Citation of this paper

Response on growth performance and chemical composition of fillet of snakehead fish (Channa sp.) fed diets composed black soldier fly larvae protein replacing fishmeal protein

Hoang Nghia Manh, Pham Thi Phuong Lan, Le Minh Tue, Nguyen Thi Thanh Thuy, Truong Thi Hoa, Vo Duc Nghia, Tran Thi Thu Suong and Nguyen Duy Quynh Tram

Hue University of Agriculture and Forestry, Hue University, Vietnam
ndqtram@hueuni.edu.vn

Abstract

This study aimed at evaluating the effect of fishmeal protein replacing by black soldier fly larvae (BSFL) protein on growth performance and chemical composition of fillet of snakehead fish (Channa sp.). A total of 1,800 fingerlings of 5.15 g were randomly located to 5 dietary treatments namely CTL, LP10, LP20, LP30 and LP40 with 3 replicates. Fish were kept in net cages 3 x 2 x 1.5 m with 20 fish/m² and fed one of 5 diets, in which fishmeal (FM) as CTL and 10%, 20%, 30% and 40% of FM protein replaced by BSFL protein. Results showed that replacing FM protein by BSFL at level of up to 30% did not affect the survival rate, growth rate and yield as well as chemical composition of snakehead fish but of 40% declined the growth performance as final weight, daily gain and fish yield. In CTL, LP10, LP20 and LP30, the final weight and daily weight gain ranged 98.3-99.2 g/fish and 1.60-1.62 g/day, respectively, were higher in LP40 (86.5 g/fish and 1.41 g/day, respectively). It suggests that up to 12.9% BSFL meal can be included in diets without adverse effect on growth performance and yield of snakehead fish.

Keywords: larval protein, net cage, snakehead fingerling

Introduction

Snakehead fish is a typical freshwater fish in many countries, including Vietnam. Snakehead fish is omnivorous and can use agricultural wastes thus it has become an object of interest to many farmers (Tuyen and Trieu 2021). Snakehead fish is easy to raise, can be reared at high density and has a high tolerance to adverse environmental conditions, so snakehead fish farming has developed nationwide. The current snakehead fish farmers used trash fish and pellet feed. Using pellet helps to supply feed more active and to reduce water pollution, as well as to mitigate the pressure on in both inland and marine natural aquatic resources. However, there is a need for reduction the pellet price (Thuy and Sinh 2015).

In addition, finding nutritionally appropriate and sustainable alternatives to fishmeal (FM) is an area of intense research, with possible alternative sources of ingredients coming from terrestrial plants, animal by-products, microalgae, macroalgae or insects, to mention some (Barroso et al 2014; Wan et al 2018). The black soldier fly (Hermetia illucens) larvae (BSFL) are considered an important candidate species to be used for animal feeds (Cammack and Tomberlin, 2017; Ngoan et al 2021). The research and industrial-scale production of BSFL as feed ingredients have been intensified in the last few years (FAO 2013; Wang and Shelomi 2017). The BSFL are also a good source of protein and limiting amino acids such as lysine and methionine, and lipids, reaching up to 30% as DW basis if grown on optimal growth media (Sealey et al 2011; Liland et al 2017; Lan et al 2022^b).

Replacement of FM with BSFL meal in aquafeeds without negative effects on growth or performance has successfully been demonstrated in some fish feeding trials (Hoa and Dung 2016; Belghit et al 2018; Dumas et al 2018; Elia et al 2018; Vongvichith et al 2020), but not in others (St-Hilaire et al 2007; Kroeckel et al 2012; Lan et al 2022^a). Hoa and Dung (2016) studied the use of black soldier fly prepupae (BSFP) meal replacement in snakehead fish (Channa micropeltes) diet and showed that the survival rate and daily gain were not significant differences between trash fish and larval diets, but FCR was lower in larval diet than in trash fish, and the quality of fish fillet was not affected. Vongvichith et al (2020) suggested that BSFL were an efficient substitute for FM in climbing perch (Anabas testudineus) feed and potentially allowed for a reduction of feed protein levels. However, replacement of dietary FM with BSFL meal reduced the growth of juvenile turbot (165–756 g BSFL meal/kg diet, Psetta maxima) and rainbow trout (300 g BSFL meal/kg diet, Oncorhynchus mykiss) (Kroeckel et al 2012; St-Hilaire et al 2007). Similarly, Lan et al (2022^a) reported that the use of BSFL replacing total trash fish reduced the productivity of Asian seabass (Lates calcarifer Bloch, 1790) in both fresh and brackish water environments. To present, in our opinion, there were not many publications on using BSFL meal in diets for snakehead fish.

Therefore, this study aimed at evaluating the effect of replacement of FM protein by BSFL protein on growth performance and chemical composition of fillet of snakehead fish (Channa sp.) cultured.

Materials and methods

Fish

Snakehead fish fingerlings size of 2.15 g in weight and 5 cm in length were bought from a local hatchery. Fish were acclimated in net cages (3 x 2 x 1.5 m) placed in an aquaculture pond at a density of 20 fish/m2 (120 fish/cage) for two weeks..

Larval meal and diet preparation

Larvae meal: BSFL were fed by tofu by-products and collected on the 9^th day after rearing. Larvae meal preparation was followed Lan et al (2022^c). Larvae were washed with water several times to remove all impurities on the body, boiled at 60°C for 30 minutes, dried at 60^oC for 48 hrs and milled into full fat BSFL meal. The proportions of ingredients and the nutritive value of the diets are presented in Table 1.

Table 1. Ingredient proportion and chemical composition of diets
Item	Treatment
Item	CTL	LP10	LP20	LP30	LP40

Ingredient (kg/100 kg)
Fish meal	32.0	28.8	25.6	22.4	19.2
BSFL meal	-	4.3	8.7	12.9	17.2
Corn meal	16.0	15.0	14.2	14.2	13.1
Rice bran	15.3	15.2	15.0	14.0	14.0
Soybean meal	15.0	15.0	15.0	15.0	15.0
Wheat meal	13.7	13.7	13.5	13.5	13.5
Premix*	4.0	4.0	4.0	4.0	4.0
CMC**	3.0	3.0	3.0	3.0	3.0
Fish oil (mL)	1.0	1.0	1.0	1.0	1.0
Total	100	100	100	100	100
Chemical composition (%) and gross energy (GE, kcal/kg DM)
DM	94.7	95.2	94.5	94.9	95.1
OM	89.1	89.2	89.3	89.2	89.4
CP	37.4	37.7	37.5	37.6	37.8
EE	7.9	7.7	7.9	8.1	8.4
CF	4.2	4.2	4.5	4.6	4.9
Total ash	10.9	10.8	10.7	10.8	10.6
GE^***	4,667	4,665	4,677	4,685	4,714
Premix vitamin-mineral; Carboxylmethyl cellulose; ^** GE (kcal/kg) = 4143 + 56 x EE + 15 x CP - 44 x ash (Ewan 1989)

All ingredients were carefully mixed according to their ratio in five diets. Then the mixtures were extruded through a 2-6 mm diameter die plate using an extruder (Binh Minh Corp., Vietnam). The mixture was dried at 45°C for 24 hrs and stored in plastic bags at room temperature prior to use.

Experimental design

The experiment was arranged in a completely randomized design with five different diets as treatment and 3 replicates (5 x 3). The treatments named as following: CTL - control diet with fishmeal; and LP10, LP20, LP30 and LP40 as replacing 10%, 20%, 30% and 40% of fishmeal protein with larval protein (LP) in the control diet. Fish were fed 3 times per day at 6h00, 11h00 and 15h00 with quantity of 5 – 7% of fish body weight. Feed of refusal (option) was collected after 1.00 hours feeding.

The experiment was lasted for 60 days, exclusive 2-week preparation time.

Measurements

Water quality parameters were measured including water temperature (°C) by HOBO logger Pendant Temp, pH, dissolved oxygen (DO) and N-NH₃ by test kit Sera (Germany), they were measured periodically and data were presented in Table 2.

Table 2. Value of environmental parameters
Parameters	Range (Min - Max)	Mean (M± SD)

Water temperature (^oC)	23.1 – 33.6	28.5 ± 1.50
pH	6.50 – 7.83	7.40 ± 0.25
DO (mg/L)	4.21 – 4.70	4.16 ± 0.15
N-NH₃ (mg/L)	0.03 – 0.09	0.06 ± 0.26

Growth performance

To determine the growth performance, ten (10) fish in each cage were randomly collected and were anesthetized with Aqui-S® at a concentration of 10 mL/m³ before weighing. Live weights of fish were recorded at beginning and every 15 days until 60 days.

Daily growth rate in weight (DWG, g/day) or in length (DLG, cm/day) was then calculated by:

DWG = (W _i – W₀)/t

DLG = (Li – L₀)/t

In which, W₀, L₀ is weight or length at the beginning and W_i, L_i is weight or length at the end of the experiment and t is the number of feeding days.

Feed conversion ratio (FCR) was a the ratio between feed intake in DM and weight gain.

Survival rate (SR, %) was determined by the ratio between survival number at the final and the initial experiment.

Yield (kg/m²) was calculated by total live weight of fish collected at terminated experiment per square meter of water surface.

Meat chemical composition At the end of the experiment, 10 fish in each cage were caught for meat sampling. The fillet of 10 fish was taken out and mixed carefully before sampling. Therefore, in each treatment 3 samples were chemically analysed.

Photo 1. The snakehead fish fed BSFL meal after 2 months

Chemical analysis

All feed samples and fish fillet samples were chemically analysed for dry matter (DM), crude protein (CP), ether extract (EE), crude fibre (CF) and total ash according to the procedures of AOAC (1990) at the Laboratory of Biotechnology of the National Institute of Animal Sciences, Ha Noi.

Data analysis

Data were presented in the form of the mean (M) and standard error of the mean (SEM). The data were statistically processed by analysis of variance (ANOVA) by the General Linear Model in Minitab v.16.2 (2010). The difference between the mean values was determined by the Tukey method at a confidence level of 95%.

Results and discussion

Survival rate

The survival rate of farmed snakehead depends greatly on the resistance of the fish, the quality of the seed and the care and management of the pond water environment. The results of the study on the effect of replacement of fishmeal with BSFL meal on the survival rate of porcupine-headed snakehead fish are shown in Figure 1.

Figure 1. The survival rate of fish in 5 treatments

As shown in Figure 1, the survival rate of snakehead fish obtained from this experiment was relatively high ranging from 86.70% to 90% and not different between treatments (p>0.05). Mortalities only occurred due to aggressive behavior and were not related to diet. Thus, it is possible to replace fish meal protein with BSFL meal protein up to 40% in the diet of snakehead fish without affecting their survival rate. This result is similar to the results of Hoa and Dung (2016) when replacing fish trash with BSFL in diets for snakehead fish (Channa micropeltes) did not affect the survival rate from 80,7 to 84%. In addition, Lan et al (2020) reported that the survival rate of snakehead fish (Channa striata) fed different crude fat and CP diets ranged 71.1-93.3%. Dung and Hien (2017) indicated the survival rate of snakehead fish given various feeding levels of 0-3% of fish body weight ranged 70.7-88%. On the other hand, Vongvichith et al (2020) studied the effect of replacement of FM by BSFL in climbing perch (Anabas testudineus) diets and found that there was no effect on the survival rate of 81.7-82.2%. Lan et al (2022^a) reported that totally replacing trashfish by BSFL did not affect the survival rate of Asian seabass (Lates calcarifer Bloch, 1790) cultured in freshwater or brackish water and the survival rate ranged 84.4-97.8%.

Growth performance and yield

At BSFL replacement levels of 10%, 20% and 30%, the final weight, final length, daily gain in weight and length, and FCR responded similar to the CTL (Table 3). However, in replacement level of 40%, these measurements were lower than in other levels (p<0.05) (Figure 2, 3, 4).

Table 3. Growth performance and yield
Items	Treatment					SEM	p- value
Items	CTL	LP10	LP20	LP30	LP40	SEM	p- value

Initial length (cm)	5.32	5.29	5.29	5.30	5.31	0.074	0.691
Final length (cm)	24.0^d	23.8^cd	23.5^b	23.6^bc	21.0^a	0.077	<0.001
Initial weight (g)	2,15	2,16	2,15	2,15	2,15	0.067	0.855
Final weight (g)	99.2^b	98.5^b	98.5^b	98.3^b	86.5^a	0.243	<0.001
DLG(cm/day)	0.31^b	0.30^b	0.31^b	0,.30^b	0.26^a	0.002	<0.001
DWG (g/day)	1.62^b	1.61^b	1.61^b	1.60^b	1.41^a	0.004	<0.001
FCR	1.68^a	1.67^a	1.69^a	1.68^a	1.73^b	0.008	0.007
Yield (kg/m²)	5.16^b	5.15^b	5.32^b	5.23^b	4.56^a	0.060	<0.001
^abc: Means in the same row without common letter are different at p<0.05

In this study, lower growth performance and yield are found at BSFL inclusion level of 17.2% as DM diet (treatment LP40), but is not significantly different between other inclusion levels. BSFL meal is one of the most studied alternative proteins in aquafeed. Several current studies examined commercially produced BSFL in aquafeeds (Barroso et al 2014; Renna et al 2017; Belghit et al 2018). Depending on fish species levels ranging from 19.5 to 100% of dietary FM could be replaced by BSFL without significant adverse effects on growth performance. Due to the substrate variation in insect rearing, differing processing technologies, fish species and developing stage, the published results are remain mostly conflicting (Henry et al 2015; Makkar et al 2014). In Nile tilapia,the first observations revealed promising effects (Devic et al 2017).


Figure 2. The effect of Black soldier fly larvae on DLG of snakehead fish

Figure 3. The effect of Black soldier fly larvae on DWG of snakehead fish	Figure 4. The effect of Black soldier fly larvae on FCR

Lower final weight and daily weight gain in LP40 probably resulted from high CF content in the diet (Table 1) due to high CF in BSFL meal. The impact of chitin is a major concern and results are often inconsistent. In turbot levels above 33% dietary BSFL reduced feed acceptance, possibly attributed to the presence of chitin (Kroeckel et al 2012). The highest tolerable inclusion level of BSFL causing no reduction of growth performance throughout this study was 18.5%. This inclusion level is slightly lower as compared to previous studies (Kroeckel et al 2012; Lock et al 2016; Magalhães et al 2017; Renna et al 2017). Several factors like fish species and varying palatability of the substituted ingredient (FM versus BSFL) have to be considered in this context. It is also suggested that the processing procedure of the insect meal could be an important factor impacting on growth response (Lock et al 2016).

Chemical composition of fillet meat

Replacing 10% to 40% FM protein by BSFL did not affect DM, CP, EE and total ash of snakehead fish fillet (p>0.05). The DM content ranged 24.4-25.8%; CP: 15.6-15.9%; EE: 5.49-5.78% and total ash 3.44-3.58% as fresh weight.

Table 4. Chemical composition of fillet (%)
Items	Treatment					SEM	p- value
Items	CTL	LP10	LP20	LP30	LP40	SEM	p- value

CP	15.9	15.7	15.7	15.7	15.6	0.267	0.918
EE	5.49	5.61	5.73	5.77	5.78	0.227	0.876
Total ash	3.44	3.51	3.55	3.57	3.58	0.144	0.957
^ab: Means in the same row without common letter are different at p<0.05

Vongvichith et al (2019) studied the effect of replacement of FM by BSFP in snakehead fish diets and reported that the CP contents of fish at harvest were not different between dietary treatments (ranged 17-18%), while the crude fat content in BSFP diet (ca. 14%) was significantly greater than in FM and FM-BSFP diets (ca. 12%). The content of total ash in BSFP diet (ca. 4.1%) was significantly lower than in the rest diets (5.4–5.7%).

Conclusion

Replacing fishmeal protein by black soldier fly larvae protein at level of up to 30% did not affect the survival rate, growth rate and yield as well as chemical composition of snakehead fish (Channa sp.). It suggests that up to 12.9% BSFL meal can be included in diets without adverse effect on growth performance and yield of snakehead fish.

Acknowledgment

The authors are grateful to acknowledge for the financial support for this research by Thua Thien Hue Department of Science and Technology under Grant No. TTH.2021-KC.26.

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