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The Effectiveness of Aerobic and Anaerobic Methods During Start-up in Biological Shrimp Pond Wastewater Treatment
Eli Hendrik Sanjaya,1,2* Aulia Amara,1 Yahya Hengky Pamungkas,1 Anugrah Ricky Wijaya,1 Aman Santoso,1 Anie Yulistyorini,3 Yu-You Li,4, 5 Hong Chen,6 and Mohd Fadhil Md Din7
1Department of Chemistry, State University of Malang (Universitas Negeri Malang), Jl. Semarang No. 5, Malang, East Java, 65145, Indonesia.
2Department of Biotechnology, State University of Malang (Universitas Negeri Malang), Jl. Semarang No. 5, Malang, East Java, 65145, Indonesia.
3Department of Civil Engineering, State University of Malang (Universitas Negeri Malang), Jl. Semarang No. 5, Malang, East Java, 65145, Indonesia.
4Department of Civil and Environmental Engineering, Tohoku University, 6-6-06 Aoba, Aramaki, Aoba-ku, Sendai, Miyagi, 980-8579, Japan.
5Graduate School of Environmental Studies, Tohoku University, 6-6-06 Aramaki Aza Aoba, Aoba-ku, Sendai, Miyagi 980-8579, Japan.
6Key Laboratory of Water-Sediment Sciences and Water Disaster Prevention of Hunan Province, School of Hydraulic Engineering, Changsha University of Science & Technology, Changsha, 410004, China.
7Centre for Environmental Sustainability and Water Security (IPASA), School of Civil Engineering, Faculty of Engineering, Universiti Teknologi Malaysia, 81310 Skudai Johor, Malaysia.
*Corresponding email: [email protected] Received 24 April 2023; Accepted 11 December 2023
ABSTRACT
One of the aquaculture wastes is shrimp pond wastewater (SPW). Generally, SPW comes from the rest of shrimp feed, shrimp dung, and died shrimp during the cultivation. As SPW is organic waste, biological wastewater treatment is the best choice. Biological method has low operating costs and environmentally friendly. There are two kind of biological methods, namely aerobic and anaerobic. Both methods have advantages and disadvantages. In this research, the performance of aerobic and anaerobic methods in treating SPW were investigated using batch experiment during start-up for 105 days. The results showed that the performance of the aerobic system in the initial phase was better than that of the anaerobic system, especially, in phase one which is the adaptation phase. At the beginning of the experiment, the performance of aerobic system is better than that of the anaerobic system. The COD, carbohydrate and protein removals in the aerobic system were 25.93%, 75.20% and 88.59%, while in the anaerobic system were 12.82%, 36.03% and 51.01%.
Overall, based on the COD removal, the performances of start-up process of both systems were quite similar. Therefore, it needs more study to investigate the aerobic and anaerobic systems for a longer duration in a continuous or semi-continuous system.
Keywords: effectiveness, aerobic and anaerobic system, shrimp pond wastewater, wastewater treatment, organic removal.
INTRODUCTION
The aquaculture industry is an industry that cultivates aquatic organisms such as fish, molluscs, crustaceans and aquatic plants in a land that has been determined and maintained for environmental quality [1], is currently growing very rapidly. Increasing population which is in line with the fisheries demand is not balanced with the amounts of marine catches [1]. Based
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on data from the Indonesian Central Bureau of Statistics, the shrimp exports were increased by 41.56% in 2021 [2].
However, one of the aquacure industry's problems is shrimp pond wastewater (SPW).
SPW commonly comes from feed that is not eaten by shrimp, shrimp excrement, and shrimp that die during the cultivation process [3]. In more detail, shrimp pond waste is all waste originally from shrimp pond disposal. The waste generated includes solids, organic matter, and dissolved metabolites such as ammonia, urea, and carbon dioxide. The sludge which comes from uneaten food pellets, excrement, and eroded pond soil is enriched with nitrogen, phosphorus and carbon [1]. In addition, waste can also come from excessive use of chemicals, careless disposal of garbage in pond areas and disturbances in pond sediments [4]. The need for high oxygen which is not accompanied by renewal of oxygen causes anoxic conditions in sediments and areas between sediments and water [5]. Nitrogen and phosphorus retention of feed in shrimp rearing was 22.27% and 9.79%, respectively, so that nutrients wasted into pond waters respectively reaching 77.73% nitrogen and 90.21% phosphorus. The high content of ammonia in SPW also causes odor [6].
Several ways to overcome SPW pollution are coagulation, advanced oxidation processes, membrane filtration processes, adsorption, dialysis, photocatalytic degradation and biological methods [4]. However, biological method is the best choice because of low capital and operating costs, energy saving and environmental friendliness [7]. Biological methods consist of two systems, namely aerobic and anaerobic.
Under aerobic conditions, microorganisms metabolize substrates by a process known as aerobic respiration [8]. Aerobic decomposition of organic matter consists of fermentation and respiration (oxidation), biosynthesis, and endogenous respiration [9][10]. The complete oxidation of the substrate under aerobic conditions is represented by Equation 1.
C6H12O6 + 6 O2 → 6 CO2 + 6 H2O Equation (1) The end products of the oxidation process consist of carbon dioxide, ammonia, energy, water, and other end products. Carbon, oxygen, hydrogen, nitrogen, and sulfur (COHNS) represents organic waste compounds [11]. Aerobic waste treatment is generally used in the treatment of waste with low organic concentrations <10,000 mg/L [12]. Microorganisms also act as adsorbents for organic substances and heavy metals, further assisting decomposition [13], however, aerobic processes produce quite a lot of sludge.
In the anaerobic process, there are four main stages in the production of methane and carbon dioxide from organic materials, namely hydrolysis, acidogenesis, acetogenesis, and metabogenesis. Hydrolysis aims to convert insoluble complex organic compounds into smaller simple molecules that can be used as an energy source. Biopolymer proteins, carbohydrates, and lipids are hydrolyzed into amino acids, simple sugars, and fatty acids, respectively, by extracellular enzymes [13]. A complex consortium of microorganisms participates in the hydrolysis of most of the bacteria among which the flora predominate are anaerobic bacteria such as Bacteroides, Clostridia, Bifidobacteria and other gram-positive and gram-negative rods. Furthermore, it is followed by some facultative anaerobes such as Streptococci and Enterobacteriaceae [14].
In the acidogenesis stage, the results of hydrolysis in the form of sugars and amino acids are fermented by bacteria into acetate, propionate, butyrate, hydrogen, carbon dioxyde, lactate, valerate, ethanol, ammonia and sulfide. Acetogenesis, consists of two groups of bacteria, namely hydrogen-producing acetogens and homoacetogens (or hydrogen-using acetogens).
While hydrogen-producing acetogens aim to catabolize certain organic acids, alcohols, and
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aromatic compounds to acetate and carbon dioxide, homoacetogens use hydrogen and carbon dioxide to form acetate [10]. Methanogenic bacteria belong to the Archaebacteria group. A large group of methanogenic bacteria have been identified. The first group consists of 33 species belonging to the family Methanobacteriaceae, Methanothermaceae, Methanococcaceae, Methanomicrobiaceae, and Methanoplanaceae. Bacteria of this family are capable of reducing carbon dioxide and hydrogen and/or formate to form methane. The second group consists of 14 species belonging to the family Methanosarcinaceae. This species utilizes acetate, methylamine and/or methanol in the formation of methane. Methanosarcina barkeri and Methanosarcina vacuolata are the most versatile because they use all known methanogenic substrates except formate [10].
Anaerobic waste treatment is generally used in waste treatment with high organic concentrations or COD levels is higher than 10,000 mg/L [12]. Waste treatment using the anaerobic method has several advantages, which are more cost-effective and energy-efficient.
Bioreactor performance is better, almost 95% of organic contaminants are converted into biogas which can be used as energy and less sludge [13].
Chemical oxygen demand (COD) is a general parameter of the level of waste contamination, namely the amount of dissolved oxygen in the waste used to oxidize the organic components in the waste. Waste is a complex matrix containing a significant concentration of solids of 350-1200 mg/l, dissolved substances and particulates with a COD of 250-1000 mg/l, microorganisms up to 109 U/mL, nutrients, heavy metals and micro pollutants [15].
The use of biological methods to reduce COD levels in SPW is a popular alternative today because of lower operational costs compared to chemical and physical waste treatment [16]. The use of microbes in the treatment of shrimp pond waste not only reduces COD levels, but can also biosorb heavy metals contained in the waste even at low concentrations [17].
Heavy metals can accumulate in human organs causing dangerous diseases such as cancer [18][19][20].
One of the shrimp ponds that has a waste problem is the shrimp pond in Prigi Trenggalek, East Java - Indonesia. This study aims to determine the effectiveness of aerobic and anaerobic methods in processing SPW from Prigi Trenggalek using a batch experiment during start-up process by analyzing the water quality parameters before and after the treatment in term of COD, carbohydrates, and protein removals.
EXPERIMENT
Chemicals and instrumentation
The tools used in this study were a waterbath shaker (Faithful), UV-Vis spectrophotometer (Thermo Fischer Scientific), pH meter, micro pipette, vortex, analytical balance, centrifuge, hot plate, oven, magnetic stirrer, thermoreactor, 500 mL Erlenmeyer (Pyrex), 10, 50 and 500 mL volumetric flask (Pyrex), 10 mL measuring cylinder (Pyrex), 50,100, and 500 mL beaker (Pyrex), test tube (Iwaki), reagent bottle, spatula, watch glass, stirring rod, 1 mL tip, knife, scissors, dry ice, serum bottle, seal bottle, gauze, wrap.
The basic material used in this research is shrimp pond wastewater taken from Prigi Trenggalek. The chemicals used were bovine serum albumin powder (Sigma Aldrick), CuSO4.5H2O powder (Merck), sodium tartrate powder (Merck), Na2CO3 powder (Merck), NaOH powder (Merck), Follin's solution (Merck), anthrone powder (Merck), 95-97% H2SO4
solution (Merck), glucose pro analysis (Merck), K2Cr2O7 powder (Merck), HgSO4 powder (Merck), AgSO4 powder (Merck), potassium hydrogen phthalate powder (C8H5KO4) (Merck), nitrogen gas, distilled water.
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Procedure
Sampling of shrimp pond wastewater
Sampling of shrimp pond wastewater was carried out in Prigi Trenggalek. Shrimp pond wastewater samples were taken from the shrimp pond outlet. Sample bottles are prepared in advance before sampling. Furthermore, samples of shrimp pond wastewater were taken using a sample bottle. Then, the sample bottle containing the shrimp pond wastewater is put into the cool box storage and move it into the refrigerator to keep the SPW.
Measurement of pH parameters
Samples of shrimp pond wastewater were taken and put into sample bottles, the pH was measured with a pH meter that had been calibrated using pH 4, 7 and 10 buffers and the pH results listed on the pH meter were recorded.
Bacterial culture
Anaerobic and aerobic bacteria obtained from PT. Tata Bestari's intuition weighed 0.6 grams of powder each. Then put it in a 500 mL Erlenmeyer for the aerobic method and seal the bottle for the anaerobic method. Add 60 mL of distilled water each and homogenize until the bacterial powder is completely dissolved. Next, the solution was incubated on a water bath shaker for 24 hours at 37oC and 100 rpm.
Treatment of shrimp pond liquid waste samples with aerobic and anaerobic methods Samples of shrimp pond wastewater that will be treated are shaken first. A sample of 300 mL of shrimp pond wastewater was put into an Erlenmeyer and a sealed bottle containing the previously made bacterial culture. For the anaerobic method, the seal bottle is added with nitrogen gas before incubation. Then, the bottle was incubated in a waterbath shaker with a temperature of 37oC and a speed of 120 rpm. The treated sample was taken as much as 40 mL every 2 times a week, while at the same time 40 mL of shrimp pond wastewater sample was added back into the Erlenmeyer after taking the mixture. If the sample test results experience an increase in parameter levels exceeding the levels prior to treatment, variations in the concentration of liquid waste are made with tap water to be added.
Testing of protein content
Pipette as much as 0.5 mL of sample and put it in a test tube. Then, 2.5 mL of biuret solution was added and vortexed. Furthermore, the sample was incubated for 10 minutes at room temperature. After incubation, 0.25 mL of Follin's reagent was added and re-incubated again for 30 minutes at room temperature. The absorbance was measured at a wavelength of 750 nm using a UV-vis spectrophotometer.
Testing of carbohydrate content
A 0.5 mL of sample was put into a test tube. A 2 mL of anthrone solution was added, then shaken until homogeneous. Then put in the bath for 8 minutes with a temperature of 100oC.
The solution is cooled in a beaker filled with tap water. The absorbance was measured at a wavelength of 630 nm using a Uv-vis spectrophotometer [21].
Testing COD content
A sample of 1.25 mL was taken and put into a test tube. 0.75 mL of high concentration digestion solution and 1.75 mL of sulfate reagent solution were added. The test tube was closed
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and shaken slowly until homogeneous. The test tube was inserted into the thermoreactor at 150oC for 2 hours. The solution is cooled down to room temperature, occasionally the test tube lid was opened to prevent gas pressure. The absorbance of the sample in the test tube was measured at a wavelength of 600 nm using a UV-Vis spectrophotometer [12][23].
RESULT AND DISCUSSION
Quality of shrimp pond inlet and outlet samples
Sampling location in the shrimp ponds of the Prigi Trenggalek area. In the shrimp pond there is a water supply from seawater with a distance of about 100 meters from the beach.
Meanwhile, the added shrimp waste is simply thrown away, channeled towards the sea without any waste treatment process. The SPW samples were tested for several parameters to determine their quality. The results of the SPW sample quality test are presented in Table 1.
Table 1. SPW Quality Test Results
Sample pH COD mg/L Proteins (mg/L) Carbohydrate (mg/L)
Inlet 6.93 94.333 180.421 137.909
Outlet 6.47 1084.333 539.895 232.455
Shrimp pond waste has an alkaline pH with a pH range of 7-9 and contains organic matter consisting of protein, carbohydrates and other inorganic materials such as nitrogen phosphorus and ammonia [24]. The inlet pH value is 6.93, this value is slightly below the seawater quality standard for marine biota based on Minister of Environment Decree No. 51 of 2004 with a predetermined pH standard, namely 7-9 [25]. Meanwhile, the pH of SPW is 6.47, lower than the standard pH of water quality.
The inlet COD level was 94.333 mg/L while the COD level in the shrimp pond oulet was 1084.333 mg/L. COD level This SPW has a fairly high COD level and is not suitable for disposal directly into the environment. The levels of carbohydrates and protein in seawater as inlet were 137.909 mg/L and 180.421 mg/L, respectively. Meanwhile, SPW carbohydrate and protein levels were 232.455 mg/L and 539.895 mg/L. This shows that there is an increase in levels of COD, carbohydrates and protein. This increase came from shrimp waste, leftover food and some dead shrimp. In addition, the high levels of these three parameters are caused by the high organic matter that is decomposed by microorganisms by utilizing dissolved oxygen in the samples [24].
Aerobic and anaerobic system performance in treating shrimp pond wastewater
The main parameter that is usually used to measure the performance of a wastewater treatment system is COD removal. In addition, removal of organic compound such as carbohydrates and proteins are also a concern in determining the performance of an organic wastewater treatment. In this study, the performance of the aerobic and anaerobic treatment system was investigated in a batch system for more than 200 days.
Comparison of the COD removal of the aerobic and anaerobic systems in treating SPW durig start-up is shown in Figure 1. At the beginning of the treatment, in phase 1, SPW as a substrate without dilution, the performance of both aerobic and anaerobic systems in the first 10 days was quite good with COD removal of around 25-68%. However, the performance of the aerobic system started to fall on day 17 and dropped on day 40. In this initial phase, the
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performance of the anaerobic system was almost the same as the aerobic system, but it was dropped on day 28. The decrease in performance until the activity is very low is probably caused by an over loading rate. Thus, variations of the SPW dilution were carried out to determine the optimum dilution.
Figure 1. The effect of substrate concentration to the cod removal in aerobic and anaerobic systems
In the second phase, the SPW substrate was diluted 4 times so that the concentration was
¼ times. In this second phase, the COD removal of the aerobic and anaerobic systems was 52.77% and 45.49%. In terms of biological wastewater treatment, these COD removal is not very good. To improve the performance, in third phase, the substrate concentration was diluted into 8 times from the original SPW. However, the performance of both aerobic and anaerobic systems decreased and drop on day 97. The next step, in the fourth, the dilution was 12x dilutions. The performance of aerobic systems is still drop to 0% of COD removal, however the anaerobic system increased just after more substrate dilutions.
To find out the performance of the two biological wastewater treatment systems holistically, carbohydrate and protein removal were also analysed. The results of the analysis of carbohydrate removal and protein removal are shown in Figures 2 and 3. In the first phase, carbohydrate removal in the aerobic system is much better than the anaerobic system. The average carbohydrate removal in the aerobic system was 75.20%, while in the anaerobic system it was only 36.03%. This low performance is possible because it is still in the adaptation phase.
In the second to third phases, it is almost the same, which is between 70% and 98%. The performance of aerobic systems was stable at about 85% and it was decreased in the anaerobic system at about 60% of protein removal in the fourth phase as shown in Figure 2.
Likewise with protein removal in both systems, in the initial phase, protein removal in the aerobic system was quite high at the average of 88.59%. This value is much better than the anaerobic system which is only around 51.01%. In the second and third phases, the performance of the anaerobic system began to improve, but was still slightly lower than that of the aerobic system. The performance of aerobic systems was stable at about 95% and it was little bit better in the anaerobic system at about 98% of protein removal in the fourth phase as shown in Figure 3.
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Figure 2. The effect of substrate concentration to the carbohydrate removal in aerobic and anaerobic systems
Figure 3. The effect of substrate concentration to the protein removal in aerobic and anaerobic systems
At the beginning of the experiment, the performance of aerobic system is better than that of the anaerobic system. It might be due to the COD concentration of the SPW which is lower than 10,000 mg/L [12]. Aerobic digestion system is suitable for the wastewater with low COD concentration. Overall, based on the COD removal, the performance of start-up process of both systems were quite similar. Therefore, it needs more study to investigate the aerobic and anaerobic systems for a longer duration in a continuous or semi-continuous system.
CONCLUSION
Generally, the performance of the aerobic system in treating the shrimp pond wastewater in phases one to three is slightly better than the anaerobic system. However, in the fourth phase,
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the anaerobic system was better than the aerobic system. As shown in phase one, there are differences in adaptability, in which the aerobic system adapts faster than the anaerobic system.
ACKNOWLEDGMENT
This research was funded by Universitas Negeri Malang through the research grant in 2022 number 18.5.60/UN.32/KP/2022.
REFERENCES
[1] V. Jegatheesan, L. Shu, and C. Visvanathan, Encycl. Environ. Heal., 2011, 123–135.
[2] Dirjend Penguatan Daya Saing Produk Kelautan dan Perikanan, “KKP | Kementerian Kelautan dan Perikanan.” https://kkp.go.id/djpdspkp/artikel/31077-naik-4-15-ekspor- kelautan-dan-perikananselama-caturwulan-i-2021-tunjukkan-tren-positif (accessed Oct.
17, 2022).
[3] A. B. Dauda, A. Ajadi, A. S. Tola-Fabunmi, and A. O. Akinwole, Aquac. Fish., 2019, 4 (3), 81–88.
[4] B. T. Iber and N. A. Kasan, Heliyon, 2021, 7 (11), e08283.
[5] C. Warwick, A. Guerreiro, and A. Soares, Biosens. Bioelectron., 2013, 41(1), 1–11.
[6] Du, H. and Shao, Z. App. Energy., 2022, 314, 118994.
[7] E. M. John, K. Krishnapriya, and T. V. Sankar, Aquac., 2020, 526, 735390.
[8] E. Yusni and T. P. Setiani, Aquasains: Jurnal Ilmu Perikanan dan Sumber Daya Perairan, 2019, 7(2), 707-714.
[9] Czekala, W., Mateusz, N., and Grzegorz P., Biores. Technol., 2023, 128813.
[10] Rocchetti, G., Alcantara, C., Bauerl, C., Garcia-Perez, J. V., Lorenzo, J. M., Lucini, L., Collado, M. C., and Barba, F. J., Food Res. Int., 2020, 136, 109534.
[11] Febri Indiani, Dyah Titisari, and Lamidi, Journal of Electronics, Electromedical Engineering, and Medical Informatics (JEEEMI), 2019, 1(1), 11–15.
[12] Feng, L., Zhao, W., Liu, Y., Chen, Y., He, S., Ding, J., Zhao, Q., and Wei, L., Chinese Chem. Lett., 2023, 34(2), 107439.
[13] Kiani, M. K. D., Parsaee, M., Ardebili, S.M.S., Reyes, I.P., Fuess, L.T. and Karimi, K., Biomass Bioenergy., 2022, 161, 106446.
[14] V. V. Ranade and V. M. Bhandari, Industrial Wastewater Treatment, Recycling and Reuse, Butterworth-Heinemann, 2014.
[15] Shankar, R., Kumar, S., Prasad, A. K., Khare, P., Varma, A. K. and Yadav, V. K., Biological wastewater treatment plants (WWTPs) for industrial wastewater: in Microbial Ecology of Wastewater Treatment Plants, Elseviers, 2021, 193-216
[16] Thapa, B.S., Pandit, S., Patwardhan, S.B., Tripathi, S., Mathuriya, A.S., Gupta, P.K., Lal, R.B. and Tusher, T.R., Sustainability., 2022, 14(14), 8379.
[17] F. Migahed, A. Abdelrazak, and G. Fawzy, Int. J. Environ. Sci. Technol., 2017, 14(6), 1169–1180.
[18] Fu, Z. and Xi, S., Toxicol. Mech. Methods, 2020, 30(3), 167-176.
[19] J. Briffa, E. Sinagra, and R. Blundell, Heliyon, 2020, 6(9), e04691.
[20] K. H. Vardhan, P. S. Kumar, and R. C. Panda, J. Mol. Liq., 2019, 290, 111197.
[21] SNI-6989.2:2019, Air dan Air Limbah-Bagian 2: Cara Uji Kebutuhan Oksigen Kimiawi (Chemical Oxygen Demand/COD) dengan refluks tertutup secara spektrofotometri,
Badan Standarisasi Nasional, 2019, accessed from
http://sispk.bsn.go.id/sni/DetailSNI/12250.
[22] Hasanah, U., Mulyati, A.H., Widiastuti, D., Warnasih, S., Syahputri, Y. and Panji, T.,
136
Journal of Science Innovare, 2020, 3(2), 35–38.
[23] Brown, H.L., Microbial Growth, in Encyclopedia of Infection and Immunity, vol 1, Elsevier, 2022, 324–335.
[24] A. Goli, A. Shamiri, S. Khosroyar, A. Talaiekhozani, R. Sanaye, and K. Azizi, J.
Environ. Treat. Tech., 2019, 7(1), 113–141.
[25] Menteri Negara dan Lingkungan Hidup, Keputusan Menteri Negara Lingkungan Hidup No 51 Tahun 2004 tentang Baku Mutu Air Laut, Lembaran Negara Republik Indonesia.
Accessed date 20 October 2021 from the ministry website https://ppkl.menlhk.go.id/website/filebox/824/191009100640Keputusan%20MENLH
%20Nomor%2051%20tahun%202004%20%20tentang%20Baku%20Mutu%20Air%20 Laut.pdf
