Joumal of Engineering and Applied Sciences 12 (Special Issue 3): 6393-6397, 2017 ISSN: 1816-949X © Medwell Journals, 2017 Fish Feed Formulation using Black Soldier Fly Pre-Pupae from Fruit Waste Foh Tze Jia and Wong Ngie Hing (Matthew) Faculty of Engineering, Computing and Seience, Swinburne Universi 193350 Kuching, Malaysia of Technology Sarawak, Abstract: Black Soldier Fly (BSF) pre-pupee production has solved many environmental problems related to manure management ancl storage. Manure mrients assimilated will convert into valuable protein-rich feedstuff In order to resolve the challenge of rapid reduction in availability of fishmeal, the snxly on the use of pre-pupae 13 its replacement is necessary. The objective of this project isto formulate fish feed wsing BSF pre-pupae 2 protein source. The experimental pre-pupae growin on fruit waste were found containing 8% protein and 22% lipid on dry matter basis. Two formulations with 10 and S0®% inclusion of pre-pupae were made, These pfe-pupae were initially sun-dried and then, combined with wheat flour (49-89%), together with vitamin and ‘mineral premix (1%) fo form pellets of 3 mm diameter. Proximate analysis results indicated that the two formulations contained protein, lipid, moisture and ash inthe range of 16-38, 3-12, $9 and 12-14%, respectively ‘This study suggests that BSF pre-pupae alone are able to substitute fishmeal as protein source in fish diet, Further research on fish feeding trials, digestibility and palatability so as mutrient retention is required to determine the applicability of BSF pre-pupae as feed on sustaining normal fish growth, Key words: Black soldier fly, pre-pupae, fish feed, proximate analysis, feeding tials, growth INTRODUCTION lac: Soldiee Fly (BSH) or Hermetiailcens are small twopical and harmless insects originating from the whole Americas. BSF adults are unlike the normal houseflies as they do not consume human food and do not spread lisease. In adition, BSF larvae are eapable of minimizing large amount of organic waste. A BSF rearing sysiem reported by Sheppard et al. (1984) had suocessflly reduced the manure amount by 50% and total nitrogen concenttation by 24%. The surprising outeome was that the pre-pupae as the by-product of bioconversion were found containing 42% protein and 35% fat (Sheppard et al, 1994) which made them potential feedstulT for many animals such as chicken, fish and swine, Early on Hale observed that she rate of weight gain inchicks fed with dried BSF larvae was higher than those fad with soybeon meal i his experiment. Results showing the high potential of BSF larvae as feed for swine were reported by Newton ef al.(1977), At present, @ rapid reduction in the availability of fishmeal and fish oil causing the price of formulated dicts to se ls been a global hot issue (Henry et a, 2015). Wilson (2002) reported that the aquaculture feeds with fishmeal ab protein source east nearly 70% of the fish production coat, In 2007, the amount of fishmeal and fish cil utilized in world agunculture food incustry was equivalent to around 17 million tons of small pelagic fFockder fish in wet weight basis, Hence, many aulhorities started to search other protein source to replace fislameal and fish oil in order to overcome the issues. Since, insects are fish natural food (Howe etal, 201 4) which rich in amino acids, lipids, vitamins and other nutrients ((terteeck and Huis, 2012), they have been considered t0 potentially replace fishmeal and fish oil, expecially, BSF pre-pupae. ‘Based on the previous studies, experiments had been conducted on several types of fish mainly channel catfish, carp, rainbow trout and tilapia (Hui et af, 2006). Fishameal with 25% substituted by BSF pre-pupae showed the best result compared to the control diets and test diet containing housefly pupae (Hilaire ef at, 2007), Furthermore, formulated diet with 33% inclusion of Hermetia meal was reported did not suppress the juvenile turbot's growth responte (Kroeckel ef al, 2012). Rana (2014) also proved that BSF larvae could replace fishmeal up to 50% in fish feed for monosex tilapia fry. ‘Nonetheless, these researchers used chopped pre-pupae to partially substitute fishmeal or live pre-pupae to feed fish, Insufficient information due to limited research on fish feed formulation with inclusion of dried BSF pre-pupae to wholly replace fishmeal in fish diets has created a knowledge gap. Corresponding Author: Foh Tze fa, Faulty of Engineering, Computing and Science, ‘Swinburne University of Technology Sarawak, 93350 Kuching, Malaysie 6393J.Eng. Applied Sei, 12 (Special Issue 3): 6393-635 ‘This study presents the utilization of BSF pre-pupae tas the only protein source in the fish feed, The research, reported aims to demonstrate how the fish diets with different inclusion levels of sundried BSF pre-pupae in combination with other necessary ingredients can be formulated. In particular, this study will shows the process of Feed pellet formulation and the analyzed results om the finished diets. MATERIALS AND METHODS Dict preparation and formulation: SunDsied (SD) pre-pupae as protein source and other ingredients ax shown in Table | were used inthe feed formulation. The larvae wore reared on fruit waste inside a biodigester and the pre-pupwe were harvested andl provides by the project supervisor. The pre-pupue were dried under the sun for three continous days until constant mass were achieved Their Dry Matters (DM) were recorded and used to calculate the actual mass of live pre-pupae required for cach diet formulation Then, the SD pre-pupae together with vitamin and mineral premix were finely grinded separately into powder form using @ mortar and pestle Whea the ingredients were ready, they ware weighed separately according to their requited quantities and snixed them thoroughly by hans, The dough like misture was then kneaded with wate, ie, 2:1 of ingredients to water by weight to obtain a desired soft conformity, Next, a sinall amcunt of dough was inserted into a syringe and being extruded out va. 3mm diameter opening to produce noodle lke long pellets. The steps were repeated fr cach diet After thet, the long pellets were dried under the sun and cut into shorter pallets manually, ie., 3-5 mm long. Lastly, the short pellets were packed into. wate impermeable bags to prevent fungus growth and they Were stored at room temperature prior to the praximate analysis Proximate analysis: Proximate composition of the live and SD pre-pupae as well as each of the formulated diets were analyzed using standard AOAC methods for protein, lipid, moisture and ash content. Protein and lipid content determination were conducted by Nabbir Laboratory (Sarawak) Sdn, Bhd. using AOAC 984.13 and 954.02, respectively. Moisture and ash content in each sample were analyzed in triplicate assays using AOAC 930.15 and 94205, respectively at the Chemical Engineering Laboratory of Swinburne, University of Technology Sarawak, Collected data from the moisture and ash rnalysis were compared using one way Analysis Of Variance (ANOVA) with p0.05 were considered to be significant. 2017 “Table: Prenton used ft diss fend ferltion Physical properties of formulated pellets Diameter: The diameter of pellets was measured using a Vernier caliper with reading error of 0.05 ram, Ten pellets were randomly selected from each formulated diets. Data recorded were subjected to ANOVA analysis (p<0.05) to evaluate whether they differed significantly between the dies. Floatability: ‘The floatability of pellets was examined using ‘300 mL. beakers, The pellets of each diet were categorized into 3-S mm in length. Thea, three pellets were randomly selected from each category. The characteristics of these pellets (floated or sunk) were observed and recorded when they were dropped into the water at time (2) equal to 0,15, 30, 45 min RESULTS AND DISCUSSION BSF pre-pupae, in DM basis were found 51% after ‘sun-drying, This result isin agreement with the research finding from Burtle, Newton ef al. (2005) who reported that the DM of pre-papae to be more than 40%, Another study (Barry, 2004) reported DM of pre-pupae ranging from 35-51% alter oven-dtying. The effect of oven-drying and suredrying on the nutritive value of food samples was examined by Ayanwale ef al. (2007). The result showed that DM of the samples did not vary significantly using different drying methods, In addition, Barry (2004) ound out that organic wastes with varying, compositions consumed by BSF larvae did not contribute to the DM of pre-pupae after bioconversion and suggested that it may be influenced by the physiological parameters of larvae, Table 2 demonstrates the results of proximate analysis for the maw ingredients. The proximate compositions for the live pre-pupae had shown lower percentage than those SD pre-pupae, except for the moisture content, i¢., approximately seven times higher. ‘The protein in SD pre-pupae had shown two times higher percentage than those live ones. Ockerman (1985) ascribed this tothe moisture content of the fresh samples, i.e, the protein content of the fresh sample decreased when its moisture content inereased. Besides that, the increase in lipid concentration for SD pre-pupae from this study was found correlated with a previous study 6394J. Eng. Applied Sei, 12 (Special sue 3): 6393-6397, 2017 ‘Teble2: Results f paint analyse fhe ries Inreien= Pueansenk 06 Live BSED sundaes BSED Whew flow rote oT sme0 1188 Lipid be 2200 : Moire 3839 or : aah 200 assy “CyDenete rene ving lower ines of ihe perenne compare tothe live BSE samples, “Table: Ress of yroxate anbse fo te fommlnted dite Formule its Pweanestnt 6 SDIGTIS SbiaT soar Protein rr 1610 39.00) Li 363 338 1200 Mojtire 07 eur ise aa ed a aor ST aad Ta denote mer fal or atenpes fe ech Fed Terai fi. "() Dance sanple tht rte aabzed st Swinburne Chania ging Lab (Siddique et al, 2012) that investigated the lipid contents im dy fish samples and showed having higher lipid contents compared tothe fresh fish samples (On the other band, the differences for the ash contents between the live and SD pre-pupae could be due to the removal of moisture tat is apt to enfance the concentration of minerals (Moms et al, 2004), The protein content in the wheat flour was presumed to be 9% based on the product packaging, yet it was found to be 12% from the analysis Table 3 shows the results of proximate analysis for the fommulated diets. Protein and lipid contents in the formulated diets were found ranging approximately 16-39 and 3-12%, respectively. SDIOTI and SD1OT2 samples were found having similar results for both contents, This has showin that the process of formulation was conducted consistently and the peroentage inelusion. of SD pre-pupae so as other ingredients were favorably controlled On the other hand, SDSOTI was having highest protein and lipid contents among the formulated dicts, ie, both nutritive values were having almest half of the values found in the SD pre-pupae ‘The moisture content ranged from 8-58% whereas the ash content ranged between 12-14% in the diets. High Inorganic residue (14%) found in SDSOT] was considered acceptable as there was diet containing a8 high as 21% ash was recorded by Rana (2014). Although, the proportion of water used was same while preparing all the diets, the results from one-way ANOVA indicated that there was statistically significant difference observed in the moisture content for the formulated diets (F,, = 45; p05). Similarly, the ash content was found significantly differ among the formulated diets (F:, ~ 21; p=0.05), Post hoe test revealed that both moisture andl ash contents ‘Table: Rel ae say ANOVA fo Pellt’sdaeter Soiree ofvorition Sum of squires of Mean spore Five paar Baweengrays 001200 OO! B88 Wain gous 10427000 - Tal os 3800 in SDSOT! were significantly different from the other two diets, In contrast, SDIOT! and SDIOT2 did not differ significantly from each other, The difference in moisture contents might be due to the proportion of wheat flour constituted in the diets, This was recorded while preparing the diets that more water is requited to bind move grounded pre-pupae with the other ingredients when using lesser amount of wheat flour (ie. the birder), ‘The proximate analysis from other studies Hilaire et al (2007), Kroeckel etal (2012) and Rana (2014), also found baving similar results compare to all the diets that were formulated in this project despite the difference in percentage inclusion of BSF pre-pupae or larvae ‘Therefore, the protein content (16-39) of the formulated diais from this project should be sufficient to ensure good ‘growth rates in some of the fish species. For instance, mullet, carp, tilapia and eafish require 28, 23-41, 30-36 and 28-40% of dietary protein, respectively. Table 4 shows that there was no significant difference observed in the diameter of pellets among the formulated diets from this project, i... Fie ~ 0.18, p>0.08. Hence, the manually made pellets in SD1OT!, SD10T2 and SDSOT1 were conridered to be consistent with 3 mm in diameter, ie, 8740.07, 28240.06 and 2.8640.05 mm. For the floatability tes, all the 3 mm long pellets were found floated on top of water surface at t = O min, However, 4 and 5 mm long pellets fom SDIOT! and SDIOT2 were found sunk to the bottom of the beaker: For 'SDSOT1, 4 mm long pellets floated while S mm long pellets sunk f the bottom. After t= 30 min, all the 3 mm long. pellets fiom SDIOTT and SD1GT2 remained floating. Inthe meantime, all the 3 and 4 mam long pellets from SDSOT1 that previously floating bad sunk to the bottom. None of the pellets were observed floating on top of water surface after t = 45 min, At the end of the floatability test, the pellets were found to remain relatively intact. ‘The difference in floatability among pellets might be due to the different inchsion of ingredients in the formulation causing the pellet’s structure, texture and viscosity varied Khater ef al, (2014) advised that high pellets physical properties must be maintained to prevent _generation of excessive amount of fine particles ard cust Their studies concluded that the pellet sizes and protein ratio were the factors affecting the flostabilty of pellets, ‘The greater the pellet sizes andthe lower the protein ratio will ower the mean Hoatability of the pellets (Khater eta 2014). Furthermore, temperature of mash conditioning and 6395J.Eng. Applied Sei, 12 (Special Issue 3): 6393-635 surface coating of pellet affecting the floatability in diets hhad also been reported (AAdeparus and Famurewa, 2011) Using mash conditioning with warm water for diet preparation and coating the surface of pellet with oil could increase the floatability of pellet CONCLUSION ‘This study presented the utilization of BSF pre-pupae as the only protein source combining with other necessary ingredients in fish feed formulation SD10 contained 10% inclusion of sun-dried pre-pupae and 89% of wheat flour whereas SDSO consisted of 508% of sun-dried pre-pupae and 45% of wheat flour. The amount of vitamin and mineral premix used (1¥6) were kept constant for both formulations. Water to feed ratio requited to knead the mixture was 1:2 by weight, Pellets of 3 mm diameter were formed extruding through a syringe tnd then dried under the sun, The compositions of raw ingredients and finished diets were analyzed using, standard AAC methods, ‘The results of proximate analysis indicated that BSP pre-pupae would be a suitable fodder a8 protein source for fish diet. The pre-pupae contained 59% protein content was found two times higher after sun-drying and the Tipid content (22%) was slightly higher than those before sum-diying. The moisture and ash contents setested in live and SD_pre-pupae were in the range of 6-49 and 2-21%, respectively. SDSO had the highest protein (39%) and lipid (12%) contents among. the formulated diets, However, SDSO had the lowest moisture content but with the highest ash content, The two trials from SD10 had shown similar resis for the protein, lipid, moisture and ash content. ‘Using a small syringe was able to produce consistent 3 mm diameter pellets requited for this study. Different length of pellets (ie,, 3-5 mm) were cut and tested for floatabilty in a $00 ml. beaker. For SD1O, most of 3 min long pellets were able to remain floated within 45 min bat the other 4 and $ mim long pellets sunk immediately to the bottom, For SDS0, only the 3 ane 4 mm long, pellets were able to float for about 30 min whereas the $ mm long ones snk instantly to the bottom, Tn short, this study has shown that the BSF pre-pupae would be a feasible alternative protein source and sustainable fodder for fish diets expecially with the increasing market price of fishmeal due to excessive demand, Incorporating agriculture and aquaculture industries with organic waste recycling using BSF biocorwersion technology will be @ great idea as the larvae not only ean reduce organic waste but also provide high quality protein souree for animals and fish. 2017 RECOMMENDATIONS Future research directions: The scope of this study was limited to presenting the proximate analysis results of the finished diets only. Hence, this section lists a number of issues to be explored for any future research, Protein and lipid cantent of wheat flour was neglected when formulating diets in this study, The actual amount of these two contents provided by sumdried BSF pre-pupae in the finished diets was therefore ‘unidentifiable. tis highly recommended to use Pearson's ‘Square Method for better feed ration formulation. Fish feeding trials can be carried out to examine the applicability of the present formulations, Digestibility and palatability $0 a3 nutrient retention analysis are requited as well to determine the most suitable formulation that results in highest growth rates of fish, Other content in BSF pre-pupae like chitin must be analyzed as its presence may reduce fish feed intake and thus affect the fish growth performance and nutrient utilization. In this situation, integration of chitin degrading enzymes into fish diets may be considered. Floatability test of pellets in this study was observed every 15 min until they were all sunk. Smaller observation time is preferable so that, the actual time and location the pellets drop from tap to bottom level of water can be recorded, Other physical charaeteristios like durability and expansion rates as well as mechanical properties regarding, crushing load and repose angle of pellets can be taken into account as well, ‘The feed pellets in this study were stored at room temperature. Other storage method ean be considered in order to compare and examine whether it affects the nutrients content in feed. Improper storage can cause insects and microorganisms infestation of feed leading to weight loss and quality loss. This damaged feed can then pose serious health hazards to fish feeding on it REFERENCES Adepanusi, BO. and LAV. Famurewa, 2011, Water temperature and surface coating effect on floatability, ‘water absorption and thickness swelling of feed. J Agric. 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