Evaluation of the replacement of fishmeal with black soldier fly larvae meal in feed for tire track eel (Mastacembelus favus) fingerling

Nguyen Huu Yen Nhi, Tran Kim Ngoc, Nguyen Thi Thuy Hang, Nguyen Thi Bich Hanh and Trinh Thi Lan

An Giang University, Vietnam National University Ho Chi Minh City, Viet Nam
nhynhi@agu.edu.vn

Abstract

Tire track eel Mastacembus favusis distributed in Asian countries (e.g., India, Lao, Pakistan, Indonesia, Sri Lanka, Thailand, China, Malaysia, Cambodia and Vietnam). It is one of the most economically important species due to delicious taste and high nutritional values so it is now being farm-raised in Vietnam to support local economies. To improve sustainable culture, a ninety day-feeding experiment was carried out to determine appropriate replacement levels of black soldier fly (Hermetia illucens) larvae meal for fishmeal in the diets of tire track eel fingerlings to reduce the utilization of fishmeal which are depended on by aqua feeds as well as animal feeds. The fish were fed one of three diets formulated with different levels of protein replacing fishmeal (0%, 25%, and 50%) with black soldier fly larvae meal (BSF) at the same energy and protein levels. Triplicate groups of 50 Tire track eel fingerlings (average weight of 4.2 g) were randomly assigned to nine identical conical 500L tanks equipped with an aeration system and substrates for shelter. Weight gains (WG), daily weight gains (DWG), specific growth rates (SGR), feed conversion ratios (FCR), and survival rates (SR) were determined at the end of the experiment. The highest growth rate of fish fed different levels of black soldier fly larvae meal was obtained in group fish fed 50% BSF. DWG of this group (0.10 g/day) was higher than those of the groups (0% and 25%) and significant different (p< 0.05). SGR of fish in the group fed the 50% black soldier fly larvae meal (1.28%/day) was similar to those of the group (25%) but was higher than the remaining group (0%). The survival rates of tire track eel were not effect by level of BSF in the diets. However, FCR was reduce when BSF increase. Most of biochemical compositions of eels were not significantly different except the lipid content that was increase when levels of BSF increase. Therefore, this study revealed that black soldier fly larvae meal can be replace for fishmeal in the diets of tire track eel and the replacement of 50% of fishmeal with BSF brought about stimulated growth and optimal FCR.

Keywords: black soldier fly larvae meal, tire track eels, optimum growth, fishmeal

Introduction

Tire track eel is one of the aquaculture species with high economic value due to its delicious taste and high nutritional value (Gupta and Banerjee 2016). Therefore, it has been raised at the households. This fish is an elongated tropical freshwater fish and distributed in many Asian countries, and commonly presents in the basins of the Mekong Rivers (Petsut and Kulabtong 2015; Rainboth 1996; Ahmad et al 2018). In Vietnam, it is found in almost all regions of the country where the water flows lightly, e.g., crevices, rock embankments, bridge feet (Khoa and Huong 1993) and is cultured in a small scale farms to support local market demand.

In aquaculture, however, feeds is considered an important factor determining the growth rates of fish. Regarding to costs, feeds often account for the highest percentage in the total costs of aquaculture, in which protein is an important nutrient component in the feed rations. In organic aqua feeds, due to the shortage of organic protein sources, fishmeal (FM) costs are often, remarkably higher (up to 50%) than these in conventional feeds for carnivorous fish such as salmonids, snakehead fish, tire track eel (Olsen and Hasan 2012, Hien 2015, Nguyen et al 2022). The replacement of a substantial amount of fishmeal components could not only reduce fishmeal contents in organic aqua feeds substantially but also improve the sustainability of aquaculture feeds in general.

Black soldier fly larvae meal (BSF) in one of the candidates for replacing fishmeal, which were researched on many fish species such as Atlantic salmon Salmo salar (Belghit et al 2019; Li et al 2020; Weththasinghe et al 2021), yellow catfish Pelteobagrus fulvidraco (Xiao et al 2018), Jian carp Cyprinus carpio (Zhou et al 2017), Pacific white shrimp Litopenaeus vannamei (Cummins et al 2017), brown trout Salmo trutta m. fario (Mikołajczak et al 2022), Nile tilapia Oreochromis niloticus (Devic et al 2018; Tippayadara et al 2021; Agbohessou et al 2021), rainbow trout Oncorhynchus mykiss (Cardinaletti et al 2019; Renna et al 2017; Józefiak et al 2019; Terova et al 2019; Melenchón et al 2020; Hoc et al 2021; Biasato et al 2022), barramundi Lates calcarifer (Katya et al 2017), turbot Psetta maxima (Kroeckel et al 2012), Japanese seabass Lateolabrax japonicus (Wang et al 2019), European sea bass Dicentrarchus labrax (Abdel-Tawwab et al 2020; Mastoraki et al 2020), Siberian sturgeon Acipenser baerii (Caimi et al 2020), African catfish Clarias gariepinus (Fawole et al 2020; Adeoye et al 2020), rice field eel Monopterus albus (Hu et al 2020), mirror carp Cyprinus carpio var. specularis (Xu et al 2020), Siberian Sturgeon Acipenser baerii (Rawski et al 2020), Dusky Kob Argyrosomus japonicus (Madibana et al 2020), Eurasian perch Perca fluviatilis (Stejskal et al 2020). Belghit et al (2019) used black soldier fly larvae meal to replace fish meal in the diets of sea-water phase of Atlantic salmon (Salmo salar) and showed that after 16 weeks the replacement of the dietary fish meal with BSF did not affect the apparent digestibility coefficients of proteins, lipids, amino acids and fatty acids, or the digestive enzyme activities. Feed intakes, daily growth rates and feed conversion rations s were also unaffected by the inclusion of BSF meal in the diets. Whole body proteins, lipids and amino acid compositions were not affected by the replacement of fish meal with BSF. In addition, the total replacement of fishmeal with BSF in the diets of sea-water Atlantic salmon was possible without negative effects on growth performances and the sensory qualities of the fillet. Xiao et al (2018) also conducted a research to find the effects of BSF (Hermetia illucens) protein as a fish meal replacement on the growth and immune index of yellow catfish (Pelteobagrus fulvidraco). After 65 days, the diet in which 25% FM protein was replaced by BSF protein resulted in the greatest growth performances and immune indexes as well as the lowest feed conversion ratio. Therefore, these above researches indicated that BSF has the potential to partially or totally replace FM in the diets of many fish species. However, at present, there are no studies in the use of BSF protein to replace FM in the diet of Tire track eels. Therefore, the study was conducted to evaluate the possibility of replacing fishmeal with black soldier fly larvae meal in the diets of tire track eels by examining the effects on growth rates, survival rates and feed efficiency of tire track eels in culture tanks. The study may contribute to the selection material sources for producing pellet feeds for this species.

Materials and methods

Experimental animals and conditions

This experiment was carried out in the experimental farm in An Giang University, Long Xuyen city, An Giang province, Vietnam. Tire track eel fingerlings were obtained from a local hatchery in Thoai Son, An Giang province, Vietnam. They were transported to the experimental unit in 0.5 m ³ plastic containers with proper aeration. All the fingerlings were soaked in a solution of 3% NaCl for 5 minutes and were then acclimatized to the laboratory conditions for two weeks prior to the experiment. During the acclimatized period they were fed the commercial feed (42% crude protein) using for snakehead fish (Uni-President Vietnam Co.,LTD, Vietnam). The fish were selected to have a relative uniform in size with average initial weights of 4.2 g/fish and initial lengths of 10.8 cm/fish.

There were nine experimental tanks. Each tank was aerated via one air stone connected to a low-pressure electrical blower (Resun GF-370, China) and equipped with substrates as plastic clusters for shelter. The experiment tanks were constructed by Quang Đat Company, Can Tho city, Viet Nam.

The experimental tanks were connected to a clear water recirculation system including nine parallel-connected composite settlement tanks with a volume of about 500L per tank. Each of the settlement tanks were connected to a sedimentation tank containing sand and stones (1- 2 mm Ø), functioning as a biological filter. The tanks were housed in an open wall and closed roof construction on a concrete foundation. The water source was running municipal tap water and was dechlorinated by aeriation for 24 hours before use. About 30% of the water was replaced twice daily.

The experiment was arranged in a completely randomized design with 9 tanks (0.5m³/tank) with 3 treatments and three replicates ( n = 3). Nine diets were formulated with different levels of black soldier fly larvae meal: 0%, 25%, and 50% with the same protein and energy levels. The protein level was around 45% and energy was about 19 kJ/g in the feed (Table 2). The experiment were conducted for 3 months with the fish density of 100 individuals/m³.

Experimental diets

Three iso-energetic (19 MJ/Kg) and protein (45% crude protein) diets were produced with fish meal, soybean meal, wheat flour, black soldier fly larvae meal, fish oil, vegetable oil, earthworm liquid, premix vitamins - minerals, and CMC (Table 1 and 2) at the levels of 0%, 25% and 50% protein from black soldier fly larvae meal replace for fishmeal. Black soldier fly larvae meal was bought from the local market, which included 55.6% crude protein, 18.5% crude fat, and 9.4% ash. Diet recipes were given in Table 1 and the chemical compositions in Table 2.

All ingredients were thoroughly mixed and then pelleted by using an electronic meat grinder (Quoc Hung company, Long Xuyen city, Vietnam) with pellet diameters and lengths in the range of 1 mm. All diets were dried by oven in 60 °C until the feed’s moisture less than 10%. Then diets were weighed and stored in sealed plastic bags in small portions at -20 °C until use.

Fish management

The tire track eels were manually fed to satiation (approximately 10% of BW) twice daily (between 7:00 - 9:00 and between 16:00 - 18:00) by using a plastic tube to put feed into the nets at the bottoms of the tanks. Each feeding was closely monitored in each tank, in which the uneaten feed was collected before next feedings, and the feeding rates were adjusted based on the consumed feed on the previous day. The amount of eaten feed was recorded during the experiment and used to calculate true feed intakes. In addition, the feeding and swimming behaviours of the fish were monitored and recorded. The water of rearing tanks was changed about 30% every day. During the experiment, water environmental factors (e.g., temperature, pH, O₂ and TAN and NO₂^-) were daily monitored and controlled. Temperature in the experimental treatments ranged from 25 to 27^oC, pH from 7.5 to 7.9, dissolved oxygen from 5.8 to 6.9 mg/L, total protein (TAN) 0.0056 - 0.01 mg/L and NO₂^- from 0.0 to 0.23 mg/L were all within the appropriate limits for the normal growth and development of tire track eels (Boyd and Pillai 1985).

Measurements and calculations

All fish were weighted and measured for weights and lengths before and after the experiment. Specific growth rates (SGR), daily weight gains (DWG), feed conversion ratios (FCR), feed conversion efficiency (FCE), and survival rates (SR) were calculated using the equations (Nguyen et al 2022):

SGR (%/day) = [(ln final Wt – ln initial Wt)/days] x 100

DWG (g/day) = (Final Wt – Initial Wt)/days

FCR = Total feed intake (g)/Total wet weight gain (g)

SR (%) = (Total number of fish harvest/Total number of fish cultured) x 100

Chemical analysis

Ingredients, feed and experimental fish sampled before allocating to the experiment and after harvesting were kept frozen (-20^oC) until analysis. Chemical compositions (e.g., crude proteins, crude lipids, crude energies, crude fibres, ash and moistures) of these samples were analysed in triplicate and measured in dry mass as described in Nguyen et al (2022). Specifically, dry matters were determined by drying in an oven at 105 ^oC until constant weight. Ash contents were determined by incineration of the samples at 560^oC for 4 hours (until constant weight). Crude proteins were calculated as 6.25 x %N and analysed by the Kjeldahl method. Crude lipids were determined by Soxhlet extraction without acid hydrolysis. Crude fibre contents were analysed using acid-base digestions (AOAC, 2000). Nitrogen-free extracts (NFE) were calculated as NFE (%) = 100 – (% protein + % lipid + % fibre + % ash). The gross energies (Kcal/Kg) were calculated by using gross energy values of 5.64 Kcal/g for crude proteins, 4.11 Kcal/g for carbohydrates, and 9.44 Kcal/g for crude fats (NRC 1993).

Statistical analysis

Statistical analyses were conducted using the general linear model procedure (GLM) of the Minitab 16.0 software. Means and standard errors were calculated. One-way ANOVA and DUNCAN tests were used to compare means between treatment groups.

Results and discussion

Growths of tire track eels with different levels of black soldier fly larvae meal

There was no significant difference in initial sizes of fish between treatments (p> 0.05, Table 3). This indicates that the initial weight of fish did not affect the growth of fish after the experiment.

There was a significant difference in growth rates of tire track eels fed different levels of black soldier fly larvae meal replacing with fish meal. The growth rate was highest in the 50% black soldier fly larvae meal diet group and the growth response of the fish to the increased levels of black soldier fly larvae meal in the diet was presented in Figure 1. In fact, the daily weight gain (DWG) of the 50% black soldier fly larvae meal group (i.e., 0.10 g/day) was significantly different from those of the 0% and 25% BSF groups. Specific growth rates (SGR) of tire track eels in the treatments using different BSF contents ranged from 1.01 to 1.28 %/day. The 50% BSF group obtained the highest SGR (1.28%/day) which was not significantly different from those of the treatment group of 25% BSF ( p > 0.05) but was significantly different (p< 0.05) from the 0% BSF group.

Figure 1. Growth response of eels to increasing level of black soldier fly larvae meal in the die

Many researches showed that the growth rates of fish increased when partly replaced fishmeal with black soldier fly larvae meal. Xiao (2018) conducted an experiment to evaluate the potential of black soldier fly larvae meal as the alternative of fishmeal in the diet of yellow catfish at 0%, 13%, 25%, 37%, 48%, 68%, 85% and 100% in a 65-day feeding trial. The study showed that compared to the control group, the growth performances of treated yellow catfish increased until the maximum of 48% of fishmeal replaced by BSF and the diet of 25% BSF replacement resulted in the greatest growth performances (e.g. the weight gain rate increased by 29.1%). Similarly, Zhou et al (2018) indicated that the growth performances of Jian carp (Cyprinus carpio var. Jian) increased when BSF replaced fishmeal until 100%.

Survival rates and feed efficiency

After the 90-day treatment period, survival rates of tire track eels were relatively high ranging from 85.3 to 97.3% and there was no significant difference in survival rates between treatment groups (p> 0.05, Table 4). Thus, the results of this study showed that diets with different black soldier fly larvae meal contents did not affect the survival rates of tire track eels. Similarly, most studies indicated that the diets with different black soldier fly larvae meal contents have had no effect on fish survival.

Siddaiah et al (2023) was conducted the experiment with five isoproteic (420 g Kg⁻¹) and isocaloric (19.5 MJ Kg⁻¹) diets by replacing fishmeal at 0% (control diet), 25% (BSF 25), 50% (BSF 50), 75% (BSF 75), and 100% (BSF 100) with BSF to assess the effects of dietary fish meal replacement with Hermetia illucens larvae meal on the production performance of snakehead (Channa striata) juveniles. The research found that the diets with different BSF levels did not affect survival rates of the fish (e.g., 100% fish alive). The results of this study are similar to those of Magalhães et al (2017) and Zhou et al (2018) on European seabass (Dicentrarchus labrax) and Jian carp (Cyprinus carpio var. Jian) where survival rates was 100% in all groups when using black soldier fly (Hermetia illucens) pre-pupae meal as a fishmeal replacement in diets up to 45% or 100% . Xiao (2018) also showed that the survival rates of yellow catfish (Pelteobagrus fulvidraco) were not affected by the levels of black soldier fly larvae meal protein as a fishmeal replacement. However, the research from Devic et al (2017) showed the SR of Nile tilapia (Oreochromis niloticus) fed the diets containing black soldier fly (Hermetia illucens) larvae meal increase when BSF in diets increase.

Feed conversion ratios (FCR) decreased gradually as black soldier fly larvae meal replacing fishmeal in diets increased from 0% to 50% (Table 4 and Figure 2). FCR was lowest in the treatment of 50% BSF (e.g., 1.23) which was significantly different from the treatment groups of 0% and 25% BSF. The results of this study are similar to those of Xiao et al (2018) on yellow catfish (Pelteobagrus fulvidraco) that FCR decreased as BSF levels of the diets increased to 48%.

Figure 2. Feed conversion rates of eels to the increasing levels of protein in the diets

Chemical compositions of tire track eels before and after the experiment

Feed has a great influence on the biochemical compositions of aquatic animals. The results of the biochemical composition analysis of tire track eels before and after the experiment are shown in Table 5.

The moisture of the fish after the experiment fluctuated between 75.2% and 76.2% and was lower than those of the fish at the beginning of the experiment (82.0%). Other biochemical components such as proteins, lipids and ash in the body of the fish after the experiment were higher than those of the fish at the beginning of the experiment (Table 5).

Lipid contents of fish after the experiment ranged from 5.51 to 7.19% and was highest in the 50% BSF group. However, there was no significant difference (p> 0.05) in the lipid contents of fish between these BSF treatment groups. Protein and ash contents in the fish bodies after the experiment also showed similar trends as the moisture contents and no significant different between the treatments (Table 5).

Conclusion

• The replacement of 50% fishmeal by black soldier fly larvae meal produced the best growth rate of tire track eels and the lowest FCR.
  
  
• Survival rates did not differ between the treatment groups.
  
  
• Lipid levels of tire tract eels increased when levels of black soldier fly larvae meal increase in the diets.

Acknowledgments

This research is funded by Vietnam National University Ho Chi Minh City (VNU-HCM) under grant number B2021-16-01.

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