ISSN: 2676-2854 (Print) 2676-2862 (Online) DOI: 10.5455/jfcom.20190722060436 Journal of Fisheries and Coastal Management 2 (2020) 56-64 Effect of pond fertilization on productivity of tilapia pond culture in Ghana Collins Prah Duodu 1, 2 *, , Daniel Adjei Boateng 2 , and Regina Esi Edziyie 2 Department of Marine and Fisheries Science, School of Biological Sciences, University of Ghana Department of Fisheries and Watershed Management, Kwame Nkrumah University of Science and Technology, Kumasi, Ghana. 1 2 ORIGINAL ARTICLE *Corresponding author E-mail: cduoduprah@gmail.com ABSTRACT Fish farmers have the desire to increase productivity but are constrained by high cost of formulated feeds. This study quantified the performance and production cost of tilapia cultured in unfertilized and fertilized ponds in six earthen ponds with sizes between 200 m2 and 300 m2. All ponds were stocked with all-male Nile tilapia (Oreochromis niloticus) fingerlings (average size = 25.4±0.56 g) at 2 individuals per m2 and fed at 3% of their body weight for the unfertilized and fertilized ponds, respectively. The results showed a comparable mean weight gain of 204.2±36.3 g and 202.9±23.8 g for the unfertilized and fertilized ponds, respectively. Similar results were obtained for the specific growth rate (1.0±0.4; 1.1±0.4 %day-1), feed conversion ratio (1.1±0.2; 1.2±0.3) and feed intake with values of 0.7±0.5; 0.7±0.3 for unfertilized and fertilized ponds respectively. Total cost of production for the unfertilized treatment was USD$ 832.00 (273.97±86.48) and the unfertilized treatment amounted to USD$ 834.72 (274.88±60.34). A profit of USD$ 319.42 (106.48±90.99) and USD$ 353.05 (120.59±98.09) were realized for the unfertilized and fertilized treatments, respectively. The results of this study indicate that farmers could increase their pond’s productivity and profit through pond fertilization and feeding with formulated feed. Keywords: Inorganic fertilizer, Tilapia, Pond culture, Cost-effectiveness. 1.0 Introduction In the past decade, fish nutrition has advanced dramatically with the development of commercial diets that promote optimal fish growth and health. Moreover, fish feed represents 50-70% of the production costs of commercial fish farming (El-Sayed, 2004). In Ghana, the use of complete commercial feed especially by pond farmers is limited due to the relatively higher cost of the feed in comparison to supplementary feeds. Consequently, pond farmers rely on low quality agro-by-products like rice bran, wheat bran, and groundnut peels etc. which are nutritionally incomplete. These by-products are lacking in essential amino acids such as methionine, lysine and contains anti-nutritional factors such as © 2020. J. Fish Coast. Mgt., Department of Fisheries and Aquatic Sciences, University of Cape Coast. All rights reserved. tannins/saponins (Tacon 1993; Annongu et al., 1996; Francis et al., 2001; Ogunji, 2004) which interfere with food utilisation and affect health and production of animals (Makkar, 1993). Farmers have the desire to increase productivity but are constrained by the high cost of formulated feeds. In pond culture, formulated feed utilization can be optimized while maximizing gains from natural food sources. Shroeder (1980) reported that natural food could account for as much as 50-70% of total available food for tilapia in pond culture even when a complete diet is provided. Studies by Green (1992) and Diana et al. (1994) showed that the growth performance of tilapia in ponds could be significantly improved by use of organic and inorganic fertilizers with formulated feeds at reduced ration. Moreover, inorganic fertilizers are reported to be hygienic and tend to produce better water Duodu et al. /Journal of Fisheries and Coastal Management 2 (2020) 56-64 57 and fish quality (Diana, 2012). Shang (1990) indicated that even though economic research is primarily important, it is often neglected by aquaculturists. This could be associated with most tilapia pond farmers in Ghana who keep little or no economic records on their farm’s operations and complain of poor returns and do not see aquaculture as a lucrative business. Yi and Diana (2008) suggested that economic analysis to determine efficiency of resource allocation and management practices is essential in aquaculture. This study was conducted to quantify the performance of fish cultured in fertilized and unfertilized ponds and fed the recommended ration of formulated feed (El-Dahhar, 2000) and to assess the costeffectiveness of using inorganic fertilizers to improve productivity of tilapia raised in ponds by use of a simple farm enterprise budget. limed at a rate of 1 kg agriculture lime per 10 m2. The pond dykes and surroundings were cleared to deny predators of hiding places. The ponds were then filled with water and allowed to stand for a day to enable suspended particles to settle before stocking. The study design followed the completely randomized design (CRD) where two treatments thus, fertilized and unfertilized treatments were assigned randomly in triplicate groups to the ponds. 2.0 Materials and Methods Ponds were fertilized with Mono Ammonium Phosphate (MAP) at 2 g m-2 and Urea at 3 g m-2 weekly; strictly according to the phytoplankton abundance, using Secchi-disk depth (25-30 cm) as a proxy for phytoplankton abundance due to its strong relationship with chlorophyll-a (Kordi et al., 2012). Fish were fed a commercial floating feed (Raanan) at rates of 3-1.5% of their body weight for the two treatments. During the 1st eight weeks of the trial, fish were given a feed that contained 2.1 Study area and pond preparation The study was conducted for 233 days at the fish farm of the Faculty of Renewable Natural Resources, Kwame Nkrumah University of Science and Technology (KNUST), in Kumasi (longitude 6.35°-6.40° and latitude 1.3°-1.35°) in the Ashanti Region, Ghana. Six ponds ranging between 200–300 m2 (Figure 1) were used. Ponds were drained, dried, de-silted and 2.2 Fingerlings and stocking All-male tilapia (Oreochromis niloticus) fingerlings were initially obtained at a size of 2 g from Crystal Lake Limited, Dodi-Asantekrom, Eastern Region, Ghana. Fish were nursed to an approximate size of 25 g before they were stocked at 2 fish m-2 (Anani et al., 2017). 2.3 Fertilizer application and feeding Figure 1: Aerial view of the study site with experimental ponds (enclosed in red border lines). 58 Effect of pond fertilization on productivity of tilapia pond culture in Ghana 33% dietary protein (2.5 mm pellet diameter), thereafter they were fed with a 30% dietary protein feed (4.5 mm pellet diameters) as per the standard practice by local fish farmers to meet the nutritional needs of tilapia at the different life stages. Fish were fed to apparent satiation and the feed rate for the next feeding determined from previous feed fed. Feeding was done twice daily between 9-10 am and between 3-4 pm. Feeding levels were adjusted after each monthly sampling. However, fish were not fed on the day they were sampled to allow recovery from stress due to seining and handling. 2.4 Fish sampling and water quality Fifty fish in each pond were sampled by seining through the ponds from one vertical end to the other. The bulk weight of the catch was determined with a weighing balance (MITSUBA model: MB-320) and the average weight calculated to monitor growth and adjust the feed levels. The growth performance and feed utilization of fish from each pond was determined by conventional methods (Agbo et al., 2011) as follows: The net yield of fish was reported as the (TDS), conductivity, temperature, and pH were collected in situ using the Hanna (HI 9828) multi parameter probe. Secchi readings were taken in each pond using a Secchi-disk every two weeks after fertilization. 2.5 Fish sales and cost-benefit analyses Fish were harvested and sold at the premises of the Faculty of Renewable Natural Resources, KNUST. Fish were put into two categories as large (260-330g) and small (200-250g) and sold per piece in accordance with the local market price. All monetary values are quoted in the United States Dollars (USD $) as at the time of the research. According to the Bank of Ghana, the average Interbank FX Rate between January 2013 and September 2013 was USD $ 1.00 = GHC 1.9421. The effectiveness and efficiency of the treatments were ascertained by developing an enterprise budget that compared the cost of production basically in terms of cost of fingerlings, fertilizer used, quantity of feed given and labour, and the revenue that accrued from fish sales. Prices of items were based on the prevailing local market price. Profit index was calculated as Where, net profit was considered as the profit accrued after all production (variable) cost has been accounted for whereas the total investment determined as only the variable cost incurred. 2.6 Statistical analyses actual biomass of fish that was harvested from the ponds at the end of the trial period. Water samples were collected fortnightly to determine the concentration of Chlorophyll-a according to the standard method described in HMSO (1983). Data for dissolved oxygen (DO) concentration, total dissolved solids Potential differences in the final weights, specific growth rates, weight gain, feed conversion ratios, feed intake, survival rates, gross yield, and net yield of the fish cultured under the two feeding treatments were evaluated using student’s t-test at p < 0.05. The same statistical procedure was used to assess potential differences in each of the Duodu et al. /Journal of Fisheries and Coastal Management 2 (2020) 56-64 59 physicochemical variables observed in the ponds used for the two treatments. All analyses were done with GraphPad Prism version 5.01 Software for Windows. 3.0 Results 3.1 Growth performance and feed utilization of Nile tilapia Growth performance was assessed by the following indicators: final weight (FW), weight gain (WG), specific growth weight (SGR), survival rate (SR), gross yield (GY) and net yield (NY) as indicated in Table 1. Generally, there was a steady increase in fish weight from January to March, but growth slowed slightly between March and April (Figure 2). Thereafter, a gradual increase in growth was recorded from May till the end of the growout period in both treatments. The SR was similar in both treatments. Nonetheless, fish weight gain was 5.7% more in the fertilized ponds compared to the unfertilized ponds. Yet, SGR in both treatments were similar. There were no significant differences between the two treatments for all the growth parameters assessed. The FCR was similar (Table 2), whereas the daily feed intake per fish revealed very low intake for both treatments although the average temperature in the treatment ponds (26.5–28.4 °C) were close to the recommended temperature (27.0 °C) (Pandit and Nakamura, 2010) for optimum growth (Table 3). Generally, all the parameters assessed for feed utilization Table 1 - Growth performance (average ± standard deviation) of Nile tilapia fed in unfertilized and fertilized earthen ponds for 233 days. Parameter Unfertilized Fertilized Initial weight (g) 25.0 ± 8.0 25.8 ± 8.5 Final weight (g) 229.1 ± 33.5 228.7 ± 23.3 Weight gain (g) 204.2 ± 27.9 202.9 ± 24.0 Specific growth rate (%/day) 1.02 ± 0.37 1.10 ± 0.35 Survival rate (%) 62.6 ± 20.3 Gross yield (kg/ha) 4935.17 ± 589.19 Net Yield (kg/ha) 4021.55 ± 677.61 N = 3 ponds for each treatment. Figure 2: Growth performance of Nile tilapia fed in unfertilized and fertilized earthen ponds. did not show any significant difference (p < 0.05) between the two treatments. 3.2 Physicochemical parameters and chlorophyll-a concentration Temperature recorded over the study period for both the unfertilized and fertilized treatments did not show any significant differences (p = 0.775 at df = 10, t-value = 0.2931). Ponds that received no fertilization recorded mean temperature of 27.8 ± 0.3°C and that for the fertilized ponds was 28.0 ± 0.3 °C over the study period (Table 4). Similarly, there was no significant difference (p = 0.698 at df = 10, t-value = 0.3984) in pH between the two treatments with both recording very narrow variations. On the other hand, DO concentrations recorded over the study period varied widely ranging from 1.8 to 7.7 mgl-1 for the unfertilized treatment and from 2.1 Table 2 - Feed utilization (average ± standard deviation) of Nile tilapia fed in unfertilized and fertilized earthen ponds for 233 days. Feeding Treatment Parameter Unfertilized Fertilized Feed conversion ratio 1.1 ± 0.2 1.2 ± 0.3 63.9 ± 13.0 Feed efficiency ratio 1.4 ± 0.9 0.9 ± 0.2 5077.08 ± 401.76 Feed fed (kg) 116.4 ± 53.0 112.0 ± 27.6 4303.27 ± 408.43 Feed intake (g/fish) 0.7 ± 0.5 0.7 ± 0.3 N = 3 ponds for each treatment. Effect of pond fertilization on productivity of tilapia pond culture in Ghana 60 Table 3 – Observed daily feed rate and temperature for tilapia cultured in unfertilized and fertilized ponds. Pond treatment Day Weight of fish (g) Daily feed/ fish (g) Observed average temperature (°C) Unfertilized 0* 16 - - 31 46 0.78 ± 0.00 27.12 ± 0.00 62 67 0.73 ± 0.11 28.38 ± 0.20 94 69 0.89 ± 0.11 28.18 ± 0.31 122 98 0.71 ± 0.37 28.36 ± 0.44 154 141 1.24 ± 1.08 28.15 ± 0.32 184 159 1.37 ± 0.49 26.94 ± 1.23 216 202 1.67 ± 0.58 27.10 ± 0.41 233 229 1.63 ± 0.48 26.45 ± 0.56 0* 16 - - 31 36 0.19 ± 0.00 27.82 ± 0.00 62 63 0.78 ± 0.18 28.60 ± 0.30 94 70 0.72 ± 0.14 28.05 ± 0.32 122 90 0.65 ± 0.20 28.34 ± 0.67 154 121 1.25 ± 0.46 28.23 ± 0.29 184 166 1.65 ± 0.43 26.83 ± 1.12 216 200 1.80 ± 0.18 27.20 ± 0.87 233 229 1.63 ± 0.48 26.20 ± 0.25 Fertilized * day of stocking to 6.2 mgl-1 for the fertilized treatment (Table 4), however, with no significant differences between treatments means (p = 0.674 at df = 10, t-value = 0.4328). Conductivity of the fertilized ponds was rather 16% higher compared to the unfertilized ponds. Secchi depth showed wide variations in both treatments and was approximately 25% clearer in the fertilized ponds than the unfertilized ponds (Table 4). Chlorophyll-a concentration was highest (p = < 0.0001 at df = 10, t-value = 6.348) and almost double in the unfertilized ponds than the fertilized treatment. The fertilized ponds however, recorded lower concentrations with a mean of 1374 ± 239 µgl-1 (Table 4). 3.3 Cost of production The costs of all inputs as well as the prices of fish were based on local market prices in Kumasi. The economics of fish production in this study indicated that the total cost of production was slightly higher (USD 834.73) for the fertilized treatment compared to the unfertilized treatment (USD 832.00). The total cost of pond preparation was marginally higher in the fertilized ponds than the unfertilized treatment. At harvest, small size (200-250 g) fish were sold at USD 1.03 and the large (> 250 g) at USD 1.29 per piece as indicated in Table 5. This resulted in a higher profit for the fertilized ponds. Table 4 – Physicochemical parameters and Chlorophyll-a concentration in unfertilized and fertilized tilapia earthen ponds during the study period. Parameter Temperature (°C) Mean ± Standard Deviation Range Unfertilized Fertilized Unfertilized Fertilized 27.9 ± 0.3 28 ± 0.3 27.0 - 28.4 26.8 - 28.6 pH 8.1 ± 0.3 7.9 ± 0.3 7.0 - 9.0 6.9 - 8.7 Dissolved Oxygen (mg/l) 4.8 ± 0.8 4.4 ± 0.6 1.8 - 7.7 2.1 - 6.2 Conductivity (µs/cm) 150 ± 21 174 ± 17 102 - 250 123 - 239 Total Dissolved Solids (mg/l) 74 ± 9 86 ± 9 51- 118 62 - 119 Secchi Depth (cm) 12.6 ± 1.6 15.7 ± 1.5 11.2 - 19.2 10.8 - 20.5 2286 ± 258 1374 ± 239 1843 - 2574 1008 - 1607 Chlorophyll-a (µg/L) N= 8; monthly averages from the three replicates for each treatment. Duodu et al. /Journal of Fisheries and Coastal Management 2 (2020) 56-64 61 Table 5 - Enterprise budget for unfertilized and fertilized feed treatments for one production cycle in Ghana. Cost and price information is in United States Dollars (USD) based on an exchange rate of USD$ 1 = GHC 1.9421 provided by the Bank of Ghana in September 2013 Unfertilised ponds Fertilised ponds ITEM (Unit) Quantity Unit price (USD) Fingerlings (/piece) 1550 0.132 204.60 1550 0.132 204.60 Pond Rent (/month) 18 3.22 57.96 18 3.22 57.96 Subtotal A Total Expenses (USD) Quantity Unit Price (USD) Total Expenses (USD) 262.56 262.56 Feed Juvenile fish (2.5mm) (kg) 31.99 1.16 37.11 23.16 1.16 26.87 Growout fish (4.5mm) (kg) 299.12 1.03 308.09 300.40 1.03 309.41 Growout fish (6.0mm) (kg) 18.15 1.03 18.69 18.21 1.03 18.76 Subtotal B 363.89 355.04 Pond Preparation and treatment Lime (kg) 35 1.03 36.05 35 1.03 36.05 MAP (kg) - - - 3 3.09 9.27 UREA (kg) - - - 4.5 0.51 2.30 Fuel for filling pond with water 6 5.15 30.90 6 5.15 30.90 Labour (per day) 180 0.77 138.60 180 0.77 138.60 Subtotal C 205.55 217.12 Total production cost (A+B+C) 832.00 834.72 Revenue from fish sales Large fish (per piece) (D) 485 1.29 625.65 458 1.29 590.82 Small fish (per piece) (E) 500 1.03 515.00 554 1.03 570.62 Reproduction (kg)* 5.23 2.06 10.77 12.78 2.06 26.33 Total revenue 1,151.42 1,187.77 Profit/Loss 319.42 353.05 Profit index 3.16 3.35 Economy of weight gain (USD.kg-1) 1.19 1.16 Return on investment (ROI) (%) 38.39 42.30 *Quantity presented in kilograms (kg) 62 Effect of pond fertilization on productivity of tilapia pond culture in Ghana 4. Discussion 4.1 Growth performance of Nile tilapia Survival rate in this experiment was generally low but was higher in the fertilized treatment compared to the unfertilized treatment. This is contrary to observations from Diana et al. (1994), who recorded a lower survival in the feed and fertilizer treatment in their experiment. The lower survival rate in this experiment could not be linked to the application of the treatments, but instead to the invasion of predators such as snakes, birds and frogs (Figure 3) which might have preyed on the fish especially at the early stages of the experiment. According to Diana et al. (1994) the presence of predators could affect the yield of fish resulting in variable survival among treatment replicates. Figure 3: Predators observed; (a) captured frog with fish in mouth during sampling (b) a bird trapped by predator net overlaying one of the experimental ponds. It was expected that fish that received the unfertilized treatment would perform better than those receiving the fertilized treatment; however, that was not the case. The growth in the fertilized treatment could be attributed to the fishes’ ability to utilize the abundant natural food available in the ponds. The unfertilized ponds rather responded poorly to the feed which resulted in many uneaten feeds observed at the surface of the pond water 30 minutes after feeding. A possible reason for the poor response could be attributed to the fish feeding on natural feed sources available in the pond as the unfertilized ponds had abundant phytoplankton levels as a result of continuous fertilization from uneaten feed, although, that was an unintended consequence (Table 4). Mud accumulation at the bottom of ponds as a result of seining during sampling led to a decrease in pond depth, dense growth of duckweed coupled with lower water levels during the dry season might have accounted for the poor response of fish to feed. The decline in tilapia growth in both treatments in April could be linked to high temperatures which made the pond water warmer thus, the fishes’ reluctance to swim up to the surface to feed. Low DO concentration at the peak of the dry season might have also caused low fish metabolism thereby depressing growth (Brett, 1979). This resulted in a lower average growth rate (1.1 ± 0.3g/day) of tilapia observed in this study compared to that reported (2.0-3.1g/ day) by other studies (Green, 1992; Diana et al., 1994). 4.2 Feed utilization and water quality The similarity in FCR of the treatments confirms the report by Hepher and Pruginin (1982) and Diana et al. (1994), who recorded similar FCR for ponds receiving feed only or feed and fertilizer input in their study. The fertilizer treatment in this study had an FCR near one suggesting that the fish benefited from natural food available in the ponds which was boosted by fertilization. Shroeder (1980) reported that natural food could account for as much as 50-70% of total available food for tilapia in pond culture even when complete diet is provided. Other authors like Green (1992), emphasized that natural productivity influenced by pond fertilization was enough to promote accelerated fish growth in tilapia pond culture. Certainly, one might expect the total quantity of feed applied to be similar in both treatments, however, feed utilized by the fertilized ponds treatment was slightly more than the unfertilized. This could be attributed to the fact that the quantity of feed given to fish in the treatments was adjusted based on the average body weight after the monthly sampling. Moreover, fish were fed to satiation and so the poor response by the fish that received the unfertilized pond treatment ended up with less feed consumed. This poor Duodu et al. /Journal of Fisheries and Coastal Management 2 (2020) 56-64 response may not be due to low DO levels since the water quality was similar for the two treatments (Table 4). A possible reason could be the availability of natural food to the fish indicated by the high chlorophyll-a levels in the no fertilizer treatment hence their poor response to the feed. One of the reasons that could account for lower feed intake in fish is poor water quality. However, since water quality in this study was generally within a favourable range for tilapia, the lower feed intake per fish per day (Table 3) could also be linked to the presence of other animals (predators) (Meena, 2014) that scare the fish away from the food by attacking them during feeding. All the water quality parameters monitored (Table 4) were in the favourable range for tilapia culture (Boyd, 1990) except for DO which varied widely. Temperature of the water in both treatments was within the optimal range of 26-30°C (Lazur, 2007). This was expected since all ponds were exposed to similar environmental conditions such as sunlight and wind (Diana et al., 1994). However, the wider variations in DO concentrations observed in both treatments could be a result of the high chlorophyll-a concentration which usually results in wider fluctuations in DO concentration within a day. Similar variation was also reported by Thakur et al. (2007). Diana et al. (1994) suggested that wide variation in DO levels could also be due to the high oxygen demand and nutrient loading on pond bottom which push DO levels extremely low. The pH range (6.89 - 8.96) recorded for all ponds in this study were similar to the range of 6.5 to 8.5 recorded by Diana et al. (1994). The lower visibility (Secchi readings) in the unfertilized ponds treatment was supported by the high chlorophyll-a levels, suggesting a high phytoplankton abundance. The higher visibility in the fertilized ponds treatment suggests a high utilization of phytoplankton by the fish. Although, natural food utilization in the unfertilized ponds were possible, continuous fertilization from uneaten feed contributed to abundant algae impeding visibility. Diana et al. (1994) reported no significant difference in Secchi depth among treatments (fertilized 63 and unfertilized ponds) and attributed this to the similar amount of light (and heat). Chlorophyll-a concentration increased in the first month of the study and showed a relatively stable concentration in the last half of the study as reported by Thakur et al. (2007). Contrary to what was reported by Thakur et al. (2007) who fertilized ponds throughout the culture period and started feeding Nile tilapia from day 80 (half way through the culture period), fertilization plus feeding in this study recorded lower chlorophyll-a concentration. Moreover, chlorophyll-a concentration differed significantly between the treatments. The lower level of chlorophyll-a was an indication of the low abundance of phytoplankton which was probably due to the fish’s reliance and effective grazing on the available phytoplankton. 4.3 Production Costs and Revenue The economic analysis (Table 5) suggests that both treatments could be profitable. This agrees with Diana et al. (1996) who observed that the 0.5 ad libitum and fertilization treatment in their experiment was the most profitable. Thakur et al. (2007) indicated that better economic returns in fertilized tilapia ponds could be attributed to improved growth performance of the fish due to the presence of natural food in ponds. The cost of production (Table 5) was slightly higher in the fertilized pond treatment and could be attributed to the cost of fertilizer input. The poor feed response by the fish fed feed without fertilization in this study contributed to a slightly lower quantity of feed fed. Another possible reason was that the quantity of feed applied to both treatments was adjusted monthly based on fish weights determined after sampling at the end of the previous month. Hence, the gross revenue and profit generated after the sale of fish did not reveal any difference between the two treatments. Even though both treatments made a profit, the fertilizer pond treatment made approximately 6% more than the unfertilized treatment. A study by Thakur et al. (2007) which assessed the culture of tilapia under fertilization plus feed compared to tilapia under feed only reported an increase in 64 Effect of pond fertilization on productivity of tilapia pond culture in Ghana net income for the former. Finally, only feed input accounted for 43.7% and 42.5% (without cost of fertilizer) to the total cost of production for the unfertilized and fertilized treatments, respectively. This suggests that when an efficient fertilization and feeding regime is applied and natural food is readily available or well utilized by fish the amount of feed required for good growth could reduce to 45% of the production cost in tilapia pond farming. Overall, this study supports the assertion by Diana et al. (1994) and Diana et al. (1996) that fertilizing tilapia ponds is profitable and produced higher yields (Thakur et al., 2007). Generally, it can be concluded that fertilizing tilapia ponds and providing feed administered strictly based on fish response to feed could have beneficial effects. The addition of inorganic fertilizer did not adversely affect the water quality. Therefore, tilapia pond farmers could increase their yield by fertilizing their ponds to enhance natural food production. 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Proceedings of the 8th International Symposium on Tilapia in Aquaculture, Cairo, Egypt, 12-14 pp. Received 12 February 2019 Accepted 06 October 2020 First Online 11 October 2020 Published Online 18 May 2021