Utilization of Silkworm (Bombyx mori) pupae meal and Javanese bird grasshopper (Valanga nigricornis) meal as alternative protein sources to replace meat and bone meal in ruminant feed: An in vitro study https://doi.org/10.12982/VIS.2026.018
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Abstract
This study aimed to evaluate the effects of replacing meat and bone meal (MBM) with insect-based protein sources, Silkworm pupae (Bombyx mori) meal (SPM), and Javanese bird grasshopper (Valanga nigricornis) meal (FGM) on rumen degradability, fermentation characteristics, total gas production, and methane emissions in vitro. Six experimental diets, each with five replicates, were tested, namely P1 (100% Napier grass), P2 (80% Napier grass + 20% MBM), P3 (80% Napier grass + 20% full-fat SPM (FSPM)), P4 (80% Napier grass + 20% defatted SPM (DSPM)), P5 (80% Napier grass + 20% full-fat FGM (FFGM)), and P6 (80% Napier grass + 20% defatted FGM (DFGM)). The result showed that the total gas and methane production were significantly lowered by insect-based protein meal and MBM (P < 0.05). In vitro dry matter degradability (IVDMD) and in vitro organic matter degradability (IVOMD) of insect-based protein meal were not significantly different from MBM and control diet (IVDMD: P = 0.87; IVOMD: P = 0.966). Insect-based protein meal and MBM reduced total volatile fatty acid (VFA) and propionate proportion while also increasing NH3-N and acetate:propionate ratio, as evidenced by P < 0.05. These results showed that insect-based proteins, particularly DSPM and FGM, had a similar digestibility to MBM, reduced methane emissions, and could effectively replace MBM in ruminant diets.
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References
Ahmed, E., Nishida, T., 2023. Optimal inclusion levels of cricket and silkworm as alternative ruminant feed: a study on their impacts on rumen fermentation and gas production. Sustainability. 15(2), 1415.
Association of Official Analytical Chemists, 2005. Official methods of analysis, 18th ed. AOAC, Washington D.C.
Cao, Y., Wang, D., Wang, L., Wei, X., Li, X., Cai, C., Lei, X., Yao, J., 2021. Physically effective neutral detergent fiber improves chewing activity, rumen fermentation, plasma metabolites, and milk production in lactating dairy cows fed a high-concentrate diet. J. Dairy Sci. 104, 5631–5642.
Erdogan, S., Kaya, M., 2016. High similarity in physicochemical properties of chitin and chitosan from nymphs and adults of a grasshopper. Int. J. Biol. Macromol. 89, 118–126.
Estetika, Y., Endrawati, Y.C., 2018. Silkworm productivities of Bombyx mori L. BS-09 race in tropic. J. Ilmu. Produksi. dan. Teknol. Has. Peternak. 6, 104–112. (In Indonesian)
Gao, M., El-Sherbiny, M., Szumacher-Strabel, M., Cieślak, A., Yanza, Y.R., Irawan, A., Xie, B., Cao, Z.J., Fusaro, I., Jalal, H., Abd El Tawab, A.M., Liu, Y.B., 2024. Cutting-edge exploration of insect utilization in ruminant nutrition—feature and future: a systematic review and meta-analysis. Front. Vet. Sci. 11, 1–15.
Herliatika, A., Permatahati, D., Pujiawati, Y., Tresia, G.E., Wina, E., 2021. Utilization of cricket (Gryllus bimaculatus), black soldier fly (Hermetia illucens), mealworm (Tenebrio molitor), and silkworm (Samia ricini dan Bombyx mori) as alternative feed protein sources. Indones. Bull. Anim. Vet. Sci. 31, 185. (In Indonesian)
Hlasny, J., 2024. A cause of bovine spongiform encephalopathy (BSE) related to the feeding of meat and bone meal (MBM) to British cows can be ruled out based on known circumstances. Sér. A. 1, 32–42.
Jayanegara, A., Gustanti, R., Ridwan, R., Widyastuti, Y., 2020. Fatty acid profiles of some insect oils and their effects on in vitro bovine rumen fermentation and methanogenesis. Ital. J. Anim. Sci. 19, 1311–1318.
Jayanegara, A., Novandri, B., Yantina, N., Ridla, M., 2017a. Use of black soldier fly larvae (Hermetia illucens) to substitute soybean meal in ruminant diet: An in vitro rumen fermentation study. Vet. World. 10, 1439–1446.
Jayanegara, A., Yantina, N., Novandri, B., Laconi, E.B., Nahrowi, Ridla, M., 2017b. Evaluation of some insects as potential feed ingredients for ruminants: Chemical composition, in vitro rumen fermentation and methane emissions. J. Indones. Trop. Anim. Agric. 42, 247–254.
Ji, H., Zhang, J.-L., Huang, J.-Q., Cheng, X.-F., Liu, C., 2015. Effect of replacement of dietary fish meal with silkworm pupae meal on growth performance, body composition, intestinal protease activity, and health status in juvenile Jian carp (Cyprinus carpio var. Jian). Aquac. Res. 46, 1209–1221.
Lähteenmäki-Uutela, A., Marimuthu, S.B., Meijer, N., 2021. Regulations on insects as food and feed: a global comparison. J. Insects as Food Feed 7, 849–856.
Makkar, H.P.S., Tran, G., Heuzé, V., Ankers, P., 2014. State-of-the-art on use of insects as animal feed. Anim. Feed Sci. Technol. 197, 1–33.
Moss, A., Jouany, J., Newbold, J., 2000. Methane production by ruminants: its contribution to global warming. Ann. Zootech. 49, 231–253.
Paulin, I.G., Purwanto, M.G., 2020. Nutritional characteristics of Teak Grasshopper (Valanga nigricornis Burmeister), Cricket (Brachytrupes portentosus L.), and Mealworm (Tenebrio molitor) as alternative food sources in Indonesia. Indones. J. Biotechnol. Biodivers. 4, 52–61.
Pengpeng, W., Tan, Z., 2013. Ammonia assimilation in rumen bacteria: a review. Anim. Biotechnol. 24, 107–128.
Purbowati, E., Rianto, E., Dilaga, W.S., Sri Lestari, C.M., Adiwinarti, R., 2014. Characteristics of the rumen fluids, type and number of ruminal microbes in Java and Ongole grade bulls. Bul. Peternak. 38, 21. (In Indonesian)
Putra, L.O., Suharti, S., Sarwono, K.A., Sutikno, S., Fitri, A., Astuti, W.D., Rohmatussolihat, R., Widyastuti, Y., Ridwan, R., Fidriyanto, R., Wiryawan, K.G., 2023. The effects of heat-moisture treatment on resistant starch levels in cassava and on fermentation, methanogenesis, and microbial populations in ruminants. Vet. World. 16, 811–819.
Rahmasari, R., Sumiati, S., Astuti, D.A., 2014. The effect of silkworm pupae (Bombyx mori) meal to substitute fish meal on production and physical quality of quail eggs (Cortunix cortunix japonica). J. Indones. Trop. Anim. Agric. 39, 180–187.
Rashmi, K.M., Chandrasekharaiah, M., Soren, N.M., Prasad, K.S., David, C.G., Thirupathaiah, Y., Shivaprasad, V., 2018. Effect of dietary incorporation of silkworm pupae meal on in vitro rumen fermentation and digestibility. Indian J. Anim. Sci. 88, 731–735.
Rashmi, K.M., Chandrasekharaiah, M., Soren, N.M., Prasad, K.S., David, C.G., Thirupathaiah, Y., Shivaprasad, V., 2022. Defatted silkworm pupae meal as an alternative protein source for cattle. Trop. Anim. Health Prod. 54.
Renna, M., Coppa, M., Lussiana, C., Le Morvan, A., Gasco, L., Maxin, G., 2022. Full-fat insect meals in ruminant nutrition: in vitro rumen fermentation characteristics and lipid biohydrogenation. J. Anim. Sci. Biotechnol. 13, 138.
Ridla, M., Asikin, N., Jayanegara, A., Samsudin, A.A., 2023. Mitigating methane emissions for rural tropical livestock through selecting appropriate grass types. Livest. Res. Rural. Dev. 35, 7.
Ridla, M., Nahrowi, 2025. Methane mitigation strategies by optimizing nutrient profiles in an eco-friendly mixture of cassava pulp and Indigofera zollingeriana branch silage with strategic protein supplementation. Adv. Anim. Vet. Sci. 13, 198–208.
Rodríguez-Rodríguez, M., Barroso, F.G., Fabrikov, D., Sánchez-Muros, M.J., 2022. In vitro crude protein digestibility of insects: a review. Insects. 13, 1–19.
Sagala, Y.G., Andadari, L., Handayani, T.H., Sholikin, M.M., Fitri, A., Fidriyanto, R., Rohmatussolihat, R., Ridwan, R., Astuti, W.D., Widyastuti, Y., Fassah, D.M., Wijayanti, I., Sarwono, K.A., 2024. The effect of silkworms (Bombyx mori) chitosan on rumen fermentation, methanogenesis, and microbial population in vitro. Vet. World. 17, 1216–1226.
Sahib, Q.S., Ahmed, H.A., Ganai, A.M., Farooq, J., Sheikh, G.G., Sheikh, I.U.D., Beigh, Y.A., 2023. Evaluation of silkworm pupae meal based calf starter diet on the performance of crossbred cattle calves. Indian J. Anim. Sci. 93, 903–906.
Samal, L., Pattanaik, A., 2014. Dairy production in India - Existing scenario and future prospects. Int. J. Livest. Res. 4, 105.
Sarwono, K.A., Kondo, M., Ban-Tokuda, T., Jayanegara, A., Matsui, H., 2019. Effects of phloroglucinol and the forage: Concentrate ratio on methanogenesis, in vitro rumen fermentation, and microbial population density. Adv. Anim. Vet. Sci. 7, 164–171.
Sarwono, K.A., Rohmatussolihat, R., Watman, M., Ratnakomala, S., Astuti, W.D., FidriyantoR., Ridwan, R., Widyastuti, Y., 2022. Characteristics of fresh rice straw silage quality prepared with addition of lactic acid bacteria and crude cellulase. AIMS Agric. Food. 7, 481–499.
Sheikh, I.U., Banday, I.M., Shaista Nissa, I.S., Bushra Zaffer, I., Bulbul, I.K., Husbandry, A., Nissa, S.S., Zaffer, B., Bulbul, K., 2018. Utilization of silkworm pupae meal as an alternative source of protein in the diet of livestock and poultry: A review. J. Entomol. Zool. Stud. 6, 1010–1016.
Siemianowska, E., Kosewska, A., Aljewicz, M., Skibniewska, K.A., Polak-Juszczak, L., Jarocki, A., Jędras, M., 2013. Larvae of mealworm (Tenebrio molitor L.) as European novel food. Agric. Sci. 04, 287–291.
Theodorou, M.K., Williams, B.A., Dhanoa, M.S., McAllan, A.B., France, J., 1994. A simple gas production method using a pressure transducer to determine the fermentation kinetics of ruminant feeds. Anim. Feed Sci. Technol. 48, 185–197.
Thirumalaisamy, G., Malik, P.K., Kolte, A.P., Bhatta, R., 2020. In vitro evaluation of graded level of Silkworm pupae (Bombyx mori) oil on methane production, fermentation characteristics, and protozoal populations. Vet. World. 13, 586–592.
Toral, P.G., Hervás, G., González-Rosales, M.G., Mendoza, A.G., Robles-Jiménez, L.E., Frutos, P., 2022. Insects as alternative feed for ruminants: comparison of protein evaluation methods. J. Anim. Sci. Biotechnol. 13, 1–8.
Ullah, R., Khan, S., Hafeez, A., Sultan, A., Khan, N.A., Chand, N., Ahmad, N., 2017. Silkworm (Bombyx mori) meal as alternate protein ingredient in broiler finisher ration. Pak. J. Zool. 49, 1463–1470.
Van Dung, D., Ba, N.X., Van, N.H., Phung, L.D., Ngoan, L.D., Cuong, V.C., Yao, W., 2013. Practice on improving fattening local cattle production in Vietnam by increasing crude protein level in concentrate and concentrate level. Trop. Anim. Health Prod. 45, 1619–1626.
Williams, S.R.O., Hannah, M.C., Eckard, R.J., Wales, W.J., Moate, P.J., 2020. Supplementing the diet of dairy cows with fat or tannin reduces methane yield, and additively when fed in combination. Animal. 14, s464–s472.
Wu, X., He, K., Velickovic, T.C., Liu, Z., 2021. Nutritional, functional, and allergenic properties of silkworm pupae. Food Sci. Nutr. 9, 4655–4665.
Xue, F., Sun, F., Jiang, L., Hua, D., Wang, Y., Nan, X., Zhao, Y., Xiong, B., 2019. Effects of partial replacement of dietary forage using kelp powder (Thallus laminariae) on ruminal fermentation and lactation performances of dairy cows. Animals. 9, 852.
Yanuartono, Nururrozi, A., Soedarmanto, I., Purnamaningsih, H., Ramadani, D., 2020. Meat bone meal as an alternative animal feed: a brief review. J. Peternak. Sriwij. 9, 35–54. (In Indonesian).