Effect of biochar on the fate and transport of manure-borne progesterone in soil

Ecological Engineering 97 (2016) 231–241

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Effect of biochar on the fate and transport of manure-borne progesterone in soil Sanaz Alizadeh a , Shiv O. Prasher a,∗ , Eman ElSayed a,b , Zhiming Qi a , Ramanbhai M. Patel a a b

Department of Bioresource Engineering, McGill University, Sainte Anne de Bellevue, H9X3V9, QC, Canada Department of Plant Protection, Zagazig University, Egypt

a r t i c l e

i n f o

Article history: Received 20 April 2016 Received in revised form 28 July 2016 Accepted 1 August 2016 Keywords: Progesterone Sorption Desorption Lysimeter Biochar Manure

a b s t r a c t The sorption affinity and desorption resistance of two types of biochars as soil amendments was evaluated in laboratory. The softwood and hardwood-biochars demonstrated a high sorption affinity for progesterone. The effective distribution coefficient (Kd eff ) of soil-soft wood-derived biochar (SBS450 ) was significantly higher (H0: P < 0.05) than soil-hardwood-derived biochar (SBH750 ), indicating its stronger sorption affinity for progesterone. Accordingly, a field-lysimeter study was conducted to elucidate the fate and transport of manure-borne progesterone in soil matrix and aquatic media in the presence of 1% softwood-biochar (BS450 ) in upper 0.1 m layer of soil. The spatial–temporal stratification of progesterone was monitored at four depths over a 46-day period where two different types of manures (swine and poultry) were applied to the topsoil of lysimeters under two treatments, soil (S) and SBS450 . The progesterone concentrations in SBS450 were significantly higher (H0: P < 0.05) and persisted for a longer period in the surface soil than in the control (S); however, the concentration was significantly lower in the deeper profile and in leachates. The results clearly showed that the application of biochar as a soil amendment can be used in alleviating the threat of pollution from manure borne progesterone. © 2016 Published by Elsevier B.V.

1. Introduction Over the last decade, several cases of biological abnormalities, including sexual, developmental and reproductive disorders, have been reported in the aquatic environment; these abnormalities are linked to the occurrence of a new paramount class of emerging contaminants with endocrine disrupting properties (Söffker and Tyler, 2012). These biologically active, organic micro pollutants are classified as Endocrine Disrupting Chemicals (EDCs). The EDCs are natural and synthetic steroid hormones, possessing the potential to modulate the functions of the endocrine system by mimicking, counteracting, altering or interfering with the metabolism and biosynthesis of endogenous hormones (Colucci et al., 2001). Androgens, estrogens, and progestin are classified as the three main categories of steroid sex hormones. Recently, advances in analytical techniques have made it possible to detect these pollutants at low concentrations in soil and water systems (Pignatello et al., 2010). However, the high chronic toxicity causing adverse long-term biological development and health issues (e.g. carcinogenicity, mutagenicity or teratogenicity) of these organic

∗ Corresponding author. E-mail address: [email protected] (S.O. Prasher). http://dx.doi.org/10.1016/j.ecoleng.2016.08.001 0925-8574/© 2016 Published by Elsevier B.V.

contaminants at concentrations as low as ng L−1 (Choi et al., 2004), has highlighted the need for studies on their fate and transport in the soil-water system. Agricultural and land management practices, such as the application of biosolids and animal manure and wastewater treatment plant discharges, frequently have been identified as the major sources of environmental female sex hormone pollutants (BarteltHunt et al., 2012; Ho et al., 2014). Poultry and liquid swine manure are the most widely applied organic fertilizers in North America. They are used to boost soil fertility; however, from the environmental and health safety perspective, these manures are major sources of bioactive levels of natural steroidal sex hormones, including 17␤-estradiol (E2), estrone (E1), testosterone and progesterone. The frequent contamination of surface water and ground water from manure-borne steroid hormones through surface runoff from agricultural fields receiving livestock and poultry manures has been reported in several studies (Jacobsen et al., 2005; Shore and Shemesh, 2003; Soto et al., 2004). The environmental occurrence and detection of estrogens, above their lowest observation effect level (LOEL), 10 ng L−1 (Shore and Shemesh, 2003), have been reported (Casey et al., 2003; Colucci et al., 2001; Jacobsen et al., 2005; Lee et al., 2003; Yu et al., 2004); however, the environmental behavior and fate of progesterone are not well-known. Progesterone has a direct reproductive role as

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an early precursor in the formation of other steroid hormones; it is produced by both sexes, with a significantly higher concentration in females. Consequently, a large fraction of this hormone is excreted by livestock, leading to significant environmental exposure (Bevacqua et al., 2011). The potential stability of progesterone is expected following the previous detection of the compound in surface water resources (Kolpin et al., 2002), however, a limited number of studies have reported on the occurrence of biologically active progesterone in surface soil and runoff from agricultural fields treated with manure. Kolpin et al. (2002) reported the contamination of 4.3% of streams in the United States with average progesterone concentrations of 0.11 ␮g L−1 and a maximum concentration of 0.199 ␮g L−1 . Detection of biologically active concentrations of progesterone, in beef cattle feedlot soil and in runoff from simulated rainfall were reported by Mansell et al. (2011). In a similar survey, Bartelt-Hunt et al. (2012) reported the presence of progesterone in both soil and manure samples. They also detected progesterone in runoff having an average concentration of 59.5 ng L−1 with a maximum concentration of 570 ng L−1 . Given the limited understanding of the environmental pathways and ecotoxicology of high-toxicityat-low-concentrations of manure-borne steroid hormones in soil and water media, there is inadequate knowledge to address the remediation of manure-borne steroid hormones in the environmental matrix. As a result, there is a pressing need to conduct detailed studies with the objective of developing feasible remediation techniques in order to reduce the environmental and biological consequences of hormonal pollution. The in-situ incorporation of carbon-rich organic amendments on contaminated soils has been deployed as a financially-feasible approach to engineer the natural process to fulfill an environmental remediation requisite (Beesley et al., 2011). Known as a byproduct of the thermo-chemical conversion of biomass and biological residues in the absence of oxygen (pyrolysis), biochar possesses an amorphous structure, containing nano-scale condensed aromatic rings with a crystalline structure and high specific surface area, which provides strong sorption sites in order to fulfill an environmental remediation of inorganic contaminants (e.g. heavy metals) and hydrophobic organic contaminants (Beesley et al., 2011). Based on the pyrolysis conditions, including different heating rates, various types of biochars with specific structural and physio-chemical properties are produced. Slow pyrolysis biochar is the final byproduct of pyrolysis at relatively low temperatures (300–500 ◦ C) with a long heating time and a heating rate less than 10 K min−1 , consisting of higher proportions of aliphatic carbons and functional groups. Whereas fast pyrolysis takes place over a short time at a high temperature (700–900 ◦ C) with a heating rate of more than 1000 K min−1 , thus resulting in a structure higher in micro porosity and with more poly aromatic carbons (Jung et al., 2013). Although, the retention capacity of biochar for different types of pesticides, including triazine and acetamiprid (Zheng et al., 2010), herbicides and heteroaromatic amines (Xiao and Pignatello, 2015), aminocyclopyrachlor, bentazone and fungicide pyraclostrobin (Cabrera et al., 2014) and endocrine disrupting compounds (Jung et al., 2013), have been determined using batch sorption-desorption experiments, there is a paucity of knowledge regarding the environmental remediation behavior of biochar under field conditions. Therefore, detailed spatio-temporal investigations of the in situ field-scale retention ability of biochar are needed. The primary objective of this study was to evaluate the feasibility of incorporation of two types of biochars (soft-wood and hard-wood derived biochars, produced at 450 ◦ C and 750 ◦ C, respectively) as a novel approach for reducing soil and water pollution of progesterone. A laboratory batch equilibrium study was conducted to determine the progesterone retention ability of these

Table 1 Physical and chemical characteristics of soil (ElSayed and Prasher, 2014), and progesterone (Colucci et al., 2001; Hao et al., 2013; Yu et al., 2004). Soil

Progesterone

Sand (%): 92.2 Silt (%): 4.3 Clay (%): 3.5 Organic matter (%): 2.97 Bulk density (kg m[3]): 1350 Hydraulic conductivity (m d−1 ): 1.67 ± 0.45* CEC (cmol kg−1 ) @ : 4.9 pH: 5.5

Molecular weight: 314.46 (g mol−1 ) Water solubility@ 20 ◦ c: 8.81 (mg L−1 ) Vapor pressure: 1.3 × 10−6 mm Hg at 25 ◦ C log KOW : 3.67-3.87 log RBA(# ): −0.7 Chemical structure:

@ * #

Cation exchange capacity. Average saturated hydraulic conductivity ± standard deviation. RBA: relative binding affinities for androgen and estrogen receptors.

biochars. To validate the lab results, a field lysimeter study was conducted to elucidate the fate and transport of the manure-borne progesterone in sandy soil. The specific objective was to study the fate and transport of progesterone in outdoor lysimeters, filled with a sandy soil, at different depths over a 46-day period, and to compare it with the lysimeters where the upper 0.1 m layer was amended with biochar, equivalent to 1% by weight, where two different types of manures (swine and poultry) were incorporated and applied with simulated rainfall. 2. Materials and methods 2.1. Analytical chemicals The analytical chemical standards for progesterone (>98% purity) was purchased from Sigma Aldrich (St. Louis, MO, USA). The physiochemical properties and the chemical structure of progesterone are summarized in Table 1. The anhydrous calcium chloride standard was provided by Science lab, Houston, Texas, USA. High performance liquid chromatography grade acetonitrile, used both as a solvent and for the mobile phase, was purchased from Fisher science. The de-ionized water was used as the mobile phase in the analytical analysis of samples, as a solvent in desorption bath equilibrium experiments and in all steps of the Standard solution preparation. 2.2. Soil characteristics The soil chosen for the laboratory batch equilibrium study was a sandy soil (with 92.2% sand content) of the Ste-Amable complex, Ferro-Humic podzol (ElSayed and Prasher, 2014), collected from the outdoor lysimeters assigned for the field evaluation. The physical characteristics of the soil are presented in Table 1. The lysimeters were located at the Macdonald campus of McGill University, SteAnne-De-Bellevue, Quebec. 2.3. Biochar characteristics Two types of biochars were provided by BlueLeaf Inc, Drummondville, Quebec, Canada. The main feedstock for the production of both biochar was spruce trees. Softwood-derived biochar (BS450 ) was produced by slow-pyrolysis of spruce tree bark, at 450 ◦ C for 2.5 h. Hard wood of spruce trees was processed under fast pyrol-

S. Alizadeh et al. / Ecological Engineering 97 (2016) 231–241 Table 2 Proximate analysis of biochar.

Particle size distribution <0.15 mm 0.15–0.18 mm 0.18-0.25 mm 0.25–0.425 mm 0.425–0.850 mm 0.85–2 mm 2–6.3 mm 6.3–9.5 mm 9.5–16 mm 16–19 mm >19 mm Hydraulic conductivity Chemical analysis Total Ash Organic Carbon Inorganic Carbon a Hydrogen/carbon (H:C) Hydrogen Oxygen Total nitrogen (N) b Butane activity

BS450

BH750

2.9 4.4 4.9 3.9 9.4 22 13.9 18.4 6.9 13.2 0 0.260 m s−1 17.30%

18.8 21.6 21.5 24.3 11.4 2.4 0 0 0 0 0 0.426 m s−1 9.80%

77% 0.50% 0.34 (molar ratio) 2.20% 3% 0.56% 9.6 g/100 g

74% 0.12% 0.55 (molar ratio) 3.40% 12.6% 0.42% 4.4 g/100 g

BS450 and BH750 are softwood-derived low-temperature pyrolysis biochar, and hardwood-derived high-temperature pyrolysis biochar, respectively. a The H/C ratio indicates the degree of carbonization of biochar. b Butane activity indicates the adsorption ability of biochar to trap butane from dry air; it represents the availability effective micro pore volume and surface area for adsorption process.

ysis at 750 ◦ C for a few minutes to produce hard-wood biochar (BH750 ). The physical, chemical and elemental characterization of each biochar was provided by Blue Leaf Inc. with the collaboration of the soil control lab of Control Laboratories Inc., Watsonville, California, USA. Saturated hydraulic conductivity was determined by constant head method described by Inzé and Van Montagu (1995). The detailed characterization of each type of biochar is presented in Table 2.

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test, and 1.5 mL of the supernatants was sub-sampled and filtered, as described earlier, and kept in the amber HPLC vials for analysis to determine the desorbed concentrations. The content of the hormone adsorbed to each treatment at sorption equilibrium (Cads s(eq) , ␮g−1 ), was calculated by the massbalance difference between the initial and analytically measured mass concentration of the hormones in the aqueous phase at adsorption equilibrium (Cads aq , mg L−1 ) of progesterone. The sorption isotherm of hormone was determined by fitting the batch equilibrium sorption data to the Freundlich sorption model: Cads s(eq) = Kf Caq 1/n

(1)

Where, Kf (mL g−1 ) and 1/n are the Freundlich adsorption coefficient and the nonlinearity coefficient, respectively. For each treatment, desorption behavior of progesterone was calculated based on the amount of hormone desorbed from adsorbed amount under equilibrium conditions. 2.5. Biodegradation of progesterone in soil The biodegradation of progesterone was investigated over 5 days under simulated field conditions to evaluate the effect of biochar amendment on the degradation pattern of progesterone. Sterile Soil samples were prepared by autoclaving at 121 ◦ C for 1 h. Four different treatments (5 g), including non-sterilized soil (non-sterile-S), non-sterilized soil amended with 1% BS450 (nonsterile-SBS450 ), sterile soil (sterile-S), and sterile soil amended with 1% BS450 (sterile-SBS450 ), in triplicates, were spiked with 1 mL of 25 mg L−1 concentration stock solution of progesterone to reach initial progesterone content of 5 mg kg−1 . The biodegradation experiment was conducted under controlled average temperatures of 25 ◦ C and a controlled humidity of 70%, under constant 24-h incandescent and fluorescent light with the intensity of 200 (␮moles m−2 s−1 ) in the growth chamber. The degradation kinetics of progesterone was explained by the first-order degradation kinetics (eq.2).

2.4. Sorption–desorption experiments

dC/dt = −kC

The sorption efficiency of both biochars was assessed through a set of laboratory batch equilibrium experiments using a modified protocol by Lee et al. (2003). Each batch included three different treatments, soil (S), soil and soft wood biochar (SBS450 ) and soil and hard-wood biochar (SBH750 ). Each soil-biochar treatment contained 1% (w/w) of biochar. Fresh stock solution of progesterone was prepared in pure HPLC-grade acetonitrile. Based on the reported sex hormone content in swine and poultry manure (Bevacqua et al., 2011), six concentrations of progesterone (0.01, 0.05, 0.1, 0.5, 1, 5 mg L−1 ) were chosen as the initial spiking concentrations. Triplicate soil samples (2 g) as a control treatment and two other biochar–containing treatments were equilibrated with 30 mL of each concentration of progesterone on a reciprocation shaker for 48 h at 25 ± 0.5 ◦ C. All three treatments were covered with aluminum foil to prevent any possible photo-degradation. After reaching the equilibrium time, all Pyrex glass tubes were centrifuged at 3230 rpm (1750 G) for 12 min and the supernatants of each treatment were transferred to another set of acid washed and sterilized Pyrex tubes. 1.5 mL of each replicate of each treatment’s aliquot were filtered by 0.22 ␮m sterile syringe filters and transferred into the amber HPLC vials and analyzed. After removing supernatants in the sorption test, 30 mL of purified Milli-Q water was added to each treatment sample in order to accomplish the desorption test. The centrifuge tubes were covered with aluminum foil again and were agitated for 48 h. The samples were centrifuged, as described in the sorption

Where C (mg kg−1 ) is the total remaining concentration of the progesterone at time t (days), and k (day−1 ) is the first-order rate constant. Accordingly, the half-life of progesterone in each treatment was calculated using Eq (3). t1/2 = 0.693/k

(2)

(3)

2.6. Instrumentation and analysis procedure All the analysis and concentration quantification of progesterone in the soil extracts and aliquots were performed by a quaternary pump LC system from Agilent 1100 series technologies (Germany) equipped with a diode array-ultraviolet detector. The detection and concentration tracing of progesterone was carried out through a Zorbax Eclipse Plus C18 column (150 × 4.6 mm) with particle size of 5 ␮m (Agilent, Santa Clara, CA). The best detector response for the identification of progesterone was at 240 nm with retention of 7.5 min. The mobile phase used in this process was a mixture of 40% of purified Milli-Q water and 60% HPLC-grade acetonitrile on volume basis, with a flow rate of 1 mLmin−1 with the injection volume of 100 ␮L for isocratic analysis. Prior to each analysis, the mobile phase was filtered using 0.45 ␮m membrane filters, and degassed. The column temperature was kept constant at 25 ◦ C during all analysis. The detection limit for the progesterone was 0.001 mg L−1 .

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Fig. 1. Schematic diagram of the lysimeter.

2.7. Field experimental setup The field experiment, investigating the influence of Soft-wood derived biochar (BS450 ) on the fate, transport and retention of the manure-borne female sex hormone, progesterone, in soil and water media was conducted using twelve PVC lysimeters (0.45 m diameter × 1 m height) located at the Macdonald Farm on the Macdonald campus of McGill University in Ste-Anne-de-Bellevue, Quebec, Canada (lat. 45◦ 24 48.8 N, long. 73◦ 56 27.9 W). Each lysimeter was sealed at the bottom with a 0.6m × 0.6 m PVC sheet, and a 0.05 m diameter drainage pipe was installed in the bottom to collect leachate (Fig. 1). The lysimeters were packed with sandy soil to a bulk density of 1350 kg m−3 (Table 1). Four 0.01 m diameter holes were drilled radially in lysimeters at 0.15, 0.35 and 0.65 m below the soil surface in order to collect soil samples. Laboratory sorption-desorption study indicated higher efficiency of BS450 in sorption of hormones as compared to BH750 , therefore BH750 was excluded in the lysimeter study. Six lysimeters were applied with poultry manure. Three lysimeters were incorporated with BS450 (SBS450 Pol) and three lysimeters were without biochar (control, soil only, S Pol). In the SBS450 Pol treatments, biochar was integrated at the recommended rate of 1% of soil (dry weight basis) in the top 0.1 m layer (Sarmah et al., 2010). Similarly, the other six lysimeters were applied with swine liquid manure

where soil treatment (control, soil only, S Swn) and biochar-soil treatment (SBS450 Swn) were used. For both types of manures, two treatments were allocated randomly. The lysimeters were kept under field conditions, sheltered by a canopy to prevent the entry of natural rainfall and perform controlled irrigation. To prevent any plant uptake, crops were not planted. Prior to the experiment, the last application of manure to these lysimeters was performed in 2007 (Kim et al., 2011). No hormone residue was found in the initial soil samples collected from these lysimeters for the current study. The meteorological data were collected from the Ste-Anne de Bellevue station of Environment Canada. During the period of the experiment (July–August 2012), the maximum, minimum and ◦ average air temperatures were 21.2, 9.8 and 15.6 C, respectively, and the average relative humidity was 66%.

2.8. Rainfall simulation To simulate natural rainfall, the lysimeters were irrigated four times on days 1, 15, 30 and 45 after the application of the manures. To simulate the spring application of manure and to imitate the worst-case scenario, the highest total amount of rainfall in May in a fifty-year period from 1962 to 2012, 173.4 mm, was applied. This amount was divided into three equal irrigations with a measured quantity of 57.8 mm for a month. The fourth irrigation applied was

S. Alizadeh et al. / Ecological Engineering 97 (2016) 231–241 Table 3 Characterization of poultry and liquid swine manure.

Dry matter (%) Organic matter (%) Density (Mg m−3 ) C/N ratio Total Nitrogen (kg Mg−3 ) Ammonia Nitrogen (N-NH4 ) (kg Mg−3 ) Phosphors (P2 O5 ) (kg Mg−3 ) Potassium (K2 O) (kg Mg−3 ) Calcium (Ca) (kg Mg−3 ) Magnesium (kg Mg−3 )

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extraction and analysis. The samples were transported to the lab, extracted and analyzed on the same day.

Poultry manure

Swine manure

26.1 17 0.622 5.8 14.7 5.25 11.7 8.4 22.1 1.5

0.1 0.6 0.977 2.8 1.1 0.45 0.4 1.3 0.3 0.1

the equal amount. The quantified amount of water was applied uniformly on the soil surface at the just ponding rate which means that water was applied in such a way that no water ponding took place on the soil surface. 2.9. Poultry and swine manures Poultry manure was provided by the Donald McQueen Shaver Poultry Complex, Macdonald campus of McGill University, Quebec, Canada. The poultry’s diet did not contain any growth promoting, or other synthetic hormones or antibiotics. Liquid swine manure was provided by the Macdonald campus swine complex located at the R. Howard Webster Centre for Teaching and Research in Animal and Poultry Science, Macdonald campus of McGill University, Quebec, Canada. Swine received several clinical veterinary treatments beyond their weaning phase, including PG600 and regumate, to control and induce the estrus, lutalyse and oxytocin for inducing and speeding up farrowing and Duplocillin LA, but the swine were not given any synthetic hormones or antibiotics. The poultry and swine manures were collected one day before initiating the field study. The characterization of both manures was conducted in AgroEnviro Lab, La Pocatière, Quebec, Canada; the data is presented in Table 3. 2.10. Manure application Poultry manure was collected and analyzed following the methods used by Andaluri et al. (2012) before application to the lysimeter. The concentration of progesterone was determined to be 9.6 ± 0.40 mg kg−1 . Considering spring as the application time, poultry manure was applied at a local recommended dose of 2.5 kg m−2 (0.4 kg per lysimeter). The manure was incorporated in the upper 0.1 m surface layer of the lysimeter. Liquid swine manure was also collected and analyzed using the same method. The concentration of progesterone was determined as 1.91 ± 0.025 mg L−1 . The manure was applied directly to the soil surface at the rate of 2.7 Liter per lysimeter on the following day. 2.11. Soil and leachate sample collection method Prior to the initiation of the experiment, all the lysimeters were brought to field capacity. After applying the manure, soil samples were collected eleven times including one day following each of the four irrigations. Samples were collected from the lysimeters’ surface layer as well as at depths of 0.15, 0.35 and 0.65 m, on day 1,2,3,7,15,16,23,30,31,45 and 46. The soil samples were stored ◦ at −24 C prior to the extraction and analysis. Sterilized amber water bottles were used to collect the leachate from the drainage pipes at the bottom of each lysimeter. During irrigation events on day 1, 15, 30 and 45, leachate was collected from each lysimeter, homogenized, and a one-liter sample was obtained for subsequent

2.12. Extraction of Hormones from Soil The extraction of hormones from the soil samples was performed based on the modified method used by Xuan et al. (2008). Soil extraction was conducted in triplicate from the collected soil samples. After thawing at room temperature, and air-drying for one day, 5 g of soil was mixed (1 min on Vortex mixer) with 5 g of anhydrous sodium Sulfate (Na2 SO4 ) and 10 mL HPLC grade acetone in 50 mL polyethylene centrifuge tubes, followed by 30 min of shaking at 250 rpm speed on a reciprocating shaker. The mixture was centrifuged at 4000 rpm for 20 min and the transparent supernatant of each tube was transferred to another clean 50 mL tube. In the second step of the extraction, another 10 mL of acetone was added to all tubes and the extraction procedure was repeated. By transferring the supernatant collected after the second step, the tubes, containing the aliquots, were centrifuged at 4000 rpm for 20 min and the resulted supernatants were transferred to 50 mL pyrex glass centrifuge tubes, and completely dried under N2 stream. They were reconstituted with 1 mL of 50/50 (v/v) acetonitrile-water solution and homogenized by sonication. The final extracts were filtered through 0.22 ␮m sterile syringe filters and transferred into 1.5 mL amber vials for HPLC analysis. 2.13. Extraction of Hormones from Leachate The leachate sample extraction was conducted as described in Stafiej et al. (2007) using a solid phase extraction (SPE) process with Oasis HLB extraction cartridges (200 mg cc−1 ,Oasis Co.Ltd, NY). One liter of each leachate sample was filtered through a 45 mm filter (Advantec, Japan) to separate the suspended soil particles and other materials. A 35% concentrated hydrochloric acid was used to acidify the water samples to pH 2. After conditioning the cartridges with 5 mL of methanol followed by 5 mL of high-purity de-ionized water, the acidified samples were passed through the cartridges at a flow rate of 20 mL min−1 . After eluting the compound with 10 mL of acetonitrile, the extract was dried under the nitrogen stream. The dried samples were dissolved in 1 mL of 50/50 (v/v) acetonitrile-water solution and were kept in the sonicator for 15 min. The final extracts were filtered by 0.22 ␮m sterile syringe filters and transferred into the amber HPLC vials and analyzed. 3. Results 3.1. Sorption-desorption behavior of progesterone in the presence of biochar The sorption isotherms of progesterone in the S, SBS450 , and SBH750 treatments are given in Fig. 2(A). The C-type isotherms of progesterone indicated the partitioning as the main sorption mechanism by distribution of the progesterone in the solid-solution phase without any specific bonding between sorbent and sorption site (Sparks, 2003). The S-type and L-type sorption isotherms of progesterone were observed in SBS450 and SBH750, respectively. The sorption equilibrium data was a good fit with the Freundlich isotherm model (0.95 < R2 < 0.99; Table 4). Results indicated that both SBS450 and SBH750 treatments demonstrated a significantly higher (H0: P < 0.05) sorption affinity for progesterone as compared to the S treatment. The sorption isotherm of progesterone in S treatment (Table 4) was linear (1/n = 1), indicating monolayer partitioning of the hormone on the sandy soil surface with a limited number of identical sites. The linear sorption isotherms of progesterone in sandy loam soil are also reported by Fang (2011). The dimensionless Freundlich constant (1/n) for progesterone in SBS450 ,

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(A) Sorebed concentration (μg/g)

12 10 SBS450

8

SBH750 6

Soil

4 2 0 0

1

2

3

4

5

6

Equilibrium concentration (Ce)(mg/L)

(B) Amount desorbed (μg mL-1)

0.07 0.06 0.05 SBS450 0.04 SBH750 0.03

Soil

0.02 0.01 0 0

2

4

6

8

10

12

Amount sorbed (μg g-1) Fig. 2. (A) Progesterone sorption isotherms for soil and soil-biochar treatments, and (B) The amount of desorbed progesterone for soil and soil-biochar treatments. Table 4 Freundlich parameters of progesterone sorption in the presence and absence of biochar. Treatment Soil +1% BS450 Soil +1% BH750 Soil without Biochar

Kf

1/n a

3.53 2.11 a 0.50 b

b

0.62 0.60 b 1.06 a

eff

R2

Kd

0.95 0.96 0.99

4.58 a 2.80 b 0.48 c

log Koc 2.42 a 2.21 a 1.44 b

Different superscript letter represent significant difference (H0: P < 0.05).

with a value of 0.624 and 0.596 in SBH750 , indicates non-linearity of sorption isotherms in the presence of biochar. This could result from the aromatic nature of biochar (Wang et al., 2016) and the presence of high percentages of carbonized fractions, 77 and 74% in BS450 and BH750 , respectively. Carbonized and non-carbonized fractions of biochar dominantly influences the sorption mechanisms and degree of nonlinearity of organic compounds by biochar (Cao et al., 2009). The carbonized fraction of biochar performs as the glassy domain function (i.e. the nonlinear sorption isotherms) where the semi-permanent nano-scale vadous zone of carbonized fraction contributes to strong adsorption sites with an internal high surface (Xia and Pignatello, 2001). In contrast, the non-carbonized fraction represents the rubbery domain function (i.e. linear sorption isotherms) (Zheng et al., 2010). To evaluate the mobility of hormones in the soil and the effect of the biochar as a soil amendment, the soil/water-organic carbon partition coefficient (Koc ) was calculated in both S and SBS450 and SBH750 treatments (Table 4). The log (Koc ) for progesterone was 1.44 in the S treatment; however, in the presence of the BS450 and BH750 the log (Koc ) values have increased indicating the significant effect of biochar on the sorption affinity of hormones. Due to the observed linearity in the sorption isotherms for soil S compared to SBS450 and SBH750 , the concentrationdependent effective sorption distribution coefficient, Kd eff

(L kg−1 ) = (Kf /Cw n−1 ) was calculated at the single solution equilibrium concentration of Cads aq = 0.5 (mg L−1 ). Likewise, a single-point organic carbon normalized partitioning coefficient (Koc ) was calculated where the Koc = (Kf /Cw n−1 )/foc at Cads aq = 0.5 mg L−1 , and foc is the organic carbon content of the soil (Sarmah et al., 2010). Significant differences (H0: P < 0.05) were observed between the effective distribution coefficient (Kd eff ) values (Table 4) between soil and soil-biochar treatments. In the presence of biochar, the sorption capacity of progesterone in soil increased noticeably. Although both SBS450 and SBH750 demonstrated high retention ability, the sorption coefficient of progesterone in SBS450 was significantly higher compared to SBH750 . The lower H/C molar ratio (0.34) for BS450 indicated high aromatic structure with more organic carbon, whereas BH750 had relatively higher H/C molar ratio (0.55). Similarly, Hao et al. (2013) correlated the increased adsorption of atrazine on corncob biochars with decreased H/C and (O + N)/C ratios. This observed negative correlation can be justified by the enhanced strong sorption sites via the increased carbon content of biochar (Salvia et al., 2014; Zheng et al., 2010). The lower H/C and O + N/C ratios could contribute to the advanced aromaticity and higher hydrophobicity (Jung et al., 2013). The positive correlation between Kf and aromaticity of biochar was reported (Kupryianchyk et al., 2016) where higher aromaticity can lead to enhancement of the interaction of organic compounds with aromatic adsorptive sites via п-п electron donor-acceptor interactions. This was observed in our sorption experiment where the highest Kf values between the three treatments was associated with the SBS450 treatment (Kf = 3.53). Accordingly, Xiao and Pignatello (2015) indicated the important role of hydrophobic interactions on the major adsorption sites provided by the aromatic carbon and hydrophobic structures of biochar. The C-20 ketone moiety in the progesterone structure (Table 1) is a triple hydrogen acceptor (Neale et al., 2009); therefore, the hydrophobic interaction between progesterone (пacceptors) and the aromatic rings (п-donors) of BS450 and BH750 can be assumed to be the way in which progesterone is strongly sorbed by biochar surface. Studies (Cabrera et al., 2014; Salvia et al., 2014) have reported that generally the higher pyrolysis temperature can lead to a higher surface area and pore volume of biochar. However, Bornemann et al. (2007) reported that higher pyrolysis temperature does not necessarily increases the surface area or micro porosity due to the loss of active components in biochar. Brown et al. (2006) also reported a significantly decreased surface area of pine biochar as the pyrolysis temperature increased. Biochar used in this study also demonstrated similar characteristics. Fast pyrolysis with high heating rate can result in biochar with low surface area and pore volume (Boateng, 2007; Zhang et al., 2004), which can be attributed to the accumulation of volatile compounds between and within particles and blockage of pore entrances (Angın, 2013). The higher volatile matter content of the BH750 (17%) than that of BS450 (6%) was consistent with the latter reported trend. Lua and Guo (1998) also associated the structural deterioration of pyrolyzed chars at high temperature (900 ◦ C) to the sintering of the mineral ash phase coupled with pore shrinkages. BS450 consists of mainly larger particles with a diameter from 2 to 19 mm, whereas BH750 contains a smaller diameter, indicating a higher heterogeneity with higher effective porosity in BS450 . Accordingly, the higher butane activity of BS450 as compare to BH750 (Table 2) indicates more available pore volume and higher surface area, and therefore a higher potential sorption affinity of BS450. Similarly, Liang et al. (2015) correlated the lower butane activity of biochar to its lower surface area. Desorption behavior of progesterone is presented in Fig. 2 (B). A statistically significant difference (H0: P < 0.05) was observed between the desorbed concentration of progesterone in the soilbiochar and soil treatments. Desorption results showed that 32% to

S. Alizadeh et al. / Ecological Engineering 97 (2016) 231–241 Table 5 Effect of biochar on dissipation and the half-life of progesterone in sterilized and non-sterilized soil. Soil treatments

Initial concentration (mg kg−1 ), t = 0.25 h

Non-sterilized soil 3.86 ± 0.15 Non-sterilized soil + 1% BS450 3.90 ± 0.22 Sterilized soil 3.85 ± 0.20 Sterilized soil + 1% BS450 3.81 ± 0.20

Final concentration (mg kg−1 ), t = 120 h

Half-life (day)

1.04 ± 0.03 1.57 ± 0.12 1.21 ± 0.11 1.83 ± 0.10

2.81a 2.97a 4.03 b 4.65 b

Different superscript letters represent significant difference (H0: P < 0.05).

55% of the sorbed amount of progesterone, at different initial spiked concentrations, was desorbed in soil treatment whereas, less than 5% of the amount of progesterone was observed to be desorbed in both SBS450 and SBH750 treatments. This indicated the strong capacity of both biochars to retain progesterone in the soil matrix over the equilibrium time. In the treatments of SBS450 and SBH750 , more effective aromatic and hydrophobic sorption sites are provided by the biochar surface with stronger interaction energies which leads to higher retention of progesterone in soil matrix. This would retard the flow of hormones in water ending up in the water bodies, and would control pollution from manure-borne hormones. Although no significant difference was found between desorption of progesterone from SBS450 and SBH750 , the results showed a marginally higher desorption potential in SBH750 compared to the SBS450 treatment. 3.2. Biodegradation of progesterone in the presence of biochar The dissipation of progesterone was determined in sterile soil and non-sterile soil to elucidate the role of microbial activity on the biodegradation process. Given the higher sorption potential of BS450 as compared to BH750 , BS450 was amended in both sterile and non-sterile soil to determine the effect of biochar on progesterone degradation. The results are presented in Table 5. The half-life of progesterone was significantly lower (H0: P < 0.05) in non-sterile treatments as compared to sterile treatments. A 73% of the initial progesterone was degraded in non-sterile-S as compared to 60% in the sterile-S over 5 days indicating the dominant role of biodegradation on fate of progesterone in soil. Soil bacteria is found to be responsible for the dissipation of progesterone in soil by providing a double bond between carbons 1 and 2, and hydroxylation at carbon 9 which leads to aromatization of the A-ring, cleavage of the B-ring, and the formation of a labile metabolite (Sparks, 2003). The calculated half-life of progesterone in non-sterile-S and non-sterile-SBS450 were calculated as 2.8 and 2.9 days (Table 5). The half-life of progesterone in sterile-S was not significantly higher than sterile-SBS450 . Therefore, there was no significant effect of biochar on the degradation of progesterone. Only 1% biochar was present in non-sterile-SBS450 . Thus, in both non-sterile SBS450 and non-sterile-S, the quantity of soil was almost equal. There would be minimal impact of biochar on degradation. However, if there were a detrimental impact of biochar on the soil biota, there would be extended half-life in non-sterile-S BS450 treatment as compared to non-sterile-S. But, the results showed no significant increase in half-life when biochar was amended and therefore, it appears that biochar does not adversely affect progesterone degrading microbes. 3.3. Fate and transport of progesterone in soil and leachate 3.3.1. Spatial-temporal movement of progesterone in soil receiving poultry manure The concentrations of progesterone in the samples collected from the lysimeters under both treatments amended with poul-

237

try manure, (S Pol and SBS450 Pol), at four depths are shown in Fig. 3. At the surface on day 1, after the first irrigation, the concentration of progesterone in soil treatment was 52.40 ± 1.30 ␮g kg−1 (average ± standard deviation); the concentration decreased to 37.51 ± 1.19 ␮g kg−1 on day 2. This rapid decrease was due to the transport of hormone to the lower depths. The concentration gradually decreased to 4.89 ± 1.17 ␮g kg−1 on day 15. The concentration continued to further decrease after irrigation was applied on day 15; after the second irrigation (day 16) less than 2% of the initial progesterone concentration remained in the soil. It appears that transport of considerable amounts of hormone occurred due to irrigation which is the result of the low sorption capacity of sandy soil, confirmed by the sorption experiment. Similarly, relatively low sorption capacity of soil with a high sand content, such as sandy loam, on hormones, than that of silt and clay loam, are reported (Casey et al., 2003; Sarmah et al., 2010). By day 30, the hormone was not detected at the surface. In the SBS450 Pol the concentration of hormone in the surface soil on day 1, after the first irrigation, was 101.24 ± 1.09 ␮g kg−1 , which was two times higher as compared to S Pol treatment. Spatio-temporal repeated measures analysis carried out using SAS (SAS Institute Inc., 2010) showed that the concentration in SBS450 Pol was significantly higher (H0: P < 0.05) as compared to S Pol treatment. This indicated that mobility of progesterone in the lysimeter was decreased, due to its higher adsorption with biochar. The observed pattern corresponded well with the sorption batch experiment result, as the highest sorption capacity of progesterone was determined in the SBS450 treatment. The highly reactive surface of biochar is promoted by the presence of primary functional groups, including hydroxyl, amino, ketone, ester, nitro aldehyde, and carboxyl groups donates at the edges of stacked carbon sheets formed during pyrolysis (Reid et al., 2013). On day 2, the concentration was 62.89 ± 3.55 ␮g kg−1 . Thus, a considerable decrease was observed. This reveals that progesterone was moved to lower layers, however, to a lesser degree as compared to the soil treatment. It gradually decreased to 24.24 ± 0.65 ␮g kg−1 on day 15 before the second irrigation. On day 16, the concentration was 19.01 ± 0.66 ␮g kg−1 . Thus, there was 21% reduction (5.23 ␮g kg−1 ) in a twenty-four-hour period. It may be noted that there was decrease of only 14.73 ␮g kg−1 (35%) in 7 days prior to the second irrigation. Thus, it suggests that irrigation causes the transport of hormones in soil. The concentration gradually decreased to 4.26 ± 0.13 ␮g kg−1 on day 30 before the third irrigation; on day 31, 1.17 ± 0.17 ␮g kg−1 was observed. Onward, it was not detected. Throughout the experiment, it was observed that at the surface, the concentrations were significantly higher in SBS450 Pol as compared to S Pol. This observed pattern supports the fact that biochar increased the adsorption of manure-borne progesterone. The sorption-desorption behavior of progesterone in the soil matrix is strongly influenced by its chemical structure, consisting of two ketone functional groups and two methyl groups and a lack of proton-accepting functional groups (Kreinberg, 2012). Similar to estrogens, progesterone has a relatively high hydrophobicity (log Kow = 3.87) (Neale et al., 2009), indicating its strong partitioning affinity for the hydrophobic surface of biochar. The strong interaction of biochar with hydrophobic organic compounds, such as progesterone, is correlated to the highly aromatic nature, high surface area, micropore volume, and the presence of an abundance of polar functional groups in biochar (Jung et al., 2013). A rapid decrease in concentration of hormones in loam soil is also reported by Ying and Kookana, (2005). As expected, progesterone concentrations decreased quickly and were not detected on day 23 in sandy soil, although it was observed in the same soil amended with biochar. Thus, the hormone would have leached to lower depths and also degraded via both biotic and abiotic transformations (Salvia et al., 2014).

S. Alizadeh et al. / Ecological Engineering 97 (2016) 231–241

100 10

a

b

a

b

S_Pol a

b

a

b

b

b

b

a

1

SBS450_Pol b a

a 0.1 a 0.01

100

Concentration (µg kg-1)

100

Soil surface

10 1

Depth 0.35 m a

a

a b

b

a

a

S_Pol

a

b b

SBS450_Pol b

a

0.1 0.01

Concentration (µg kg-1)

Concentration (µg kg-1)

1000

10 1

Depth 0.15 m a

b

a

b

a b

a

S_Pol SBS450_Pol

a

b

b

ab

0.1 a 0.01

100

Concentration (µg kg-1)

238

Depth 0.65 m

10

S_Pol SBS450_Pol

1 0.1 0.01

Fig. 3. Concentration of progesterone in the S Pol and SBS450 Pol treatments at four sampling depths of lysimeters receiving poultry manure (log scale used for Y axis plotting), different letters in bars for a day represent significant difference (H0: P < 0.05).

At 15 cm depth, after the first irrigation on day 1 and day 2, the concentrations in S Pol were 31.63 ± 1.29 ␮g kg−1 and 36.34 ± 0.28 ␮g kg−1 progesterone; however, only 14.30 ± 2.08 ␮g kg−1 and 15.72 ± 0.33 ␮g kg−1 of hormones were detected in the lysimeters with the biochar-soil treatment. This is probably due to the fact that a larger quantity of progesterone leached from the upper profile in S Pol, compared to SBS450 Pol. The leaching would have continued until day 2 in S Pol as the concentrations marginally increased. The high sand content of the soil (more than 92%) would potentially lead to less sorption, resulting in more mobilization of progesterone in the aqueous phase, causing more leaching to lower depths. The concentrations were significantly higher (H0: P < 0.05) in soil as compared to SBS450 Pol at this depth. The concentration progressively decreased to 9.34 ± 0.79 ␮g kg−1 and 2.20 ± 0.40 ␮g kg−1 after 7 days in S Pol and SBS450 Pol, respectively. In both treatments, the concentration continued to decrease; however, progesterone was not detected on day 23 in the soil treatment. In SBS450 Pol, progesterone was detected on day 23, although only a trace amount, which indicates that the hormone would have been leaching slowly from the biochar applied at the surface layer. Until day 15, the concentrations were significantly higher in the soil treatment as compared to the SBS450 Pol treatment at this depth; at later stages, the concentrations were quite low. At 35 cm depth, progesterone concentration in S Pol was 14.77 ± 1.11 (␮g kg−1 ) on day 1. Although the concentration slightly decreased to 10.45 ± 0.47 (␮g kg−1 ) on day 2, it progressively increased to 23.14 ± 0.79 (␮g kg−1 ) on day 15. Considering the relatively high concentrations of progesterone in soil at 15 cm depth for 15 days, water with these high concentrations of progesterone would have leached to lower depths, and as a result, high concentrations were detected at 35 cm depth in S Pol. In contrast, progesterone concentrations were quite low at this depth in the SBS450 Pol treatment. It may be expected given higher adsorption in the upper profile where biochar was amended, resulting in less transport of progesterone to lower depths. Thus, clear effects of biochar application in surface soil were evident at 35 cm depth. At 65 cm depth, progesterone was not observed in soil treatment until day 16 (2.75 ± 0.26 ␮g kg1 ), which further decreased to 1.08 ± 0.04 ␮g kg−1 on day 23. The concentration at 35 cm depth

Table 6 Spatio-temporal repeated measures analysis of progesterone concentrations at four depths over time in lysimeters applied with poultry and swine manure.

Treatment Depth Time Treatment x depth Treatment x time Treatment x depth x time *

Poultry Manure

Swine manure

*

*

*

*

*

*

*

*

*

*

*

*

Significant (H0: P < 0.05).

progressively increased and was at peak on day 15; therefore, it may be expected that there may be leaching from this depth to lower depths in the following days. Progesterone was not detected onward as the concentrations at the upper depth diminished and minimal leaching would have occurred; a small quantity of progesterone present at 65 cm might have degraded with time. No progesterone was found in the soil samples collected from the depth of 65 cm in the SBS450 Pol treatment in the entire 46 days of the study period. The spatial-temporal concentration profiles of both treatments demonstrated a rapid and greater leaching in soil as compared to SBS450 Pol. Progesterone remained at higher concentrations in the upper most profile of biochar-amended soil for an extended period of time. Contrary to this, the concentrations in the lower profiles were higher in the soil treatment for a longer period as compared to BS450 Pol. These trends are reflected in the statistical analysis of progesterone concentrations at four depths for nine measuring events using repeated measures; results revealed a significant (H0: P < 0.05) effect of treatment, depth, time and their interactions (Table 6). 3.3.2. Temporal presence of progesterone in leachate from lysimeters receiving poultry manure The initial sampling event took place on the first day of the experiment after the application of manure and the first irrigation event and repeated after each irrigation event on day 15, 30 and 45. The concentration of manure-borne progesterone, the volume of drain flow and mass of progesterone in leachate, collected after each irrigation, are presented in Table 7. The concentration of pro-

S. Alizadeh et al. / Ecological Engineering 97 (2016) 231–241 Table 7 Concentration of progesterone in leachate, leachate volume, and progesterone mass. Treatment

Day 0 Day 15 Concentration (␮g L-1 )

Day 30

S Pol SBS450 Pol

72.9 ± 2.2a 34.8 ± 8.0b Volume (L) 6.6 ± 0.9 4.9 ± 1.0 Mass (␮g) 480.0 ± 60.5a 167.1 ± 19.8b Concentration (␮g L-1 ) 117.5 ± 13.1a 46.2 ± 2.9 Volume (L) 6.3 ± 0.9 6.0 ± 2.5 Mass (␮g) 735.9 ± 35.8a 278.2 ± 117.4b

40.9 ± 4.4a 16.2 ± 3.7b

18.4 ± 3.8a 1.9 ± 0.0 b

4.2 ± 1.5 3.5 ± 0.4

1.9 ± 0.2 2.8 ± 1.3

174.3 ± 71a 57.6 ± 18.2b

34.3 ± 7.7 a 5.3 ± 2.5b

52.3 ± 0.2a 18.7 ± 1.4

9.9 ± 0.8a 1.8 ± 1.3

5.5 ± 1.1 3.7 ± 1.5

3.1 ± 2.0 2.1 ± 1.1

287.3 ± 58.0a 68.1 ± 25.0b

31.9 ± 23.1a 4.3 ± 4.8b

S Pol SBS450 Pol S Swn SBS450 Swn S Swn SBS450 Swn S Swn SBS450 Swn

The values are average ± standard deviation. Different superscript letters show significant difference between treatments (H0: P < 0.05).

gesterone and the volume of drain flow varied with treatment and decreased with time. The highest concentration of progesterone in leachate samples (72.9 ± 2.2 ␮g L−1 ) was measured after the first irrigation event from S Pol treatment. The highest volume of drain flow was collected after the first irrigation in S Pol treatment and therefore highest amount of progesterone (480 ± 60.5 ␮g) was leached. Although the highest concentration of progesterone was similarly observed in leachate under SBS450 Pol treatment after first irrigation event; the concentration was half of the concentration observed in S Pol, and it was significantly lower (H0: P < 0.05). The progesterone mass in SBS450 Pol was 65% less than that from S Pol (H0: P < 0.05). The progesterone concentration decreased over time in both S Pol and SBS450 Pol treatments. In S Pol, 56% and 25% of the concentration from first irrigation were observed on day 15 and day 30 events; the corresponding progesterone mass was 36% and 7%.

Concentration (µg kg-1)

3.3.3. Spatial-temporal movement of progesterone in soil receiving swine manure The spatial-temporal distributions of progesterone concentrations in lysimeters applied with liquid swine manure, at four sampling depths including the surface, 15, 35 and 65 cm over 46 days under the two treatments, S Swn and SBS450 Swn are presented in Fig. 4. The progesterone distribution in the lysimeters was found to be similar to the hormone distribution observed in the lysimeters receiving poultry manure. A descending trend over time was observed for progesterone concentrations at surface soil under both treatments. On day 1, 50.65 ± 3.48 ␮g kg−1 was observed in S Swn, whereas 74.37 ± 4.36 ␮g kg−1 was observed in SBS450 Swn treatment. On day 2, the detected progesterone concentrations were 24.94 ± 0.74 ␮g kg−1 and 55.67 ± 2.08 ␮g kg−1 in the respective treatments. It showed that in one day the concentration decreased to about 50% of the concentration on day 1 in the soil treatment; however, it decreased to only about 75% in SBS450 Swn. This trend reveals that biochar adsorbed the hormone to a greater extent as compared to soil. Our sorption experiment results are in agreement with observed patterns. The estimated Kd eff for the SBS450 treatment (4.58 L kg−1 ) was 9 times larger than Kd eff for the S treatment (Table 4) confirming the enhanced retention capac100

1000

Soil Surface S_Swn

100 10

Therefore, there was considerable decrease in concentration as well as the progesterone mass with time. In SBS450 Pol concentrations were 47% and 5%, respectively; the progesterone mass was 34% and 3%, respectively. When progesterone mass was compared between treatments, only 33% and 16% progesterone mass was observed on day 15 and day 30 in SBS450 Pol as compared to the mass observed in S Pol; these were significantly lower (H0: P < 0.05; Table 7). Thus biochar amendment can reduce pollution from drain flow. Hormone was not detected after the fourth irrigation on day 45 in both treatments. The laboratory study showed greater sorption of progesterone with biochar amendment, and similar effect was observed in lysimeter soil. As a result, there was reduced concentration of progesterone in leachate which translated into significantly lower mass of progesterone in SBS450 Pol.

ab

a

b

b a

b

SBS450_Swn

b

a

b

b

a

1

b

a

0.1

a

Concentration (µg kg-1)

S Pol SBS450 Pol

0.01

a

a b

S_Swn

a

b b

b

SBS450_Swn

a

0.01

Concentration (µg kg-1)

Concentration (µg kg-1)

a

a

b 0.1

b

a

a b

b

S_Swn

a b

a

SBS450_Swn

1 b 0.1

100

Depth 0.35 m b

1

10

Depth 0.15 m a

a a a

a

0.01

100

10

239

10

Depth 0.65 m S_Swn SBS450_Swn

1

0.1

0.01

Fig. 4. Concentration of progesterone in the S Swn and SBS450 Swn treatments at four sampling depths of lysimeters receiving swine manure (log scale used for Y axis plotting), different letters in bars for a day represent significant difference (H0: P < 0.05).

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ity of sandy soil for progesterone. The concentrations gradually decreased to 4.12 ± 0.14 ␮g kg−1 and 26.58 ± 0.103 ␮g kg−1 on day 15. Similar to the poultry manure applied lysimeters, there was a considerable decrease in concentrations after irrigation observed on day 16 in swine manure applied lysimeters. Progesterone was not found in soil treatment, although it was present on day 23 in SBS450 Swn. The hormone concentration was always higher at the surface soil in SBS450 Swn (Fig. 4) and persisted for a longer period due to higher adsorption. Considering the partitioning and pore–filling as major processes for the organic compound association in low H/C materials such as biochar with crumpled polyaromatic sheets (Salvia et al., 2014), the preliminary phase of sorption of progesterone in the presence of biochar is potentially a function of the gradual intra-particle diffusion mechanics (Zheng et al., 2010). As in the case of poultry manure, in the swine manure applied lysimeters, the concentration of progesterone increased from 28.12 ± 1.34 ␮g kg−1 on day 0–37.24 ± 1.29 ␮g kg−1 on day 1 in soil treatment at depth 15 cm. However, the concentration decreased from 16.38 ± 3.9 ␮g kg−1 to 9.61 ± 0.86 ␮g kg−1 in SBS450 Swn. This was probably due to more leaching in the soil treatment and more adsorption to biochar in the upper profile. The concentrations gradually decreased until day 23 in both treatments; afterward progesterone was not detected as it would have degraded. At 35 cm depth, the concentrations were 14.71 ± 0.93 ␮g kg−1 and 5.4 ± 0.86 ␮g kg−1 in S Swn and SBS450 Swn on day 1. A slight decrease in the concentration was observed on day 2; however, it gradually increased in the soil treatment until day 7 due to leaching from the profile above. The concentration decreased until day 23, beyond which the hormone was not detected. In SBS450 Swn, progesterone was only about 10% of the concentration in S Swn treatment on day 0. The concentration was consistently low over time. At 65 cm, hormone was observed in S Swn, not in the SBS450 Swn treatment. Overall, the hormone transport trends were similar as those of the poultry manure treatments. Statistical analysis also showed similar trends as poultry manure (Table 6). 3.3.4. Temporal presence of progesterone in leachate from lysimeter receiving swine manure The concentrations, leachate volume and mass of progesterone in both S Swn and SBS450 Swn treatments are given in Table 7. The progesterone in leachate samples receiving liquid swine manure was similar to the results from poultry manure treatments. In S Swn treatment, the highest progesterone concentration 117.5 ± 13.1 ␮g L−1 and mass of 735.9 ± 35.8 ␮g were measured on day 1 after the first irrigation. The corresponding values in SBS450 Swn treatment were 46.2 ± 2.9 ␮g L−1 and 278.2 ± 117.4 ␮g; these were significantly lower than the values in S Swn treatment (H0: P < 0.05). The differences between treatments are likely due to the enhanced retention capacity of soil for progesterone by biochar application. Similarly, the greater log Koc values associated with SBS450 (Table 4) confirmed the minimized mobility of the hormone in the presence of biochar. After the second irrigation on day 15, 287.3 ± 58.0 ␮g, equivalent to 40% of the initial detected hormone mass was quantified in leachate from S Swn, whereas the leachate from SBS450 Swn contained only one fourth (68.1 ± 25.0 ␮g) the amount. On day 30, after the 3rd irrigation 31.9 ± 23.0 ␮g hormone was found in leachate from S Swn; however, less than 5 ␮g of progesterone was quantified in leachate from SBS450 Swn treatment. Hormone was not detected in leachate from the 4th irrigation on day 45. The results show that biochar can act as a filter to restrict the flow of manure borne hormones to water bodies. These results are consistent with the sorption/desorption results as a significantly less amount of progesterone was desorbed from biochar treatments when compared to soil without biochar. Therefore, biochar application in the surface soil may be consid-

ered as an economical and effective method for minimizing manure borne hormones transport via surface and drainage flow from agricultural soils. 4. Conclusions The application of the two, tested biochars demonstrated a high sorption affinity for progesterone and a strong resistance for desorption than soil. In both sorption and desorption batch equilibrium experiments, BS450 showed better remediation capacity of hormonal pollution compared to BH750 ; BS450 was used in the field lysimeter study. The results from the lysimeter study demonstrated that considerable amounts of progesterone leached from surface soil profile to the lower profile in soil treatment. However, more hormones were retained for an extended period in the upper soil profile, when amended with biochar, and the concentrations were significantly lower in the lower soil profile as well as in the leachate from biochar treatment as compared to the soil treatment. Thus, it appears that amendment of 1% BS450 in the 0.1 m surface layer significantly increased the retention of progesterone in topsoil (Ho: P < 0.05) and thus mitigates the pollution threat to water bodies. However, the impact of the presence of hormones in the surface layer at elevated levels over an extended period needs to be evaluated. From an environmental and agricultural perspective, the outcome of this study can be seen as a proof of the need for further field-scale applications of biochar as a novel, clean and feasible remediation technique. However, detailed, long-term studies are required to investigate the competitive sorption ability of biochar in the simultaneous presence of different chemicals in the soil media. References Andaluri, Gangadhar, suri, PRominder P.S., Kumar, Kuldip, 2012. Occurrence of estrogen hormones in biosolids, animal manure and mushroom compost. Environ. Monit. Assess. 184, 1197–1205. Angın, Dilek, 2013. Effect of pyrolysis temperature and heating rate on biochar obtained from pyrolysis of safflower seed press cake. Bioresour. Technol. 128, 593–597. Bartelt-Hunt, Shannon L., Snow, Daniel D., Kranz, William L., Mader, Terry L., Shapiro, Charles A., van Donk, Simon J., Shelton, David P., Tarkalson, David D., Zhang, Tian C., 2012. Effect of growth promotants on the occurrence of endogenous and synthetic steroid hormones on feedlot soils and in runoff from beef cattle feeding operations. Environ. Sci. Technol. 46, 1352–1360. Beesley, Luke, Moreno-Jiménez, Eduardo, Gomez-Eyles, Jose L., Harris, Eva, Robinson, Brett, Sizmur, Tom, 2011. A review of biochars’ potential role in the remediation, revegetation and restoration of contaminated soils. Environ. Pollut. 159, 3269–3282. Bevacqua, Christine E., Rice, Clifford P., Torrents, Alba, Ramirez, Mark, 2011. Steroid hormones in biosolids and poultry litter: a comparison of potential environmental inputs. Sci. Total Environ. 409, 2120–2126. Boateng, A.A., 2007. Characterization and thermal conversion of charcoal derived from fluidized-bed fast pyrolysis oil production of switchgrass. Ind. Eng. Chem. Res. 46, 8857–8862. Bornemann, Ludger C., Kookana, Rai S., Welp, Gerhard, 2007. Differential sorption behaviour of aromatic hydrocarbons on charcoals prepared at different temperatures from grass and wood. Chemosphere 67, 1033–1042. Brown, Roberta A., Kercher, Andrew K., Nguyen, Thanh H., Nagle, Dennis C., William, William P., 2006. Production and characterization of synthetic wood chars for use as surrogates for natural sorbents. Org. Geochem. 37, 321–333. Cabrera, A., Cox, L., Spokas, K., Hermosín, M.C., Cornejo, J., Koskinen, W.C., 2014. Influence of biochar amendments on the sorption-desorption of aminocyclopyrachlor, bentazone and pyraclostrobin pesticides to an agricultural soil. Sci. Total Environ. 470, 438–443. Cao, Xinde, Ma, Lena, Gao, Bin, Harris, Willie, 2009. Dairy-manure derived biochar effectively sorbs lead and atrazine. Environ. Sci. Technol. 43, 3285–3291. ˇ Casey, Francis X.M., Larsen, Gerald L., Hakk, Heldur, Simunek, Jirí, 2003. Fate and transport of 17í-estradiol in soil-water systems. Environ. Sci. Technol. 37, 2400–2409. Chanil, Jung, Kwang, Junyeong Park, Lim, Hun, Park, Sunkyu, Heo, Jiyong, Her, Namguk, Oh, Jeill, Yun, Soyoung, Yoon, Yeomin, 2013. Adsorption of selected endocrine disrupting compounds and pharmaceuticals on activated biochars. J. Hazard. Mater. 263, 702–710. Choi, Seul Min, Yoo, Sun Dong, Lee, Byung Mu, 2004. Toxicological characteristics of endocrine-disrupting chemicals: developmental toxicity, carcinogenicity, and mutagenicity. J.Toxicol. Environ. Health B 7, 1–23.

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