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Water Air Soil PollutDOI 10.1007/s11270-011-0864-z Characterization of Swine Wastewater by ToxicityIdentification Evaluation Methodology (TIE) C. Alejandra Villamar & Teresa Cañuta &Marisol Belmonte & Gladys Vidal Received: 25 January 2011 / Accepted: 13 June 2011 # Springer Science+Business Media B.V. 2011 Abstract Since swine wastewater is used by farmers (around 100%) and total nitrogen (95.5%). This for soil fertilization, evaluation of toxic compounds or finding suggests that part of the toxicity comes from micro-contaminants of separate streams is required.
anionic compounds, such as chlorine.
This paper uses the toxicity identification evaluation(TIE) procedure for the physicochemical and ecotox- Keywords Acute toxicity. Daphnia magna .
icological characterization of swine wastewater. To Swine wastewater. TIE distinguish the most important toxic compounds, aphysicochemical characterization and phase I-TIEprocedure were performed. The acute toxic effect of swine wastewater and treated fractions (phase II-TIE)were evaluated using Daphnia magna determining Intensive swine production, due to its input demand 48-h LC50. Results show a high level of conductivity and the introduction of its residue (e.g., swine (23.5 μS cm−1), which is explained as due to the wastewater) into the soil, has resulted in soils concentration of ions, such as ammonium (NH +– saturated with nutrients (e.g., N, P, K, organic matter), N 1.6 g L−1), sulfate (SO 2− 397.3 mg L−1), and heavy metals (e.g., Cu and Zn), hormones, antibiotics chlorine (Cl−1,230.0 mg L−1). The acute toxicity of (tetracycline, sulfamines and β-lactamase), and salts the swine wastewater was evaluated on D. magna (e.g., NaCl), which are mobile and can reach (48-h LC50=3.4%). Results of the different water subterranean waters producing environmental impacts treatments indicate that anionic exchange treatments (Burton and Turner ; Moral et al. could reduce 22.5% of swine wastewater's acute Swine wastewater is a mixture of feces (45%), toxicity by reducing chlorine (to around 51%) and urine (55%), and washing water, which are character- conductivity (8.5%). On the other hand, cationic ized by a high content of organic matter (14.2–26.3 g exchange treatment increased acute toxicity on D.
BOD5L−1), nutrients (2.4–6.0 gL−1 of total nitrogen, magna (% RT= −624.4%), by reducing NH +– and 0.6–1.4 gL−1 of total phosphorus), ions (2.3–5.1 g Cl− L−1, 1.5–3.8 g NH +– N L−1 and 0.03–0.07 g N L−1), and other compounds (Blanes-Vidal et C. A. Villamar : T. Cañuta : M. Belmonte : G. Vidal (*) al. ; Moral et al. ; Moral et al. Provolo Engineering and Biotechnology Environmental Group, and Martínez-Suller The toxic effect, Environmental Science Center EULA—Chile, University expressed as median lethal concentration (LC50), of swine wastewater on aquatic cladocer organisms, such P.O. Box 160-C, Concepción, Chilee-mail: as Daphnia magna, demonstrate its high toxicity at Water Air Soil Pollut low concentrations of this waste (LC50=1–>10%; De indicator organism and toxicity identification evalua- la Torre et al. ). Some earlier studies have related tion (TIE) as the evaluation tool.
the toxicity of Daphnia spp. with ionic compoundslike those found in swine wastewater, whose toxicitymechanisms on algae and cladocers have not been 2 Materials and Methods studied (Pretti et al. ). They also speculated thatthe toxicity on D. magna would be related with 2.1 Swine Wastewater enzymatic inhibition and membrane disruption.
Cationic compounds, such as NH + Swine wastewater was obtained from a small piggery um), under certain conditions (pH>8.0, temperature, farm located in southern Chile. Swine wastewater was and ammonium concentration) can be transformed sampled after the primary settling treatment and was into deionized compound, such as NH3 (or ammonia), transported in polyethylene containers and immedi- which is toxic to aquatic organisms (Arauzo and ately stored at approximately 4°C to avoid degrada- Valladolid ; Blanes-Vidal et al. ; Leung et al.
tion. When the bioassays could not be executed A study of the effect of treated swine within 48 h, samples were filtered through a 0.45- wastewater (24-h LC on the reproduction and m pore size nitrocellulose membrane and adapted to longevity of Daphnia carinata and Moina austral- the assay's temperature.
iensis found that D. carinata and M. australiensisadults can tolerate swine wastewater concentrations in 2.2 Toxicity Testing the order of 2.8 and 8.8 mg L−1 of NH3, respectively,while juveniles can only tolerate concentrations in the Acute toxicity was determined by exposing D. magna order of 2.2 and 7.5 mg L−1 of NH3, respectively juveniles (<24 h) to each sample for 48 h. At the end (Leung et al. ). However, ammonium presents of exposure, mortality, defined as lack of organism acute toxicity on D. magna at values of 48-h LC ≥ mobility when the vessel was shaken, was recorded.
1.5 mg L−1 of NH +– N (Martins et al. ). In Organisms were obtained from in-house cultures species of Ceriodaphnia dubia, the chronic toxicity maintained according to guidelines given by USEPA caused by the NH + depends on the oxidation time of ). Before the experiment, cultures and animal and NO3 , which affects the reproduction of exposures were conducted at 20±2°C in a photope- these crustaceans (Dave and Nilsson ).
riod of 16-h light/8-h dark and fed three times per Additionally, the toxicity of anionic compounds, week with a recharge of water each 48 h, permitting such as Cl− has been studied. In the case of parthenogenetic conditions. Dilution and control Daphnia spp., the acute toxicity of this anion has water was moderately hard, reconstituted water been studied at concentrations > 0.005 mg L−1 prepared according to USEPA (The solutions (AQUIRE ; Pretti et al. It has also been were not renewed and the organisms were not fed reported that the formation of organochlorides during the experiments. The 24-h median lethal (AOX) can produce acute toxicity in organisms concentrations (LC50) were calculated using the such as D. magna at concentrations of 24-h EC50 of Spearman–Karber method (Finney ; ). Esti- 2.2 mg AOX L−1 (Emmanuel et al. ). The mation of the samples' theoretical toxicity was based presence of metallic salts, such as CuCl2, CdCl2, and on the chemical analysis of the individual potential FeCl2, can present greater acute toxicity for D.
toxicants. Therefore, to express the relative toxicity of magna (24-h LC50=0.08–172.0 mg L−1) due to the a certain compound, acute toxic units (TUtheorical) for synergic effect between chloride and the metals, this compound were calculated as: nominal com- contrasting with salts like that present lower toxicity pound concentration in test solution divided by LC50.
(24-h LC50= 240.0–1020.0 mg L−1) (Martins et al.
Then, assuming that these toxicants were the real causative agents, they were determined according to The objective of this study is to provide a the concentration-addition model of Anderson and physicochemical and ecotoxicological characteriza- Weber ). If no interaction between toxicants is tion of swine wastewater using D. magna as the expected, then the observed sample toxicity should be Water Air Soil Pollut equal to the theoretical toxicity of the toxicant with (electrical conductivity) were evaluated using electro- the greatest effect. The sensitivity of the testing is des. The total alkalinity (TA) was determined by 0.05 mg Hg L−1 (USEPA ).
titration (Vidal and Diez ). In this study,ammonia (NH – 3 N) concentration was calculated for 2.3 Toxicity Identification Evaluation each test according to Anthonisen et al. ( The baseline test, the pH/adjustment test (pH 3/ad andpH 11/ad), the graduated pH test (pH 6.9/grad and 3 Results and Discussion pH 8.9/grad), and the pH adjustment/aeration test(pH 3/aer, pH 7.5/aer, and pH 11/aer) were performed 3.1 Physicochemical and Ecotoxicological as described by Norberg-King et al. (The other Characterization of the Swine Wastewater fractionations were based on procedures described byVan Sprang and Janssen (The EDTA (ethyl- Table shows the results obtained from the physico- enediaminetetraacetic acid) concentrations (Titriplex chemical characterization of swine wastewater. High III, p.a., Merck) in the dilution water and effluent values were observed for electrical conductivity sample were based on the 24-h 10% lethal concen- (13.8–25.6 μS cm−1), alkalinity (TA, 11.4–12.0 g tration (LC10) on D. magna and were conducted at a CaCO3L−1), organic matter (COD, 16.2–24.9 g O2 final EDTA of 300 mg L−1. For the pH adjustment/ L−1; BOD5, 4.5–9.8 g O2L−1), nutrients (TN, 1.9–2.5 g filtration test (pH 3/fil, pH 7.5/fil, pH 11/fil), 2.5 × N L−1), and ions (NH +– N, 0.7–2.0 gL−1; SO4 , 25-cm glass columns were filled with synthetic 271.9–503.1 mg L−1, and Cl−, 1,210–1,250 mg L−1).
hydrophobic filter wool (Diprolab MR). Additional These values are characteristic for this type of column tests consisted in an anion (20 g, amberlite discharge coming from the fattening phase (Blanes- IRA-440, Merck), a cation (20 g, amberlite IR-120, Vidal et al. Moral et al. Moral et al. ; Merck), and an 8- to 20-mesh activated carbon (15 g, Provolo and Martínez-Suller ). Moral et al. Sigma). Baseline toxicity tests were performed in relates the high electric conductivity in swine waste- duplicate, whereas TIE manipulations were not. ThepH was adjusted and readjusted by adding dropwise Table 1 Physicochemical and toxicological characterization pro-analysis HCl and NaOH of different normalities (D. magna) of the swine wastewater (0.1, 1.0, and 2.0 N; p.a., Merck).
For all manipulations, the 24-h LC verted to toxic units (TUfractionation=100÷24-h LC50) and were compared with the baseline toxicity test (TUbaseline). The toxicity reduction percentage (%TR) due to the applied fractionation was calculated as %TR ¼ 100  1  TUfractionation 2.4 Analytical Methods COD (chemical oxygen demand), BOD5 (biological oxygen demand), ammonium (NH +– gen (TN), total phosphorus (TP), sulfate (SO 2− EC electrical conductivity, COD chemical oxygen demand, chloride (Cl−) were determined according to the 5 biological oxygen demand, TA total alkalinity, TN total nitrogen, NH + – N ammonium, NH3-N ammonia TP total protocols established in Standard Methods (APHA- phosphorus, SO −2 sulfate, Cl− chlorine, 48 h LC50 median AWWA-WPCF ). Parameters such as: pH and EC Water Air Soil Pollut water to the high salt (e.g., Cl−, p<0.001) and protein for the cationic exchange fraction, where it increased content (e.g., NH + 4 , p < 0.001), which is present in the to 12. This result can be explained by the nature of diet for animals in the final growth stage, thus this fraction, which resulted in an increase in the explaining the results with respect to ionized com- (non-retained or available) OH- groups during this pounds (Martínez-Suller et al. Moral et al. test. A reduction of 51% in Cl− was observed in the The pH was between the values 7.2 and 7.5. Moral anionic exchange test without any evidence of et al. () indicate that a pH> 7.9 favors the changes in the pH (7.5). In the cationic exchange 3. The NH3 N concentration in this test, NH4 N was reduced in 100%. It is important to study fluctuated between 5.5 and 15.9 mg L−1, which indicate that the cationic exchange fraction had the is considered to be toxic for D. magna (48-h LC50, greatest variation in physicochemical characteristics 2.9–6.9 mg L−1) according to Pretti et al. ( of the swine wastewater, presenting an increase in pH Indeed, the results of the ecotoxicological assays with and total elimination of NH +– N. In contrast, SO4 D. magna found acute toxicity of 48-h LC50=3.3– increased (<55%) in all the treatments except for 3.7%. These values on average are 1.94 times the anionic exchange, due to the chemical composition of values reported by De la Torre et al. (for D.
the anionic resin. Reyes et al. ) reported similar magna (48-h LC50=1.8±0.2%) for pig slurry (e.g., results in this test for kraft cellulose effluent.
fattening farm). While the reproductive swine farmsamples have acute toxicity values (48-h LC50) 3.3 Phase II: Reduction of the Toxicity of Swine between 3% and >10% (De la Torre et al. ).
Wastewater in D. magna 3.2 Phase I: Toxicity Identification Evaluation Table shows the effect of the different TIE fractions of Swine Wastewater for the acute toxicity on D. magna (48 h) of the swinewastewater. For the cationic exchange fraction, an Table shows the physicochemical characterization of acute toxicity was observed to increase seven times the swine wastewater before and after fractioning with (48-h LC50=0.48%). In contrast, when comparing the the TIE technique. In all the tests, the pH did not fractions corresponding to pH 11 (aeration/filtration/ change (7.3–7.4) with respect to the control, except adaptation) with the control, toxicity increased only Table 2 Physicochemical characteristics for the different treated fractions from swine wastewater using TIE methodology Cationic exchange EDTA ethylenediaminetetraacetic acid Water Air Soil Pollut Table 3 48-h LC50 and TU Median lethal concentration on D. magna values below 95% of confidence intervalfor swine wastewater using Cationic exchange Aeration (pH 3.0) Aeration (pH 7.5) Aeration (pH 11.0) Filtration (pH 3.0) Filtration (pH 7.5) Filtration (pH 11.0) Adaptation (pH 3.0) TU toxic units, TU standardized toxic units standardized with respect 1.1–1.5 times. Additionally, toxicity diminished in organisms such as Daphnia spp. On the other hand, 22.5% for the anionic exchange fraction (48-h LC species of D. magna exposed to NH4 are observed to 4.45%), 11% in the activated carbon test (48-h LC50= be less sensitive to Cl− (48-h LC50 = 1.5– 3.83%), and 7% in the EDTA test (48-h LC50= 492.0 mg L−1; Mangas-Ramírez et al. When 3.66%). The results presented in Table demonstrate comparing the values obtained in the present study that the reduction in NH +– N in the cationic exchange with those reported in the literature, D. magna is fraction provokes an increase in toxicity (Table observed to be more sensitive to Cl− than to NH +– which is reflected in a higher TU value (208.33). In 50: the average value of NH4 contrast, the diminishment in the toxic units in the 4.4 mg L−1 and the average value of Cl− is 43.1± anionic exchange fraction (TU, 22.47) would be 3.5 mg L−1 which could explain the acute toxicity. It related with the 51% reduction in Cl−, showing the is important to note that an increase in pH in the greater sensitivity of D. magna to anionic compounds.
swine can increase the formation of NH3-N, provok- In the fractions corresponding to pH 11 (aeration/ ing an increase in toxicity for ammoniac compounds.
filtration/adaptation), a slight increase in toxic units Figure shows the percentage of TR generated by (TU, 43.29, 34.72, and 33.78, respectively) was each fraction of the TIE (Fig. ) and the removal observed and attributed to the formation of NH efficiency percentage for NH4 N and Cl− in each test due to the pH conditions in the swine wastewater.
(Fig. ). It can be observed in Fig. a that toxicity Table shows the different LC50 values for several (percentage of TR) was reduced in the following tests: cladocer organisms that were reported in the literature anionic exchange (22.5%), activated carbon (9.9%), to be produced by compounds such as Cl−, NaCl, and EDTA (5.7%). However, an increase in TR N, and NH3 N. On the other hand, Cl− is values, between 4.6% and 624.4%, was observed for observed to have high acute toxicity for Daphnia spp.
the other fractions and especially in the cationic between 0.005 and 0.16 mg L−1 exchange fraction. Thus, NH4 N was removed in (AQUIRE and 48-h LC50 between 0.12 and the anionic exchange (48.7%), cationic exchange 0.15 mg L−1 (Pretti et al. where the least toxic (100%), activated carbon (87.2%), and EDTA is salt (24-h LC50=1,020.0–3,240.0 NaCl mg L−1) (87.2%) fractions, while Cl− was removed in all the (Sarma et al. ; Martins et al. ). These results fractions except for the fraction pH 7.5/aeration where indicate that chloride is more bio-available for it increased (−7.1%).
Water Air Soil Pollut Table 4 Salinity and ammonium effect on aquatics organisms Range concentration Sarma et al. Martins et al. () Ceriodaphnia dubia Dave and Nilsson Ceriodaphnia dubia Mangas-Ramírez et al. () Mangas-Ramírez et al. () Thus, the increase in swine wastewater toxicity is observed to be related to the reduction of Cl− and thepresence of NH +– N. First, a diminishment of toxicity Using the TIE technique, the swine wastewater was is observed when Cl− and NH +– N are reduced at the characterized for each test, finding that the acute same time. The reduction of Cl− in the anionic toxicity for D. magna increased seven times in the exchange fraction would provoke the diminished cationic exchange fraction. From the estimation of TR toxicity, and an increase in toxicity would result from (percent), Cl− (605.0 ±21.2 mg L−1) was found to be the presence of 78% Cl− in the cationic exchange the compound provoking this acute toxicity in the even with a total reduction of NH +– anionic exchange. The increase in toxicity obtained in results indicate that NH +– N in swine wastewater acts the cationic exchange test (624%) is related to the as a Cl− immobilizing compound, diminishing toxic- retention of NH +– N (0.04±0.01 mg L−1) and a ity even when the ammonium is more toxic than Cl− greater bio-availability of Cl− in its ionic form, when the pH changes.
causing the acute toxicity for D. magna.
Activated C.
-40-600 -610 -620 -630 -640 -650 -140 -120-100 -80 -60 -40 -20 40 60 80 100 120 140 Removal Efficiency (%)
Fig. 1 a Toxicity reduction (TR) percentage for D. magna (48 h) for swine wastewater resulting Phase I TIE fractionation techniques.
b Removal efficiency of NH +– N (black bar), Cl− (white bar) for each test Water Air Soil Pollut This work was supported by CONICYT/ Mangas-Ramírez, E., Sarma, S. S. S., & Nandini, S. (2002).
PBCT (Grant TPI-01) and Innova Bío-Bio (Grant 07-PC S1- Combined effects of algal (Chlorella vulgaris) density and 198). The authors thank Mr. C. Contreras from Sucesion Yanine ammonia concentration on the population dynamics of for use of their facilities in the realization of this study.
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