The using of larvae Black Soldier Fly Hermetia illucens as a substitute for soybean meal for the production of functional eggs high in lauric acid

Muhamad Alfian, Sumiati and Tuti Suryati

Department of Animal Nutrition and Feed Technology, Faculty of Animal Science, IPB University, Bogor 16680, Indonesia
sumiati@apps.ipb.ac.id

Abstract

Black Soldier Fly Larvae (Hermetia illucens) aged 15 days have a crude protein content of 36.6%. Apart from that, it is also rich in AMP (antimicrobial peptide) and has a high lauric acid content (49.18%) which can function as an antibacterial. This study aimed to analyze and evaluate the effectiveness of using BSF larvae or Hermetia illucens larvae as a substitute for soybean meal protein in artificial diets to produce functional chicken eggs high in lauric acid, and good egg production performance. The feeding trial used three feed treatments, namely BSF 0 (0% BSF larvae), BSF6.7 (6.7% BSF larvae, replacing 30% soybean meal), and BSF 13(13% BSF larvae, replacing 60% soybean meal). The experimental design used was a completely randomized design (RAL) with three treatments, five replications, and each replication consisting of 10 Lohman chickens. The results showed that the use of 13% BSF larvae, replacing 60% soybean meal protein, had a significant effect (p>0.05 on feed consumption, HDP value, egg mass, diet conversion, egg white weight, haugh unit value, and egg yolk lauric acid content, but did not affect (p>0.05) egg weight. The results of the research can be concluded that the substitution of soybean meal protein by BSF larvae protein to the level of 30-60% can increase daily egg production levels by 2.34% - 8.21%, egg mass value 1.25 - 6.7 g/head and can reduce the FCR value by 0.06 - 0.16 compared to the and produces functional eggs high in lauric acid with a percentage stored in egg yolk of 0.89% - 1.28%

Keywords: Hermetia illucens, Lohman brown chicken, production performance, egg quality

Introduction

The need for food of animal origin is increasing every year. Estimates from the World Food and Agricultural Organization (FAO) state that by 2040 food production of animal origin must be increased by 60-70% to meet the needs of the world community, including Indonesia. On the other hand, the need for food of animal origin that is healthy and contains organic compounds that are good for health should be a concern, because seeing the increasing prevalence of degenerative diseases is now becoming an increasingly worrying condition. One of these degenerative diseases is cardiovascular disease, which is the most dangerous disease because it can cause lifelong disability and even death.

Based on the Data and Information Center of the Indonesian Ministry of Health in 2018, showed that the prevalence of cancer reached 1.8%, heart disease at 1.5%, diabetes mellitus at 8.5%, hypertension at 34.1%, and stroke at 10.9%. The increase in the incidence of degenerative diseases is thought to increased the consumption of animal products containing bad cholesterol (Salter, 2013), but in reality, many studies have been conducted to dispel this assumption. The results of the study showed that total fat, saturated fatty acids (SFA), and monounsaturated fatty acids (mono-unsaturated fatty acids, MUFA) contained in animal products were not related to the risk of coronary heart disease, stroke, and cardiovascular disease (Siri-Tarino et al 2010).

Prevention of degenerative diseases in general is closely related to the intake of nutrients from food. Recent studies showed that several food products of animal origin that are of good quality contain fatty acids that have been proven to prevent heart disease, cancer, and diabetes (Brayden and Bairdn 2013), Some of the foods of animal origin such as eggs have complete nutritional content, its protein content is high. High levels make eggs an important source of animal protein besides meat, fish, and milk. Another unique thing, according to Zaheer (2015), is that the nutritional quality of eggs can be improved depending on the feed source consumed by chickens to increase production and produce functional food, one of which is by providing lauric acid or medium chain fatty acid (ALRM) which has 12 carbon atoms. Lauric acid has many benefits in the human body, including acting as an antiviral, antiprotozoal, and antibacterial which can dissolve viral membranes in the form of lipids so that it disrupts immunity and inactivates viruses (Roopashree et al 2021).

BSF larvae (Hermetia illucens) have great potential as an alternative feed ingredient as a source of protein in diets for laying hens in the layer phase because its protein content is quite high namely 40% - 50%, including essential amino acids. Apart from that, BSF larvae can be used as fatty acids according to the opinion of Spranghers et al (2017) that fatty acids can be produced from the use of BSF larvae, where BSF larvae have the highest lauric acid (C12:0) compared to other fatty acid components. Use larvae flour. BSF in diets can replace fish meal up to 100% (Sugiyono et al 2015). Research regarding the effect of giving BSF larvae as a substitute for soybean meal protein on the quality of chicken eggs high in lauric fatty acid as a functional food needs to be carried out to increase the use of larvae as a prospective feed source in the future. This research aimed to examine the effectiveness of substituting soybean meal protein with BSF larvae protein or Hermetia illucens L larvae to produce functional eggs high in lauric acid and produce good production performance.

Materials and method

This research was conducted from September 2022 to October 2022. Chicken rearing tests were carried out in the Field Laboratory (Cage C) of the Faculty of Animal Science, IPB University. This research was conducted on 150 laying hens of the Lohmann Brown type (average weight = 1.2–1.5 kg) at the age of 29 weeks. Each feed treatment consisted of 50 heads (5 replications with 10 heads/replication). Chickens are placed in battery or individual drums measuring 22 cm x 40 cm x 40 cm filled with one chicken. The feed used during this research was feed prepared based on the nutrient needs of laying hens, including corn, bran, dried larvae, CGM, fish meal, CPO oil, soybean meal, premix, DCP, Nacl, and CaCO3. Dried BSF larvae are made using the opening method from Harlystiarini, (2019). The composition and formulation of diets are in Table 1. Making dry BSF larvae after being harvested is treated by thawing frozen larvae, as well as washing the larvae until they are clean. The larvae are then steamed (steam) for 45 minutes at a temperature of approximately 90℃. After that, the larvae are dried using an oven at a temperature of 60℃ for 12-24 hours.

Procedure

This research was carried out using a completely randomized design (RAL), with 3 treatments and 5 replications with 10 chickens for each replication. The treatments used in this research are as follows:

BSF0: Control diet (diet without giving BSF larvae).
BSF6.7: Diet containing 6.7% larvae BSF (30% substitute for soybean meal protein).
BSF13: Diet containing 13% larvae BSF (60% protein substitution soybean meal).

Maintenance of laying hens, laying hens are placed in group battery cages (3 treatments x 5 replications x 10 chickens) and labeled according to the randomization, then the laying hens are immediately given drinking water with added vitamins to reduce stress. Maintenance is carried out for 6 weeks, starting from 28 weeks to 34 weeks of age. The adaptation and feeding trial period was carried out in the first two weeks. Providing a ratio of 108 g/head/day with larvae given separately and drinking water given ad libitum. Treatment diets are given twice a day, at 08.00 a.m. and 15.00 p.m. Every day the temperature and humidity of the cage, feed consumption, egg weight, daily egg production, egg mass, and feed conversion are recorded. Egg collection is carried out every day at 09.00 a.m, then weighing and counting the number of eggs obtained.

Measured variables and data analysis

Observed variables include feed consumption (g ), Feed Conversion Ratio (FCR), Hen day production (%), Egg mass production (g), Egg weight (g), and Egg yolk Fatty Acids (%). Data was analyzed using analysis of variance (ANOVA), If there are significantly different results then do Duncan's test. The data obtained was analyzed using software for Windows, SPSS 25 version.

Resultados and discussions

Chicken Performance

The performance of Lohman Brown laying hens aged 30-34 weeks during the study is presented in Table 3. Based on the results obtained, it is known that the use of BSF larvae protein as a substitute for soybean meal protein in diets has a significant effect (p<0.05) on feed consumption, daily egg production, egg mass, and feed conversion ratio. but did not show a real effect (p>0.05) on egg weight.

Figure 1. Correlation of hen day production and the replacing soya bean meal with BSFL

The feed consumption of laying hens in this study showed results that had a significant effect (p<0.05) on the three treatments. Treatment BSF0 (control) or without giving BSF larvae was significantly different from treatment BSF13 giving 13% larvae flour, but not significantly different from treatment BSF6.7 giving 6.7% larvae flour. The diet consumption value for laying hens in this study is lower than the standard consumption value for Lohman Brown laying hens aged 30 - 34 weeks, namely 108 g head^-1 day^-1 (Lohman, 2020). Consumption of different diets showed that there are differences in both quality, digestibility value, and form of diets given. The main cause of increased diet consumption in treatments BSF6.7 and BSF13 is the provision of BSF dry larvae in whole form with a larger particle size and a bright yellow color. so that by giving BSF larvae there is a level of sensory assessment of livestock in the diet because the palatability level of the diet is influenced by the physical characteristics of the feed such as particle size, color, taste smell which influence feed consumption (Setyawan et al 2020). Another assumption is that the value of feed consumption increases along with the level of substitution given, giving BSF larvae in the diet can speed up the digestive system, according to statement Manangkot (2014), BSF larvae have several enzymes, namely protease, amylase, lipase which are useful for the digestive process. food ingredients, so that the diet will be digested quickly if the feed provided contains these enzymes and results in a higher feed consumption value.

Egg weight in the study ranged from 54.58 - 55.30 g item-1, lower than the Lohman standard which ranged from 55.60 - 56.70 g (Lohman, 2020). Based on Table 3, the addition of BSF larvae to the diet had no significant effect (p>0.05) on egg weight. According to Leeson and Summers (2005), the weight of the eggs produced is correlated with the nutrient content of the feed provided, especially protein and amino acid intake. BSF larvae contain complete protein and essential amino acids and can be used as a feed ingredient for preparing diets (Harlystiarini, 2017) so replacing the soybean meal protein source by 30% and 60% with BSF larvae protein did not affect the weight of the eggs produced in this study.

Based on Figure 1, the average value of hen day production (HDP) in the study was 78.72 - 86.64%. From these average results, the substitution of soybean meal by BSF larvae showed results that had a significant effect (p<0.05) on the hen day production value, where the higher the level of BSF larvae substitution, the better the daily egg production level. The high value of hen day production in treatments BSF6.7 and BSF13 was because the treatment diets had a higher fat content than the control ratio BSF0, the high-fat contribution in these treatments resulted from the provision of BSF larvae. The fat content in the ratio of laying hens will contribute to an increase in plasma cholesterol which is a precursor of thyroid hormone in the ovaries, in line with the statement of Juneo et al (2018), thyroid hormone functions for the reproductive system of laying hens because it can modulate metabolism and development of the ovarian function, other studies show that plasma cholesterol produced from fat can contribute to the synthesis of the hormones progesterone and estradiol, which hormones will increase follicle development and produce better egg production (Miller et al 2007). However, there are two sources of body cholesterol, namely, endogenous cholesterol resulting from the synthesis in the body's cells and exogenous cholesterol resulting from feed consumption which is then absorbed by the digestive tract into the intestinal lymphatic tract (Piliang and Djojosoebagio, 2006). Attivi et al (2022) stated that dried BSF larvae contain 25% crude fat. Therefore, the higher the provision of BSF larvae in the diet, the better the zootechnical performance of poultry, especially for regulating reproductive function and improving egg-laying performance.

The average egg mass production during this study ranged from 43.09 – 50.69 g/head Based on Table 1, the substitution of soybean meal with BSF larvae in the diet showed a significant effect on egg mass production (p<0.05). The average egg mass production value (egg mash) in this study was lower than the standard egg mass for Lohman laying hens aged 30 weeks, which was 56.9 g/head (Lohman, 2020). According to Sh et al (2012), low egg mass production values are positively correlated with low egg production values, while egg mass production is the product of daily egg production (hen day production) by egg weight. In addition, the value of egg mass is influenced by the protein content and quality of the diet. Adequate and balanced protein and amino acid content

Chemical Quality of Egg Yolk

Data on the chemical quality of egg yolk in this study is the lauric acid content in egg yolk. The synthesis of lauric acid in egg yolk is influenced by the content of the diet containing lauric acid and consumed by chickens. One source of lauric acid used in this research comes from BSF larvae which is used as a substitute for soybean meal. The effect of using BSF larvae in the ratio of the lauric acid content in egg yolks can be seen in Table 4.

The administration of BSF larvae as a substitute for soybean meal protein in this study showed a significant effect (p<0.05) on the lauric acid content in egg yolk. The lauric acid content of treatments BSF6.7 and BSF13 was 0.07% and 0.09%, while the control treatment (BSF0) without BSF larvae did not contain lauric acid in egg yolk. The lauric acid deposited in the egg yolk in treatments BSF6.7 and BSF13 was produced from the use of BSF larvae and the higher the level of BSF larvae use, the higher the lauric acid content deposited in the egg yolk. This is because the process of egg biosynthesis begins with the formation of ova in the ovaries which begins with the formation of follicles, one of the constituent ingredients of which is synthesized from fatty acids. This is in line with the opinion of (Heuel et al 2021) that the composition of diet fat has an influence. on the fatty acid profile content of egg yolk.

(BSF0) control feed, (BSF6.7) feed containing 30% larvae BSF replacing the use of soybean meal protein (BSF13) Feed containing 60% larvae BSF replacing soybean meal protein. Numbers followed by different letters in the same column indicate there is a significant difference (p<0.05).

BSF6.7 treatment feed contained 75.1mg of lauric acid, while egg yolk contained 0.96mg. So the percentage of feed lauric acid deposited in egg yolk was 1.28%, and in the BSF13 treatment, it was 0.89%. This study’s results are higher than Heuel et al (2021), namely 0.39 – 0.41% below 1% of the lauric acid consumption given. This is different from the transfer rate of other saturated fatty acids such as myristic and palmitate which can reach 30 - 100% into the egg yolk. The low level of transfer of lauric acid content in egg yolk can be caused by the shorter chain length (MCFA) so that it is used more for other functions, especially energy metabolism, in the chicken's body (de novo enterohepatic lipogenesis). This is in line with the opinion of Ravindran et al (2016) showed that in broiler chickens, the absorption capacity of saturated fatty acids (SFA) decreased with increasing chain length. However, on the contrary, when the chain length is shorter, the absorption will be higher, which will later be used to repair damaged cell membranes, supported by the opinion of Miyagawa et al (2018) stated that lauric acid (ALRM) a smaller molecular size, a lower melting point, is liquid at room temperature, and a lower energy content (8.4 compared to 9.2 Kcal/g). This causes the metabolism of lauric acid to be very easily digested and easily absorbed by the body first and the rest of the body's metabolism will be synthesized in the liver via the portal vein which will be deposited into the ovaries for the formation of egg yolk.

Conclusion

Substitution at a BSF larval protein level of 6.7% - 13% equivalent to 30 and 60% soybean meal:

• Increase daily egg production levels up to 2.34% - 8.21%, egg mass values 1.25 – 6.7 g/head and can reduce FCR values up to 0.06 - 0.16 compared to controls
  
  
• Produces functional eggs high in lauric acid with a percentage stored in egg yolk of 0.89% - 1.28%

References

USDA United States Department of Agriculture 2000 Egg Grading Manual [Internet]. (diunduh 2023 july 8). United States Department of Agriculture, United States. Tersedia dari: https://naldc.nal.usda.gov/ download/CAT83788953/PDF.

Attivi K, Mlaga K G, Agboka K, Tona K, Kouame Y A E and Lin H 2022 Effect of fish meal replacement by Black Soldier Fly (Hermetia illucens) larvae meal on serum biochemical indices, thyroid hormone and zootechnical performance of laying chickens. Journal of Applied Poultry Research,31(3): 100275.

Brayden D J,Baird A W 2013 Opportunities for drug-delivery research in nutraceuticals and functional foods. Therapeutic Delivery. 4(3): 301±305.

Harlystiarini H, Mutia R, Wibawan I W T and Astuti D A 2019 In vitro antibacterial activity of black soldier fly (Hermetia illucens) larva extracts against gram-negative bacteria. BuletinPeternakan, 43(2):125-129. doi:10.21059/buletin.v43i2.428 33

Heuel M, Kreuzer M, Sandrock C, Leiber F, Mathys A, Gold M and Terranova M 2021 The transfer of lauric and myristic acid from black soldier fly larval lipids to egg yolk lipids of hens is low. Lipids, 56(4), 423-435. doi:10.1002/lipd.12304.

Leeson S J D, Summer 2005 Commercial Poultry Nutrition. (Edisi ke-3rd). University Book. University Guelph. Guelph, Ontario, Canada.

Lohmann, Tierzucht 2020 Lohmann Brown-Lite Layers.Germany (DE): Cuxhaven.

Manangkot H J, Rondonuwu S J, Pinontoan O R, Najoan M and Rumokoy J M 2014 Black soldier fly larvae manure degradation as fish meal replacer in native chicken ration. Lucrări Ştiinţifice-Seria Zootehnie, 62, 139-142.

Miller W L, Geller DH and Rosen M 2007 Ovarian and adrenal androgen biosynthesis and metabolism. Androgen Excess Disorders in Women: Polycystic Ovary Syndrome and Other Disorders, 19-33.

Miyagawa Y, Mori T, Goto K, Kawahara I, Fujiwara-Tani R, Kishi S, Sasaki T, Fujii K, Ohmori H and Kuniyasu H 2018 Intake of medium-chain fatty acids induces myocardial oxidative stress and atrophy. Lipids in Health and Disease, 17(1). doi : 10.1186/s12944-018-0908-0.

Piliang W G, Djojosoebagio S 2006 Fisiologi Nutrisi Volume ke-1. Bogor (ID): IPB Press.

Ravindran V, Tancharoenrat P, Zaefarian F and Ravindran G 2016 Fats in poultry nutrition: Digestive physiology and factors influencing their utilization. Animal Feed Science and Technology, 213:1–21. https://doi.org/10.1016/j.anifeedsci.2016.01.012

Roopashree P G, Shetty S S and Kumari N S 2021 Effect of medium chain fatty acid in human health and disease. Journal of Functional Foods, 87, 104724. doi : 10.1016/j.jff.2021.104724

Salter A M 2013 Impact of consumption of animal products on cardiovascular disease, diabetes, and cancer in developed countries. Animal Frontiers. 3(1):20-27 doi:10.2527/af.2013-0004

Setyawan A E E, sudjarwo E, Widodo H S and Prayogi 2020 Pengaruh penambahan limbah teh dalam pakan yterhadap penampilan produksi telur burung puyuh . Jurnal Ilmu- Ilmu Peternakan 23 (1) :7-10. //jiip.ub.ac.id/

Sh R, El-Ghar A, Debes A, Ghanem H and Ali O2012 Prediction of breeding values and selection for egg mass in a closed flock of White Leghorn. Int J Livest Prod. 32(1):63-74.

Silva, Juneo F, Ocarino N M and Serakides R 2018 Thyroid hormones and female reproduction. Biology of reproduction 99(5), 907-921. doi:10.1093/biolre/ioy115

Siri-Tarino P W, Sun Q, Hu F B and Krauss R M 2010 Saturated fatty acids and risk of coronary heart disease: modulation by replacement nutrients. Current atherosclerosis reports, 12, 384-390. doi: 10.1007/s11883-010-0131-6

Spranghers T, Ottoboni M, Klootwijk C, Ovyn A, Deboosere S, De Meulenaer B, Michiels J, Eeckhout M, De Clercq P and De Smet S 2017 Nutritional composition of black soldier fly (Hermetia illucens) prepupae reared on different organic waste substrates. J. Sci. Food Agric. 97, 2594–2600. doi: 10.1002/jsfa.8081

Sugiyono N, Elindratiningrum and Primandini Y 2015 Determinasi energi metabolis dan kandungan nutrisi hasil samping pasar sebagai potensi bahan pakan lokal ternak unggas. Jurnal Agripet. 15 (1): 41-45. doi: 10.17969/agripet.v15i1.2298

Zaheer K 2015 An updated review on chicken eggs: production, consumption, management aspects and nutritional benefits to human health. Food and Nutrition Sciences, 6(13), 1208. doi:org/10.4236/fns.2015.613127