determination of the Mineralization Rates of Study Materials .................41
C O 2 E evolution ...................................................... ................. 44
Study 3: Glass House Study 1st Application ................................. ................ 49
Bahiagrass Dry M atter Production ...................................... .......................... 49
Bahiagrass Root Dry M atter Production .................................. ............... 49
V isual Q quality R eating .......... .................................. .....................50
Bahiagrass Chem ical Com position.................................. ........................ 52
L teaching E valuation ........ ........ .. ........................ ................. ..... .........56
Study 4: Glass House Study 2nd Application................................. ............... 61
Bahiagrass Dry Matter Production ..... .................... ...............61
Bahiagrass Root Dry M atter Production .................................. ............... 62
Bahiagrass Quality Rating......... .................. ...................... ... 63
Bahiagrass Chem ical Com position.................................. ........................ 65
Leaching Evaluation ........ ............................... .. .. ........ ... ........ ..... 69

5 CON CLU SION S .................................... .. ... ........ ...... ..... ...... 74

L IT E R A T U R E C IT E D ............................................................................ ....................76

BIO GRAPH ICAL SK ETCH .................................................. ............................... 86
















LIST OF TABLES


Table page

2-1 Chemical composition of the two biosolids ............. ..............................................6

2-2 Inorganic and organic P soluble in water in different amendments.........................19

4-1 Heavy metal concentrations of biosolids in relation to USEPA ceiling
concentration limits for land application (on a dry weight basis). .........................40

4-2 Percentage of N mineralized in 256 days in relation to biosolids type and the
resulting soil pH and EC ............................................... .............................. 43

4-3 Analysis of variance (ANOVA) table used for the determination of statistical
differences in the analysis for effects of biosolids and rate source on total CO2
ev olv ed .............................................................................4 8

4-4 Bahiagrass stolon dry matter production after 112 days.......................................50

4-5 Visual rating during Fall 2004 comparing the biosolids at different rates ..............51

4-6 Analysis of variance (ANOVA) table used for the determination of statistical
differences in the analysis for effects of biosolids and rate source on total dry
m matter accum ulation. ........................ ....... .. .. ........ ...............52

4-7 Total N uptake by bahiagrass from biosolids in first 112 days.............................54

4-8 Quantity of P applied by each material and percentage of P uptake by bahiagrass.55

4-9 Total NH4-N and NO3-N content in leachates from organic materials ....................57

4-10 Concentration of NO3-N and NH4 -N in leachate as affected by the incubation
p e rio d ......................................................................... 5 8

4-11 Leachate pH values from 7 to 112 days after treatment application......................60

4-12 Leachate EC values from 7 to 112 days after treatment application.....................60

4-13 Bahiagrass root dry matter production 112 days after reapplication of the
m materials. ............................................................................62









4-14 Visual rating during Spring 2005 comparing N sources at different rates after
biosolid reapplication. .......................... ........................ .. ........... ............ 63

4-15 Total N uptake by bahiagrass from biosolids in 112 days after reapplication of
the m materials. .........................................................................66

4-16 Quantity of P applied by each material and percentage of P uptake by bahiagrass
after reapplication of the m aterials............................................... ......... ...... 68

4-17 Total NH4-N and NO3 -N content in leachates from the different organic
m materials after reapplication ......................................................... ............. 70

4-18 Concentration of N03-N and NH4 -N in leachate as affected by biosolid
reapplication. ..........................................................................70

4-19 pH measurements in the leachate from 7 to 112 days after reapplication of the
m ate ria ls .......................................................................... 7 2

4-20 EC measurements in the leachate from 7 to 112 days after reapplication of the
m ate ria ls .......................................................................... 7 3















LIST OF FIGURES


Figure page

4-1 Data represent total nitrogen, phosphorus and potassium. A. Total Kjeldahl
Nitrogen, B. Phosphorus and C. Potassium concentration ofbiosolids (on a dry
w eight basis)................................. .. ................................ ..........37

4-2 Data represent total calcium, magnesium, iron and manganese. (A) Ca, (B) Mg,
(C) Fe and (D) Mn concentrations ofbiosolids (on a dry weight basis) ................38

4-3 Data represent CO2 evolved from biosolids at weeks 1(A), 2(B), 8 (C), 16 (D),
24(E) and 33(F), respectively, at 90 Kg N ha-1. Data represent means and
standard error of three replicates. Means with the same letter are not different at
P > 0 .0 5. ........................................................... ................ 4 6

4-4 Data represent CO2 evolved from biosolids at weeks 1(A), 2(B), 8 (C), 16 (D),
24(E) and 33(F), respectively, at 180 Kg N ha-l. Data represent means and
standard error of three replicates. Means with the same letter are not different at
P > 0.05. .............................................................................47

4-5 Data represent total CO2 evolved from different biosolids at 90 Kg N ha-1 (A)
and 180 Kg N ha- (B). Data represent means and standard error of three
replicates. Means with the same letter are not different at P>0.05 ........................48

4-6 The effect of nitrogen rate on cumulative forage dry matter accumulation over
1 1 2 d a y s ........................................................................ 5 1

4-7 The effect of nitrogen source and rate on cumulative bahiagrass tissue dry matter
accum ulated over 112 days. ............................................ ............................. 64















Abstract of Thesis Presented to the Graduate School
of the University of Florida in Partial Fulfillment of the
Requirements for the Degree of Master of Science

EVALUATION AND CHARACTERIZATION OF ORGANIC WASTE PRODUCTS
AS NUTRITIONAL SOURCES FOR BAHIAGRASS


By

Caroline Borges Reis Hamilton

May 2006

Chair: Jerry B. Sartain
Co-chair: Jack E. Rechcigl
Major Department: Soil and Water Science

Biosolids can be used as nutrient sources in agricultural and horticultural areas. The

objectives were to chemically characterize biosolid materials relative to their nutritional

value and potential toxic environmental impact; determine the mineralization rates of the

materials relative to their ability to supply nutritional fertility and compare the biosolids

in a glasshouse setting relative to their potential for inducing growth response of

bahiagrass in a typical flatwoods soil. The materials used in this study were limed slurry,

limed cake, black kow, black hen, disney compost, milorganite, n-viro and baltimore

pellets. All materials were characterized for N, P, K, Ca, Mg, Fe, Mn and heavy metals

(Cd, Zn, Cu, Pb, Mo and As) using four replications. Milorganite contained the highest N

content followed by Black hen and Baltimore pellets. Limed slurry contained more than

2% N and P; limed cake, limed slurry and N-viro had adequate amounts of Ca while all

the sources had adequate amounts of Mg. Milorganite and baltimore pellets contained the









highest levels of Fe (39500 and 39000 ppm, respectively) while baltimore pellets and

black hen had the largest amount of Mn (850 and 550, respectively). All the biosolids

tested contained EPA acceptable heavy metals concentrations. For the mineralization

study, the materials (90 kg N ha-1 and 180 kg N ha-1) were mixed with an uncoated white

sand (1710g) and a surface layer of Arredondo fine sand (90 g) and placed into

incubation lysimeters. A CO2 trap of 1N NaOH was placed in the head space of the

lysimeters to estimate microbial decomposition rate. Limed slurry had the highest percent

mineralization at both N rates, followed by Milorganite and Baltimore pellets. Based on

CO2 evolved, the system was biologically active. Biosolids appear to have satisfactory

nutritional values and mineralization rates to support plant growth. For the glasshouse

study, bahiagrass was established by sodding in tubs using a typical flatwood soil from

Pine Acres area for a growth period of 112 days. Materials were surface applied

following establishment at rates of OKg ha-1, 90Kg ha-1 (1.74g N) and 180Kg ha-1 (3.48g

N) on a dry weight basis. Ammonium sulfate was used as a mineral N source. Parameters

estimated were bahiagrass root and tissue dry matter production, bahiagrass tissue

chemical composition (N and P) and leachate composition (pH, EC, N and P analysis).

Biosolids induced bahiagrass forage production similar to AS and superior to the

untreated control. Tissue N and P increased with increasing rates of application; limed

slurry and N-viro induced N uptake similar to AS. None of the biosolids contributed as

much N to the leachate as AS. No detectable P was found in leachates through out the

study, including after reapplication. Therefore, biosolids used in this study have the

potential for use in agriculture as a supplemental source of fertilizer and soil conditioner

not representing a potential toxic environmental impact.














CHAPTER 1
INTRODUCTION

Manures have been used as fertilizer for centuries, but crop fertilization with

manure has received renewed attention in recent years as concern for water pollution

potential from excess manure application has increased. Most people commonly view

biosolids as waste materials, but they can also be used as nutrient sources in agriculture

and horticultural areas .With proper treatment and processing, biosolids have the

potential to be used as fertilizers.

Biosolids are generated when solids accumulate during domestic sewage

processing. Wastewater residuals are produced wherever the population is concentrated

enough to require a centralized domestic wastewater treatment facility. These treatment

plants continuously generate sewage sludge that must be disposed of by one of several

means. Sewage sludge becomes biosolids when it undergoes pathogen control treatment

to meet federal and state sewage sludge regulatory requirements, followed by land

application to beneficially recycle it. During the past decades, some disposal practices

have been restricted. Landfill space is scarce, sittings of incineration facilities are difficult

and certain types of surface waters can be adversely impacted by excess nutrient load and

contaminants from treated effluents. Because of the wide range of environmental and

societal problems associated with municipal wastewater and sludge disposal,

municipalities have a need to find new ways to dispose of or, make better use of these

materials. For coastal cities in particular, regulations requiring elimination of ocean

disposal of sludge have precipitated the need for other management alternatives.









Fortunately, these materials have the potential for use in agriculture. The simultaneous

increase in the demand for water, coupled with tighter regulation over the safety of

sludge, widens an opportunity to use treated effluent for irrigation, and to use treated

sludge as a supplemental source of fertilizer and soil conditioner.

In the quest for more appropriate utilization of the biosolids on agricultural lands,

the following studies were conducted with the following objectives:

1. Chemically characterize several biosolids materials relative to their nutritional
value and potential toxic environmental impact.

2. Determine the mineralization rates of the materials relative to their ability to supply
required nutritional fertility and/or their potential for supplying excess loading of
identified hazardous heavy metals.

3. Compare the materials in a glasshouse setting relative to their potential for inducing
growth response of bahiagrass grown in a typical flatwoods soil.














CHAPTER 2
LITERATURE REVIEW

Manures and Biosolids in General

The most widely used organic amendments are sewage sludge and compost, which

contain high organic matter, N and P, making them suitable for agricultural purposes

(Usman et al., 2004). Biosolids are widely used as a soil conditioner and inexpensive

source of nutrients within agriculture (Webber et al., 1996; Petersen et al., 2003).

Biosolids provides short-term input of plant available nutrients and stimulation of

microbial activity, and it contributes to long-term maintenance of nutrient and organic

matter pools (Petersen et al., 2003). Land application of the residual biosolids (sludge)

from municipal wastewater treatment has increased in recent decades to become a

common disposal method in North America and Europe (National Research Council

2002). Where biosolids are used on agricultural cropland or forestland, the economical

benefits of improving soil structure and adding nutrients for plant uptake can partially

offset, the additional processing and transportation costs necessary to implement such a

program (Bastian, 1986; Kelty et al., 2004). The main alternative disposal methods of

landfilling and incineration have no such offsetting benefits. A third widely used disposal

method ocean dumping was banned in the US in 1992 (National Research Council 2002).

Biosolids application has been used more commonly in agriculture and wasteland

reclamation than in forestry (Kelty et al., 2004).

Manure is an important "commodity" that can enhance plant growth and reduce the

necessity for application of mineral fertilizer. Manure contains a range of compounds that









have rapid or intermediate N mineralization characteristics, or that are strong

immobilizers of N. Manures vary compositionally depending upon source (Serna and

Pomares, 1991; Van Kessel et al., 2000) and in vitro incubation experiments indicate that

the organic C and N are typically less mineralizable in cattle (Bos taurus) manure than in

swine (Sus scrofa) and poultry (primarily chicken, Gallus gallus domesticus) manures

(Serna and Pomares, 1991; Saviozzi et al., 1993; Van Kessel et al., 2000).

The dairy industry is under increasing pressure to more efficiently use and recycle

nutrients and to minimize detrimental losses to the environment. One of the critical

nutrient issues involves land application manure. Over-application of nutrients can lead to

contamination of surface or ground water while under-application results in reduced crop

yield. Soils in some regions of the United States are replete in P and manure application

is P-based in these areas, however, application rates have more typically been dictated by

N requirements. Information required to estimate the appropriate rate of manure

application includes: crop nutrient requirements, soil nutrient supply, and manure nutrient

supply (Van Kessel et al., 2002).

Manure N is an important resource that could be used more efficiently in crop

production. However, manure contains a wide variety of N compounds of varying

complexity that makes estimation of plant N availability difficult (Van Kessel et al.,

2000). Nitrogen mineralization is a relatively slow microbial process that is affected by

factors such as amendment composition, soil type, temperature, pH, aeration, and

moisture (Mikkelsen et al. 1995; Van Kessel et al., 2002). Organic N availability is

estimated to be 35%, 12% and 2% of the initial organic N in the first through to the third

year after dairy manure application (Van Kessel et al., 2002). Although fixed values are









generally used, estimates of mineralized organic N from cow manure in the lst year of

application are highly variable, and range from 0% to 50% (Serna and Pomares 1991;

Kirchmann and Lundvall 1993; Paul and Beauchamp 1994; Van Kessel et al., 2002).

According to Van Kessel et al. (2002) when manure-amended soils were incubated

for 56 days, all of the NH4+-N was depleted; NO3- -N concentrations increased during this

time period whereas NO2- -N did not accumulate in these samples and concentrations

were always negligible (<1% of total inorganic N). Overall, net mineralization ranged

from -29.2% to 54.9%. But if the average mineralizability of 12.8% was used to

determine crop-available N for a manure application, and the N from manure was

actually immobilized, the crop would have insufficient N. Alternatively, if the true net

mineralization was the other extreme, inorganic N would be added in excess and this

could have detrimental environmental and economical implications. Accurate estimates

of manure N composition and availability are an essential part of an efficient nutrient

utilization plan.

Biosolids

One main characteristic of biosolids is that they are very variable in composition

and do not form a group of fertilizers with a consistent nutrient content. As an example,

we have the chemical composition of two different types of biosolids in Table 2-1. The

fertilizer value of biosolids can be significant, but varies considerably depending on

origin and processing prior to application (Smith et al., 1998; Petersen et al., 2003).The

pools of dissolved nutrients in biosolids are typically small, and plant uptake must await

mineralization of organic constituents or, for P, dissolution of precipitates. This makes

information about the extent and temporal dynamics of nutrient release an important

aspect of waste characterization (Petersen et al., 2003).









Phosphorus content ranges generally from 34 to 57 g P205 Kg-1 dry matter

(Sommellier et al., 1996; Quilbe et al., 2005), but this content, as well as speciation

(mineral/organic form), depends on original effluent composition and types of treatment

(raw, digested or composted sludges). Surface runoff is the major cause of phosphorus

loss from cultivated fields, and this process was shown to be closely linked to sediment

transport (Sharpley et al., 1992; Quilbe et al., 2005). Nitrogen in biosolids is mainly in

organic forms, while mineral forms are generally in low concentration and are mainly

represented by NH4+-N. It was shown that, in biosolids, this element can be volatilized or

rapidly mobilized by runoff and leaching (Gangbazo et al., 1995; Quilbe et al., 2005). In

contrast, nitrate transfers occur only after a long-term nitrification process in soil (Garau

et al., 1986; Serna and Pomares, 1992; Quilbe et al., 2005).

Table 2-1: Chemical composition of the two biosolids
Parameter Anaerobically Digested Limed Sludge

Dry matter (%) 60.4 50.5
pH 7.6 12.4
Organic matter (g kg-1 dry matter) 328 172
Organic Carbon (g C kg-1 dry matter) 164 87
C/N ratio 11.1 8.9
Total Nitrogen (g N kg-1 dry matter) 15 19
Ammonium N (g N-NH4 kg-1 dry 2.4 0.2
matter)
Total Phosphorus (g P205 kg-1 dry 78 22
matter)

Quilbe et al., 2005

Pellets are a more highly processed material that requires greater initial production

costs but has important advantages over liquid or dewatered forms. Most importantly, the

heating process used to produce pellets substantially reduces microbial populations,

including pathogenic species, which changes the biosolid material from class B to class A

in US Environmental Protection Agency (EPA) regulations (National Research Council









2002). A consequence for land application is that precaution must be taken to limit access

to land for up to 1 year where class-B biosolids have been spread, whereas no such

requirement exists for applying class-A biosolids (Kelty et al., 2004). In addition,

transportation costs are lower for pellets than for liquid sludge, because the moisture

content is reduced by heat treatment to 10% or less. For use of liquid sludges (90%-95%

water), the moisture content of the material is often reduced for transportation and then

rewatered at the site of application, but the dewatered form still has a moisture content of

70%-85% (Tchobanoglous and Burton 1991; Kelty et al., 2004). Cake (dewatered)

sludges lose a substantial proportion of their inorganic soluble NH4-N in the liquid phase

during the dewatering process. Therefore, the type of sludge, and hence the form in

which the N is present in the soil, will affect the N mineralization dynamics of the sludge

amended soil (Smith et al., 1998; Smith et al., 2004). This is important from both the

estimation of N requirement for crop nutrition and the potential for N03-N leaching

(Smith et al., 2004).

Since biosolid production is increasing, more and more attention is being given to

anaerobic digestion as a Waste Activated Sludge (WAS) stabilization process. This

interest is due to the capacity of anaerobic digestion to reduce the amount of organic

solids and also to the formation of biogas which is a renewable energy source (Valo et al.,

2004). In anaerobic digestion of WAS, hydrolysis is considered to be the limiting step.

Indeed, after aerobic treatment in a wastewater treatment plant, the main part of the

sludge's organic matter is enclosed in bacterial flocs which reduce its availability to

anaerobic microorganisms (Valo et al., 2004).









Manure

Application of animal manures to agricultural fields is a widely used method of

increasing soil organic matter and fertility (Khaleel et al., 1981; Loecke et al., 2004).

Most solid livestock manures can be applied directly to crop fields or piled for

composting. Composting manure can reduce field application costs by increasing bulk

density and reducing volume. Composting can also increase application uniformity due to

a reduction in particle size, and decrease amounts of viable weed seeds (Wiese et al.,

1998; Loecke et al., 2004) and phytotoxic substances (Tiquia and Tam, 1998; Loecke et

al., 2004) contained in manure or manure bedding mixtures.

However, with composting, there are potentially greater production and

environmental costs associated with extra handling and possible losses of nutrients.

Nitrogen losses during composting of animal manures have ranged from 200 to 700g kg-1

of total N (Martins and Lewes, 1992; Rao Bhamidimarri and Pandey ,1996; Eghball et

al., 1997; Tiquia et al., 2002; Loecke et al., 2004). Lower N-use efficiency of compost is

typically attributed to increased humification of the compost relative to its feedstock fresh

manure (Loecke et al., 2004). Sewage sludge stabilization through composting is a

successful way to reduce the negative effects of the unstable organic matter in the soil

(Sanchez-Monedero et al., 2004; Bernal et al., 1998a), and increase the agricultural value

of the sludge as a consequence of the organic matter humification during the process.

Furthermore, the reduction in size of the wastes during stabilization reduces the costs of

transportation (Sanchez-Monedero et al., 2004). Composting manure is a useful method

of producing a stabilized product that can be stored or spread with little odor or fly-

breeding potential. The other advantages of composting are that it kills pathogens and

weed seeds, and improves the handling characteristics of manure by reducing manure









volume and weight. However, composting has some disadvantages that include nutrient

and C loss during composting, the cost of land, equipment, and labor required for

composting, and odor associated with composting (Eghball, 2002).

Bahiagrass (Paspalum notatum)

Bahiagrass (Paspalum notatum Flugge.) was introduced from Brazil in 1914. It was

first used as a pasture grass on the sandy soils of the southeastern United States

(Trenholm et al., 2003). Bahiagrass (Paspalum notatum Flugge.), a warm-season

perennial, is grown throughout Florida and in the Coastal Plain and Gulf Coast regions of

the southern United States. Bahiagrass is adapted to climatic conditions throughout

Florida and can be grown on upland well-drained sands as well as the moist, poorly-

drained flatwoods soils of peninsular Florida. In Florida, bahiagrass is used on more land

area than any other single pasture species, covering an estimated 2.5 million acres. Most

of this acreage is used for grazing with some hay, sod, and seed harvested from pastures

(Chambliss, 2000).

Bahiagrass is relatively easy and inexpensive to establish. It produces moderately

well on soils with very low fertility, yet has good response to fertilization (Chambliss,

2000). Bahiagrass is very deeply rooted, but tolerant of flooding. It is able to maintain

sod even under conditions of extremely low soil fertility and to continuously cycle

nutrients while building up a store of nutrients and carbon reserves. Bahiagrass is

commonly grown for a number of years prior to incorporation. During this time, it is able

to accumulate a large amount of dry matter and N. Incorporation of bahiagrass sod

improves soil tilth and slightly increases soil organic matter (Chambliss, 2000). It is

likely that the presence of growing sod increases the total microbial biomass of the soil.

This microbial biomass may present an additional source of organic N available for









mineralization. Concentration of N in bahiagrass aboveground herbage varies

substantially according to fertilization practices and generally ranges betweenl5-25 g kg-1

(Johnson, 2004). Much of the total biomass of bahiagrass is composed of non-herbage

components such as rhizomes, and roots. The N concentration of these parts is much

lower than it is in the forage. The N concentration of the residue also depends on

fertilization regime.

Nitrogen is a limiting growth factor for many of the forage species grown on sludge

amended soils. Much work has been undertaken to determine the nutrient benefits of

sewage sludge derived N (Epstein et al., 1978; Parker and Sommers, 1983; Coker et al.,

1987; Smith et al., 1998; Hallett et al., 1999; Smith et al., 2004). In particular, Smith et al.

(1998) categorized different types of sludge according to whether they showed high NO3-

N production potential (liquid digested and lagooned liquid undigested sludge), initial

immobilization of native soil N followed by limited N03-N production (dewatered

undigested sludge), or high resistance to mineralization and N03-N production (air dried

digested sludge) (Smith et al., 2004). The concentration of N will determine both the rate

and total quantity of N released.

Organic Matter and Microbial Activity

Franco-Hernandez et al., 2003 observed that the application of biosolids to soil will

increase its organic matter. An increase in organic matter increases infiltration of water,

increases CEC, improves soil structure and prevents erosion. Approximately 28% of

organic C of the biosolids mineralized within 42 days if no priming effect was considered

(Kuzyakov et al., 2000; Franco-Hernandez et al., 2003). Franco-Hernandez et al. (2003)

observed that production of CO2 increased when biosolids were added to soil, and there

was no difference in the production of CO2 between the different types of biosolids added









to the soil. Application of biosolids treated with Ca(OH)2 and manure treatment, reduced

the production of CO2 compared to the other types of biosolids treatments (Franco-

Hernandez et al., 2003). In order to determine microbial activity, Sanchez-Monedero et

al. (2004) utilized the specific respiration activity, which represents the amount of CO2

mineralized per unit of biomass carbon per day, which was calculated by dividing the

daily amount of CO2 evolved by the amount of biomass carbon, and expressed as mg CO2

per gram of biomass per day. This parameter has been successfully used to detect

disturbance or stress of the soil microbial biomass due to external inputs of organic

matter or the presence of toxic substances as heavy metals that were usually reflected as

an increase of specific respiration (Sanchez-Monedero et al., 2004).

Pascual et al. (1997) demonstrated that the addition of urban organic wastes

(municipal solid wastes, sewage sludge and compost) to the soil increases the values of

biomass carbon, basal respiration, biomass C/total organic C ratio, and metabolic quotient

(qCO2), indicating the activation of soil microorganisms (Usman et al., 2004). The

organic carbon mineralization during the experiment was recorded as cumulative CO2-

evolution (g kg-1 soil). The addition of sewage sludge and compost led to an increase in

organic carbon mineralization compared to unamended soil (Usman et al., 2004). The

cumulative amount of C mineralized increased with increasing application rate of sewage

sludge and compost. Usman et al., 2004 observed that the highest organic carbon

mineralization was found for sewage sludge. High loss of organic C by mineralization in

sewage sludge amended soil could have been due to the high microbial activity as a

consequence of the high concentration of dissolved organic C introduced with sewage

sludge. Dissolved organic C is the most important source of energy for microorganisms









(Usman et al., 2004). Whereas the evaluation of the size and activity of soil microbial

biomass could give useful information about deviation of soil environment from its

equilibrium, a combination of these two parameters as in the case of the specific

respiration (qCO2), that expresses the amounts of C02-C produced per unit biomass and

time, could be a more sensitive indicator of environmental changes, both permanent

(stress) and temporary (disturbance). This parameter has been successfully used to detect

disturbance or stress of the soil microbial biomass due to external inputs of organic

matter (Anderson and Donsch, 1990; Sanchez-Monedero et al., 2004) or the presence of

toxic substances as heavy metals (Brookes and McGrath, 1984 and Sanchez-Monedero et

al., 2004) that were usually reflected as an increase of qCO2.

Franco-Hernandez et al. (2003) observed that the higher pH in the soil at the onset

of the incubation when biosolids treated with Ca(OH)2 was added might have affected

microbial activity directly or indirectly through the formation of NH3. It might be

worthwhile to follow the salinity and sodicity in soil when biosolids are applied more

than once. High sodicity and salinity are known to inhibit plant growth and affect soil

processes (Franco-Hernandez et al., 2003). Application of biosolids treated with Ca(OH)2

might have the additional effect of increasing pH in acid soils but it might inhibit

mineralization in more alkaline soils (Franco-Hernandez et al., 2003).

Microbial biomass is a dynamic component of soil organic matter and therefore of

particular interests as a source of, or competitor for, plant nutrients (Petersen et al., 2003).

Smith (1998) concluded that the N flow through the microbial biomass would in many

cases be able to supply all the N required for crop growth. Petersen et al. (2003) observed

that microbes catalyze mineralization processes and therefore presumably multiplied









during sludge degradation. Addition of a fresh sewage sludge mixture caused the highest

soil qCO2 increase, from 27 to 210 mgCO2 g-1 day 1, after 3 days of incubation

(Sanchez-Monedero et al., 2004). After the initial increase, the specific respiration tended

towards the values of the control, indicating the pattern of the soil to recover its initial

equilibrium status (Sanchez-Monedero et al., 2004). Sanchez-Monedero et al. (2004)

observed that the highest qCO2 obtained for the soils amended with the less stabilized

sewage sludge mixtures implied that the newly developed soil microbial biomass

mineralized a larger amount of the added organic C, per unit biomass C than the soil

amended with mature compost. In fact, the carbon mineralization during incubation of

soil amended with the fresh mixture was 20% of the added C, whereas the C

mineralization did not exceed 5% when the mature compost was added (Sanchez-

Monedero et al., 2004).

Nitrogen Mineralization

In 1994, the U.S. broiler industry produced 7 billion broilers and generated about

10 million Mg of litter (Georgia Agricultural Statistics Service, 1995), which is a mixture

of chicken excreta and bedding material. Assuming an average concentration of 30g N

Kg-1, this litter contained 300,000 Mg of N, a valuable resource for fertilizing crops

(Gordillo et al., 1997). In a study involving manures, sewage sludge, and soil

amendments, Douglas and Magdoff (1991) found a good relationship (r2=0.82) between

the fraction of organic N mineralized and N released by Walkely-Black acid-dichromate

digestion. According to Gordillo et al. 1997, nitrogen mineralization from broiler litter

started immediately after the litter was incorporated into the soil, with inorganic N

accumulating very rapidly thereafter and on average, approximately 50% of the total N

released was mineralized during the first 24 hours. In addition, other easily mineralizable









N compounds, such as urea and creating, are present in fresh chicken manure (Gordillo et

al., 1997) and may also be present in broiler litter.

Composted dairy manures are a major source of N in organic farming where the use

of manufactured chemicals is prohibited (Hadas et al., 1994). The composting process

reduces the viability of weed seeds in manures, and suppressive effects of composts on

soil pathogens have also been found; thereby, the need for pesticides for the following

crop is reduced. The rate of net N mineralization of composts in the soil must be known

to optimize their use, minimize hazards of NO3 contamination, and predict the amount of

additional N needed during the growth period for optimal crop production (Hadas et al.,

1994). Castellanos and Pratt (1981) showed that composted manures released

considerably less available N than fresh manures. Hadas and Portnoy (1994) showed that

differences in the composition of the composts demonstrated the general problem that

composts are not a defined product, even if they are all produced from cattle manure.

Yard waste compost materials that are well cured during production, or that have

had additional curing time in the soil, appear to be able to release N at amounts and rates

similar to moderately fertile reference soils. Compost amendment levels of 500 kg total N

ha-1 appears to provide similar cumulative amounts of N release as the granitic subsoils

under these experimental conditions. Application of about 1000 kg total N ha-1 is

estimated to approach the cumulative N release amounts of the reference granitic topsoil.

The rates of long-term N release also approach those of the selected reference soil

materials, depending on compost type. Initial periods of immobilization appear to occur

for uncured or fibrous yard waste compost materials. Use of such materials could

potentially reduce initial plant establishment and growth, although such materials may









function well when used as surface mulch applications that decompose more slowly

(Claassen et al., 2004).

Net nitrogen mineralization rates from co-composted materials are much higher

than from yard waste composts initially, although the long-term N release rates were

more similar. Sizable initial releases of available N may occur from co-composts, which

may potentially allow leaching losses to watersheds or may facilitate weedy invasion

(Claassen et al., 2004). Approximately 1 to 7% of the total N contained in the yard waste

composts was released during the incubation period, suggesting that the yard waste

composts have the potential to continue to mineralize N for many years after application

to field soils. In contrast, the co-compost materials released about 27%, providing much

more short term available N, but less potential for continued N release (Claassen et al.,

2004). Immature composts may be high in ammonium, which is phytotoxic to plants.

Low availability of N and other macronutrients limits growth, if the feedstock for

composting is carbonaceous or otherwise low in nutrients (Barker, 2001). Franco-

Hernandez et al., 2003 observed that N mineralization was increased following

amendment with limed biosolids.

In the past, nitrogen availability has been assessed on the basis of either anaerobic

or aerobic soil incubation (Petersen et al., 2003). Nitrogen release by short-term

anaerobic incubation was largely of microbial origin (Petersen et al., 2003). However, net

N mineralization in laboratory systems without plants may not give a quantitative

estimate of plant N availability in the field, where crop roots compete with microbial

immobilization processes (Petersen et al., 2003) and, probably, field observations provide

a more realistic picture of nutrient availability. The mineralization of the nitrogen









contained in organic residuals depends on a variety of factors, including the type and

form of the residuals. Aerobically digested sludge had higher rates of N mineralization

than anaerobically digested sludge (19-50% versus 16-41%) after 16 weeks of incubation

at 300C (Adgebidi et al., 2003).

Composted sewage sludge exhibits a lower N mineralization rate compared to non-

composted sludge. Tyson and Cabrera observed organic mineralization rates of 0.4-5.8%

and 25.4-39.8% for composted and non-composted poultry litter, respectively, after 8

weeks of incubation at 250C (Adgebidi et al., 2003). Nitrogen immobilization, attributed

to relatively wide C/N ratios, which occurs when the nitrogen content of the sludge is not

adequate enough to meet the demands of the soil microbial community, has also been

reported for composts (Adgebidi et al., 2003). As the decomposition of the sludge occurs,

carbon evolved as carbon dioxide (C02), and the C/N ratio is gradually diminished,

enhancing nitrogen availability. The physical and chemical conditions prevailing in a

decomposition medium also affect its mineralization rates in poultry litter (Adgebidi et

al., 2003).

Phosphorus Mineralization

Most Florida forages are grown in sandy, infertile soils, and many mineral

deficiencies observed in grazing species can be related to the characteristics of soils of

the region (Tiffany et al., 2001).Municipal biosolids (sludge) have been used as fertilizer

for some time often due to their high phosphorus (P) content, which promotes growth of

agricultural crops (Tiffany et al., 2001). Mineral uptake by plants is affected by biosolid

load rate and chemistry, soil type and pH, and plant genetics. (Tiffany et al., 2001).

Today, dried, heat treated, anaerobically digested, pathogen-free, biosolids are widely









marketed. These biosolids contain varying amounts of mineral elements which could

prove beneficial, if they improve soils and forage nutritive value without creating

toxicity. (Tiffany et al., 2001).

Approximately 1.85 x 108 Kg of P enters the environment in the form of animal

manure per year in the USA (Wodzinski and Ullah, 1996; Parham et al., 2002). Sound

waste management in ecosystems or watersheds demands prudent consideration of P

inputs (Pierzynski et al., 1994). Unfortunately biosolids and manure-borne P reactions

have not received nearly the study that waste-bound N has. We do know that (as with N)

calculations based on total waste-P concentrations to determine allowable P loads to a

soil can be grossly over-conservative, as all the (total) P may not be bioavailable or

soluble enough to leach. Phosphorus in biosolids and manures exists in a variety of forms

(e.g. McCoy et al., 1986; Pierzynski et al., 1994) resulting in reported bioavaliabilities (to

plants) ranging from 10% to 100% of that of soluble fertilizer P (e.g. de Haan, 1980;

Gestring and Jarrel, 1982; Sikora et al., 1983; MacCoy et al., 1986). Most agricultural

best management practices for animal wastes are now based on providing sufficient

nitrogen (N), in a timely manner, to meet crop N requirements at a realistic yield.

Documented problems with nitrate-N (N03-N) contamination of ground waters in areas

with high animal densities have been the driving force behind this approach. However, a

number of emerging environmental issues, such as the eutrophication of surface waters

by phosphorus (P) in runoff; the fate of trace elements, antibiotics, pesticides, and growth

hormones in wastes; and the effects of pathogens in wastes on human and animal health

have forced us to re-evaluate the N-based management of animal wastes (Sims and Wolf,









1994). Franco-Hernandez et al., 2003 observed that phosphorus mineralization occurred

but the application of biosolids did not increase rate of mineralization.

Inorganic phosphate (Pi) partakes in many abiotic processes, such as sorption to

soil particles and precipitation. Phosphorus is also an essential nutrient and as such is

involved in biological reactions in the soil (Petersen et al., 2003). Biotic and abiotic

processes occur simultaneously and interactively, and both aspects should be considered

in discussions of soil P dynamics (Petersen et al., 2003). The concentration of inorganic

phosphate in the soil solution is low and crops to a large extent draw upon P associated

with the solid phase, which complicates the assessment of P availability to the crop

(Petersen et al., 2003). Petersen et al. (2003), observed that inorganic N accounted for

only a small fraction of total N in both sludges anaerobicallyy digested sludge and

activate sludge), and the crop N uptake observed thus implies a significant net N

mineralization.

The continued application of fertilizers and manures in many areas has resulted in

the buildup of soil P concentrations above those required for optimum plant growth,

because of these elevated concentrations the potential for P loss is increased. This loss is

a function of, but not exclusively, topography, soil type, soil test phosphorus (STP)

concentration, and soil hydrology (McDowell and Sharpley, 2001). Both the properties of

the biosolids (P forms, solubility) and the soil being amended (P fixing capacity)

determine the potential environmental hazard of such accumulated P (Anjos et al., 2000).

Leaching of Phosphorus

Areas with intensive livestock farming often have soils enriched with P due to the

addition or disposal of manure (Withers et al., 2001). In areas of Europe and North

America, P-rich soils are common (Fixen, 1998; Skinner and Todd, 1998). Until about 15









years ago, there was a common misconception within the literature that P loss via

subsurface pathways was negligible as P is strongly sorbed to soil and thus, only surface

pathways were of concern (Sharpley and Menzel, 1987). However in free-draining soils

and those where drainage is enhanced, P can be lost in environmentally significant

quantities (Heckrath et al., 1995; Hesketh and Brookes, 2000). A comparison of inorganic

and organic P soluble in water in different amendments is showed in table 2-2:

Table 2-2: Inorganic and organic P soluble in water in different amendments
Fraction From Dairy Dairy Poultry Poultry Superphosphate
manure compost manure compost P (%)
P (%) P (%) P (%) P (%)
Water Inorganic 51 15 26 21 100
Organic 12 1 8 1 ----

Total Inorganic 63 92 84 87 100
_Organic 25 5 14 11 ----
(Derived from Macdowell, R.W. and A.N. Sharpley. 2003)

MacDowell and Sharpley (2003) noted that for high-flow lysimeters, more P was

leached from dairy compost and poultry manure amended soils compared with mineral

superphosphate and dairy manure. In both cropped and grassland situations, it has been

shown that increasing soil P is associated with higher concentrations of P in drainage

water than in low P soils (Sharpley et al., 1996; Sims et al., 1998; Sharpley et al., 2000).

Measures to reduce the potential for P enrichment to runoff include manure treatment,

composting, peletizing, and transport to areas with a nutrient deficit (Sharpley et al.,

1998; Sharpley et al., 2000).

Phosphorus Runoff

Phosphorus loss in runoff can cause eutrophication of streams and lakes resulting in

surface water quality problems. An important component in managing P losses to the

environment is understanding crop P uptake from various nutrient sources. Plant P









availability may differ from one organic P source to another as a result of different

origins (e.g. municipal waste, dairy, swine, poultry) or variations in management (e.g.,

storage time or temperature, chemical treatments, manure separation, inclusion of

bedding) (Ebeling et al., 2003). Knowing the differences in P availability among P

sources will guide application rate decisions so that crop P requirements are satisfied

while minimizing soil P accumulation and runoff losses (Ebeling et al., 2003).

Beneficial use of biosolids as a soil amendment is based on their potential to

positively alter soil properties such as plant nutrient availability, water holding capacity,

tilth (physical conditions of soil related to tillage, seed bed and rooting media), and cation

exchange capacity (related to enhanced soil organic matter status) (Camberato et

al.,1997). However, due to their high nutrient content and other chemicals such as trace

elements, sewage sludges may create potential health and environmental problems that

might question their use in agriculture. In particular, mobilization of P and N in runoff

after sludge spreading is likely to contribute to eutrophication of downstream surface

waters (Quilbe et al., 2005).

The wastewater treatment process involves a primary settling step to separate heavy

organic solids from sewage. A secondary biological process uses cycling anaerobic and

aerobic conditions to remove P from plant effluent. The solids from primary settling and

the waste biomass from secondary treatment are digested anaerobically to decompose and

stabilize the solids. The digested solids are thickened by use of a gravity belt thickener

and organic polymer addition prior to being recycled to agricultural land (Ebeling et al.,

2003).









Data from a greenhouse and field study conducted by Goss and Stewart (1979)

indicated that feedlot manure produced higher crop utilization efficiency than

superphosphate fertilizer. They suggested that the microbial activity in the soil increased

manure P availability, reduced luxury P consumption by plants, and extended the time

period of adequate P availability levels (Ebeling et al., 2003). Ebeling et al.(2003) studies

indicated that P sources with relatively high soluble P or bioavailable P concentrations

(CaHPO4 and biosolids) provided higher levels of P for plant uptake in poorly buffered

systems such as sand media.

Manure Management

Manure management recommendations must account for site vulnerability to

surface runoff, macropore leaching, and erosion, as well as soil P content and saturation,

because not all soils and fields have the same potential to transfer P by surface runoff and

leaching to surface waters. As a result, threshold soil P levels should be indexed against P

transport potential with lower values for P source areas than for areas not contributing to

water export (Sharpley et al., 2000).

When applied to the soil, inorganic P (MCP and DCP) and ammoniated phosphate

remain localized around the point of application unless cultivation or other mixing

processes occur (James et al., 1996).Kissel et al. (1985) described the chemistry of P in

calcareous soils as a continuum of adsorption, formation of amorphous calcium

phosphate compounds, and gradual formation of crystalline phosphate compounds (James

et al., 1996). In soils exposed to manure over extended periods, P enrichment occurs

below the surface 20-cm layer (James et al., 1996). Dalal (1977) concluded that the

accumulation of organic P in soils was primarily a result of microbial activity and the

organic P may become available to plants by mineralization (James et al., 1996). James et









al.(1996) concluded that long-time manure disposal on land increased the concentration

of extractable inorganic P in subsoil layers as deep as 210cm and that manuring increased

extractable organic P concentrations markedly in the 0 to 30 cm layer but this organic P

dissipated to background levels within 2 to 3 years after manuring ceased. According to

Parham et al (2002) animal manure-P is relatively more mobile but less available for

plants than inorganic fertilizer-P. Long-term application of cattle manure did not result in

excessive accumulation of P in the surface 0-30cm soils, but promoted microbial

activities and P cycling in soil. James et al. (1996) observed that organic P and inorganic

P from heavy manuring do not threaten soil or groundwater quality under deep calcareous

soils if manure application is limited by acceptable N-loading rates. Wastewater

treatment and sludge processing methods markedly influence biosolids P mobility (Elliott

et al., 2002). Lu and O'Connor (1999) observed that sludge materials when treated with

Fe and Al to remove P from the corresponding liquid wastes, thereby containing total Fe

and Al of 10% or more may increase soil P sorption when applied to the land, especially

when the sludge is applied to soils with low native P adsorption capacity such as Myakka

sand soils. Lu and O'Connor (2001) observed that increases in P sorption were

correlated with increases in oxalate-extractable Fe and Al contents of amended soils.

Although temporary, the increased retention of P effected by biosolids applications can

have important implications. Phosphorus in biosolids containing (or tailored to contain)

abundant Fe and/or Al can be expected to behave as a slowly available P source, and to

be less subject to excessive leaching losses than completely soluble sources. This can be

very important in many areas of Florida dominated by soils that sorb P poorly and allow

extensive P leaching.









Soils common to most of the USA contain sufficient P-sorptive capacity (Al and Fe

oxides) to prevent P leaching and to mask inherent differences in the P solubilities of

biosolids materials. Recent Florida legislation requires P-based biosolids application

rates in watersheds associated with P-sensitive water bodies (Sec. 373.4595 FL Statutes).

Compared with N-based nutrient management, a P-based approach dictates lower waste

application rates (Elliott et al., 2002). Eghball (1999) found that application of beef cattle

feedlot manure or compost increased the soil surface (0-15cm) pH while N application as

NH4N03 reduced the pH (from 6.4 to 5.6). The increase in soil pH with manure and

compost application was attributed to a beef cattle diet that contains approximately 15g

CaCO3 kg1.

Application of manure with an increased N: P ratio at rates based on crop N

requirements would more closely match crop uptake of P and reduce the potential for soil

P accumulation (Sharpley et al., 2000). Controls over agricultural P inputs are required to

prevent soil P build-up, causing increased background losses; better management of fresh

P inputs is required to avoid incidental losses of P that occur when storm events follow

applications of manures to the soil surface and more precise land management is required

on naturally dispersive soils in runoff producing areas (Sharpley et al., 2000).

Micronutrients

Moral et al. (2002) studies indicated that sewage sludge applications can be useful

to increasing the available micronutrients (Fe, Mn, Cu and Zn) concentrations in

calcareous soils. They suggested that metal availability in the calcareous soils was closely

related to texture due to their influence in the organic matter dynamics. The dynamics of

the metal fraction are dependent upon the stabilization of the organic matter (composted

vs non-composted), rate of amendment applied (non linear increments with rate), and also









the texture of the soils (loamy vs sandy soils) (Moral et al., 2002). In addition, Kuo

(1990) observed that sewage sludge mineralization increased the soluble organic

compounds, especially low molecular weight organic acids in soils, that could mobilize

not only the exogenous micronutrients from the sewage sludge but also the non-available

metal fraction in the soil.

Heavy Metal Additions

Unlike sewage sludge application, where land application is limited based on

allowable metal loadings by EPA 503 regulations (U.S. EPA, 1993), regulations

governing livestock and poultry manure by-products are generally governed at the state

level, based on total N and/or P loading (Bolan et al., 2004). The U.S. Environmental

Protection Agency (EPA) Part 503 rule regulates nine metals in sewage sludge that is to

be land applied. These include As, Cd, Cu, Hg, Mo, Ni, Pb, Se, and Zn (U.S. EPA, 1993).

U.S. EPA Part 503 risk assessment indicates that other metals do not pose potential health

or environmental risks at land application sites at typical application rates (Bolan et al.,

2004). It has often been observed that metals added through organic amendments, such as

sewage sludge and manure by-products, accumulate in the surface layer, indicating a

strong retention by the amended soil (Adriano, 2001; Bolan et al., 2004).

Distribution of metals in sewage sludge and manure by-products and their

subsequent redistribution in soils amended with these materials have generally been

investigated by selective sequential chemical extraction techniques (Bolan et al., 2004).

The principal metal forms in manure and manure-treated soils are soluble, exchangeable,

adsorbed, organic-bound, oxide-bound, and precipitated (Bolan et al., 2004).

Concentrations of heavy metals recovered from dry biosolids were lower than 100 mgKg-

1 except for zinc (Zn) and chromium (Cr) on materials treated with lime. (Franco-









Hernandez et al., 2003). Applications rates to soil should be limited so that heavy metals

do not accumulate and end up into the food chain or groundwater, that pathogens can not

infect people or that concentrations of toxic compounds become dangerous (Franco-

Hernandez et al., 2003). Whereas the risk associated with the presence of heavy metals

and persistent organic pollutants can only be controlled by limiting the amounts of sludge

applied to the soil, the amount of pathogens can be reduced by sludge stabilization

treatment prior to addition to the soil (Sanchez-Monedero et al., 2004).

Sanchez-Monedero et al. (2004) observed that the bioavailable DTPA-extractable

Cu in the soil amended with any of the organic mixtures (38% sewage sludge + 62% of

cotton waste in four composting periods: initial, during the thermophilic phase after 21

days of composting, the end of the active phase after 49 days of composting and mature

compost after 109 days of composting) was on average 14% lower in the soil amended

than the control soil, indicating a positive effect of the organic matter on the

immobilization of this heavy metal in the soil. The opposite effect was shown for Zn, as

all the soils amended with the organic mixtures showed higher concentration of the

available metal (on average about 39% higher than the control soil). They also observed

that the amount of heavy metals in both soil and organic materials were under the

maximum levels accepted by the current legislation. The increase in total metal content

following a single sludge addition was negligible (<1 mg kg-1) for Cu, Ni, Cd, Cr and

Pb). In the case of Zn, amendment of soil with the mature compost caused an increase of

total content of 2.9 mg kg1.

Organic wastes such as sewage sludge and compost increase the input of carbon

and nutrients to the soil. However, sewage sludge-applied heavy metals, and organic









pollutants adversely affect soil biochemical properties (Usman et al., 2004). It is evident,

that the heavy metals introduced with sewage sludge or compost cause accumulation of

organic matter and decrease the turnover rate of organic matter, presumably because of

inhibitory effects on the microbial biomass (Usman et al., 2004). Moreno et al, (1999)

found that the addition of cadmium-contaminated sewage sludge compost to the soil

decreased microbial biomass C and stimulated the metabolic activity of the microbial

biomass (Usman et al., 2004).The addition of sewage sludge to soils could affect the

potential availability of heavy metals (Usman et al., 2004). Mineralization of sludge

organic matter may release heavy metals into more soluble forms that may harm sensitive

crops and microbes (Usman et al., 2004). The availability of heavy metals in the sewage

sludge and soils treated with sewage sludge depends on many factors such as the

properties and amount of heavy metals, the partitioning of heavy metals between solution

and solid phase, and soil characteristics (Usman et al., 2004). It is widely known that the

bioavailability of heavy metals in soil is strongly influenced by the amount and the

quality of organic matter that can react with the heavy metals, forming complexes and

chelates of varying stability (Usman et al., 2004). Soil organic matter is quite effective in

retaining heavy metals. Heavy metal-organic associations can occur both in soil solution

and at the solid surfaces of either native soil constituents or any added material (e.g.,

Biosolids) (Usman et al., 2004). In a heavy metal-polluted soil, Kiikila et al. (2002)

studied the effect of biosolids as organic immobilizing agents and observed that the

exchangeable Cu concentration decreased (Usman et al., 2004). On the other hand, the

heavy metal could be completed by dissolved organic matter (DOC), which enhances

leaching in the field (Usman et al., 2004).









Numerous laboratory and fields studies have demonstrated that most trace elements

(including common industrial metal elements) are relatively immobile in soils and once

added will remain in the layer of incorporation. Therefore, problems associated with trace

elements entering ground water beneath land where sludge has been applied is unlikely

(Yingming and Corey, 1993). Results of field studies demonstrated that crops grown on

sludge-amended soils benefited from the plant nutrients in sewage sludge, but would not

accumulate As, Cr, Cu, Pb, Hg, Ni, or Zn in amounts sufficient to be harmful to

consumers. Depending upon the soil conditions, however, there was a potential, for Cd,

molybdenum (Mo), and Se to be taken up by crops in amounts harmful to humans (Cd)

and animals (Cd, Mo, and Se) who consumed the affected harvest (Logan and Chaney,

1983). Cadmium is one of the most controversial of the contaminant elements. Unlike Cr,

Cu, Zn or Se it plays no known role in plant or animal metabolism (Zhang et al., 2000).

Neither total nor available heavy metal concentrations were increased in the soil by

sewage sludge compost application, in fact, the composting process reduces the heavy

metal availability in the raw material, possibly due to adsorption on or completing by

humic substances (Korboulewsky et al., 2002). Available B is strongly associated with

organic matter in soil. Highly organic materials like compost may have a large portion of

the B they contain in the available and potentially toxic forms (Zhang et al., 2000).

Copper sulfate (CuSO4) is a potentially beneficial feed additive in swine (Sus

scrofu domesticus ) and poultry (Gallus gallus domesticus) production, but while

supplementing copper (Cu) levels in swine production, concern has been expressed over

elevated Cu concentrations (up to 1000 mg Cu kg-1) in the manure resulting from this

practice (Miller et al., 1986). Studies using sequential metal extraction techniques on









sewage sludges and Cu in manures have suggested sulfides, carbonates, and organic

complexes of Cu as major chemical forms (Miller et al., 1986). Calibration of sequential

extractants against standard Cu minerals allowed Stover et al. (1976) to infer that sulfides

may be an important Cu form in sewage sludge (Miller et al., 1986). Miller et al. (1986)

concluded that Cu in swine manure is primarily bound to organic, contrary to earlier

findings, and that sludges also contains predominately alkali-soluble Cu, although

differences in origin and composition of the waste have been shown to be important.

Miller et al. (1986) further concluded that drying of manure prior to chemical extraction

substantially increases the solubility of Cu.

Environmental Aspects

Continuous heavy manuring may pose environmental problems, such as odor,

pollution of ground and surface water due to leaching and run-off of organic and

nutrients, and soil accumulation of heavy metals (Hsu et al., 2001). The composting of

organic wastes is mainly a biological decomposition process of organic materials,

resulting in a net loss of total organic matter (OM) and a concentration of inorganic

constituents (Hsu et al., 2001).The main products of the composting process are fully

mineralized materials such as CO2, H20, mineral ions, stabilized OM (mostly humic

substances), and ash. Composting reduces the volume and weight of the raw material

resulting in a stable product that can be applied to soil as a valuable fertilizer and soil

amendment (Hsu et al., 2001). In cases where the potential toxic metal concentrations of

compost are high, the leachability of metals associated with compost is of concern.

Studies have shown that the chemical form of an element is more important than the total

concentration in determining its leachability into ground water (Hung Hsu et al., 2001).

According to Hung Hsu et al. (2001), during composting, the major portions of Cu, Mn,









and Zn were in the organically-bound, solid particulate and organically completed

fractions, respectively. Metal distributions in different chemical fractions were generally

independent of composting age and, thus, independent of respective total metal

concentrations in the composts. Jackson et al. (2003) observed that trace elements Ni, Cu,

and As were found to be readily soluble in poultry litter and that more than 70% of Ni

and Cu was in the cationic form, or bound in relatively labile complexes that dissociated

to the cationic species, therefore trace metals from poultry litter are expected to be readily

sorbed by soil mineral phases on land application of poultry litter. It is becoming

increasingly important for producers to closely monitor manure nutrient application and

adjust fertilizer application based on manure inputs (Van Kessel et al., 2000).














CHAPTER 3
MATERIALS AND METHODS

This research consisted of three separate studies conducted at the University of

Florida, Gainesville, Florida. The first study was conducted on an array of sludge

materials to determine their nutritional value and potential toxic environmental impact in

the spring of 2004 from February through April. The second study was conducted in the

laboratory on the same sludge materials to determine the mineralization rates in the

spring of 2004 from April through December. The third study was conducted in a

glasshouse on bahiagrass established from sod during the fall of 2004 from September

through January 2005. A second growth response evaluation was conducted during the

spring of 2005 from March through July to document the nutrients build up in the soil

and to assess their nutrient availability to bahiagrass.

Study 1: Chemically Characterize the Materials Relative to Their Nutritional Value
and Potential Toxic Environmental Impact.

This portion of the project the organic materials were chemically characterized for

the following elements: N, P, K, Ca, Mg, Fe, Mn and heavy metals (Cd, Zn, Cu, Pb, Mo

and As), using four replications for each material. The materials evaluated were: Limed

Slurry (Anaerobically digested, lime stabilized slurry biosolid (Class B)); Limed Cake

(Lime-stabilized cake biosolids (Class B)); Black Kow (Composted cow manure (Class

A)); Black Hen ( Composted Poultry manure (Class A)); Disney Compost (Composted

biosolid) (Class A)); Milorganite (Anaerobically digested sewage sludge (Class A));









Tarpon Springs N-Viro (Aerobically digested (Class A)); Baltimore pellets

(Anaerobically digested sludge (Class A)).

Materials were prepared for analysis for Ca, Mg, K, Fe, Mn and P using the ashing

method described by Hanlon et al. (1994). Five grams of the dried material were placed

in 50ml pyrex beakers. Samples were ashed in a furnace at 5500C for 5 hours. Samples

were then moistened using a small amount of deionized water and 3.0 M Nitric acid.

They were then placed on a hot plate and dried. Samples were placed back in the furnace

and heated at 4500C for an additional 10 minutes. Samples were allowed to cool and 1.0

ml of 5.0 M HC1 was added. The sample was scraped from the beaker using an angle-

edged rubber policemen, decanted into a 50 ml volumetric flask, and diluted to 50 ml

using deionized water.

Calcium, Mg, K, Fe and Mn were determined using a Varian Atomic Absorption

Spectrophotometer (Varian Analytical Instruments, Sugar Land, TX). Phosphorus was

determined using the Molybdate Blue method (Hanlon et al., 1994). Nitrogen was

determined by TKN, Total Kjeldahl Nitrogen, method described by Horneck and Miller

(1998). A two grams tissue sub-sample was placed in a 50 ml digestion tube to which 1.5

g of digested catalyst (Kel -Pak 1.0g K2SO4, 0.3g CuSO4) and 3mL of 35N H2SO4 were

added. The mixture was mixed using vortex shaker and heated on a Tecator Digestion

System 40-1016 Digestor (Tecator AB, Hoganas, Sweden) at 3700C for 3 hours. Once

the solution cooled, it diluted to 50 ml using deionized water. Total N was determined

using Alpkem RFA-300 auto analyzer. For heavy metals the materials were digested

according to Jackson (1958) as presented by Fiskell (1965). One half gram of the

materials was transferred to a digestion tube, 25 ml of water and 2ml of concentrated









HNO3 was added and evaporated to dryness on a hot plate. Ten ml of 30% hydrogen

peroxide was added and the solution was evaporated to near dryness at 900C. This

treatment was repeated until no effervesces occurred on addition of H202. Three drops of

concentrated H2SO4 and 10 ml of HF were added. Samples were placed on the block

digester (2000C) and the solution evaporated to dryness. Finally, 15 ml of concentrated

HNO3, 2 ml of concentrated H2SO4, and 5 ml of HCLO4 were added and samples were

continuously heated until strong fumes of SO3 were produced. Samples were allowed to

cool, 25 ml of water was added, the sides of the containers were rinsed and policed with a

polyethylene rod to complete the extraction of the metals. Solution was transferred to a

50 ml volumetric flask and diluted to volume (Page et al., 1982). Copper and Zn were

determined by atomic absorption and Cd, Pb, Mo and As were determined by ICP (Page

et al., 1982).

Study 2: Determination of the Mineralization Rates of the Materials

The mineralization rates of the materials was determined in order to evaluate their

potential for supplying nutrients over time. Incubation lysimeters (30 cm section of 7.5cm

diameter PVC tubing fitted with fiberglass mat across the lower end, held in place by a

PVC cap) were used to determine the mineralization rates.

The materials were applied at rate equivalent to 90 kg N ha-1 and 180 kg N ha-1

respectively, on a dry weight basis. Uncoated white sand (1710 g) and a surface layer (0

to 5cm depth) of Arredondo fine sand (90 g) were mixed with the materials and placed in

the incubation lysimeters. The mixture was brought to 10% moisture (approximately 80%

water holding capacity). A CO2 trap of IN NaOH was placed in the head space of the

incubation lysimeter to estimate microbial decomposition rate. The CO2 trap was

replaced and titrated with IN HC1 weekly to estimate CO2 evolution. After 7, 14, 28, 56,









84, 112, 140, 168, 208 and 263 days each lysimeter was leached with 0.02 N citric acid

and ammoniacal-N, nitrate-N and P were determined on the leachate. Lysimeters were

maintained at near optimum moisture, but to induce leaching an additional /2 pore

volume of excess water (ca. 500 ml) was added every leachate day for a total of 10

leachates. These leachates were designed to represent the natural removal of the

mineralized N. The leached volume was recorded and an aliquot was taken for NH4,

NO3-N and P analysis (Horneck and Miller, 1998).

Study 3: Glass House Study

This study was set up to compare the biosolids in a glasshouse setting relative to

their potential for inducing growth response of bahiagrass in a typical flatwoods soil. The

study was conducted at the University of Florida's Turfgrass Envirotron, Gainesville,

Florida. Bahiagrass (Paspalum notatum) was established by sodding during the fall of

2004 in a glass house in tubs (0.19m2) using a typical flatwood soil from Pine acres area

(Candler fine sand sandy, siliceous, uncoated, hyperthermic Typic Quartzipsamments)

for a growth period of 112 days. Materials (Limed cake, Limed slurry, Disney compost,

Baltimore pellets and N-viro) were surface applied following bahiagrass establishment at

rates of 0 kg N ha-1, 90 kg N ha-1 (1.74 g N) and 180 kg N ha-l (3.48 g N) on a dry weight

basis. A standard inorganic N source, Ammonium sulfate, was used for comparison. The

parameters estimated were: bahiagrass root dry matter production, bahiagrass quality

rating, bahiagrass chemical composition and leaching evaluation (pH, EC, N and P

analysis). The experiment was arranged in RCB design with 4 replicates.

Glasshouse conditions were maintained relatively constant by means of computer

integration of overhead fans, an exhaust fan, a wall length humidifier, and automatic roof

window. Leachates were collected after 7, 14, 28, 56, 84 and 112 days. Clipping for dry









matter production were taken as needed for a total of 7 collections. Stolon samples were

collected at termination and washed free of soil. Tissue and stolon samples were dried at

70C for 48 hours. Tissue samples were ground to 2 mm mesh and ashed (Hanlon et al.,

1994c). Samples were analyzed for P by spectrophotometry using the molybdenum-

reduced molybdophosphoric blue color method, in a sulfuric acid system (Hanlon et al.,

1994a). Leachates were collected at 7, 14, 28, 56, 84 and 112 days. The leachate volume

was recorded and an aliquot was taken for pH, EC, P, NH4 and NO3 analysis (Hanlon et

al., 1994). A second growth response evaluation was conducted during the spring of 2005

from March through July to document the nutrients build up in the soil and to assess their

nutrient availability to bahiagrass. The same tubs with the established stand of bahiagrass

were employed. Statistical analysis for all studies was performed using SAS for analysis

of variances (SAS Institute, 1987). Duncan's Multiple Range Test (DMRT) was used to

identify differences among means.














CHAPTER 4
RESULTS AND DISCUSSION

Study 1: Chemical Characterization of Organic Materials Relative to Their
Nutritional Value and Potential Toxic Environmental Impact.

Most biosolids contain 40 g kg-1 to 70 g kg-1 of N, 1 to 20 g kg-1 P205 and very

little K 0.001 to 10 g kg-1. The stabilization process used to treat wastewater alters the

nutrient composition of the resulting biosolids (Muchovej et al., 2001). In this study

Milorganite contained the highest N content (59 g kg-1) followed by Black hen and

Baltimore pellets with 31.5 g kg-1. Limed slurry contained 23 g kg-1, Disney compost 16

g kg-1 and Limed cake 12.9 g kg-1. The materials N-viro and Black kow contained the

lowest content of total kjeldahl nitrogen (TKN) with 1.8 and 3.5 g kg-1, respectively (Fig

4-1A.). Concentrations of 16 g Kg-1 N are usually sufficient to increase mineralization

under aerobic conditions (Sartain, personal communication). Nitrogen availability

depends largely on microbial activity which in turn depends on soil conditions (e.g.,

moisture and temperature) and the nature of the biosolid. Conditions favorable for

conversion of N from organic to inorganic forms are roughly the same as those favorable

for crop growth. For waste materials, it is generally estimated that 40 to 50% of the

organic N will become available during the year following application (Kidder, G.,

2001).

Black hen contained the highest P content (27.3 g kg-1) followed by Baltimore

pellets (25.6 g kg-1), Milorganite and Limed slurry (22 g kg-1), Disney compost (11.7g kg-

1) and Limed cake (9. Ig kg-1). Black kow and N-viro had the lowest P content, 3.7 and









2.6g kg-1, respectively (Fig 4-1B.). Materials were generally very low in total K, ranging

from 5.6 g kg-1 (Limed slurry) to 0.7 g kg-1 (Baltimore pellets). The only exception to this

trend was Black hen which contained 46 g kg-1 of K (Fig 4-1C.). The materials Limed

cake, N-viro and Limed slurry contained 2.4, 1.2 and 1.1 g kg-1 of Ca and 6.9, 6.5 and 6.6

g kg-1 of Mg, respectively, suggesting that these materials could be used as liming

sources (Fig 4-2A. and 4-2B.). Waste lime is generally fine and reacts rapidly with soil,

giving quick increases in soil pH, especially if cultivated. These materials can often be

obtained at little or no cost to the farmer or rancher (Kidder, G., 2001). Muchovej et al.,

2001, suggested that lime stabilized biosolids contained lower N, P, and metal

concentrations, but higher Ca concentrations than digested biosolids, due to the large

amount of lime added to the material. Milorganite (39.5 and 0.21 g kg-1), Baltimore

pellets (39 and 0.9 g kg-1) and Disney compost (14.5 and 0.9 g kg-1) contained the highest

concentrations of Fe and Mn, respectively (Fig 4-2C. and 4-2D.). Iron and Mn hydrouss

oxides forms) adsorb metals in the biosolid and in amended soils, thus being able to

reduce phytoavailability of many trace elements (for example Cadmium). Nitrogen and P

receive the most attention in management of organic wastes because of their importance

in both plant nutrition and water contamination. However, all the other nutrient elements

are also returned to the soil in organic waste applications and can contribute to improved

crop performance (Kidder,G. 2001).Based on their N and P compositions, these biosolids

can be used as low-grade N and P fertilizer and as a source of calcium, especially the

lime-stabilized residuals.













Milorganite

N-viro

Baltimore pellets

Disney compost

Black hen

Black kow

Limed cake

Limed slurry


I I


.6


Total Kjeldahl Nitrogen (g Kg-1)


Milorganite

N-viro

Baltimore pellets

Disney compost

Black hen

Black kow

Limed cake

Limed slurry


nil


Total Phosphorus (g Kg-1)


Fig 4-1: Data represent total nitrogen, phosphorus and potassium. A. Total Kjeldahl
Nitrogen, B. Phosphorus and C. Potassium concentration of biosolids (on a
dry weight basis).


o ,


H


-R


E3


-I










C.

Milorganite

N-viro

Baltimore pellets

Disney compost

Black hen

Black kow

Limed cake

Limed slurry

0 10 20 30 40 50
Total Potassium (g Kg"1)


Fig 4-1: Continued

A.

Milorganite

N-viro

Baltimore pellets

Disney compost

Black hen

Black kow

Limed cake

Limed slurry

0.0 0.5 1.0 1.5 2.0 2.5 3.0

Total Calcium concentration (g kg-1)


Fig 4-2: Data represent total calcium, magnesium, iron and manganese. (A) Ca, (B) Mg,
(C) Fe and (D) Mn concentrations ofbiosolids (on a dry weight basis).














Milorganite

N-viro

Baltimore pellets

Disney compost

Black hen

Black kow

Limed cake

Limed slurry


I I


Total Magnesium concentration (g kg"1)


Milorganite

N-viro

Baltimore pellets

Disney compost

Black hen

Black kow


Limed cake

Limed slurry


Total Iron concentration (g kg"1)


Fig 4-2: Continued


1-












Milorganite

N-viro

Baltimore pellets

Disney compost

Black hen

Black kow


Limed cake

Limed slurry

0.0 0.1


I I


I I


I I I I 1 I
0.2 0.3 0.4 0.5 0.6 0.7
Total Manganese concentration (g kg"1)


0.8 0.9 1.0
0.8 0.9 1.0


Fig 4-2: Continued


Table 4-1: Heavy metal concentrations of biosolids in relation to USEPA ceiling
concentration limits for land application (on a dry weight basis).


Arsenic Cadmium Lead Copper
---------------------------------------mg Kg-----


Zinc Molybdenum


Cumulative pollutant
loading rates
Ceiling
Concentration Limits
Limed slurry
Limed cake
Black kow
Black hen
Disney compost
Baltimore pellets
N-viro
Milorganite
NL: No limit (EPA,


41

75

3.98
6.97
2.29
15.60
22.50
5.86
37.20
4.34
1994).


39

85

0.72
0.51
0.06
0.24
0.71
17.90
0.38
2.83


300

840

6.79
15.30
2.86
1.55
15.60
202
16.90
37.20


1,500 2,800

4,300 7,500


317
197
26
672
291
393
177
232


330
198
138
641
296
1010
92
640


According to EPA document 40 CFR Part 503 biosolids application rates should be

based on certain heavy metals ceiling concentration limits. All materials in this study

contained concentrations below the ceiling concentration limits and below the cumulative

pollutant loading rates. These materials do not represent a potential toxic environmental


Biosolids


NL

75

8.14
5.93
1.48
3.54
15.30
8.94
7.20
9.02


E3









risk (Table 4-1). Similar results were observed by Adjei et al.(2002). The use of these

materials in the production of crops for human consumption when practiced in

accordance with existing federal guidelines and regulations, present negligible risk to the

consumer, to crop production, and to the environment (NRC 1996). Franco-Hernandez et

al., 2003 reported concentrations of heavy metals recovered from dry biosolids were

lower than 0.1 g kg-1 except for zinc (Zn) and chromium (Cr) in materials treated with

lime. Similar results were observed by Adjei et al. (2002) and Sigua et al.(2005), where

concentrations of As, Cd, Cu, Pb, Mo, Ni and Zn of sewage sludge were far below the

national USEPA health risk limits. Since the level of trace metals in the biosolid

materials that were used in this study were below the USEPA risk limits, it is not

expected that their levels will be harmful on soil and forage quality once they were

applied for agricultural purposes (e.g. forage and pasture establishments).

Study 2: Determination of the Mineralization Rates of Study Materials

Nitrogen mineralization is a relatively slow microbial process that is affected by

factors such as amendment composition, soil type, temperature, pH, aeration, and

moisture (Van Kessel et al., 2002; Mikkelsen et al. 1995). The stabilization process used

to treat wastewater alters the nutrient composition of the resulting biosolids. The rate of

nutrient release, or mineralization, is also affected by the process. Mineralization values

ranging from 41 to 50% of the organic N from aerobically-digested sewage sludge and 23

to 41% of the organic N from anaerobically-digested sludge have been reported during

the initial crop season (Muchovej et al., 2001). The N release characteristics of the

materials in this study differed widely. Limed slurry had the highest percent of N

mineralized when applied at 180 kg N ha-1 with 81% of the total N being mineralized

(Table 4-2). Lindemann et al. (1984) observed total mineralization rates ranging from









56% to 72% for sewage sludge after 32 weeks of incubation. Mineralization rates of all

materials increased when applied N was increased, except for limed cake. When applied

at 180 Kg N ha-1, limed cake increased pH to 11.6 and EC to 2811 iS cmQ1, which

resulted in reduced CO2 evolved implying that microbial activity was reduced (Fig 4-5).

The lower rate of net N mineralization in the first week, in the presence of compost,

such as black kow, black hen and disney compost could be partly due to N assimilation

by the growing microbial biomass, that was stimulated by both compost and soil organic

C. Composted sewage sludge exhibits a lower N mineralization rate compared to non-

composted sludge (Adegbidi et al., 2003). Baltimore pellets had the second highest

percent of N mineralized at 90 Kg N ha- (41%) while Milorganite had the second highest

percent of N mineralized at 180 Kg N ha'(58%). Based on the variable mineralization

rates of all materials relative to rate applied there was no definite trend, each material has

a specific pattern of N release based on its composition and type of treatment received

(Table 4-2). Adegbidi et al. (2003) observed that for sewage sludges, N mineralization

rates of composts are specific to the materials utilized and to the experimental conditions

and that on average non-composted organic residuals had higher mineralization rates than

composted organic residuals.

Leachates were analyzed for water soluble P and no detectable P was found in the

leachates through all 263 days of incubation. Unlike the case with nitrogen, leaching of P

to ground water has not traditionally been a major transport process. In, many soils, high

P-sorbing oxide components keep leachate P levels well below eutrophication thresholds

(0.01 to 0.05 mg L1, Sims et al., 1998).









It is very important to observe the pH values when biosolids are being applied to

the soil, because the soil pH also influences the rate of mineralization. The pH values in

the columns ranged from 6.2 to 11.6, with that of the untreated control soil around pH

6.6. Limed materials and manure application (Black kow and Black hen) increased soil

pH, while composted materials application, like Baltimore pellets, resulted in slightly

lower soil pH values (Table 4-2).

Table 4-2: Percentage of N mineralized in 256 days in relation to biosolids type and the
resulting soil pH and EC.
Biosolids Total N Total N %N pH EC
applied (Kg Mineralized mineralized (itS/cm)
N ha-1) (mg)
Limed cake 90 20 25 9.3 498
180 3 2 11.6 2811
Limed slurry 90 44 55 7.2 359
180 130 81 7.2 612
Black kow 90 5 6 7.2 198
180 11 7 7.2 311
Black hen 90 12 15 7.0 287
180 20 12 7.0 466
Disney compost 90 15 19 6.8 188
180 24 15 6.8 260
Baltimore pellets 90 31 39 6.2 208
180 65 41 6.2 336
N-viro 90 21 27 8.3 2409
180 30 19 8.7 3456
Milorganite 90 29 36 6.7 201
180 92 58 6.7 338

N-Viro application at 90 kg N ha-1 and 180 kg N ha-lincreased the EC in the soils

up to 2409 iS cm-1 and 3456 iS cm-1, respectively (Table 4-2) Usman et al. (2004) found

that the addition of biosolids raised the soil salinity level more than the addition of

compost and also that the salinity increased with increasing application rate of organic

wastes. Wong et al. (2001) found that the EC of soil increased due to the addition of

biosolids.









CO2 Evolution

A CO2 trap of 1N NaOH was placed in the head space of the incubation lysimeter

to estimate microbial decomposition rate. The CO2 trap was replaced and titrated with 1N

HC1 weekly to estimate CO2 evolution. The amount of CO2 evolved was used as an

indicator that the materials were being mineralized and microbial activity was occurring

in the system.

An interaction between biosolids and rate was observed at P>0.05 (Table 4-3). The

intensive microbial activity during the first week is shown by the rate of CO2 evolution

that was enhanced in the presence of compost (Fig 4-3A. and 4-4A.). Similar results were

observed by Hadas et al., 1994. The effect of the added organic materials on CO2

evolution decreased rapidly with time, and in the first two weeks was greater in limed

slurry and milorganite at 90 kg N ha-1 and 180 kg N ha-l, whereas the organic materials

contributed very little to soil respiration after 16 weeks. At 90 kg N ha- and 180 kg N ha-

1 there was a distinctive difference between CO2 evolved from biosolids at week 1 and

week 2. At week 8 these difference was not noticeable anymore, as we observe in figure

4-3C. and 4-4C., there was no difference between biosolids. At week 16, we begin to

observe differences between biosolids once more, however now the differences are

related to the rapid decrease in CO2 evolved from some biosolids, like limed slurry. By

week 33, very little CO2 evolution was taking place (Fig. 4-3 and 4-4). We observed that

the materials milorganite, limed slurry, limed cake and N-viro showed the same amount

of CO2 evolved during the first week at 90 kg N ha- and 180 kg N ha- (Fig 4-3A and 4-

4A). With milorganite and limed slurry at 180 kg N ha- the quantity of CO2 being

evolved was so high that it neutralized all of base solution, thus these data do not show

the correct amount of CO2 evolved for those materials at 180 kg N ha- during the first









week of the study. Therefore, if more base solution had been added to the lysimeters

containing these treatments, we would probably had have a much higher amount of CO2

evolved from them, consequently the difference between rates, 90 kg N ha-1 and 180 kg

N ha-1, for those materials would be much higher in figures 4-3A and 4-4A. In the case

of N-viro and Limed cake at 180 kg N hal-, N-viro increased the EC values (3456 [S cm

1) while Limed cake produced a large increase in pH (11.6), this high EC and pH

probably inhibited microbial activity consequently the CO2 evolved at 180 kg N ha-1 was

the same as at 90 kg N ha-1.

The cumulative CO2 evolution represents the sum of measured weekly quantities of

CO2 evolved. Overall, CO2 production increased as the rate of applied biosolid increased.

Black kow produced the highest amount of total CO2 evolved at 90 kg N ha-1 with 3646

mg of CO2 column', followed by Disney compost and Baltimore pellets, 3536 and 2947

mg CO2 column', respectively (Fig. 4-5). Black hen, N-viro and limed cake produced

the lowest CO2 evolved with 2624, 2433 and 1978 mg CO2 column'1 respectively (Fig. 4-

5). Because a true estimate was not obtained for Milorganite and Limed slurry, as

explained above, they are not included in this discussion, even though those materials

would probably have produced the highest CO2 evolved at 180 kg N ha-1 we can not

make this assertion. Comparing all other biosolids when applied at 180 kg N ha-1, Black

kow produced the highest amount of CO2 evolved (5802 mg CO2 column-) followed by

Disney compost with 5017 mg CO2 column1 (Fig. 4-5). These findings are in agreement

with those reported by Franco-Hernandez et al. (2003) who observed that production of

CO2 increased as the rate of biosolids added increased.















b

=lf
M-d
E-Hd
C
_e

Ia

f


0 50 100 150 200 250 300 350 400 450 500

CO2 evolved (mg wk-1)


Milorganite
N-Viro
Baltimore Pellets
Disney Compost

Black Hen
Black Kow
Limed Slurry
Limed Cake


D. Week 16
bec


ba

Sba
: dc
f ba

de


M ilorganite
N-Viro
Baltimore Pellets

Disney Compost
Black Hen

Black Kow

Limed Slurry
Limed Cake


B. Week 2
M ilorganite

N-Viro
Baltimore Pellets
U,
2 Disney Compost
o
o Black Hen
m
Black Kow
Limed Slurry
Limed Cake


fb

:Mf
dc


fd


n a


M ilorganite
N-Viro
Baltimore Pellets
Disney Compost
Black Hen
Black Kow

Limed Slurry
Limed Cake


0 50 100 150 200 250 300 350 400 450 500

CO2 evolved (mg wk1)


E. Week 24
flbc

ee3 bac
4 bac
:E ba


a
3-bc
--bc


50 0 1 150 200 250 300 350 400 450 500

CO, evolved (mg wk-1)


M ilorganite
N-Viro

Baltimore Pellets

DisneyCompost
Black Hen

Black Kow

Limed Slurry
Limed Cake


F. Week 33


-4a

3 b

ocb
3b
fib


3cb
IC


0 50 100 150 200 250 300 350 400 450 500

CO2 evolved (mg wk1)


0 50 100 150 200 250 300 350 400 450 500

CO2 evolved (mg wk-)


Fig 4-3: Data represent CO2 evolved from biosolids at weeks 1(A), 2(B), 8 (C), 16 (D),
24(E) and 33(F), respectively, at 90 Kg N ha-1. Data represent means and standard error
of three replicates. Means with the same letter are not different at P>0.05.


A. Week 1


0 50 100 150 200 250 300 350 400 450 500

CO, evolved (mg wk1)


C. Week 8


M ilorganite
N-Viro
Baltimore Pellets

Disney Compost
Black Hen
Black Kow

Limed Slurry
Limed Cake


E b
f ba
,ba

,ba
,ba


---Iba

a













A. Week 1


D. Week 16


Milorganite

N-Viro

Baltimore Pellets

SDisney Compost
o
U Black Hen

Black Kow

Limed Slurry

Limed Cake


a

3d

b

l b

a

C

a

3d


Milorganite

N -Viro

Baltimore Pellets

Disney Compost

Black Hen

Black Kow

Limed Slurry

Limed Cake


f ba

flbc

dc

ba

dc

a
d

m dc


0 50 100 150 200 250 300 350 400 450 500

CO2 evolved (mg wk1)


0 50 100 150 200 250 300 350 400 450 500

CO2 evolved (mg wk1)


B. Week 2


E. Week 24


~ b

=B4f

C
ic

Ib

d

me

a

--f


0 50 100 150 200 250 300 350 400 450 500

CO2 evolved (mg wk1)


Milorganite
N-Viro
Baltimore Pellets
Disney Compost
Black Hen
Black Kow
Limed Slurry
Limed Cake


= cd
,--4 cbd
cbd
fb


", a
,-Icb
Pd


0 50 100 150 200 250 300 350 400 450 500

CO2 evolved (mg wk1)


-a

: 3' c
C
fl ba

bc

bc

Sba

f ,,c


Milorganite
N-Viro
Baltimore Pellets

Disney Compost

Black Hen

Black Kow
Limed Slurry

Limed Cake


F. Week 33


=P b




cb

3ed
---I ed
la

3e
3 ed


0 50 100 150 200 250 300 350 400 450 500

CO2 evolved (mg wk-1)


0 60 100 160 200 260 300 360 400 460 600

CO2 evolved (mg wk1)


Fig 4-4: Data represent CO2 evolved from biosolids at weeks 1(A), 2(B), 8 (C), 16 (D),
24(E) and 33(F), respectively, at 180 Kg N ha-1. Data represent means and
standard error of three replicates. Means with the same letter are not different
at P> 0.05.


Milorganite

N-Viro

Baltimore Pellets

Disney Compost

Black Hen

Black Kow

Limed Slurry

Limed Cake


C. Week 8


Milorganite

N-Viro


Baltimore Pellets

Disney Compost
Black Hen

Black Kow

Limed Slurry

Limed Cake










N-viro Icb


Baltimore pellets

v Disney compost
U)
C Black hen

Black kow

Limed cake

A. 90Kg N ha-


Cb




-ccb

*


0 1000 2000 3000 4000 5000 6000 7000
Total CO2 evolved (mg column"1)


N-viro

Baltimore pellets

Disney compost

Black hen

Black kow

Limed cake

B. 180Kg N ha.1


cd


I cb


ed


0 1000 2000
0 1000 2000


3000 4000 5000 6000 7000


Total CO2 evolved (mg column"1)
Fig 4-5: Data represent total CO2 evolved from different biosolids at 90 Kg N ha-1 (A)
and 180 Kg N ha-' (B). Data represent means and standard error of three
replicates. Means with the same letter are not different at P>0.05.

Table 4-3: Analysis of variance (ANOVA) table used for the determination of statistical
differences in the analysis for effects of biosolids and rate source on total CO2
evolved.
Source DF F value Po.o5 > F
Biosolid 6 122.69 <0.0001
Rate 1 137.99< 0.0001
Biosolid x rate 6 20.92<0.0001
Replication 2
Error 26
Total 41


: l a

e









In summary, the amount of CO2 evolved was a good indicator that the materials

were being mineralized, and according to the N mineralization rates observed in this

study, biosolids have the ability to supply nutritional fertility to plants.

Study 3: Glass House Study 1st Application

Bahiagrass Dry Matter Production

All sources of N produced a higher level of dry matter than the unfertilized control.

Dry matter production varied within biosolid and rates (90 kg N ha-1 and 180 kg N ha-1).

Dry matter accumulation was greater with the 180 kg N ha-1 compared with the 90 kg N

ha-'(Fig 4-6A. and 4-6B.). At 90 kg N ha-1 ammonium sulfate induced the highest dry

matter accumulation (250.7 g m-2) followed by Limed slurry (239.3 g m-2) and N-viro

(229 g m-2) (Fig 4-6A.). At 180 kg N ha-1 limed slurry induced highest dry matter

accumulation (346.1 g m-2) followed by ammonium sulfate (323.8 g m-2) and N-Viro

(318.1 g m-2) (Fig 4-6B.). Limed cake and disney compost induced the lowest dry matter

accumulation at 90 kg N ha-1 and 180 kg N ha-l. Nitrogen is the most limiting nutrient for

grass production on Florida's spodic sandy soils as evidenced by the lowest cumulative

yield for the nonfertilized control compared with the N-fertilized treatments (Adjei et al.,

2002).

Bahiagrass Root Dry Matter Production

At the end of the experiment, a 45.6cm2 diameter x 15cm depth sample was taken

from each tub to estimate differences in stolon growth between biosolids and applied

rates (90 kg N ha-1 and 180 kg N ha-1). Stolon dry weight did not differ between biosolids

or rates (90 kg N ha-1 and 180 kg N ha-1) after 112 days (Table 4-4). Ammonium sulfate

stolon dry weight did not differ from biosolids. Generally, when biosolids are applied as a

sole N source at normal application rates the release of N is not sufficient to induce an









optimum turf grass response relative to quality or growth. Biosolids application in this

study were based on normal application rates, therefore the N release after 112 days of

application was probably not sufficient to induce bahiagrass stolon dry matter production

to the point that differences between biosolids could be observed.

Table 4-4: Bahiagrass stolon dry matter production after 112 days
Biosolids Kg N ha-1 stolon weight ns

--------------g --------------
Limed cake 90 9.74
180 10.81
Limed slurry 90 10.62
180 11.73
Disney compost 90 11.58
180 10.03
Baltimore pellets 90 10.31
180 12.94
N-viro 90 10.98
180 10.16
Ammonium sulfate 90 12.57
180 13.05
control 0 10.34
ANOVA P-value 0.4
CV 21
ns: not significant at 0.05.

Visual Quality Rating

Bahiagrass quality was rated visually on the basis of color, uniformity, and density

on a 1 to 9 scale, where 1 was dead, 5.5 or above was acceptable, and 9 was superior

quality. Bahiagrass quality was different from the control for all biosolids at 90 kg N ha-1

and 180 kg N ha-1 (Table 4-5). All biosolids produced acceptable quality turfgrass but

none of the biosolids produced superior quality. Ammonium sulfate is an inorganic

source of N used for a comparison with the biosolids. Limed slurry and N-viro at 180 kg

N ha-1 produced a turf quality comparable and slightly higher than the one observed when

ammonium sulfate was applied (Table 4-5). While stolon growth and visual quality were







51


not affected, these data suggest that application rates (90 kg N ha-1 and 180 kg N ha1)

may have affected dry matter accumulation.

Table 4-5: Visual rating during Fall 2004 comparing the biosolids at different rates.
Biosolids kg N ha-1 Visual Quality Ratings
Scale: 1 to 9

Limed cake 90 7.0c
180 7.4ba
Limed slurry 90 7.1b
180 7.6a
Disney compost 90 6.7c
180 7.4ba
Baltimore pellets 90 6.7c
180 7.1b
N-viro 90 7.2ba
180 7.4ba
Ammonium sulfate 90 7.2ba
180 7.3ba
control 0 5. d
ANOVA P- value <0.0001
CV 3.7
* Means with the same letter are not different at 0.05.

A. 90kg N ha-1

300

a
cC 250 b b
E d d
-- e
200-




S100-
I f





S50
0 g








Limed cake Limed slurry Disney Baltimore N-viro Ammonium Control
compost pellets sulfate

Fig 4-6: The effect of nitrogen rate on cumulative forage dry matter accumulation over
112 days.










B. 180 Kg N ha1

400
a
350 c b

0 300 d
= e f
0
2 250

S200



E 100

50

0 -
Limed cake Limed slurry Disney Baltimore N-viro Ammonium Control
compost pellets sulfate

*Means with same letter are not different at 0.05 by Duncan test.

Fig 4-6: Continued

Table 4-6: Analysis of variance (ANOVA) table used for the determination of statistical
differences in the analysis for effects of biosolids and rate source on total dry
matter accumulation.
Source DF F value Po.o5 > F
Biosolid 6 17.10 <0.0001
Rate 1 33.96< 0.0001
Biosolid x rate 5 7.95<0.0001
Replication 3
Error 35
Total 50


Bahiagrass Chemical Composition

Total N Uptake: Nitrogen uptake by the crop is important when biosolids are land

applied because it is the means by which N is removed to prevent N03-N leaching to the

ground water. The uptake of N by bahiagrass (Table 4-7) increased with increasing

biosolid application rate (90 kg N ha-1 vs 180 kg N ha-1), consequently, applying biosolid

increased plant uptake of N compared to the control (Table 4-7).Limed slurry,









ammonium sulfate and N-viro induced the highest N uptake at 90 kg N ha-1 and 180 kg N

ha-1. N-Viro, at 90 kg N ha-1 and 180 kg N ha-l, response was probably related to the fact

that this material is slow-release form of N, as showed in the previous study, allowing

more time for bahiagrass uptake. These results are in agreement with the N

mineralization data and CO2 evolution data showed at previously study, but when

observing the CO2 evolution data, we see that N-viro (90 kg N ha-1 and 180 kg N ha-1)

had one of the lowest CO2 evolved at week 1, probably because the conditions, high pH

(8.3 and 8.7) and EC (2409 iS cm-1 and 3456 iS cm-1), created in the soil when N-viro

was applied inhibited microbial activity at first, however after week 8 N-viro showed an

increase in CO2 evolution, suggesting an increase in microbial activity, increasing N

mineralization and ultimately increasing N available for bahiagrass uptake.

N-Recovery: Mineralization rates of approximately 40%, 15%, and 8% have also

been estimated for waste-activated sludge, anaerobically-digested sludge, and composted

sludge, respectively (Muchovej et al., 2001). At 90 kg N ha-1, the apparent nitrogen

recovery was slightly higher when inorganic fertilizer, ammonium sulfate, was applied,

(35%) compared with Limed slurry (33%) and N-viro (27%). At 180 kg N ha-1, limed

slurry surpassed the inorganic fertilizer inducing a 30% nitrogen recovery by bahiagrass

followed by N-viro that induced a 24 % apparent nitrogen recovery (Table 4-7). The

apparent nitrogen recovery of biosolid N ranged from 12 to 33%, which is lower than the

47 to 85% ANR of fertilizer N by forage grasses reported in previous studies (Lynch et

al., 2004). A higher recovery of N in the grass-soil system is desirable and would not

only increase N application efficiency, thereby reducing fertilizers costs to producers, but

would also reduce NO3- and NH4+ losses to the environment.









Table 4-7: Total N uptake by bahiagrass from biosolids in first 112 days.
Biosolids Kg N ha-1 Total N uptake % N recovered
---------- g of N m2-
Limed cake 90 3.80 de 21
180 5.20 b 19
Limed slurry 90 4.80 bcd 33
180 7.15 a 30
Disney compost 90 2.95 ef 12
180 5.10b 18
Baltimore pellets 90 3.90 cde 22
180 5.10b 18
N-viro 90 4.30 cd 27
180 6.20 a 24
Ammonium sulfate 90 4.95 be 35
180 6.70 a 27
control 0 1.95 f
ANOVA P- value <0.001
CV 14.6
biosolid x rate Pr>F 0.12
* Means followed by the same letter are not different at 0.05 by Duncan test.
Total P Uptake: All applications were N based, so each material had a different

quantity of P applied depending on concentration of P in each material (Table 4-8).

Phosphorus uptake is a product of dry matter accumulation and tissue P concentration.

Phosphorus uptake increased with increasing rate of biosolids. All biosolids had an effect

on bahiagrass P uptake (Table 4-8). Bahiagrass P uptake in the treated tubs was greater

than the untreated control (Table 4-8). This suggests that P in the biosolids became

available for plant absorption within a relatively short period of time (112 days). At 90

Kg N ha-1, limed slurry induced the highest P uptake, differing from all treatments,

followed by ammonium sulfate and N-viro (Table 4-8). Disney compost supplied the

lowest quantity of P differing from all treatments at P< 0.05. These results are in

agreement with the P findings in the biosolids prior to application and based on the total

amount of P applied. Ammonium sulfate does not contain P, however when applied a









significant drop in pH is observed (from 6.0 to 5.0). It is suspected that this pH drop

caused a release of P forms in the soil that were previously unavailable for uptake by the

grass.

Table 4-8: Quantity of P applied by each material and percentage of P uptake by
bahiagrass
Biosolids Kg Quantity Quantity of P uptake Tissue P % Recovery
N ha-1 of P applied (*) concentration of applied P
material by each
applied material
on each
tub
-----g-- --g tub- -- --- ---g Kg--

Limed cake 90 135 1.22 0.14 ef 3.80 11.5
180 270 2.45 0.20 abcd 3.90 8.2
Limed slurry 90 76 1.68 0.19 bcde 4.20 11.3
180 151 3.34 0.25 a 3.70 7.5
Disney 90 109 1.28 0.12 f 4.00 9.4
compost 180 218 2.55 0.18 cde 3.90 7.1

Baltimore 90 55 1.40 0.16 def 3.90 11.4
pellets 180 110 2.80 0.20 abcd 3.90 7.1
N-viro 90 967 2.54 0.17 de 3.90 6.7
180 1933 5.08 0.23 abc 3.80 4.5
Ammonium 90 8 -- 0.18 cde 3.80
sulfate 180 17 -- 0.24 ab 3.80
Control 0 -- 3.80
ANOVA P- <0.0001
value
CV 20.98
biosolid x Pr>F 0.9229
rate
(*)Means with the same letter are not different at 0.05 by Duncan test.

In previous studies, biosolid fertilizer rates as low as 4.5 to 9 t ha-1 improved

bahiagrass P concentrations to about 3 g kg-1 (Tiffany et al., 2001). Kincheloe et al.

(1987) and Rechcigl et al. (1992) showed that adequate production of bahiagrass is

achieved with a tissue P concentration of approximately 2.0 g kg-1. Increases in biosolids









loading rate did not increase percentage P removal in bahiagrass over 112 days of

biosolids application (Table 4-8). Although, as observed in table 4-8, the amount of P

applied with each biosolid was different since the application was N based, the percent

recovery calculation was based in each P load separately. Bahiagrass recovery of P was

higher with limed cake, followed by baltimore pellets and limed slurry, all at 90 Kg N ha-

1. A low percentage of P was recovered in bahiagrass harvested over 112 days. Johnson et

al., 2004, found that a low percentage of wastewater or poultry litter sources of P was

recovered in forage harvested over four years.

Leaching Evaluation

Total N in Leachate: The leachate was analyzed for NH4 NO3-, and P to assess N

and P movement in soils amended with wastes. At 90 Kg N ha-1, limed slurry contributed

the highest quantity of inorganic N to the leachate. In excess of 11.04% of the N applied

as limed slurry was found in the leachate, followed by 7.56% of the applied N for

ammonium sulfate and 1.10% of the N applied as disney compost. At 180 Kg N ha-1,

ammonium sulfate contributed the highest quantity of N to the leachate with 15.70% of

the N applied, followed by limed slurry with 10.31% and Disney compost with 3.60%

(Table 4-9). Ammonium sulfate is commonly used in turf fertilization. Therefore,

ammonium sulfate was used to establish a comparison between inorganic and organic

sources of N. Besides limed slurry at 90 Kg N ha-1, none of the biosolids contributed as

much N to the leachate as ammonium sulfate, suggesting these biosolids can be used to

reduce the threat of environmental contamination (Table 4-9). Leachate N03-N

concentration was influenced by loading rate (90 Kg N ha-1 and 180 Kg N ha-1) and the

length of time after the initial application. As expected, N03-N concentration increased

with increasing level of applied biosolids and decreased with time after application









(Table 4-9, 4-10). Leachate collected on day 84 did not contain detectable N03-N (Table

4-10). The NH4-N data are presented in table 4-9 and 4-10. In general, the concentration

of NH4-N was several fold lower than the N03-N concentration. By day 56, there was no

detectable levels of NH4-N in the leachate (Table 4-10). Most of the materials produced

maximum levels of leached NH4 within 7 days (Table 4-10). As for NO3, microbial

activity had to mineralize the materials to soluble forms, becoming available for plant

uptake and runoff. According to Van Kessel et al. (2002) when manure-amended soils

were incubated for 56 days, all of the NH4+-N was depleted; NO3- -N concentrations

increased during this time period. Nitrogen in sludge is mainly in organic forms, while

mineral forms are generally in low concentration and are mainly represented by

ammonium nitrogen.

Total P in Leachate: No detectable P was found in the leachates through all the

leachate collections, 7 to 112 days. Biosolids and manure-borne P reactions have not

been researched as much as waste bond N. We do know that (as with N) calculations

based on total waste-P concentrations to determine allowable P loads to a soil can be

grossly over-conservative, as all the (total) P may not be bioavailable or soluble enough

to leach (Pierzynski et al., 1994). Soils common to most of the USA contain sufficient P-

sorptive capacity (Al and Fe oxides) to prevent significant P leaching and to mask

inherent differences in the P solubilities of biosolids materials, therefore leaching of P

appears to be quite small for most biosolids, even when applied to meet crop N

requirements on sandy soils with limited P-sorbing capacity (Elliott et al., 2002).

Table 4-9: Total NH4-N and N03-N content in leachates from organic materials
Biosolids Kg Total Total NO3 Total N in % of N applied
N ha-1 NH4 in in leachate (7 leached
leachate leachate through 112













Limed cake


Limed slurry


Disney
compost

Baltimore
pellets
N-viro

Ammonium
sulfate
Control


--mg -- --mg --
90 8.40 6.30


180
90
180


9.20
12.70


42.90
188.10


64.80 302.60


90 5.00


180
90
180
90
180
90
180
0


37.40
4.50
17.30
5.80
6.87
29.70


22.80
96.10
4.35
13.10
19.10
117.40
110.60


173.80 380.80


4.95


3.75


Table 4-10: Concentration of N03-N and NH4 -N in leachate as affected by the


incubation period
Biosolids Kg 7 days
N


14 days 28 days 56 days 84 days


mg NO3 NH4 NO3 NH4 NO3 NH4 NO3 NH4 NO3 NH4
Limed cake 90 3 4 3 5 0 0 0 0 0 0
180 12 4 18 5 12 0 1 0 0 0
Limed 90 45 9 74 4 69 0 1 0 0 0
slurry 180 46 43 157 22 97 0 3 0 0 0
Disney 90 11 3 9 2 3 0 0 0 0 0
compost 180 25 9 47 29 23 0 2 0 0 0
Baltimore 90 2 2 2 2 1 0 0 0 0 0
pellets 180 3 10 5 7 6 0 0 0 0 0
N-viro 90 5 4 8 2 6 0 0 0 0 0
180 12 2 27 4 66 1 12 0 0 0
Ammonium 90 32 20 62 8 17 2 0 0 0 0
sulfate 180 64 127 162 35 148 12 7 0 0 0
Control 0 1 2 1 2 2 0 0 0 0 0

pH and EC in Leachate: The relationship between pH and EC and incubation

period at different rates of biosolid application is presented in table 4-11 and 4-12. The

leachate pH ranged from 5.3 to 6.6, with the untreated control around 6.0. Limed

materials increased the pH while ammonium sulfate reduced soil pH to about 5.3. The


days)
------mg -------
14.70


52.10
200.80
367.40
27.80
133.50
8.85
30.40
24.90
124.27
140.30
554.60
8.70


0.34
1.25
11.04
10.31
1.10
3.60
0.01
0.62
0.93
3.32
7.56
15.70









incubation period affected the pH of the leachate, with or without biosolid. Interestingly,

the leachate pH increase observed in the mineralization study for limed cake (9.3 at 90

Kg N ha-1 and 11.6 at 180 Kg N ha-1) and N-viro (8.3 at 90 Kg N ha-1 and 8.7 at 180 Kg

N ha-)) was not observed in the glasshouse study. This difference is probably due to the

interaction between the biosolids and the turf and to the biosolids to soil rates. In the

incubation study the biosolids were applied based on a dry weight basis while in the

glasshouse study the biosolids were surface applied on an area basis. On the other hand,

in the incubation study the absence of the turf meant that all elements that were

mineralizing from the biosolids were free, to be adsorbed by the soil and to change in the

soil characteristics, like pH. When biosolids were applied in the glass house study, the

same mineralization was occurring, but now, interaction between the biosolid and the turf

was taking place, nutrients were also being absorbed by the turf, ions attached to the

surface of root hairs, such as H+ ions, may exchange with those held on the soil surface,

keeping the pH from increasing as much as they increased without the turf.

Electrical conductivity (EC, an estimate of soluble salts) increased for soils

receiving biosolids. N-Viro applied at 180 Kg N ha-1 produced the highest EC from 7

days through 112 days going from 1571 uS cm-1 to a peak of 3227 uS cm-1 at 56 days,

from there decreasing to 1141 uS cm-1 at 112 days (Table 4-12). Electrical conductivity

of the leachate from ammonium sulfate treated soils peaked at 14 days at 2270 uS cm1.

N-Viro at 90 Kg N ha-1 produced a peak EC at 56 days at 2072 uS cm-1. All the other

treatments differ from the control, increasing the EC when compared to the control.










Table 4-11: Leachate pH values from 7 to 112 days after treatment ap location.
Biosolids Kg 7 days 14 days 28 days 56 days 84 112 days
N days
ha-
pH p pH ppH pH pH
Limed cake 90 5.9 bac 6.0 ba 6.3 ba 6.4 a 6.1 a 6.3 bac
180 6.1 ba 6.0 bac 6.2 bac 6.1 ba 6.3 a 6.5 ba
Limed 90 5.8 bac 6.0 bac 6.2 bac 6.4 a 6.2 a 6.4 bac
slurry 180 6.2 ba 5.9 bac 6.2 ba 6.4 a 6.4 a 6.4 bac
Disney 90 5.8 bac 5.8 bc 6.2 ba 6.2 ba 6.2 a 6.3 bac
compost 180 5.7 bc 5.7 c 6.0 bc 6.0 b 6.2 a 6.3 bac
Baltimore 90 5.9 bac 5.9 bac 6.2 bac 6.1 ba 6.2 a 6.2 c
pellets 180 5.8 bac 5.9 bac 6.2 bac 6.2 a 6.1 a 6.3 bc
N-viro 90 6.3 a 6.2 a 6.3 ba 6.4 ba 6.3 a 6.6 a
180 6.2 ba 5.9 bac 5.9 bc 6.3 ba 6.2 a 6.5 a
Ammonium 90 6.0 bac 5.7 c 6.2 bac 6.0 ba 6.1 a 6.3 bac
sulfate 180 5.5 c 5.3 d 5.8 c 6.0 b 5.8 b 5.9 d
Control 0 6.0 ba 6.1 ba 6.4 a 6.3 ba 6.2 a 6.3 bac
(*) Means with same letter are not different at 0.05
(**) Interactions effects between biosolids and rates are not significant; therefore
comparisons were made across all biosolids at both rates.
Table 4-12: Leachate EC values from 7 to 112 days after treatment application.
Biosolids KgNha-1 7 days 14 days 28 days 56 days 84 days 112 days
EC EC EC EC EC EC
Limed cake 90 414 f 409 d 408 d 534 b 430 b 379 b
180 464 ef 484 d 497 d 593 e 451 b 413 b
Limed slurry 90 571 ef 771 c 721 bc 521 b 413 b 362 b
180 702 e 975 c 847 c 546 g 409 f 370 d
Disney compost 90 585 ef 593 dc 528 cd 649 b 410 b 377 b
180 712e 935 c 857 c 769 c 416 e 375 c
Baltimore pellets 90 437 ef 455 d 432 d 547 b 405 b 355 b
180 477 ef 535 d 550 d 649 d 406 g 350 f
N-viro 90 1267 c 1712 a 2039 a 2072 a 1136 a 745 a
180 1571b 2161 b 2987 a 3227 a 1668 a 1141 a
Ammoniumsulfate 90 993 d 1036 b 751 b 642 b 399 b 326 b
180 1938a 2270 a 1649 b 934 b 449 c 360 e
Control 0 381f 405 d e 434 d d 585 b f 417 b d 361 be
(*) Means with same letter are not different at 0.05
(**) Rates 90 Kg N ha-1 and 180 Kg N ha-1 statistics analyzed separately from 14 to 112
days.
Because the EC was measured from the soil leachate, these results indicate

movement of salt from surface applied biosolid and inorganic fertilizer (ammonium

sulfate) applications. Kingery et al. (1994) found higher EC values in soil under fescue









pastures with a long-term history of poultry litter application compared to pastures

receiving no poultry litter.

Study 4: Glass House Study 2nd Application

A second growth response evaluation was conducted during the spring of 2005

from March through July to document the nutrients build up in the soil and to assess their

nutrient availability to bahiagrass. Biosolids were reapplied to the same tubs with the

established stand of bahiagrass was employed.

Bahiagrass Dry Matter Production

After reapplication of treatments the same experimental methodologies were

employed through 112 days. All sources of N produced a higher dry matter mass than the

unfertilized control. Bahiagrass dry matter response to N source varied within sources

and rates (90 kg N ha-1 and 180 kg N ha-1). Dry matter accumulation was greater with the

180 kg N ha-1 compared with the 90 kg N ha-'(<0.0001). At 90 kg N ha-1 Limed slurry

provided the highest dry matter accumulation (355 g m-2) followed by Limed cake (340

g m-2) and Ammonium sulfate (325 g m-2) (Fig 4-7A.). At 180 kg N ha-1 Limed slurry

provided highest dry matter accumulation (575g m-2) followed by Ammonium sulfate

(475 g m-2) and Baltimore pellets (468 g m-2) (Fig 4-7B.).The increased bahiagrass dry

matter response to biosolids compared to the unfertilized control is probably related to

the nutrients, especially N (most limiting nutrient), incorporated to the soil and its

availability to the grass after mineralization. Statistical comparisons were made to

compare the bahiagrass dry matter production after first application and after

reapplication. At 0.05 probability (two tailed test), the F value was 2.05; when comparing

the MSE values of the first application with the reapplication the F value was 3.70, which









was higher than 2.05, consequently the response variation for both studies are different,

therefore a comparison between both studies was not made.

Bahiagrass Root Dry Matter Production

A sample was taken from each tub at termination to evaluate the influence of

biosolids on stolon growth. Stolon dry weight differed between N-sources and rates (90

kg N ha-1 and 180 kg N ha-1) 112 days after reapplication (Table 4-13). Stolon growth of

bahiagrass receiving baltimore pellets at 180 kg N ha-1 was greater than the stolon growth

of the unfertilized control. This might be because baltimore pellets has a high P content,

as shown in study 1, therefore bahiagrass receiving baltimore pellets contained the

highest tissue P concentration (Table 4-17) and P influence in root growth is evidenced as

shown in Tisdale and Nelson, 1999. Stolon growth in response to all the other biosolids

did not differ from the unfertilized control.

Table 4-13: Bahiagrass root dry matter production 112 days after reapplication of the
materials.
Biosolids Kg N ha-1 root weight
------------ g ------------
Limed cake 90 9.95 ba*
180 8.68 b
Limed slurry 90 9.55 b
180 8.78 b
Disney compost 90 12.01 ba
180 9.61 b
Baltimore pellets 90 12.00 ba
180 13.20 a
Ammonium sulfate 90 9.70 ba
180 10.30 ba
control 0 8.7 b
ANOVA P-value 0.004
CV 20
Means with the same letter are not different at 0.05.
Statistical comparisons were made to compare the bahiagrass root dry matter

production after first application and after reapplication. At 0.05 probability (two tailed









test), the F value was 2.05; when comparing the MSE values of the first application with

the reapplication the F value was 1.04, which was lower than 2.05, therefore there was no

difference between the root harvests (first and second application), thus the parameter

root weight was analyzed as a combined analysis. When analyzed combined, bahiagrass

root growth receiving baltimore pellets at 180 kg N ha-1, remained greater when

compared to the root growth of the unfertilized control and the other biosolids.

Bahiagrass Quality Rating

Bahiagrass quality was rated visually on the basis of color, uniformity, and density

on a 1 to 9 scale, where 1 was dead, 5.5 or above was acceptable, and 9 was superior

quality. Bahiagrass quality was different from the control for all treatments at 90 Kg N

ha1 and 180 Kg N ha1 (Table 4-14).

Table 4-14: Visual rating during Spring 2005 comparing N sources at different rates after
biosolid reapplication.
Biosolids kg N ha-1 Visual rating means
Scale: 1 to 10

Limed cake 90 7.20 f
180 7.60 d
Limed slurry 90 7.40 e
180 7.70 c
Disney compost 90 7.00 h
180 7.80 a
Baltimore pellets 90 7.10 g
180 7.60 d
Ammonium sulfate 90 7.60 d
180 7.75 b
control 0 5.40 i
ANOVA P- value <0.0001
CV 0
*Means with the same letter are not different at 0.05.
All the treatments exhibited acceptable quality after reapplication but none of the

treatments exhibited superior quality. Disney compost and ammonium sulfate at rate 180

Kg N ha-1 produced a higher turf quality than that of other treatments (Table 4-14).








64



Forage production often requires significant inputs of lime, N fertilizer, and less


frequently of P and K fertilizers; all the N-sources used in this experiment to induce


bahiagrass growth contained from 59 g kg'1 to 13 g kg1 N, as shown in study 1.


400.0

350.0

300.0

250.0

200.0

150.0

100.0

50.0-

0.0


a


b


c


d


e


-~ -


Limed cake Limed slurry Disney compost Baltimore
pellets


A. 90kg N

700.0


S600.0-
E
0)
500.0-
c
.o
400.0
E

o 300.0


C 200.0*


a 100.0.


0.0.


Ammonium
sulfate


Limed cake Limed slurry Disney compost Baltimore pellets Ammonium Control
sulfate

B. 180 KgN ha-1
*Means with same letter are not different at 0.05 by Duncan test.


Fig 4-7: The effect of nitrogen source and rate on cumulative bahiagrass tissue dry matter
accumulated over 112 days.


Control









Bahiagrass Chemical Composition

Total N Uptake: Nitrogen applied to soils in mineral fertilizers and biosolids, and

also indigenous soil organic N, is subject to a complex series of interrelated biochemical

and physical processes which collectively form the nitrogen cycle. Nitrogen applied as

nitrate may be subject to leaching; therefore nitrogen uptake by the crop is important

when biosolid land application is used because it is the means by which N is removed to

prevent N03-N leaching to the ground water. The uptake of N for bahiagrass forage after

reapplication (Table 4-15) increased with increasing biosolid application rate (90 kg N

ha-1 and 180 kg N ha-1), Pr>F 0.0004, consequently, biosolid application increased plant

uptake of N compared to the control (Table 4-15). Limed slurry and Limed cake supplied

the highest level of N to the plant at 90 kg N ha-l while Limed slurry and AS supplied the

highest level of N at 180 kg N ha-'.Limed slurry response at 90 kg N ha-l was probably

related to the fact that its N was readily available at 90 kg N ha-l, as shown in the

previous study limed slurry mineralized 55% of the N applied, while limed cake

mineralized 25% of the N applied, being readily available for bahiagrass uptake. Both of

those materials are limed materials, which resulted in a soil pH close to neutrality, which

probably increased the N availability for plant uptake. Limed slurry at 180 kg N ha-1, had

the highest percentage of N being mineralized in the incubation study (81%), therefore N

was readily available for plant uptake. Ammonium sulfate at 180 kg N ha-l, reduced the

pH (4.2) in the system in the first 14 days. Statistical comparisons were made to compare

the N uptake by bahiagrass after first application and after reapplication. At 0.05

probability (two tailed test), the F value was 2.05; when comparing the MSE values of

the first application with the reapplication the F value was 2.10, which was higher than

2.05, consequently the response variation for both studies are different, therefore a









comparison between both studies was not made. The apparent nitrogen recovery (ANR)

of biosolid N by bahiagrass was calculated by difference as: ANR (%) = [(N uptake for

biosolid N uptake for unfertilized control)/ N reapplied] x 100.The recovery of biosolid

N is recorded in table 4-15. Recovery of biosolid N ranged from 44.5 to 39% for Limed

slurry N and 44.2 to 32.2% for Baltimore pellets was recovered in the harvested grass,

which is related to the ANR of N-sources (47 to 85%) by forage grasses reported in

previous studies (Lynch et al., 2004). This high percentage N recovery might be related

to residual effect from previous application. Muchovej and Rechcigl (1997) estimated a

minimum recovery rate from biosolids by bahiagrass crop of 70% in 2 years with the

remaining 30% becoming available with additional time.

Table 4-15: Total N uptake by bahiagrass from biosolids in 112 days after reapplication
of the materials.
Biosolids Kg N ha-1 Total N uptake % N recovered
---------- g ofN m2---2
Limed cake 90 5.00 c 35.0
180 7.35 b 31.0
Limed slurry 90 5.35 c 39.0
180 9.70 a 44.5
Disney compost 90 2.85 e 10.3
180 4.70 dc 15.8
Baltimore pellets 90 4.75 dc 32.2
180 7.70 b 44.2
Ammonium 90 4.55 dc 29.9
sulfate 180 8.00 b 34.8
control 0 1.95 e
ANOVA P- value <0.0001
CV 20.02
Biosolid x rate Pr>F 0.0837
* Means followed by the same letter are not different by Duncan test.

The recovery of biosolid N reported here shows a good relationship with the

recovery estimated by Muchovej and Rechcigl (1997), however these results are









estimated within 7 months after the first application and within 3 months after

reapplication of the materials.

Total P Uptake: At 90 kg N ha-lapplication of limed slurry resulted in the highest

P uptake by bahiagrass, differing from all N-sources, followed by baltimore pellets and

limed cake (Table 4-16). Application of disney compost resulted in the lowest P uptake

by bahiagrass differing from all N-sources at P< 0.05. At 180 kg N ha-1, limed slurry was

the material that induced the highest P uptake by bahiagrass differing from all N-sources

followed by baltimore pellets and limed cake. Disney compost induced the lowest P

uptake by bahiagrass differing from all N-sources (Table 4-16). Ammonium sulfate is a

material that has no P on its composition; however this material caused a drop in soil pH

at first, going from 6.2 to 4.2. This pH drop might have caused a release of P forms that

before were unavailable for uptake by the grass. Comparing tables 4-8 and 4-16,

ammonium sulfate was the only N-source that did not enhance the total P uptake by

bahiagrass after reapplication, implicating that the increase in P uptake by bahiagrass

when AS was added was related to the pH. Ebeling et al., 2003, suggested that P sources

with relatively high soluble P or bioavailable P concentrations (CaHPO4 and biosolids)

provided high levels of P for plant uptake in poorly buffered systems such as sand media,

which has little capacity of converting soluble P to less available forms by reaction with

or sorption by Fe, Al, Mg, or Ca compounds.

All applications were N based, so each material had a different quantity of P

applied depending on concentration of P in the material (Table 4-16). Phosphorus uptake

is a product of dry matter accumulation and tissue P concentration. Statistical

comparisons were made to compare the bahiagrass P uptake after first application and









after reapplication. At 0.05 probability (two tailed test), the F value was 2.05; when

comparing the MSE values of the first application with the reapplication the F value was

2.15, which was higher than 2.05, consequently the response variation for both studies

are different, therefore a comparison between both studies was not made. Bahiagrass P

uptake in the tubs that received biosolids application was greater than the untreated

control (Table 4-16). This suggests that P in the biosolids became partly available for

plant absorption within a relatively short period of time (112 days).

Table 4-16: Quantity of P applied by each material and percentage of P uptake by
bahiagrass after reapplication of the materials
Biosolids Kg N Quantity of Quantity P uptake Tissue P % Recovery
ha-1 material of P (*) concentration of applied P
applied on applied
each tub by each
material
-------g---- -g tub-1- ----g---- ----g Kg-1----
Limed cake 90 135 1.22 0.25 de 4.00 20.5
180 270 2.45 0.37 bc 4.10 15.1
Limed slurry 90 76 1.68 0.28 cde 4.20 16.7
180 151 3.34 0.48 a 4.30 14.4
Disney 90 109 1.28 0.15 f 3.90 11.7
compost 180 218 2.55 0.25 de 4.00 9.8
Baltimore 90 55 1.40 0.26 de 4.30 18.6
pellets 180 110 2.80 0.43 ab 4.80 15.4
Ammonium 90 8 -- 0.19 ef 3.10
sulfate 180 17 -- 0.30 cd 3.40
Control 0 -- 3.60
ANOVA P-value <0.0001
CV 20.37
Biosolids x Pr>F 0.38
rate
(*)Means with the same letter are not different at 0.05.
Repeated use of biosolids may result in either increased soil P accumulation or P

surface water runoff which could pose potential eutrophication problems for sensitive









water bodies such as the Lake Okeechobee Basin (Adjei and Rechcigl, 2002; Reddy and

Flaig, 1995; Walker and Havens, 1995).

Leaching Evaluation

Total N in Leachate: The leachate was analyzed for NH4 NO3-, and P to assess N

and P movement in soils after biosolids reapplication. At 90 kg N ha-1, ammonium sulfate

induced the highest total N content in leachate at 1.26% of the N applied followed by

limed slurry with 1.18% of N. At 180 kg N ha-1, ammonium sulfate produced the highest

total N in the leachate with 4.19% of the N applied, followed by limed slurry with 1.01%

and limed cake with 0.24%. Ammonium sulfate was the only inorganic fertilizer used in

this study in order to establish a comparison between inorganic fertilizer and biosolids.

None of the biosolids analyzed supplied as much N to the leachate as ammonium sulfate,

suggesting that these biosolids do not represent a hazardous to the water table when used

as fertilizer (Table 4-17). When dividing the MSE for first application with the MSE for

reapplication the F value was 0.16, which is lower than 2.05, therefore there was no

difference between the total N in leachate (first and second application), thus, the

parameter total N in leachate was combined for analyzes. Overall, there was a difference

in total N in leachate when compared to the first application, Pr>F was 0.0011, which

was lower than 0.05. The total N in leachate after reapplication was lower than in the first

application leachate. This could be explained by the bahiagrass N uptake after

reapplication, which increased compared with the first application, so more N is being

removed from the soil preventing NO3-N leaching to the ground water. The concentration

of NO3-N in the leachate from the different N-sources was influenced by loading rate (90

kg N ha-1 and 180 kg N ha-1) and the length of time after the initial application. Nitrate-N

concentration increased with increasing level of applied biosolids and decreased with










time after application (Table 4-17, 4-18). Leachate collected on day 28 did not contain

detectable N03-N (Table 4-18).

Table 4-17: Total NH4-N and NO3 -N content in leachates from the different organic
materials after reapplication
Biosolids Kg N Total NH4 Total NO3 Total N in % of N applied
ha-1 in leachate in leachate leachate (7 leached
through 112
days)


Limed cake 90
180
Limed slurry 90
180
Disney 90
compost 180

Baltimore 90
pellets 180
Ammonium 90
sulfate 180
Control 0


---mg ---
6.75
5.25
14.75
22.75
2.00
0.00
1.00
0.00
22.00
138.00
0.00


---mg --- -------mg -------
0.00 6.75
3.00 8.25
5.75 20.5
12.50 35.25
0.00 2.00


0.25
0.00
0.00
0.00
8.00
0.00


0.25
1.00
0.00
22.00
146.00
0.00


Table 4-18: Concentration of NO3-N and NH4 -N in leachate as affected by biosolid
reapplication.
Biosolids Kg 7 days 14 days 28 days 56 days 84 days
N
ha-1
----------------------------------mg--------------------------------------
NO3 NH4 NO3 NH4 NO3 NH4 NO3 NH4 NO3 NH4
Limed cake 90 0 7 0 0 0 0 0 0 0 0
180 0 4 3 2 0 0 0 0 0 0
Limed 90 6 15 0 0 0 0 0 0 0 0
slurry 180 10 20 3 3 0 0 0 0 0 0
Disney 90 0 2 0 0 0 0 0 0 0 0
compost 180 0 0 0 0 0 0 0 0 0 0
Baltimore 90 0 1 0 0 0 0 0 0 0 0
pellets 180 0 0 0 0 0 0 0 0 0 0
Ammonium 90 0 20 0 2 0 0 0 0 0 0
sulfate 180 5 107 3 32 0 0 0 0 0 0
Control 0 0 0 0 0 0 0 0 0 0 0


0.39
0.24
1.18
1.01
0.11
0.01
0.06
0.00
1.26
4.19
0.00









The NH4-N data are presented in table 4-17 and 4-18. In general, the concentration

of NO3-N was several fold less than the NH4-N concentration. By day 28 all the materials

had no detectable NH4 being leached (Table 4-19). All NH4 was readily available in

soluble forms because most of the materials produced their leached NH4 peaks at 7 days

(Table 4-18). As for NO3, microbial activity had to occur in order to mineralize the

materials and break them into soluble forms, becoming available for plant uptake and

runoff.

Total P in Leachate: No detectable P was found in the leachates through all the

leachate collections, 7 to 112 days. Unlike N, leaching of P has not traditionally been

viewed as a major ground water problem. In many soils, abundant P-sorbing oxide

components keep leachate P levels well bellow eutrophication thresholds (Elliot at al.,

2002). Peterson et al., (1994) concluded that there is no need to worry about P leaching to

ground water because leaching was practically zero. Certainly, areas with shallow ground

water and course-textured soils of low P-sorbing capacity were not taken under

consideration. Studies have reported that leaching of biosolids P is minor or negligible

(Elliot et al., 2002; Sui et al., 1999; Peterson et al., 1994). Then, for most locations,

restricting biosolids application rates to the P needs of the crops would normally be

unnecessary to minimize leaching concerns (Elliot et al., 2002).

pH and EC: The relationship between pH and EC of the leachate and incubation

period at different rates of biosolid after reapplication is presented in table 4-19 and 4-20.

The pH values in the leachate ranged from 4.2 to 7.2, with the untreated control around

6.3. Limed materials increased the pH from 6.3 to 7.2 while ammonium sulfate reduced

soil pH from 6.3 to 4.2 at days. The incubation period had a marked effect on the pH of









the leachate, with or without biosolids. Similar to the first application of the biosolids, the

pH increase observed in the incubation study was not observed here as well, most likely

for the same reason stated in the previous study. Eghball (1999) found that application of

beef cattle feedlot manure or compost increased the soil surface (0-15cm) pH while N

application as NH4N03 reduced the pH (from 6.4 to 5.6).

Table 4-19: pH measurements in the leachate from 7 to 112 days after reapplication of
the materials
Biosolids Kg 7 days 14 days 28 days 56 days 84 days 112 days
(*) N
(**) ha-
------------------------------ --------------------------------
Limed cake 90 6.5 a 6.5 ab 6.0 ab 6.8 b 6.8 b 7.2 a
180 6.7 a 6.8 a 6.9 a 7.1 a 7.2 a 7.2 a
Limed 90 6.4 ab 6.3 b 6.5 cd 6.3 cd 6.5 c 6.6 cd
slurry 180 6.5 a 6.5 ab 6.6 bc 6.6 bc 6.7 bc 6.9 b
Disney 90 6.1 c 6.3 b 6.4 cd 6.3 cd 6.5 c 6.5 d
compost 180 6.1 c 6.2 b 6.4 cd 6.3 cd 6.5 c 6.5 d
Baltimore 90 6.0 c 6.2 b 6.4 cd 6.3 cd 6.5 c 6.5 d
pellets 180 6.0 c 6.2 b 6.5 cd 6.3 cd 6.4 cd 6.6 d
Ammonium 90 5.6 d 6.2 b 6.3 cd 6.3 cd 6.6 bc 6.7 c
sulfate 180 4.2 e 5.1 c 5.7 e 5.7 e 6.1 d 6.2 d
Control 0 6.2 bc 6.3 b 6.4 cd 6.5 bcd 6.6 bc 6.7 c
(*) Means with same letter are not different at 0.05 by Duncan test.
(**) Interactions effects between biosolids and rates are not significant; therefore
comparisons were made across all biosolids at both rates.

Electrical conductivity (EC, an estimate of soluble salts) levels increased for soils

receiving biosolids. Leachate of soils receiving ammonium sulfate reached a peak at 7

days with 910 uS cm-1 at 90 Kg N ha-1 and 1450 uS cm-1 at 180 Kg N ha-l. All other N-

sources differed from the control, increasing the EC when compared to the control.

Because the EC was measured from the soil leachate, these results indicate movement of

salt from surface applied biosolid and inorganic fertilizer (ammonium sulfate)

applications, as shown by Eghball (2002).










Table 4-20: EC measurements in the leachate from 7 to 112 days after reapplication of
the materials
Biosolids
(*) Kg N
(**) ha'1 7 days 14 days 28 days 56 days 84 days 112 days
------------------ -----------------------------------
Limed cake 90 574 cd 509 abc 534 a 561 a 424 a 541a
180 675 bcd 579 ab 549 a 560 a 416 a 476 ab
Limed slurry 90 589 cd 377 bc 418 b 452 bc 335 b 413 bc
180 768 bc 473 abc 457 ba 494 b 368 ab 429 abc
Disney 90 576 cd 439 abc 464ba 503 b 319b 419 bc
compost 180 837 b 488 abc 455 ba 501 b 368 ab 445 abc
Baltimore 90 490 d 374 bc 407 b 418 c 299 b 342 c
pellets 180 554 cd 364 c 393 b 429 c 320 b 374 bc
Ammonium 90 910 b 389 bc 387 b 420 c 303 b 359 bc
sulfate 180 1450 a 629 a 433 b 486 b 326 b 377 bc
Control 0 467 d 379 bc 410 b 422 c 315 b 359 bc
(*) Means with same letter are not different at 0.05 by Duncan test.
(**) Interactions effects between biosolids and rates are not significant; therefore
comparisons were made across all biosolids at both rates.














CHAPTER 5
CONCLUSIONS

(i). Nutritional value and potential toxic environmental impact: All the

materials contained sufficient N and P to support mineralization and nutrient release for

bahiagrass growth. Limed slurry, N-viro and limed cake contained higher Ca

concentration, due to the large amount of lime added to these materials. Based on their N

and P compositions, these biosolids can be used as low-grade N and P fertilizer and as a

source of calcium, especially the lime-stabilized residuals. All the materials used in this

study were of class A or class B in terms of USEPA's pathogens and pollutant

concentration limit, not exceeding the ceiling and loading concentration limits, thus not

representing a potential toxic environmental impact.

(ii) Mineralization: Biosolids N is an important resource that could be used more

efficiently in crop production. Percentage N mineralized from biosolids in 263 days of

incubation ranged from 2 to 81% of their total N content. All biosolids increased

mineralization rates when N applied was increased (180 kg N ha-1) except for limed cake,

that when applied at 180 kg N ha- increased pH to 11.6 and EC to 2811 uS cm-1, which

may have inhibited microbial activity in those columns thus reducing mineralization.

Intensive microbial activity during the first week was shown by the rate of CO2 evolution

that was enhanced in the presence of biosolids. Overall, CO2 production increased as the

rate of applied biosolid increased.

(iii) Biosolids potential for inducing growth response of bahiagrass: The

potential use of biosolids as an turf grass amendment is important to producers and









environmentalists, since their use may improve forage nutritive value and at the same

time be a means for biosolids disposal. Most of the biosolids used in this study, specially

the lime-stabilized materials, induced bahiagrass forage production at a similar rate and

to the same extent as the inorganic ammonium sulfate fertilizer. All N-sources gave better

forage production when compared to the untreated control. After reapplication the large

cumulative dry matter production obtained for N-sources indicate that biosolids can

provide sufficient in season and residual plant available N to support forage crops. Tissue

N and P increased with increasing rate of biosolids. Limed slurry and N-viro induced N

uptake by bahiagrass to the same extent as the inorganic fertilizer. Besides limed slurry at

90 kg N hal-, none of the biosolids contributed as much N to the leachate as ammonium

sulfate, suggesting these biosolids can be used to reduce the threat of environmental

contamination. As for P, no detectable P was found in the leachates throughout both

study periods, including after reapplication.

There is much to be learned about the use of biosolids on agricultural crops.

Society will benefit from wise application of this material and thus from the recycling of

a valuable source.
















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BIOGRAPHICAL SKETCH

Caroline Reis was born in Uberlandia, Minas Gerais, Brazil, in April of 1979. After

graduating from high school, she attended Federal University of Uberlandia, where she

graduated in 2003 with a bachelor's degree in agronomy and where she was influenced

by Dr. Korndorfer's interest in soil science and fertilizer.

Upon graduation, Caroline applied to the University of Florida where she met Dr.

Jerry Sartain. She was accepted by Dr. Sartain into his program and will graduate with a

M.S. degree in soil and water science in 2006.