Replacing the unsustainable and wild-caught fishmeal with field cricket (Gryllus bimaculatus) meal in Catla (Catla catla) fry diet: Effect for growth, in vivo digestibility, carcass composition histopathological alterations and disease tolerance

Replacing the unsustainable and wild-caught fishmeal with field cricket (Gryllus bimaculatus) meal in Catla (Catla catla) fry diet: Effect for growth, in vivo digestibility, carcass composition histopathological alterations and disease tolerance | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Replacing the unsustainable and wild-caught fishmeal with field cricket (Gryllus bimaculatus) meal in Catla (Catla catla) fry diet: Effect for growth, in vivo digestibility, carcass composition histopathological alterations and disease tolerance G. S. Champika Perera, M. R. Afridin, A. M.A.N. Adikari, P. P.M. Heenatigala, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3193929/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 06 Oct, 2023 Read the published version in Aquaculture International → Version 1 posted 7 You are reading this latest preprint version Abstract Insect meal has been identified as one of the cutting-edge alternatives to substitute fishmeal (FM) in aquafeeds. However, the potential of FM replacing with field cricket meal (FCM) related to Calta catla fry stage has yet to be researched. Therefore, an eight-week experiment was designed to evaluate the performance of C. catla fry (0.22 ± 0.04 g/fish). The protein content of the FM of the control feed (0FCM) was replaced with 35% (35FCM), 70% (70FCM), and 100% (100FCM) of FCM. All the fish were hand-fed by respective research diets 5% of the body weight twice daily. Results showed that FCM could replace wild-caught fishmeal (67.1 % crude protein) without adversely affecting growth performance, carcass composition, liver functions, and disease tolerance. Though in vivo protein digestibility significantly decreased ( P <0.05) when the FCM percentage increased in the feed, it did not affect the performance of the fish. Therefore, FCM could be used to prepare fish-free feed in the C. catla fry stage. However, optimum technical know-how should be identified to certify the accessible and economically viable mass-scale production of FCM. insect meal sustainable aqua feed fishmeal substitutes Figures Figure 1 Figure 2 Figure 3 Introduction Fishmeal is supposed to be the key protein component (Barlow 2003 ) in conventional aquafeed formulations due to its superior nutrient qualities (Schipp 2008 ; Mallison 2017 ; Chapman and Miles 2018 ). However, soaring prices (dos Santos 2016 ), food security issues (Chan et al. 2019 ), unsustainability (Ghamkhar and Hicks 2020 ), and the impacts of climate change (van Huis and Oonincx 2017 ) have forced the industry to find alternative ingredients. Moreover, a considerable percentage of world fish production has still been used to produce fishmeal instead of human consumption (FAO 2020 ; Auchterlonie 2018 ; SEAFISH 2016 ). Simultaneously, per capita fish consumption is increasing (Ye 1999 ). Therefore, alternative ingredients are explored to replace the fishmeal in aquafeeds, and numerous innovative feedstuffs are being researched to find novel ingredients (Gasco et al. 2018 ). It has been predicted that world aquaculture production will be increased by using alternative protein sources without increasing the inclusion level of fishmeal (Olsen and Hasan 2012 ). Furthermore, the inclusion level of fishmeal in some fish feeds has been sharply reduced (Kok et al. 2020 ; Tacon et al. 2011 ). It has been predicted that world aquaculture production will be increased due to the usage of alternative protein sources without increasing the inclusion level of fishmeal (Olsen and Hasan 2012 ). According to recent research results, insect meal has been identified as a highly potential source to replace fishmeal in aquafeeds (Nogales-Mérida et al. 2019). Insect meal is comparable to fishmeal in terms of adequate protein, amino acid, fatty acid, vitamin and mineral contents with high digestibility (Nogales-Merida et al. 2019 ; Wang et al. 2005 ; Akinawa and Ketiku 2000 ) and high feed conversion efficiency (Katayama et al. 2008 ). Besides, it is economically practical due to its short life cycle, and lower production cost (Taufek et al. 2017 ). More importantly, insect production is environmentally sustainable due to its low carbon footprint (Henry et al. 2015 ). Furthermore, the insect culture does not compete for space or resource complement (FAO 2013 ). G. bimaculatus is a frequently cultured (van Huis 2020 ) edible cricket species (Hanan et al. 2022 ; Hwang et al. 2019 ) with a high nutrient value (Ngonga et al. 2020 ). Notably, the applicable species has been frequently researched to find the replacement potential of the fishmeal in aquafeeds with encouraging results (Fan et al. 2023 ; Perera and Bhujel 2022 ; Peh et al. 2021 ; Taufek et al. 2017 ). However, the scope of the above studies has been limited to a few fish species. Therefore, further research is needed to fill the gaps in the uncovered study areas of the various fish species. C. catla is an Indian Carp species with a fast growth rate and high market demand, especially in the South Asian region (Srivastava et al. 2013 ) due to its taste and nutrition (Shahzad et al.2020; Das and Das 2015 ). The statistics of 2018 reveal that the global production of C. catla was 3041.3 thousand tonnes, and it ranked sixth in the major species production in world aquaculture, possessing 5.6 percent of the global fish production (FAO 2020 ). Limited research results are available in reference to fishmeal replacement by insect meal related to the researched fish. Therefore, this experiment was conducted to find out the fishmeal replacement potential of the fry stage of C. catla by FCM. Materials and Methods The experiment was conducted in the Aquaculture Research Center of the head office of National Aquatic Resources Research and Development Agency (NARA), Crow Island, Mattakkuliya, Colombo 15, Sri Lanka, from September to November 2022. Experimental setup Twelve fibreglass aquaria of each 160 L were occupied to stock C. catla fry. Each aquarium had a cylindroconical column initiated with a 146 cm diameter at the bottom to facilitate the faecal matter collection. Those tanks were filled with conditioned freshwater to 30.5 cm in depth. Each aquarium was aerated by using a SHOWFOU BS-232 air compressor (SHOWFOU ELECTRIC MACHINE CO. LTD, Kaohsiung City, Taiwan). Air temperature and light intensity were maintained at 27.6 ± 0.4 0 C and 998 ± 56 Lux, respectively. The photoperiod was maintained as 12 hours (light)/12 hours (dark) using fluorescent tube bulbs as a light source during the experimental period. Illumination was regulated by a SMART SENSOR AS803 digital lux meter (ARCO Electronics Ltd, Dong Guan City, China). Purchasing and conditioning of C. catla fry A total of thousand C. catla fry (initial weight, 0.22 ± 0.04 g/fish) were procured from Carp Breeding Center, National Aquaculture Development Authority, Udawalawa, Sri Lanka. Randomly selected 360 fish among those fish were stocked in the aquarium mentioned above at a density of 164 /m 3 (30 fish per aquarium) and were acclimatised for a week. The control feed containing 41.7 ± 0.2% crude protein was fed twice daily at 5% of the body weight during the above week. Water height was maintained at 30.5 cm, and water quality parameters were maintained as pH; 6.9 ± 0.2, dissolved oxygen; 6.8 ± 0.1 mg/l, ammonia; 0.34 ± 0.03 mg/l, alkalinity; 67.2 ± 0.4 and water temperature; 28.5 ± 0.0 0 C. Testing the biochemical composition of FM and FCM Before preparing the feeds, the biochemical composition of the ingredients was analysed; proximate composition, pepsin digestibility, and amino acid profiles of FM and FCM were tested. Testing the proximate composition of the FM, and FCM The standard protocol of AOAC ( 1990 ) was followed to test the moisture, ash, fibre, crude protein, and crude lipid and the below tests were conducted at the Head Office of NARA, Colombo 15, Sri Lanka. Moisture content was tested by Air Oven Method utilising the above-mentioned hot air steriliser. Ash content was determined by incineration in a Hobersal HD 230 muffle furnace (Hobersal Furnaces and Ovens Technology, Barcelona, Spain). The crude fibre was tested following Weende Method operating Behrotest EXR-6 apparatus (Behr Labor-Technik, Dusseldorf, Germany). The crude protein was estimated according to the Micro-Kjeldahl Method using RAYPA MBCM-40 protein digestor (RAYPA, Barcelona, Spain) and RAYPA DNP-3000 distiller (RAYPA, Barcelona, Spain). As recommended by Ritvanen et al. ( 2020 ), the nitrogen-to-protein conversion factor for field cricket meal was considered as five (5.0). The crude lipid was determined by Soxhlet Method using FOSS Soxtec 2043 apparatus (FOSS Scino (Suzhou) Co. Ltd, Suzhou, China). The proximate composition of the FM and FCM were tested; the results are mentioned in Table 1 . Table 1 Proximate composition of the FM, and FCM (%, dry weight basis) Parameter FM FCM Dry matter content 94.3 91.3 Protein 67.1 b 58.0 a Lipid 8.4 8.2 Moisture 5.7 a 8.7 b Ash 14.7 b 4.8 a Fiber 1.4 a 9.2 b NFE 2.6 a 11.1 b Energy (KJ/g) 19.7 a 24.4 b Testing the pepsin digestibility of FM, and FCM FM and FCM samples were sent to the Nutrition Laboratory of Ceylon Grain Elevators PLC, Colombo 15, Sri Lanka. The pepsin digestibility was detected following the protocol of AOAC 971.09 (1999), as mentioned in Table 2 . Testing the amino acid profile of FM and FCM Amino acid compositions of FM and FCM were analysed using biochrom 30 + amino acid analyser (Biochrom Ltd, Cambridge, United Kingdom) following the standard protocol of AOAC 994.12 (2005). The values are mentioned in Table 3 . Table 2 Comparison of amino acid profiles of FM and FCM used in the experimental diets (g/100g, dry weight basis). Amino Acid Fishmeal (Tuna) Field cricket meal EAA Arginine 3.58 ± 0.04 a 4.64 ± 0.05 b Histidine 0.65 ± 0.03 a 0.93 ± 0.02 b Isoleucine 1.84 ± 0.17 a 2.47 ± 0.10 a Leucine 3.71 ± 0.64 a 4.85 ± 0.05 b Lysine 4.39 ± 0.01 a 4.29 ± 0.15 a Methionine 1.59 ± 0.09 a 1.18 ± 0.05 a Threonine 2.25 ± 0.06 a 2.67 ± 0.06 a Valine 2.41 ± 0.05 a 3.55 ± 0.07 b NEAA Alanine 3.05 ± 0.02 a 4.92 ± 0.03 b Aspartic Acid 4.80 ± 0.01 a 5.52 ± 0.07 a Cystine 1.00 ± 0.00 a 0.90 ± 0.00 b Glutamic acid 7.49 ± 0.12 a 8.02 ± 0.12 a Glycine 3.93 ± 0.01 a 3.58 ± 0.06 a Proline 2.80 ± 0.07 a 4.30 ± 0.01 b Serine 2.16 ± 0.04 a 2.88 ± 0.07 b Notes: All values are Mean ± SE, calculated from three replicates. a,b Means with different letters are significantly different (P < 0.05) from each other Phenylalanine, Tryptophan, and Tyrosine were not detected. Test diet preparation Three iso-protein (41.3 ± 0.2% crude protein) (Gandotra et al. 2014 ) and iso-caloric (19.2 ± 0.3 kJ GE/g) test diets were prepared according to the formulas as outlined in Table 4 . The crude protein content of the FM in the control diet (0FC) was replaced by FCM at 35%, 70%, and 100% in 35FC, 70FC, and 100FC diets, respectively. The FCM was purchased from Cricket Fit Limited, Thailand, and other ingredients were purchased from local suppliers. Ingredients were mixed well, and warmed distilled water was included in the mixture to form a duff. It was passed through a Sherry UH-B12MEC-B meat mincer (Sherry Bakery Equipment Suppliers (Pvt) Limited, Malabe, Sri Lanka). After transferring to the trays, the feeds were dried at 60 0 C for 24 hours in a GEMMY YCO-010 hot air steriliser (Gemmy Industrial Corporation, Taipei, Taiwan). Finally, drying constantly, cooled feeds were transferred to sealed bags and stored in a refrigerator. Table 3 The formulation and biochemical composition of the test diets (%, dry weight basis) Ingredients 0FC 35FC 70FC 100FC Fish meal 25.0 16.3 7.5 0.0 Field cricket meal 0.0 10.1 20.3 29.0 Soybean meal 42.0 42.4 43.5 43.0 Rice polish 16.5 15.2 8.7 10.0 Corn 6.5 6.5 12.5 11.0 Fish oil 5.0 4.5 2.5 2.0 Vitamin-mineral premix 1 2.0 2.0 2.0 2.0 Carboxyl methyl cellulose 2.0 2.0 2.0 2.0 Dicalcium phosphate 0.5 0.5 0.5 0.5 Chromium oxide 0.5 0.5 0.5 0.5 Dry matter content 90.3 90.5 90.4 90.5 Crude protein 41.7 41.1 41.7 40.8 Crude lipid 10.8 10.9 10.4 10.8 Moisture 9.7 9.5 9.6 9.5 Ash 10.3 9.7 8.5 7.8 Fiber 5.0 5.3 5.7 6.5 NFE 22.6 23.5 26.2 24.6 Energy (KJ/g) 18.5 19.1 19.3 19.8 1 Composition of Vitamin-mineral mixture per 1Kg- Vitamin A-1,000,000 IU, Vitamin D3- 100,000 IU, Vitamin E- 10,000 IU, Vitamin C- 10,000 mg, Vitamin K- 800 mg, Vitamin B1- 1500 mg, Vitamin B2- 1200 mg, Vitamin B6- 750 mg, Vitamin B12- 20 mg, Pantothenic Acid- 3000 mg, Niacin- 2150 mg, Folic Acid- 300 mg, Inositol − 25,000 mg, Biotin- 25 mg, Selenium- 30 mg, Iron- 20,000 mg, Zinc- 32,000 mg, Copper- 2,000 mg, Cobalt- 150 mg, Iodine- 325 mg, Magnesium- 6,000 mg, Potassium- 100 mg, Sodium- 5.9 mg, Manganese- 1500 mg Rearing and feeding After conditioning the C. catla fry for one week, as mentioned in 2.2, The bodyweights of the randomly selected five fish in each treatment were measured and recorded on the first day of the experiment using RADWAG AS 220.R2 chemical balance (RADWAG Balances & Scales, Radom, Poland). The fish were hand-fed at 5% of the body weight (Gandotra et al. 2014 ) twice daily at 09.00 and 15.00 hours. The body weights of the randomly selected five fish in each treatment were measured weekly, and feed budgets were adjusted. Unused feed particles were collected from siphoning after 15 minutes of feeding and dried in the above-mentioned oven at 55 C 0 . Finally, the body weights of all fish in each treatment were measured after 56 days (8 weeks) using the same balance. The below-mentioned parameters related to growth performance were calculated based on the following equations. $$Survival rate=(Final fish number÷Initial fish number)\times 100$$ $$Weight gain \left(WG\right)=Final weight \left(g\right)-Initial weight \left(g\right)$$ $$Daily weight gain \left(DWG\right)=Weightgain \left(g\right)÷Numberof days$$ $$Relative weight gain \left(RWG\right)=Weight gain \left(g\right)÷Initialweight \left(g\right)\times 100$$ $$Specific growth rate \left(SGR\right)=(Ln weight \left(final\right)-Ln weight (initial)÷number of days\times 100$$ $$Feed Conversion Ratio \left(FCR\right)=Total feed consumeed÷Total weight of product produced$$ $$Protein intake \left(PI\right) per fish=Protein quantity in the diet \left(\%\right)\times Feed intake per fish÷100$$ $$Digestible protein intake \left(DPI\right)=Protein digestibility fraction\times Protein intake \left(PI\right)$$ $$Protein efficiency ratio \left(PER\right)=Wetweight gain/protein intake$$ Water quality management Two-thirds of the water volume was changed from each aquarium every week. The excreta was siphoned out daily, and then refilling the tank to a 30.5 cm level. Total ammonia was tested weekly according to the standard methods of APHA (2017). Dissolved oxygen was measured daily using the Winkle method. The pH and water temperature were measured daily using HANNA edge H12020-01 pH meter (HANNA Instruments, Woonsocket, USA). Water quality parameters were maintained among the optimum ranges as water temperature 27.6 ± 0.4 0 C, pH 6.93 ± 0.25, dissolved oxygen 8.6 ± 0.4 mg/l total ammoniacal nitrogen 0.136 ± 0.048 mg/l, and total alkalinity 69.7.9 ± 2.1. Testing of in vivo digestibility Faecal matter was collected one hour prior to the next feeding at the cone bottoms of each aquarium. The collected faecal matter was filtered immediately with Whatman #1 filter paper and dried in a GEMMY YCO-010 hot air oven (GEMMY Industrial Corporation, Taipei, Taiwan) for 12 hours at 60 0 C. All the faecal residues from each aquarium were pooled, and they were frozen after labelling until further use. The crude protein contents of the feed and faecal matter were tested following AOAC ( 2003 ). The chromic oxide contents of the feed and faecal residues were determined according to the Acid-Digestion Method (Furukawa and Tsukahara 1966 ). The apparent digestibility content of protein (ADP) was calculated using the equation of Cho and Slinger ( 1979 ) below. $$ADP \left(\%\right)=100-100\times (\% chromin oxide in the feed\times \% protein in the faeces)÷(\% chromic oxide in the faeces\times \% protein in the feed)$$ Liver and gill testing for histopathological studies Randomly selected one fish from each aquarium was euthanised exposing 250 mg/L of MS-222 for 10 minutes (AVMA 2007 ). After sacrificing, fish were preserved in 10% neutral buffered formalin (Mumford 2004 ). Samples were sent to the Department of Veterinary Pathobiology, Faculty of Veterinary Medicine and Animal Science, University of Peradeniya, Sri Lanka and histological analysis was done following Hematoxylin and Eosin Staining (NSH 2001). Challenge Test against Aeromonas hydrophila A 7-day long bacterial challenge test was conducted to evaluate the efficacy of experimental diets against disease resistance in fish infected with the pathogenic bacterium Aeromonas hydrophila . A. hydrophila was originally sourced from diseased Koi carp ( Cyprinus rubrofuscus) and was cultured in Tryptic Soy Broth (TSB, High media, India) at 28°C for 24 hours and identification of bacterial isolates was made by 16S ribosomal RNA sequencing (Macrogene Korea). A. hydrophila was cultivated at 28°C for 24 h in Tryptic Soy Agar (TSA, High media, India) and a single colony was chosen to incubate in TSB for 24 hours at 27°C to use for the challenge test. The pellet was extracted and suspended in sterile phosphate-buffered saline solution after centrifuging the A. hydrophila cultured broth for 10 min at 3000g at 4°C. The lethal dose (LD 50 ) of A. hydrophila was determined by injecting 0.1 ml of 24 hour live bacteria culture intraperitoneally with different concentrations (OD values 0.5, 0.7, 0.9 at 600 nm) into the fish. The mortality of challenged fish was recorded daily for up to seven days. LD 50 was evaluated as a 0.67 OD value with 7.27 × 10 19 CFU mL − 1 cell density and was used in the bacterial challenge test according to McFarland standards. Ten fish from each replicate were randomly selected at the end of the feeding trial to evaluate the fish resistance against bacterial infection. A total of 0.1 mL A. hydrophila (7.27 × 10 19 CFU mL − 1 ) was injected intraperitoneally into fish. The rest of the fish in each treatment was IP injected with 0.1 mL of saline solution as a control. The water in the experimental tanks was unchanged, and no water circulation was provided during the test period. The challenged fish was observed daily to record any aberrant clinical signs and mortalities. If dead fish were observed, they were removed immediately. The pathological samples such as skin mucus, liver, gills and kidney were immediately removed and stored under − 80°C for further analysis. The bacterium was re-isolated from the fish organs to ensure that the mortalities were induced by A. hydrophila . Carcass composition testing At the end of the experiment, all the remaining fish were euthanised following the above-mentioned method and dried in a GEMMY YCO-010 hot air oven (GEMMY Industrial Corporation, Taipei, Taiwan) for 24 hours at 100 0 C. All the samples were ground and stored in air-tight zipper bags for proximate analysis following the standard protocol, as mentioned in 2.3.1. Statistical analysis All the measurements were repeated in triplicate. Data were processed and compiled using MS Excel. Treatments were compared using one-way ANOVA followed by Tukey's post hoc test after confirming the normality of the data and homogeneity of the variance by the Levene test. Statistical analysis was completed using SPSS software version 22.0. All the means were expressed as the mean ± standard error at P < 0.05 significance level. Results 3.1 Testing the biochemical composition of FM and FCM FM's crude protein and ash contents are significantly higher ( P < 0.05) than FCM's. However, moisture, fiber, NFE, and energy levels of FCM are higher than that of FM while the dry matter content and lipid are statistically similar in both ingredients. However, the profile of the EAAs of FCM is richer than FM; i.e., four out of eight tested EAAs are higher in FCM than in FM. Simultaneously, no differences were observed when comparing FM's isoleucine, lysine, methionine, and threonine contents with those of FCM. 3.2 Growth performance and feed utilisation of the fish All the research diets were well accepted by C. catla fry during the experimental period. Table 5 shows the growth performance, feed utilisation, and survival of the fish fed by various inclusion levels of FCM. The results of the present study reveal that the growth performance and survival of C. catla fry were not affected by the different inclusion levels of FCM and FM could be totally replaced by FCM. Table 4 Growth performance, feed utilisation, and survival of C. catla fry during the experimental period. Parameter 0FC 35FC 70FC 100FC P value Initial weight (g) 0.15 ± 0.01 0.17 ± 0.00 0.18 ± 0.00 0.17 ± 0.00 0.26 Final weight (g) 0.37 ± 0.03 0.39 ± 0.02 0.45 ± 0.00 0.45 ± 0.00 0.01 WG (g) 0.22 ± 0.03 0.23 ± 0.02 0.28 ± 0.01 0.28 ± 0.01 0.07 DWG (mg/day) 4.30 ± 0.70 4.70 ± 0.30 6.00 ± 0.00 5.70 ± 0.30 0.59 RWG 146.55 ± 22.24 137.65 ± 13.76 159.77 ± 4.90 174.16 ± 21.37 0.50 SGR 2.87 ± 0.14 2.73 ± 0.07 2.77 ± 0.03 2.88 ± 0.16 0.74 Survival (%) 67.67 ± 3.76 73.33 ± 14.53 85.67 ± 4.33 85.67 ± 4.33 0.32 FCR 2.22 ± 0.45 2.48 ± 0.19 1.85 ± 0.49 1.78 ± 0.31 0.35 PI 0.19 ± 0.02 0.23 ± 0.01 0.21 ± 0.01 0.20 ± 0.03 0.47 DPI 10.78 ± 1.37 10.27 ± 0.23 8.31 ± 0.79 6.76 ± 0.91 0.06 PER 1.16 ± 0.20 0.99 ± 0.07 1.30 ± 0.04 1.48 ± 0.09 0.34 3.3 Pepsin digestibility of FM and FCM and in Vivo digestibility of the feed Table 6 shows that the research feed's in vivo digestibility sharply decreased ( P < 0.05) when the inclusion level of FCM was increased. The in vivo digestibility of 70FC and 100FC diets significantly differed ( P < 0.05) from the FM-included control. However, the 35FC diet did not differ from the control diet. Figure 1 further confirmed the decreasing trend of the applicable parameter when the replaced percentage of FCM was increased. Simultaneously, Table 2 indicates that the pepsin digestibility of FCM is significantly lower than FM's. Table 5 Pepsin digestibility of FM, and FCM Ingredient Pepsin Digestibility (%) FM 93.1 a FCM 71.2 b Table 6 In vivo digestibility of C. catla fry during the experimental period Parameter 0FC 35FC 70FC 100FC In vivo Digestibility (%) 56.1 ± 5.0 b 45.0 ± 1.6 ab 38.5 ± 2.5 a 33.4 ± 1.2 a Varying in vivo digestibility of C. catla fry with the replacing percentage of FCM 3.4 Analysis of the carcass composition of the fish The proximate analysis of the carcasses confirmed that protein, lipid, moisture and fiber contents were statistically similar ( P < 0.05) with the inclusion of the FCM. In contrast, the ash content of the fish increased non-significantly ( P < 0.05) from 0FC to 70FC, and it was significantly higher in 100FC than in the control (0FC). Simultaneously, the fish's initial protein composition at the experiment's beginning was similar to the treated fish. However, the lipid, moisture and fiber contents of the researched fish have considerably higher than the initial fish sample. The ash content of the treated fish increased from 0FC to 70FC when compared with the ash content of the initial sample. However, the ash contents of the initial sample and 100FC were statistically similar. Table 7 Carcass composition of C. catla fry at the end of the experiment. Parameter Initial Sample 0FC 35FC 70FC 100FC Protein 75.94 ± 3.23 66.38 ± 5.05 75.07 ± 0.17 75.19 ± 0.17 72.46 ± 0.10 Lipid 06.37 ± 0.23 a 24.36 ± 0.44 b 21.21 ± 1.86 b 21.36 ± 1.49 b 26.33 ± 0.10 b Moisture 62.20 ± 0.33 a 78.71 ± 0.63 b 79.32 ± 0.39 b 79.65 ± 0.20 b 85.41 ± 4.15 b Ash 19.72 ± 1.01 c 08.58 ± 1.09 a 09.90 ± 0.21 ab 09.36 ± 1.58 ab 15.42 ± 1.31 bc Fiber 02.20 ± 0.30 b 00.54 ± 0.03 a 00.62 ± 0.05 a 00.50 ± 0.02 a 00.63 ± 0.01 a 3.5 Histopathological analysis of the liver and gill of the fish The histopathological architecture of the gill and liver tissues showed normal morphology without anomalies. Livers were red-pink, and whitish or pale-coloured livers were not observed among all the dissected treatment fish. Moreover, alterations in the liver tissues, like cloudy swelling and vacuolar degeneration, were not seen. The gill tissues did not observe histopathological changes in lamella and epithelia, hyperplasia, and desquamation. Represented histological structure of liver (a) and gill (b) of C. catla fry 3.6 Challenge Test against Aeromonas hydrophila The LD50 value was predicted as 0.65 using the mortality rates after 24 hours (Table 8 ) and Fig. 2 . When the concentration of bacteria was increased, the mortality rate also increased. Table 9 shows that the inclusion of the FCM in the feed did not affect adversely in terms of disease resistance. However, the survival of the control fish which were not injected with A. hydrophila . Table 8 Mortality rates of C. catla fry at different concentrations of A. hydrophila Absorbance Mortality rate after (nm) 24 hours (%) 0.0 0 0.5 30 0.6 40 0.7 60 0.8 70 0.9 90 Determining the LD 50 value of C. catla fry Table 9 Mortality rates of C. catla fry at 0.67 nm concentration of A. hydrophila Treatment Control 0FCM 35FCM 70FCM 100FCM P value Survival Rate (%) 0.0 ± 0.0 b 79.4 ± 13.7 a 68.9 ± 12.6 a 61.3 ± 1.8 a 75.0 ± 00.0 a 0.04 Discussion Insect meal has been materialised as a potential alternative to fishmeal with promising results. Therefore, replacing fishmeal with insect meal is an area of research thrust in aqua nutrition. Simultaneously, FCM has been identified as one of the ideal insect meals among many edible insects due to its better performance in previous research on Oreochromis niloticus (Perera et al. 2023 ; Hanan et al. 2022 ; Perera and Bhujel 2021 ), Clarias gariepinus (Taufek et al. 2018 and 2017 ), Channa striata (Prachom et al. 2023 ; Prachom and Suharman 2022 ), and Poecilia reticulata (Perera and Bhujel 2022 ). However, the effects of the FCM inclusion in C. catla feed are yet to be explored. Simultaneously, fishmeal has been totally replaced by silkworm pupae included diet in C. catla fingerling (Hasan 1991 ). The results of this experiment confirmed that FCM could totally replace fishmeal in terms of growth performance. The EAA composition of the FCM is comparable to FM (Table 2 ), and the results agree with the previous research (Perera and Bhujel 2022 ; Taufek et al. 2017 ; Wang et al. 2005 ). Lysine and methionine are the key EAAs responsible for fish growth, and lower levels of the above EAA contents in feed reduce fish growth feed efficiency (NRC 2011). Moreover, soybean meal has been included as a plant-based ingredient in a high percentage (42%-43.5%) of each treatment diet (Table 1 ). The previous research results have proved that soybean meal is a better plant-based ingredient in C catla feed in terms of weight gain (Srivastava et al. 2013 ) because of its higher nutritive values when compared with the other plant-based protein sources (Eyo1991). Based on the above facts, it is suggested that FCM could replace FM when the optimum feed formulations with adequate nutrition are available. Fish fed by all the treatment diets have shown statistically similar survival rates. Therefore, the fish accepted all the research feeds well, and the applicable feeds supplied adequate nutrition. Moreover, as mentioned above, FCM is rich in EAAs, and the EAA composition of both FM and FCM is sufficient. Literature confirms that the given feed's balanced amino acid profile decreases the fish's mortality (Aragao et al. 2007 ). In vivo protein digestibility is significantly higher in 0FC diet-fed fish compared to 70FM and 100FM diet-fed fish. Figure 1 exhibited a decreasing trend in the in vivo protein digestibility when the FCM level was gradually increased. The pepsin digestibility comparison (Table 5 ) confirms that the digestibility of FCM is significantly lower than FM's. The lower digestibility of insect meals is attributed to the availability of chitin (Gasco et al. 2019 ). However, PI has been increased non-significantly in FCM-included diet-fed fish compared to the control diet-fed fish (Table 4 ). Previous studies also confirmed that when the insect meal was introduced into the diet, the PI of the applicable fish increased non-significantly (Hammed et al. 2023 ). Digestible protein intake (DPI) increases when the PI increases (DPI = PI× Protein Digestibility Fraction). Therefore, it could be hypothesised that although the In vivo digestibility decreased in the fish fed with FCM highly included diets (70FCM and 100FCM), it did not affect the growth performance because the DPI was not significantly different. However. in vivo protein digestibility was low in all the treatment diet-fed fish (33.4–45.0%) and even in the control (56.1). Simultaneously, lower apparent digestibility coefficients (47–58%) of protein (ADCs) were obtained when microparticulate diets were fed to the nursery stage of fish in the previous research, and the ADC was significantly higher(76–86%) when live feeds ( Artemia ) were fed (Johnson et al., 2009). Most studies confirmed that the fry stage of fish prefers and performs better when provided with zooplankton than with the formulated diets (Kadhar et al. 2014 ). Many freshwater fish depend on zooplankton as their nursery feed ranging from one week to one or more months, and contrary, feeding the fish with formulated diets results in lower ingestion and poor digestibility (Conceicao et al. 2011 ). Moreover, C. catla exhibits a high selection of zooplankton during its fry-to-fingerling stage (Ahmed et al. 2000 ). On the contrary, inadequate supply, uncertainties in the quality, the potential to carry diseases, and the labour-intensive hatching process force the farmers to use alternatives to the live feeds (Perera et al. 2022 ). Therefore, farmers rear C. catla and other Indian carp with formulated feed in cultured earthen ponds. It may be assumed that the farmers could benefit from rearing the C. catla fry with the FCM-included diet in those ponds. Results of the body carcass further confirmed that the FCM inclusion in C. Catla fry feed did not adversely affect the growth of the fish. Protein, lipid, moisture, and fiber contents were similar in all fish carcasses. Therefore, the fish fed by different research diets have obtained adequate nutrition through the FCM-included diets. However, Seenappa and Devaraj ( 1995 ) have found that the growth of C. catla fingerling increased when the ash content in the carcass was high. In contrast to the above results, growth performance was not significantly different in 100FCM and 0FCM feed-fed fish though the ash content of the 100FCM fed-fish was higher than that of 0FCM fed-fish. Furthermore, many studies have confirmed that feeding fish with insect meal-based diets had no impact on the nutrient content of the fish and on contrary to the above findings and, some studies have reported that when the lipid percentage of the insect meal was increased, carcass lipid level was also increased (Jannathulla et al. 2022 ). Defatted/fat reduced FCM of 8.2% fat was used for this experiment, and it was comparable with the fat content in the FM used, which was 8.4%. Therefore, no significant effect on the carcass lipid composition. It may assume that the issue of accumulation of the higher lipid levels in fish carcasses when feeding with insect meal-based diets can be overcome by using the defatted or fat reduced insect meals. However, further research is required to resolve the above contradictory results. Basically, health status and environmental stress can be expressed using histopathological studies (Sultana et al., 2016 ). Stress conditions create histopathological alterations in the gills of fish, and the gill structure of the fish is an indicator of the water quality (Rankin et al. 1982 ). Moreover, the literature of the previous research confirmed that the deficit of nutrients, i.e., pantothenic acid and vitamin C, create alterations in the gills and, therefore, it could be inferred that the availability of the nutrients and feeding affects the morphology of the gills of fish (Strzyzewska et al. 2016 ). Gills with normal morphology (Fig. 2 ) suggest that feeding the fish with any level of FCM did not adversely affect the optimum feeding requirement of the fish. Fish ingest pollutants indirectly from feed, which consolidate in the tissues (Mohamed 2009 ) and toxic and harmful substances in feed ingredients cause fatty liver in fish ( www.linkedin.com ). Moreover, high carbohydrate content in the feed also creates fatty liver in fish (Huang et al. 2022 ). Histopathological results of the livers indicated that supplementation FCM did not affect the liver functions of the C. catla fry. High fishmeal-added diets have shown high disease resistance and total immunoglobulin level in fish compared to fish fed with fishmeal-replaced diets with alternative plant-based protein sources (Khosravi et al. 2015 ). However, disease tolerance is statistically similar in all the fish though the values are numerically high in FCM-included diet-fed fish. Similarly, previous research based on housefly ( Musca domestica ) pupae, melon fly ( Bactrocera cucurbitae) , Japanese oak silkmoth ( Antheraea yamamai ), silkworm ( Bombyx mori ) and Black soldier fly ( Hermetia illucens ) larvae confirmed that insect meals enhance the immunostimulation and disease resistance in fish due to the availability of chitin in the insect meal (Wang et al. 2022 ; Ido et al. 2019 ). Furthermore, Taufek et al. ( 2018 ) have reported that FCM could improve innate immunity and disease tolerance in African catfish due to the availability of chitin and unknown bioactive substances. Conclusions Insect meal has been explored as a sustainable and promising ingredient to substitute wild-caught fishmeal in aquafeeds. Results of this experiment further confirmed that high-quality fishmeal could be totally replaced by FCM in C. catla fry diet in terms of growth performance without adversely affecting carcass composition, liver functions and disease resistance. However, in vivo protein digestibility for FCM was lower than fishmeal, and it did not affect the concerned parameters in this experiment. The practical usage of insect meals has been limited to a few countries due to higher prices and unavailability. Thus, suitable techniques must be identified to produce cost-effective insect meals on a mass scale. Declarations Authors contribution G.S.C.P : Conceptualisation, designing, formal analysis, and original draft writing; M.R.A : Performing the experiment and data collection; A.M.A.N.A : Supervision and lab testing; P.P.M.H : Draft writing; K.L.W.T.M .: Sampling and lab testing; S.B.K.D: Lab testing . Funding This experiment was funded by the National Aquatic Resources Research and Development Agency (NARA), Sri Lanka. Declaration of interest statement The authors reported no conflict of interest. Acknowledgements The authors are grateful to the staff of the Regional Research Center, Panapitiya, Waskaduwa, Sri Lanka of NARA, staff of the IARAD, the head office of NARA, and Dr K.K. Asanka Sanjeewa, Senior Lecturer, University of Sri Jayawardenepura, Sri Lanka.. References Ahmed ZF, Wahab MA, Haq MS, Miah MAH (2000) Evaluation of food selection of Catla catla (Hamilton) fingerling by determining Electivity Index grown in earthen ponds in Bangladesh. 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Afridin","email":"","orcid":"","institution":"University of Sri Jayawardenapura","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"M.","middleName":"R.","lastName":"Afridin","suffix":""},{"id":221296303,"identity":"c1c69509-7081-4a53-8c35-8eb45ed2277f","order_by":2,"name":"A. M.A.N. Adikari","email":"","orcid":"","institution":"National Aquatic Resources Research and Development Agency (NARA)","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"A.","middleName":"M.A.N.","lastName":"Adikari","suffix":""},{"id":221296304,"identity":"f6042857-a12c-4d55-aedc-98b70290d190","order_by":3,"name":"P. P.M. Heenatigala","email":"","orcid":"","institution":"National Aquatic Resources Research and Development Agency (NARA)","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"P.","middleName":"P.M.","lastName":"Heenatigala","suffix":""},{"id":221296305,"identity":"72c0aa4f-bdf8-4ea3-9595-7f958c3809c7","order_by":4,"name":"K. L.W.T. Maduka","email":"","orcid":"","institution":"National Aquatic Resources Research and Development Agency (NARA)","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"K.","middleName":"L.W.T.","lastName":"Maduka","suffix":""},{"id":221296306,"identity":"74202653-df58-4f44-b16e-b62cbf94a588","order_by":5,"name":"S.B.K. Dunusinghe","email":"","orcid":"","institution":"National Aquatic Resources Research and Development Agency (NARA)","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"S.B.K.","middleName":"","lastName":"Dunusinghe","suffix":""}],"badges":[],"createdAt":"2023-07-22 07:59:23","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-3193929/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-3193929/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s10499-023-01288-0","type":"published","date":"2023-10-06T09:09:47+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":40723348,"identity":"47192e9e-f2c5-47d1-9e73-5f42e74a9cb9","added_by":"auto","created_at":"2023-07-28 14:11:59","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":45128,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eVarying \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003ein vivo\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e digestibility of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eC. catla\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e fry with the replacing percentage of FCM\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-3193929/v1/d260dc9c25943d0b3caeeb58.png"},{"id":40723356,"identity":"bf111376-8329-42a6-b4b4-6f136c30e24b","added_by":"auto","created_at":"2023-07-28 14:12:00","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":248025,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eRepresented histological structure of liver (a) and gill (b) of C. catla fry\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-3193929/v1/ffd5bc36bc6c856081a4ae8b.png"},{"id":40723351,"identity":"b62e98af-71e3-4ce4-bb93-b9c5783a4caf","added_by":"auto","created_at":"2023-07-28 14:11:59","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":17690,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eDetermining the LD 50 value of C. catla fry\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-3193929/v1/ede1bf221e9fe78c95c9a035.png"},{"id":57334257,"identity":"1867e058-64be-4274-b283-9b4e52b40bc5","added_by":"auto","created_at":"2024-05-29 09:09:53","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1409072,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3193929/v1/ed057503-42da-471c-9ae4-fa696746666e.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Replacing the unsustainable and wild-caught fishmeal with field cricket (Gryllus bimaculatus) meal in Catla (Catla catla) fry diet: Effect for growth, in vivo digestibility, carcass composition histopathological alterations and disease tolerance","fulltext":[{"header":"Introduction","content":"\u003cp\u003eFishmeal is supposed to be the key protein component (Barlow \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2003\u003c/span\u003e) in conventional aquafeed formulations due to its superior nutrient qualities (Schipp \u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Mallison \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Chapman and Miles \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). However, soaring prices (dos Santos \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2016\u003c/span\u003e), food security issues (Chan et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), unsustainability (Ghamkhar and Hicks \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), and the impacts of climate change (van Huis and Oonincx \u003cspan citationid=\"CR80\" class=\"CitationRef\"\u003e2017\u003c/span\u003e) have forced the industry to find alternative ingredients. Moreover, a considerable percentage of world fish production has still been used to produce fishmeal instead of human consumption (FAO \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Auchterlonie \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; SEAFISH \u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Simultaneously, per capita fish consumption is increasing (Ye \u003cspan citationid=\"CR85\" class=\"CitationRef\"\u003e1999\u003c/span\u003e). Therefore, alternative ingredients are explored to replace the fishmeal in aquafeeds, and numerous innovative feedstuffs are being researched to find novel ingredients (Gasco et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIt has been predicted that world aquaculture production will be increased by using alternative protein sources without increasing the inclusion level of fishmeal (Olsen and Hasan \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). Furthermore, the inclusion level of fishmeal in some fish feeds has been sharply reduced (Kok et al. \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Tacon et al. \u003cspan citationid=\"CR76\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). It has been predicted that world aquaculture production will be increased due to the usage of alternative protein sources without increasing the inclusion level of fishmeal (Olsen and Hasan \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2012\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAccording to recent research results, insect meal has been identified as a highly potential source to replace fishmeal in aquafeeds (Nogales-M\u0026eacute;rida et al. 2019). Insect meal is comparable to fishmeal in terms of adequate protein, amino acid, fatty acid, vitamin and mineral contents with high digestibility (Nogales-Merida et al. \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Wang et al. \u003cspan citationid=\"CR83\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Akinawa and Ketiku \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2000\u003c/span\u003e) and high feed conversion efficiency (Katayama et al. \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). Besides, it is economically practical due to its short life cycle, and lower production cost (Taufek et al. \u003cspan citationid=\"CR77\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). More importantly, insect production is environmentally sustainable due to its low carbon footprint (Henry et al. \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Furthermore, the insect culture does not compete for space or resource complement (FAO \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2013\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cem\u003eG. bimaculatus\u003c/em\u003e is a frequently cultured (van Huis \u003cspan citationid=\"CR82\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) edible cricket species (Hanan et al. \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Hwang et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) with a high nutrient value (Ngonga et al. \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Notably, the applicable species has been frequently researched to find the replacement potential of the fishmeal in aquafeeds with encouraging results (Fan et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Perera and Bhujel \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Peh et al. \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Taufek et al. \u003cspan citationid=\"CR77\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). However, the scope of the above studies has been limited to a few fish species. Therefore, further research is needed to fill the gaps in the uncovered study areas of the various fish species.\u003c/p\u003e \u003cp\u003e \u003cem\u003eC. catla\u003c/em\u003e is an Indian Carp species with a fast growth rate and high market demand, especially in the South Asian region (Srivastava et al. \u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e2013\u003c/span\u003e) due to its taste and nutrition (Shahzad et al.2020; Das and Das \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). The statistics of 2018 reveal that the global production of \u003cem\u003eC. catla\u003c/em\u003e was 3041.3 thousand tonnes, and it ranked sixth in the major species production in world aquaculture, possessing 5.6 percent of the global fish production (FAO \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eLimited research results are available in reference to fishmeal replacement by insect meal related to the researched fish. Therefore, this experiment was conducted to find out the fishmeal replacement potential of the fry stage of \u003cem\u003eC. catla\u003c/em\u003e by FCM.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cp\u003eThe experiment was conducted in the Aquaculture Research Center of the head office of National Aquatic Resources Research and Development Agency (NARA), Crow Island, Mattakkuliya, Colombo 15, Sri Lanka, from September to November 2022.\u003c/p\u003e \u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eExperimental setup\u003c/h2\u003e \u003cp\u003eTwelve fibreglass aquaria of each 160 L were occupied to stock \u003cem\u003eC. catla\u003c/em\u003e fry. Each aquarium had a cylindroconical column initiated with a 146 cm diameter at the bottom to facilitate the faecal matter collection. Those tanks were filled with conditioned freshwater to 30.5 cm in depth. Each aquarium was aerated by using a SHOWFOU BS-232 air compressor (SHOWFOU ELECTRIC MACHINE CO. LTD, Kaohsiung City, Taiwan). Air temperature and light intensity were maintained at 27.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4 \u003csup\u003e0\u003c/sup\u003eC and 998\u0026thinsp;\u0026plusmn;\u0026thinsp;56 Lux, respectively. The photoperiod was maintained as 12 hours (light)/12 hours (dark) using fluorescent tube bulbs as a light source during the experimental period. Illumination was regulated by a SMART SENSOR AS803 digital lux meter (ARCO Electronics Ltd, Dong Guan City, China).\u003c/p\u003e \u003cp\u003e \u003cb\u003ePurchasing and conditioning of\u003c/b\u003e \u003cb\u003eC. catla\u003c/b\u003e \u003cb\u003efry\u003c/b\u003e\u003c/p\u003e \u003cp\u003eA total of thousand \u003cem\u003eC. catla\u003c/em\u003e fry (initial weight, 0.22\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04 g/fish) were procured from Carp Breeding Center, National Aquaculture Development Authority, Udawalawa, Sri Lanka. Randomly selected 360 fish among those fish were stocked in the aquarium mentioned above at a density of 164 /m\u003csup\u003e3\u003c/sup\u003e (30 fish per aquarium) and were acclimatised for a week. The control feed containing 41.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2% crude protein was fed twice daily at 5% of the body weight during the above week. Water height was maintained at 30.5 cm, and water quality parameters were maintained as pH; 6.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2, dissolved oxygen; 6.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1 mg/l, ammonia; 0.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03 mg/l, alkalinity; 67.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4 and water temperature; 28.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0 \u003csup\u003e0\u003c/sup\u003eC.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eTesting the biochemical composition of FM and FCM\u003c/h2\u003e \u003cp\u003eBefore preparing the feeds, the biochemical composition of the ingredients was analysed; proximate composition, pepsin digestibility, and amino acid profiles of FM and FCM were tested.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eTesting the proximate composition of the FM, and FCM\u003c/h2\u003e \u003cp\u003eThe standard protocol of AOAC (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e1990\u003c/span\u003e) was followed to test the moisture, ash, fibre, crude protein, and crude lipid and the below tests were conducted at the Head Office of NARA, Colombo 15, Sri Lanka. Moisture content was tested by Air Oven Method utilising the above-mentioned hot air steriliser. Ash content was determined by incineration in a Hobersal HD 230 muffle furnace (Hobersal Furnaces and Ovens Technology, Barcelona, Spain). The crude fibre was tested following Weende Method operating Behrotest EXR-6 apparatus (Behr Labor-Technik, Dusseldorf, Germany). The crude protein was estimated according to the Micro-Kjeldahl Method using RAYPA MBCM-40 protein digestor (RAYPA, Barcelona, Spain) and RAYPA DNP-3000 distiller (RAYPA, Barcelona, Spain). As recommended by Ritvanen et al. (\u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), the nitrogen-to-protein conversion factor for field cricket meal was considered as five (5.0). The crude lipid was determined by Soxhlet Method using FOSS Soxtec 2043 apparatus (FOSS Scino (Suzhou) Co. Ltd, Suzhou, China).\u003c/p\u003e \u003cp\u003eThe proximate composition of the FM and FCM were tested; the results are mentioned in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eProximate composition of the FM, and FCM (%, dry weight basis)\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"3\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eParameter\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFM\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eFCM\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDry matter content\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e94.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e91.3\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eProtein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e67.1\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e58.0\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLipid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e8.2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMoisture\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5.7\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e8.7\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAsh\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e14.7\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.8\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFiber\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.4\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e9.2\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNFE\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.6\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e11.1\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEnergy (KJ/g)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e19.7\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e24.4\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eTesting the pepsin digestibility of FM, and FCM\u003c/h2\u003e \u003cp\u003eFM and FCM samples were sent to the Nutrition Laboratory of Ceylon Grain Elevators PLC, Colombo 15, Sri Lanka. The pepsin digestibility was detected following the protocol of AOAC 971.09 (1999), as mentioned in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eTesting the amino acid profile of FM and FCM\u003c/h2\u003e \u003cp\u003eAmino acid compositions of FM and FCM were analysed using biochrom 30\u003csup\u003e+\u003c/sup\u003e amino acid analyser (Biochrom Ltd, Cambridge, United Kingdom) following the standard protocol of AOAC 994.12 (2005). The values are mentioned in Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eComparison of amino acid profiles of FM and FCM used in the experimental diets (g/100g, dry weight basis).\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"3\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAmino Acid\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFishmeal (Tuna)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eField cricket meal\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEAA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eArginine\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.58\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.64\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHistidine\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.65\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.93\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIsoleucine\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.84\u0026thinsp;\u0026plusmn;\u0026thinsp;0.17\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.47\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLeucine\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.71\u0026thinsp;\u0026plusmn;\u0026thinsp;0.64\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.85\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLysine\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4.39\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.29\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMethionine\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.59\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.18\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eThreonine\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eValine\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.41\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.55\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNEAA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAlanine\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.92\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAspartic Acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4.80\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5.52\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCystine\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.90\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGlutamic acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e7.49\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e8.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGlycine\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.93\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.58\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eProline\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.80\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.30\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSerine\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.16\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.88\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cem\u003eNotes: All values are Mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SE, calculated from three replicates.\u003c/em\u003e \u003csup\u003e\u003cem\u003ea,b\u003c/em\u003e\u003c/sup\u003e\u003cem\u003eMeans with different letters are significantly different (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) from each other\u003c/em\u003e\u003c/p\u003e \u003cp\u003e \u003cem\u003ePhenylalanine, Tryptophan, and Tyrosine were not detected.\u003c/em\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eTest diet preparation\u003c/h2\u003e \u003cp\u003eThree iso-protein (41.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2% crude protein) (Gandotra et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2014\u003c/span\u003e) and iso-caloric (19.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3 kJ GE/g) test diets were prepared according to the formulas as outlined in Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e. The crude protein content of the FM in the control diet (0FC) was replaced by FCM at 35%, 70%, and 100% in 35FC, 70FC, and 100FC diets, respectively. The FCM was purchased from Cricket Fit Limited, Thailand, and other ingredients were purchased from local suppliers.\u003c/p\u003e \u003cp\u003eIngredients were mixed well, and warmed distilled water was included in the mixture to form a duff. It was passed through a Sherry UH-B12MEC-B meat mincer (Sherry Bakery Equipment Suppliers (Pvt) Limited, Malabe, Sri Lanka). After transferring to the trays, the feeds were dried at 60\u003csup\u003e0\u003c/sup\u003eC for 24 hours in a GEMMY YCO-010 hot air steriliser (Gemmy Industrial Corporation, Taipei, Taiwan). Finally, drying constantly, cooled feeds were transferred to sealed bags and stored in a refrigerator.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe formulation and biochemical composition of the test diets (%, dry weight basis)\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIngredients\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0FC\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e35FC\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e70FC\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e100FC\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFish meal\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e25.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e16.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e7.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eField cricket meal\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e10.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e20.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e29.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSoybean meal\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e42.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e42.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e43.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e43.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRice polish\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e16.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e15.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e8.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e10.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCorn\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e6.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e6.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e12.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e11.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFish oil\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e5.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e4.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e2.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eVitamin-mineral premix\u003csup\u003e1\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e2.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCarboxyl methyl cellulose\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e2.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDicalcium phosphate\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.5\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eChromium oxide\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.5\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDry matter content\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e90.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e90.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e90.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e90.5\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCrude protein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e41.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e41.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e41.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e40.8\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCrude lipid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e10.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e10.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e10.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e10.8\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMoisture\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e9.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e9.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e9.5\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAsh\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e10.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e8.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e7.8\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFiber\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e5.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e5.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e5.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e6.5\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNFE\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e22.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e23.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e26.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e24.6\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEnergy (KJ/g)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e18.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e19.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e19.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e19.8\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"5\"\u003e\u003csup\u003e1\u003c/sup\u003eComposition of Vitamin-mineral mixture per 1Kg- Vitamin A-1,000,000 IU, Vitamin D3- 100,000 IU, Vitamin E- 10,000 IU, Vitamin C- 10,000 mg, Vitamin K- 800 mg, Vitamin B1- 1500 mg, Vitamin B2- 1200 mg, Vitamin B6- 750 mg, Vitamin B12- 20 mg, Pantothenic Acid- 3000 mg, Niacin- 2150 mg, Folic Acid- 300 mg, Inositol \u0026minus;\u0026thinsp;25,000 mg, Biotin- 25 mg, Selenium- 30 mg, Iron- 20,000 mg, Zinc- 32,000 mg, Copper- 2,000 mg, Cobalt- 150 mg, Iodine- 325 mg, Magnesium- 6,000 mg, Potassium- 100 mg, Sodium- 5.9 mg, Manganese- 1500 mg\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eRearing and feeding\u003c/h2\u003e \u003cp\u003eAfter conditioning the \u003cem\u003eC. catla\u003c/em\u003e fry for one week, as mentioned in 2.2, The bodyweights of the randomly selected five fish in each treatment were measured and recorded on the first day of the experiment using RADWAG AS 220.R2 chemical balance (RADWAG Balances \u0026amp; Scales, Radom, Poland). The fish were hand-fed at 5% of the body weight (Gandotra et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2014\u003c/span\u003e) twice daily at 09.00 and 15.00 hours. The body weights of the randomly selected five fish in each treatment were measured weekly, and feed budgets were adjusted. Unused feed particles were collected from siphoning after 15 minutes of feeding and dried in the above-mentioned oven at 55 C\u003csup\u003e0\u003c/sup\u003e. Finally, the body weights of all fish in each treatment were measured after 56 days (8 weeks) using the same balance. The below-mentioned parameters related to growth performance were calculated based on the following equations.\u003cdiv id=\"Equa\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equa\" name=\"EquationSource\"\u003e\n$$Survival rate=(Final fish number\u0026divide;Initial fish number)\\times 100$$\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Equb\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equb\" name=\"EquationSource\"\u003e\n$$Weight gain \\left(WG\\right)=Final weight \\left(g\\right)-Initial weight \\left(g\\right)$$\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Equc\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equc\" name=\"EquationSource\"\u003e\n$$Daily weight gain \\left(DWG\\right)=Weightgain \\left(g\\right)\u0026divide;Numberof days$$\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Equd\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equd\" name=\"EquationSource\"\u003e\n$$Relative weight gain \\left(RWG\\right)=Weight gain \\left(g\\right)\u0026divide;Initialweight \\left(g\\right)\\times 100$$\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Eque\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Eque\" name=\"EquationSource\"\u003e\n$$Specific growth rate \\left(SGR\\right)=(Ln weight \\left(final\\right)-Ln weight (initial)\u0026divide;number of days\\times 100$$\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Equf\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equf\" name=\"EquationSource\"\u003e\n$$Feed Conversion Ratio \\left(FCR\\right)=Total feed consumeed\u0026divide;Total weight of product produced$$\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Equg\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equg\" name=\"EquationSource\"\u003e\n$$Protein intake \\left(PI\\right) per fish=Protein quantity in the diet \\left(\\%\\right)\\times Feed intake per fish\u0026divide;100$$\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Equh\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equh\" name=\"EquationSource\"\u003e\n$$Digestible protein intake \\left(DPI\\right)=Protein digestibility fraction\\times Protein intake \\left(PI\\right)$$\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Equi\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equi\" name=\"EquationSource\"\u003e\n$$Protein efficiency ratio \\left(PER\\right)=Wetweight gain/protein intake$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003eWater quality management\u003c/h2\u003e \u003cp\u003eTwo-thirds of the water volume was changed from each aquarium every week. The excreta was siphoned out daily, and then refilling the tank to a 30.5 cm level. Total ammonia was tested weekly according to the standard methods of APHA (2017). Dissolved oxygen was measured daily using the Winkle method. The pH and water temperature were measured daily using HANNA edge H12020-01 pH meter (HANNA Instruments, Woonsocket, USA). Water quality parameters were maintained among the optimum ranges as water temperature 27.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4 \u003csup\u003e0\u003c/sup\u003eC, pH 6.93\u0026thinsp;\u0026plusmn;\u0026thinsp;0.25, dissolved oxygen 8.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4 mg/l total ammoniacal nitrogen 0.136\u0026thinsp;\u0026plusmn;\u0026thinsp;0.048 mg/l, and total alkalinity 69.7.9\u0026thinsp;\u0026plusmn;\u0026thinsp;2.1.\u003c/p\u003e \u003cp\u003e \u003cb\u003eTesting of\u003c/b\u003e \u003cb\u003ein vivo\u003c/b\u003e \u003cb\u003edigestibility\u003c/b\u003e\u003c/p\u003e \u003cp\u003eFaecal matter was collected one hour prior to the next feeding at the cone bottoms of each aquarium. The collected faecal matter was filtered immediately with Whatman #1 filter paper and dried in a GEMMY YCO-010 hot air oven (GEMMY Industrial Corporation, Taipei, Taiwan) for 12 hours at 60 \u003csup\u003e0\u003c/sup\u003eC. All the faecal residues from each aquarium were pooled, and they were frozen after labelling until further use. The crude protein contents of the feed and faecal matter were tested following AOAC (\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2003\u003c/span\u003e). The chromic oxide contents of the feed and faecal residues were determined according to the Acid-Digestion Method (Furukawa and Tsukahara \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e1966\u003c/span\u003e). The apparent digestibility content of protein (ADP) was calculated using the equation of Cho and Slinger (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e1979\u003c/span\u003e) below.\u003cdiv id=\"Equj\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equj\" name=\"EquationSource\"\u003e\n$$ADP \\left(\\%\\right)=100-100\\times (\\% chromin oxide in the feed\\times \\% protein in the faeces)\u0026divide;(\\% chromic oxide in the faeces\\times \\% protein in the feed)$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eLiver and gill testing for histopathological studies\u003c/h2\u003e \u003cp\u003eRandomly selected one fish from each aquarium was euthanised exposing 250 mg/L of MS-222 for 10 minutes (AVMA \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). After sacrificing, fish were preserved in 10% neutral buffered formalin (Mumford \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2004\u003c/span\u003e). Samples were sent to the Department of Veterinary Pathobiology, Faculty of Veterinary Medicine and Animal Science, University of Peradeniya, Sri Lanka and histological analysis was done following Hematoxylin and Eosin Staining (NSH 2001).\u003c/p\u003e \u003cp\u003e \u003cb\u003eChallenge Test against\u003c/b\u003e \u003cb\u003eAeromonas hydrophila\u003c/b\u003e\u003c/p\u003e \u003cp\u003eA 7-day long bacterial challenge test was conducted to evaluate the efficacy of experimental diets against disease resistance in fish infected with the pathogenic bacterium \u003cem\u003eAeromonas hydrophila\u003c/em\u003e. \u003cem\u003eA. hydrophila\u003c/em\u003e was originally sourced from diseased Koi carp (\u003cem\u003eCyprinus rubrofuscus)\u003c/em\u003e and was cultured in Tryptic Soy Broth (TSB, High media, India) at 28\u0026deg;C for 24 hours and \u003cem\u003eidentification\u003c/em\u003e of bacterial isolates was made by 16S ribosomal RNA sequencing (Macrogene Korea).\u003c/p\u003e \u003cp\u003e \u003cem\u003eA. hydrophila\u003c/em\u003e was cultivated at 28\u0026deg;C for 24 h in Tryptic Soy Agar (TSA, High media, India) and a single colony was chosen to incubate in TSB for 24 hours at 27\u0026deg;C to use for the challenge test. The pellet was extracted and suspended in sterile phosphate-buffered saline solution after centrifuging the \u003cem\u003eA. hydrophila\u003c/em\u003e cultured broth for 10 min at 3000g at 4\u0026deg;C.\u003c/p\u003e \u003cp\u003eThe lethal dose (LD\u003csub\u003e50\u003c/sub\u003e) of \u003cem\u003eA. hydrophila\u003c/em\u003e was determined by injecting 0.1 ml of 24 hour live bacteria culture intraperitoneally with different concentrations (OD values 0.5, 0.7, 0.9 at 600 nm) into the fish. The mortality of challenged fish was recorded daily for up to seven days. LD\u003csub\u003e50\u003c/sub\u003e was evaluated as a 0.67 OD value with 7.27 \u0026times; 10\u003csup\u003e19\u003c/sup\u003e CFU mL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e cell density and was used in the bacterial challenge test according to McFarland standards.\u003c/p\u003e \u003cp\u003eTen fish from each replicate were randomly selected at the end of the feeding trial to evaluate the fish resistance against bacterial infection. A total of 0.1 mL \u003cem\u003eA. hydrophila\u003c/em\u003e (7.27 \u0026times; 10\u003csup\u003e19\u003c/sup\u003e CFU mL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) was injected intraperitoneally into fish. The rest of the fish in each treatment was IP injected with 0.1 mL of saline solution as a control. The water in the experimental tanks was unchanged, and no water circulation was provided during the test period. The challenged fish was observed daily to record any aberrant clinical signs and mortalities. If dead fish were observed, they were removed immediately. The pathological samples such as skin mucus, liver, gills and kidney were immediately removed and stored under \u0026minus;\u0026thinsp;80\u0026deg;C for further analysis. The bacterium was re-isolated from the fish organs to ensure that the mortalities were induced by \u003cem\u003eA. hydrophila\u003c/em\u003e.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eCarcass composition testing\u003c/h2\u003e \u003cp\u003eAt the end of the experiment, all the remaining fish were euthanised following the above-mentioned method and dried in a GEMMY YCO-010 hot air oven (GEMMY Industrial Corporation, Taipei, Taiwan) for 24 hours at 100 \u003csup\u003e0\u003c/sup\u003eC. All the samples were ground and stored in air-tight zipper bags for proximate analysis following the standard protocol, as mentioned in 2.3.1.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eStatistical analysis\u003c/h2\u003e \u003cp\u003eAll the measurements were repeated in triplicate. Data were processed and compiled using MS Excel. Treatments were compared using one-way ANOVA followed by Tukey's post hoc test after confirming the normality of the data and homogeneity of the variance by the Levene test. Statistical analysis was completed using SPSS software version 22.0. All the means were expressed as the mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard error at \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05 significance level.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e\n \u003ch2\u003e3.1 Testing the biochemical composition of FM and FCM\u003c/h2\u003e\n \u003cp\u003eFM\u0026apos;s crude protein and ash contents are significantly higher (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) than FCM\u0026apos;s. However, moisture, fiber, NFE, and energy levels of FCM are higher than that of FM while the dry matter content and lipid are statistically similar in both ingredients. However, the profile of the EAAs of FCM is richer than FM; i.e., four out of eight tested EAAs are higher in FCM than in FM. Simultaneously, no differences were observed when comparing FM\u0026apos;s isoleucine, lysine, methionine, and threonine contents with those of FCM.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec16\" class=\"Section2\"\u003e\n \u003ch2\u003e3.2 Growth performance and feed utilisation of the fish\u003c/h2\u003e\n \u003cp\u003eAll the research diets were well accepted by \u003cem\u003eC. catla\u003c/em\u003e fry during the experimental period. Table \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e shows the growth performance, feed utilisation, and survival of the fish fed by various inclusion levels of FCM. The results of the present study reveal that the growth performance and survival of \u003cem\u003eC. catla\u003c/em\u003e fry were not affected by the different inclusion levels of FCM and FM could be totally replaced by FCM.\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\n \u003ctable id=\"Tab4\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eGrowth performance, feed utilisation, and survival of \u003cem\u003eC. catla\u003c/em\u003e fry during the experimental period.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eParameter\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e0FC\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e35FC\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e70FC\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e100FC\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP\u003c/em\u003e value\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eInitial weight (g)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.15\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.17\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.18\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.17\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.26\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eFinal weight (g)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.37\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.39\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eWG (g)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.22\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.28\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.28\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.07\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eDWG (mg/day)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4.30\u0026thinsp;\u0026plusmn;\u0026thinsp;0.70\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.59\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eRWG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e146.55\u0026thinsp;\u0026plusmn;\u0026thinsp;22.24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e137.65\u0026thinsp;\u0026plusmn;\u0026thinsp;13.76\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e159.77\u0026thinsp;\u0026plusmn;\u0026thinsp;4.90\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e174.16\u0026thinsp;\u0026plusmn;\u0026thinsp;21.37\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.50\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSGR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.87\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.73\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.77\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.88\u0026thinsp;\u0026plusmn;\u0026thinsp;0.16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.74\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSurvival (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e67.67\u0026thinsp;\u0026plusmn;\u0026thinsp;3.76\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e73.33\u0026thinsp;\u0026plusmn;\u0026thinsp;14.53\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e85.67\u0026thinsp;\u0026plusmn;\u0026thinsp;4.33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e85.67\u0026thinsp;\u0026plusmn;\u0026thinsp;4.33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.32\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eFCR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.22\u0026thinsp;\u0026plusmn;\u0026thinsp;0.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.48\u0026thinsp;\u0026plusmn;\u0026thinsp;0.19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.85\u0026thinsp;\u0026plusmn;\u0026thinsp;0.49\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.78\u0026thinsp;\u0026plusmn;\u0026thinsp;0.31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.35\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePI\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.19\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.21\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.20\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.47\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eDPI\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e10.78\u0026thinsp;\u0026plusmn;\u0026thinsp;1.37\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e10.27\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e8.31\u0026thinsp;\u0026plusmn;\u0026thinsp;0.79\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6.76\u0026thinsp;\u0026plusmn;\u0026thinsp;0.91\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.06\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePER\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.16\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.30\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.48\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.34\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003cp\u003e\u003cstrong\u003e3.3 Pepsin digestibility of FM and FCM and\u003c/strong\u003e \u003cstrong\u003ein Vivo\u003c/strong\u003e \u003cstrong\u003edigestibility of the feed\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eTable \u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e shows that the research feed\u0026apos;s \u003cem\u003ein vivo\u003c/em\u003e digestibility sharply decreased (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) when the inclusion level of FCM was increased. The \u003cem\u003ein vivo\u003c/em\u003e digestibility of 70FC and 100FC diets significantly differed (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) from the FM-included control. However, the 35FC diet did not differ from the control diet. Figure \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e further confirmed the decreasing trend of the applicable parameter when the replaced percentage of FCM was increased. Simultaneously, Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e indicates that the pepsin digestibility of FCM is significantly lower than FM\u0026apos;s.\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\n \u003ctable id=\"Tab5\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003ePepsin digestibility of FM, and FCM\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eIngredient\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePepsin Digestibility (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eFM\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e93.1\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eFCM\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e71.2\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003cdiv class=\"gridtable\"\u003e\n \u003cdiv align=\"left\" class=\"colspec\"\u003e\u003cbr\u003e\u003c/div\u003e\n \u003ctable id=\"Tab6\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 6\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003e\u003cem\u003eIn vivo\u003c/em\u003e digestibility of \u003cem\u003eC. catla\u003c/em\u003e fry during the experimental period\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eParameter\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e0FC\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e35FC\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e70FC\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e100FC\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eIn vivo\u003c/em\u003e Digestibility (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e56.1\u0026thinsp;\u0026plusmn;\u0026thinsp;5.0\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e45.0\u0026thinsp;\u0026plusmn;\u0026thinsp;1.6\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e38.5\u0026thinsp;\u0026plusmn;\u0026thinsp;2.5\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e33.4\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003cp\u003e\u003cstrong\u003eVarying\u003c/strong\u003e \u003cstrong\u003ein vivo\u003c/strong\u003e \u003cstrong\u003edigestibility of\u003c/strong\u003e \u003cstrong\u003eC. catla\u003c/strong\u003e \u003cstrong\u003efry with the replacing percentage of FCM\u003c/strong\u003e\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec17\" class=\"Section2\"\u003e\n \u003ch2\u003e3.4 Analysis of the carcass composition of the fish\u003c/h2\u003e\n \u003cp\u003eThe proximate analysis of the carcasses confirmed that protein, lipid, moisture and fiber contents were statistically similar (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) with the inclusion of the FCM. In contrast, the ash content of the fish increased non-significantly (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) from 0FC to 70FC, and it was significantly higher in 100FC than in the control (0FC). Simultaneously, the fish\u0026apos;s initial protein composition at the experiment\u0026apos;s beginning was similar to the treated fish. However, the lipid, moisture and fiber contents of the researched fish have considerably higher than the initial fish sample. The ash content of the treated fish increased from 0FC to 70FC when compared with the ash content of the initial sample. However, the ash contents of the initial sample and 100FC were statistically similar.\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\n \u003ctable id=\"Tab7\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 7\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eCarcass composition of \u003cem\u003eC. catla\u003c/em\u003e fry at the end of the experiment.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eParameter\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eInitial Sample\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e0FC\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e35FC\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e70FC\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e100FC\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eProtein\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e75.94\u0026thinsp;\u0026plusmn;\u0026thinsp;3.23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e66.38\u0026thinsp;\u0026plusmn;\u0026thinsp;5.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e75.07\u0026thinsp;\u0026plusmn;\u0026thinsp;0.17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e75.19\u0026thinsp;\u0026plusmn;\u0026thinsp;0.17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e72.46\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eLipid\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e06.37\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e24.36\u0026thinsp;\u0026plusmn;\u0026thinsp;0.44\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e21.21\u0026thinsp;\u0026plusmn;\u0026thinsp;1.86\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e21.36\u0026thinsp;\u0026plusmn;\u0026thinsp;1.49\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e26.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMoisture\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e62.20\u0026thinsp;\u0026plusmn;\u0026thinsp;0.33\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e78.71\u0026thinsp;\u0026plusmn;\u0026thinsp;0.63\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e79.32\u0026thinsp;\u0026plusmn;\u0026thinsp;0.39\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e79.65\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e85.41\u0026thinsp;\u0026plusmn;\u0026thinsp;4.15\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAsh\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e19.72\u0026thinsp;\u0026plusmn;\u0026thinsp;1.01\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e08.58\u0026thinsp;\u0026plusmn;\u0026thinsp;1.09\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e09.90\u0026thinsp;\u0026plusmn;\u0026thinsp;0.21\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e09.36\u0026thinsp;\u0026plusmn;\u0026thinsp;1.58\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15.42\u0026thinsp;\u0026plusmn;\u0026thinsp;1.31\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eFiber\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e02.20\u0026thinsp;\u0026plusmn;\u0026thinsp;0.30\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e00.54\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e00.62\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e00.50\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e00.63\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec18\" class=\"Section2\"\u003e\n \u003ch2\u003e3.5 Histopathological analysis of the liver and gill of the fish\u003c/h2\u003e\n \u003cp\u003eThe histopathological architecture of the gill and liver tissues showed normal morphology without anomalies. Livers were red-pink, and whitish or pale-coloured livers were not observed among all the dissected treatment fish. Moreover, alterations in the liver tissues, like cloudy swelling and vacuolar degeneration, were not seen. The gill tissues did not observe histopathological changes in lamella and epithelia, hyperplasia, and desquamation.\u003c/p\u003e\n \u003cdiv id=\"Sec19\" class=\"Section3\"\u003e\n \u003ch2\u003eRepresented histological structure of liver (a) and gill (b) of C. catla fry\u003c/h2\u003e\n \u003cp\u003e\u003cstrong\u003e3.6 Challenge Test against\u003c/strong\u003e \u003cstrong\u003eAeromonas hydrophila\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eThe LD50 value was predicted as 0.65 using the mortality rates after 24 hours (Table \u003cspan class=\"InternalRef\"\u003e8\u003c/span\u003e) and Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e. When the concentration of bacteria was increased, the mortality rate also increased. Table \u003cspan class=\"InternalRef\"\u003e9\u003c/span\u003e shows that the inclusion of the FCM in the feed did not affect adversely in terms of disease resistance. However, the survival of the control fish which were not injected with \u003cem\u003eA. hydrophila\u003c/em\u003e.\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\n \u003ctable id=\"Tab8\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 8\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eMortality rates of \u003cem\u003eC. catla\u003c/em\u003e fry at different concentrations of \u003cem\u003eA. hydrophila\u003c/em\u003e\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eAbsorbance\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMortality rate after\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e(nm)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e24 hours (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e60\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e70\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e90\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec20\" class=\"Section2\"\u003e\n \u003ch2\u003eDetermining the LD 50 value of C. catla fry\u003c/h2\u003e\n \u003cdiv class=\"gridtable\"\u003e\n \u003ctable id=\"Tab9\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 9\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eMortality rates of \u003cem\u003eC. catla\u003c/em\u003e fry at 0.67 nm concentration of \u003cem\u003eA. hydrophila\u003c/em\u003e\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eTreatment\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eControl\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e0FCM\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e35FCM\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e70FCM\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e100FCM\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eP\u003c/em\u003e value\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eSurvival Rate (%)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e79.4\u0026thinsp;\u0026plusmn;\u0026thinsp;13.7\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e68.9\u0026thinsp;\u0026plusmn;\u0026thinsp;12.6\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e61.3\u0026thinsp;\u0026plusmn;\u0026thinsp;1.8\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e75.0\u0026thinsp;\u0026plusmn;\u0026thinsp;00.0\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.04\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003cdiv id=\"Sec23\" class=\"Section3\"\u003e\u003cbr\u003e\u003c/div\u003e\n\u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eInsect meal has been materialised as a potential alternative to fishmeal with promising results. Therefore, replacing fishmeal with insect meal is an area of research thrust in aqua nutrition. Simultaneously, FCM has been identified as one of the ideal insect meals among many edible insects due to its better performance in previous research on \u003cem\u003eOreochromis niloticus\u003c/em\u003e (Perera et al. \u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Hanan et al. \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Perera and Bhujel \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), \u003cem\u003eClarias gariepinus\u003c/em\u003e (Taufek et al. \u003cspan citationid=\"CR78\" class=\"CitationRef\"\u003e2018\u003c/span\u003e and \u003cspan citationid=\"CR77\" class=\"CitationRef\"\u003e2017\u003c/span\u003e), \u003cem\u003eChanna striata\u003c/em\u003e (Prachom et al. \u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Prachom and Suharman \u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), and \u003cem\u003ePoecilia reticulata\u003c/em\u003e (Perera and Bhujel \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). However, the effects of the FCM inclusion in \u003cem\u003eC. catla\u003c/em\u003e feed are yet to be explored. Simultaneously, fishmeal has been totally replaced by silkworm pupae included diet in \u003cem\u003eC. catla\u003c/em\u003e fingerling (Hasan \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e1991\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe results of this experiment confirmed that FCM could totally replace fishmeal in terms of growth performance. The EAA composition of the FCM is comparable to FM (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e), and the results agree with the previous research (Perera and Bhujel \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Taufek et al. \u003cspan citationid=\"CR77\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Wang et al. \u003cspan citationid=\"CR83\" class=\"CitationRef\"\u003e2005\u003c/span\u003e). Lysine and methionine are the key EAAs responsible for fish growth, and lower levels of the above EAA contents in feed reduce fish growth feed efficiency (NRC 2011). Moreover, soybean meal has been included as a plant-based ingredient in a high percentage (42%-43.5%) of each treatment diet (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The previous research results have proved that soybean meal is a better plant-based ingredient in \u003cem\u003eC catla\u003c/em\u003e feed in terms of weight gain (Srivastava et al. \u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e2013\u003c/span\u003e) because of its higher nutritive values when compared with the other plant-based protein sources (Eyo1991). Based on the above facts, it is suggested that FCM could replace FM when the optimum feed formulations with adequate nutrition are available.\u003c/p\u003e\u003cp\u003eFish fed by all the treatment diets have shown statistically similar survival rates. Therefore, the fish accepted all the research feeds well, and the applicable feeds supplied adequate nutrition. Moreover, as mentioned above, FCM is rich in EAAs, and the EAA composition of both FM and FCM is sufficient. Literature confirms that the given feed's balanced amino acid profile decreases the fish's mortality (Aragao et al. \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2007\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e \u003cem\u003eIn vivo\u003c/em\u003e protein digestibility is significantly higher in 0FC diet-fed fish compared to 70FM and 100FM diet-fed fish. Figure\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e exhibited a decreasing trend in the \u003cem\u003ein vivo\u003c/em\u003e protein digestibility when the FCM level was gradually increased. The pepsin digestibility comparison (Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e) confirms that the digestibility of FCM is significantly lower than FM's. The lower digestibility of insect meals is attributed to the availability of chitin (Gasco et al. \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). However, PI has been increased non-significantly in FCM-included diet-fed fish compared to the control diet-fed fish (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). Previous studies also confirmed that when the insect meal was introduced into the diet, the PI of the applicable fish increased non-significantly (Hammed et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Digestible protein intake (DPI) increases when the PI increases (DPI = PI× Protein Digestibility Fraction). Therefore, it could be hypothesised that although the \u003cem\u003eIn vivo\u003c/em\u003e digestibility decreased in the fish fed with FCM highly included diets (70FCM and 100FCM), it did not affect the growth performance because the DPI was not significantly different.\u003c/p\u003e\u003cp\u003eHowever. \u003cem\u003ein vivo\u003c/em\u003e protein digestibility was low in all the treatment diet-fed fish (33.4–45.0%) and even in the control (56.1). Simultaneously, lower apparent digestibility coefficients (47–58%) of protein (ADCs) were obtained when microparticulate diets were fed to the nursery stage of fish in the previous research, and the ADC was significantly higher(76–86%) when live feeds (\u003cem\u003eArtemia\u003c/em\u003e) were fed (Johnson et al., 2009). Most studies confirmed that the fry stage of fish prefers and performs better when provided with zooplankton than with the formulated diets (Kadhar et al. \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Many freshwater fish depend on zooplankton as their nursery feed ranging from one week to one or more months, and contrary, feeding the fish with formulated diets results in lower ingestion and poor digestibility (Conceicao et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). Moreover, \u003cem\u003eC. catla\u003c/em\u003e exhibits a high selection of zooplankton during its fry-to-fingerling stage (Ahmed et al. \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2000\u003c/span\u003e). On the contrary, inadequate supply, uncertainties in the quality, the potential to carry diseases, and the labour-intensive hatching process force the farmers to use alternatives to the live feeds (Perera et al. \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Therefore, farmers rear \u003cem\u003eC. catla\u003c/em\u003e and other Indian carp with formulated feed in cultured earthen ponds. It may be assumed that the farmers could benefit from rearing the \u003cem\u003eC. catla\u003c/em\u003e fry with the FCM-included diet in those ponds.\u003c/p\u003e\u003cp\u003eResults of the body carcass further confirmed that the FCM inclusion in \u003cem\u003eC. Catla\u003c/em\u003e fry feed did not adversely affect the growth of the fish. Protein, lipid, moisture, and fiber contents were similar in all fish carcasses. Therefore, the fish fed by different research diets have obtained adequate nutrition through the FCM-included diets. However, Seenappa and Devaraj (\u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e1995\u003c/span\u003e) have found that the growth of \u003cem\u003eC. catla\u003c/em\u003e fingerling increased when the ash content in the carcass was high. In contrast to the above results, growth performance was not significantly different in 100FCM and 0FCM feed-fed fish though the ash content of the 100FCM fed-fish was higher than that of 0FCM fed-fish. Furthermore, many studies have confirmed that feeding fish with insect meal-based diets had no impact on the nutrient content of the fish and on contrary to the above findings and, some studies have reported that when the lipid percentage of the insect meal was increased, carcass lipid level was also increased (Jannathulla et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Defatted/fat reduced FCM of 8.2% fat was used for this experiment, and it was comparable with the fat content in the FM used, which was 8.4%. Therefore, no significant effect on the carcass lipid composition. It may assume that the issue of accumulation of the higher lipid levels in fish carcasses when feeding with insect meal-based diets can be overcome by using the defatted or fat reduced insect meals. However, further research is required to resolve the above contradictory results.\u003c/p\u003e\u003cp\u003eBasically, health status and environmental stress can be expressed using histopathological studies (Sultana et al., \u003cspan citationid=\"CR75\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Stress conditions create histopathological alterations in the gills of fish, and the gill structure of the fish is an indicator of the water quality (Rankin et al. \u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e1982\u003c/span\u003e). Moreover, the literature of the previous research confirmed that the deficit of nutrients, i.e., pantothenic acid and vitamin C, create alterations in the gills and, therefore, it could be inferred that the availability of the nutrients and feeding affects the morphology of the gills of fish (Strzyzewska et al. \u003cspan citationid=\"CR73\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Gills with normal morphology (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e) suggest that feeding the fish with any level of FCM did not adversely affect the optimum feeding requirement of the fish.\u003c/p\u003e\u003cp\u003eFish ingest pollutants indirectly from feed, which consolidate in the tissues (Mohamed \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2009\u003c/span\u003e) and toxic and harmful substances in feed ingredients cause fatty liver in fish (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e\u003ca href=\"http://www.linkedin.com\" target=\"_blank\"\u003ewww.linkedin.com\u003c/a\u003e\u003c/span\u003e\u003cspan address=\"http://www.linkedin.com\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e). Moreover, high carbohydrate content in the feed also creates fatty liver in fish (Huang et al. \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Histopathological results of the livers indicated that supplementation FCM did not affect the liver functions of the \u003cem\u003eC. catla\u003c/em\u003e fry.\u003c/p\u003e\u003cp\u003eHigh fishmeal-added diets have shown high disease resistance and total immunoglobulin level in fish compared to fish fed with fishmeal-replaced diets with alternative plant-based protein sources (Khosravi et al. \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). However, disease tolerance is statistically similar in all the fish though the values are numerically high in FCM-included diet-fed fish. Similarly, previous research based on housefly (\u003cem\u003eMusca domestica\u003c/em\u003e) pupae, melon fly (\u003cem\u003eBactrocera cucurbitae)\u003c/em\u003e, Japanese oak silkmoth (\u003cem\u003eAntheraea yamamai\u003c/em\u003e), silkworm (\u003cem\u003eBombyx mori\u003c/em\u003e) and Black soldier fly (\u003cem\u003eHermetia illucens\u003c/em\u003e) larvae confirmed that insect meals enhance the immunostimulation and disease resistance in fish due to the availability of chitin in the insect meal (Wang et al. \u003cspan citationid=\"CR84\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Ido et al. \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Furthermore, Taufek et al. (\u003cspan citationid=\"CR78\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) have reported that FCM could improve innate immunity and disease tolerance in African catfish due to the availability of chitin and unknown bioactive substances.\u003c/p\u003e"},{"header":"Conclusions","content":"\u003cp\u003eInsect meal has been explored as a sustainable and promising ingredient to substitute wild-caught fishmeal in aquafeeds. Results of this experiment further confirmed that high-quality fishmeal could be totally replaced by FCM in \u003cem\u003eC. catla\u003c/em\u003e fry diet in terms of growth performance without adversely affecting carcass composition, liver functions and disease resistance. However, \u003cem\u003ein vivo\u003c/em\u003e protein digestibility for FCM was lower than fishmeal, and it did not affect the concerned parameters in this experiment. The practical usage of insect meals has been limited to a few countries due to higher prices and unavailability. Thus, suitable techniques must be identified to produce cost-effective insect meals on a mass scale.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthors contribution\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eG.S.C.P\u003c/strong\u003e: Conceptualisation, designing, formal analysis, and original draft writing; \u003cstrong\u003eM.R.A\u003c/strong\u003e: Performing the experiment and data collection; \u003cstrong\u003eA.M.A.N.A\u003c/strong\u003e: Supervision and lab testing; \u003cstrong\u003eP.P.M.H\u003c/strong\u003e: Draft writing; \u003cstrong\u003eK.L.W.T.M\u003c/strong\u003e.: Sampling and lab testing; \u003cstrong\u003eS.B.K.D:\u0026nbsp;\u003c/strong\u003eLab testing\u003cstrong\u003e.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis experiment was funded by the National Aquatic Resources Research and Development Agency (NARA), Sri Lanka.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDeclaration of interest statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors reported no conflict of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors are grateful to the staff of the Regional Research Center, Panapitiya, Waskaduwa, Sri Lanka of NARA, staff of the IARAD, the head office of NARA, and Dr K.K. Asanka Sanjeewa, \u0026nbsp;Senior Lecturer, University of Sri Jayawardenepura, Sri Lanka..\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003e\u003cspan\u003eAhmed ZF, Wahab MA, Haq MS, Miah MAH (2000) Evaluation of food selection of \u003cem\u003eCatla catla\u003c/em\u003e (Hamilton) fingerling by determining Electivity Index grown in earthen ponds in Bangladesh. Pak J Biol Sci 3(6):1061\u0026ndash;1068\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003e\u003cspan\u003eAkinawa O, Ketiku AO (2000) Chemical composition and fatty acid profile of edible larva of \u003cem\u003eCirinal forda\u003c/em\u003e (westwoo). Afr J Biomedical Res 3:93\u0026ndash;96\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003e\u003cspan\u003eAOAC (1990) Official Methods of Analysis of Association of Official Analytical Chemists, 15th edn. 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FAO, Rome\u003c/span\u003e\u003c/li\u003e\n\u003c/ol\u003e\n\u003cp\u003e\u003cstrong\u003eWeb References\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003ewww.linkedin.com/pulse/why-fish-gets-liver-diseases-michelle-wang/ Accessed \u0026nbsp;04 July 2023.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"","identity":"aquaculture-international","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"","snPcode":"10499","submissionUrl":"https://submission.nature.com/new-submission/10499/3","title":"Aquaculture International","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"","reportingPortfolio":"VoR Journals","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"insect meal, sustainable aqua feed, fishmeal substitutes","lastPublishedDoi":"10.21203/rs.3.rs-3193929/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3193929/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eInsect meal has been identified as one of the cutting-edge alternatives to substitute fishmeal (FM) in aquafeeds. However, the potential of FM replacing with field cricket meal (FCM) related to\u003cem\u003e Calta catla\u003c/em\u003e fry stage has yet to be researched. Therefore, an eight-week experiment was designed to evaluate the performance of \u003cem\u003eC. catla\u003c/em\u003e fry (0.22 ± 0.04 g/fish). The protein content of the FM of the control feed (0FCM) was replaced with 35% (35FCM), 70% (70FCM), and 100% (100FCM) of FCM. All the fish were hand-fed by respective research diets 5% of the body weight twice daily. Results showed that FCM could replace wild-caught fishmeal (67.1 % crude protein) without adversely affecting growth performance, carcass composition, liver functions, and disease tolerance. Though \u003cem\u003ein vivo\u003c/em\u003e protein digestibility significantly decreased (\u003cem\u003eP\u003c/em\u003e\u0026lt;0.05) \u0026nbsp;when the FCM percentage increased in the feed, it did not affect the performance of the fish. Therefore, FCM could be used to prepare fish-free feed in the\u003cem\u003e C. catla\u003c/em\u003e fry stage. However, optimum technical know-how should be identified to certify the accessible and economically viable mass-scale production of FCM.\u003c/p\u003e","manuscriptTitle":"Replacing the unsustainable and wild-caught fishmeal with field cricket (Gryllus bimaculatus) meal in Catla (Catla catla) fry diet: Effect for growth, in vivo digestibility, carcass composition histopathological alterations and disease tolerance","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2023-07-28 14:11:53","doi":"10.21203/rs.3.rs-3193929/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Major revision","date":"2023-08-03T15:44:32+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2023-08-02T03:53:13+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"e2c24270-1d1e-4af2-b3e3-bca3a15a297e","date":"2023-07-25T09:38:14+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2023-07-25T09:19:14+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2023-07-25T09:09:20+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2023-07-25T04:40:57+00:00","index":"","fulltext":""},{"type":"submitted","content":"Aquaculture International","date":"2023-07-22T07:53:47+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"","identity":"aquaculture-international","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"","snPcode":"10499","submissionUrl":"https://submission.nature.com/new-submission/10499/3","title":"Aquaculture International","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"","reportingPortfolio":"VoR Journals","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"aee2ab17-ae4b-4076-981e-0a7e8c69a920","owner":[],"postedDate":"July 28th, 2023","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2024-05-29T09:09:47+00:00","versionOfRecord":{"articleIdentity":"rs-3193929","link":"https://doi.org/10.1007/s10499-023-01288-0","journal":{"identity":"aquaculture-international","isVorOnly":true,"title":"Aquaculture International"},"publishedOn":"2023-10-06 09:09:47","publishedOnDateReadable":"October 6th, 2023"},"versionCreatedAt":"2023-07-28 14:11:53","video":"","vorDoi":"10.1007/s10499-023-01288-0","vorDoiUrl":"https://doi.org/10.1007/s10499-023-01288-0","workflowStages":[]},"version":"v1","identity":"rs-3193929","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3193929","identity":"rs-3193929","version":["v1"]},"buildId":"7rjqhiLT3MXkJMwkYKINL","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}