i ASSESSMENT OF REARING TECHNIQUE FOR THE BLACK SOLDIER FLY AND TERMITE COLLECTION TECHNIQUE FOR USE BY SMALLHOLDER POULTRY AND FISH FARMERS IN GHANA. BY HETTIE ARWOH BOAFO (10129957) THIS THESIS IS SUBMITTED TO THE UNIVERSITY OF GHANA LEGON, IN PARTIAL FULFILMENT OF THE REQUIREMENT FOR THE AWARD OF PhD IN ENTOMOLOGY DEGREE UNDER THE INSECT SCIENCE PROGRAMME* UNIVERSITY OF GHANA NOVEMBER, 2020 ⃰ JOINT INTERFACULTY INTERNATIONAL PROGRAMME FOR THE TRAINING OF ENTOMOLOGISTS IN WEST AFRICA: DEPARTMENT OF ANIMAL BIOLOGY AND CONSERVATON SCIENCES (FACULTY OF SCIENCE) AND THE CROP SCIENCE DEPARTMENT (COLLEGE OF AGRICULTURE AND CONSUMER SCIENCE), UNIVERSITY OF GHANA, LEGON University of Ghana http://ugspace.ug.edu.gh ii DECLARATION This is to certify that this thesis is the result of original research undertaken by Boafo Hettie Arwoh of the African Regional Postgraduate Programme in Insect Science, University of Ghana, under the supervision of Prof. Vincent Eziah, Dr Eric Cofie Timpong - Jones, and Dr Maxwell Billah. This research has not been included in any thesis or dissertation submitted to any other institution for a degree or any qualification. Authors whose works were used have been duly referenced/ recognised. ………………………….. Date: 2 September 2022 Boafo Hettie Arwoh (10129957) ………………………….. Date: 2 September 2022 Prof. Vincent Eziah (Supervisor) Department of Crop Science, University of Ghana, Legon Date: 2 September 2022 Dr Eric Cofie Timpong-Jones (Supervisor) Department of Animal Science/ LIPREC, University of Ghana, Legon Date: 2 September 2022 Dr Maxwell Billah (Supervisor) Department of Animal Biology and Conservation Science University of Ghana http://ugspace.ug.edu.gh iii ABSTRACT Insects offer a cheap source of protein for especially smallholder poultry and fish farmers. Insects contain about 40-70 % protein, 35 % fat, and other micro-nutrients. The black soldier fly (BSF) larvae and termites are promising insects that can replace the animal protein component of livestock feed. The full adoption of their use by farmers is however slow due to insufficient scientific data on the production of BSF larvae and termite collection techniques. In this study, the existing production techniques for BSF larvae and collection of termites were evaluated to recommend the most efficient for adoption by smallholder poultry and fish farmers. Six organic substrates (pito mash, millet porridge mash, pig manure, chicken manure, fruit waste, and waste from roots and tubers) known to be suitable for BSF larvae production were evaluated for their suitability as oviposition attractants and larval development. The substrates were first exposed outdoors to measure the quantity of eggs laid on them by naturally occurring BSF females. The quality of the substrate(s) as larval rearing media was also tested by placing a standard amount of egg mass to measure the individual and total weights of prepupae obtained, total number, and development time. The nutritional profile of the prepupae and the substrates were also determined. Furthermore, the production of BSF larvae under natural oviposition (in garden bins) was assessed by varying the rate of loading substrate (pito mash) and the quantities of the substrate on the overall prepupal harvest. The substrate used significantly influenced the quantity of eggs laid and the development of the resulting prepupae but the substrates most favourable for larval development were not the most favoured by gravid BSF for oviposition. In the oviposition tests, millet porridge mash was the most attractive substrate whereas only a few eggs were recovered from the other substrates. All substrates allowed the successful development of larvae but pig manure was more productive than the others. The crude protein content of the larvae ranged between 35 - 43%, with the shortest development time of 16 days. Applying small quantities of substrates at a constant rate (10 kg University of Ghana http://ugspace.ug.edu.gh iv per week) in garden bins produced higher prepupal yields than larger quantities (20 kg). Unlike BSF larvae, termites cannot be easily produced but are obtained from chippings of mounds or by trapping using containers with filled organic matter. The commonly used termite collection method was assessed to quantify the amount of termites harvested with commonly used organic matter. Furthermore, indigenous knowledge on the use of termites as poultry feed in Ghana and factors affecting its use were assessed. Containers filled with the four commonly used organic matter (mango seed, maize cobs, dried cow dung, yam peels, and their mixtures) were placed on trails of termites to quantify the daily harvest. Surveys were also conducted in four regions in Ghana to collect information, by the administration of questionnaires, on the use of termites as poultry feed, termite species collected, species not used, and collection methods. Samples of termite species mentioned were collected and identified to the genus level. Twenty- three per cent and 19% of farmers mentioned that termites are always or often used to feed poultry whereas 11% never use termites. A binomial regression analysis showed that termite use was affected by region, sex, education, farm size, and income. Termites collected belonged to eight genera, the main ones being Macrotermes, Trinervitermes, and Odontotermes. Five collection methods are used to obtain termites and involve either breaking mounds or using containers as traps. Collection methods vary with species and region and the abundance of termite genera varies with season. Farmers identified some species as poisonous to poultry. A Kruskal-Wallis test showed that there were significant differences in the quantity of termites collected using different substrates for both Odontotermes species and Macrotermes species. A mixture of corn cobs and yam peels yielded the highest dry weight harvest of 14.8 g/day in Macrotermes species. Likewise, the mixture of mango seed and cow dung gave the highest average yield of 19.40 g/day dry weight of Odontotermes species. Termites and black soldier fly larvae are important in indigenous poultry production because they are a readily available and cheap protein source for local farmers. University of Ghana http://ugspace.ug.edu.gh v DEDICATION I dedicate this thesis to my dad, Mr J.K Boafo, who has always encouraged me to become a scholar, and to my mother and siblings, for their prayers and support. University of Ghana http://ugspace.ug.edu.gh vi ACKNOWLEDGMENT To God be the glory for how far He has brought me in this life. I acknowledge the contribution of Dr Marc Kenis of CABI, Switzerland, who contracted me for this research work, and for his immense contribution in sharing experience and linking me up to other professionals in the field, without him, I would not have completed my PhD programme. Also, to Dr Victor Attuquaye Clottey of CABI- WAC, Ghana for giving me this opportunity and for encouraging me. I am most indebted to the Swiss Agency for Development and Cooperation and the Swiss National Science Foundation, in the framework of the Swiss Program for Research on Global Issues for Development (R4D) for funding my entire PhD programme. My profound appreciation goes to my supervisors, Dr Eric Timpong-Jones, Prof. Vincent Eziah, and Dr Maxwell Billah, for taking the time to direct and make suggestions for the research work. I thank the poultry farmers of the four regions, especially Afa Musa of Tonjin for assisting me with the testing of termite collection methods. To my friends, Adom Médétissi, Osei Kwaku, Eli Dzikunu, Dr Richard Minkah, Dr Aubin Amagnide, Dr Nancy Aweh, Dr Charlemagne Gbemavo, Jeffery Edue of CABI-WAC, Dr Owusu Fordjour and Dr Shaphan Chia for their contributions and encouragement. Finally, to my sister Esther Boafo, the Library assistant turned entomologist, for assisting me in carrying out my research work, I say God bless you! University of Ghana http://ugspace.ug.edu.gh vii TABLE OF CONTENT DECLARATION...................................................................................................................... ii ABSTRACT ............................................................................................................................ iii DEDICATION.......................................................................................................................... v ACKNOWLEDGMENT ........................................................................................................ vi TABLE OF CONTENT ......................................................................................................... vii LIST OF TABLES .................................................................................................................. xi LIST OF FIGURES ............................................................................................................... xii LIST OF PLATE.................................................................................................................. xiii LIST OF ABBREVIATIONS .............................................................................................. xiv CHAPTER ONE ...................................................................................................................... 1 1.0 General Introduction ......................................................................................................... 1 1.1 Background to study ......................................................................................................... 1 1.3 Broad Objective ................................................................................................................ 5 1.4 Specific Objectives ........................................................................................................... 5 CHAPTER TWO ..................................................................................................................... 6 2.0 Literature Review .............................................................................................................. 6 2.1 Edible Insects .................................................................................................................... 6 2.2 The use of insects as Animal Feed ................................................................................... 8 2.3 Nutritional value of edible insects .................................................................................... 9 2.4 Safety of insects as food and feed .................................................................................. 11 2.5 Farming Edible Insects ................................................................................................... 14 2.6 Advantages of farming insect ......................................................................................... 16 2.7 Nutritional requirements of insects ................................................................................. 18 2.8 The black soldier fly ....................................................................................................... 20 2.9 The life cycle of black soldier flies, Hermetia illucens .................................................. 21 2.10 The effect of diet (substrate) on growth, development, and nutrient composition of black soldiers fly flies ........................................................................................................... 23 2.11 Substrates Suitable for rearing black soldier fly larvae ................................................ 25 2.12 Termites ........................................................................................................................ 26 2.13 Termites as food and feed ............................................................................................. 28 University of Ghana http://ugspace.ug.edu.gh viii 2.14 Collection of termites ................................................................................................... 29 2.15 Poisonous termites ........................................................................................................ 30 CHAPTER THREE ............................................................................................................... 32 3.0 Evaluate black soldier fly larvae rearing systems ......................................................... 32 3.1 Introduction .................................................................................................................... 32 3.2 Hypothesis 1: The substrates most attractive for black soldier fly (BSF) oviposition are also those that are most suitable for larval development. ..................................................... 34 3.2.1 Methodology ............................................................................................................ 34 3.2.2 Determining the substrate(s) that can maximize oviposition by female black soldier flies .................................................................................................................................... 36 3.2.3 Determining the substrate(s) appropriate for enhanced larvae development ........... 37 3.2.4 Data analysis ............................................................................................................ 39 3.2.5 Results ...................................................................................................................... 41 3.2.5.1 Proximate analysis of substrates ........................................................................ 41 3.2.5.2 Determine the substrate(s) appropriate for enhanced larval development ......... 44 3.2.6 Discussion ................................................................................................................ 47 3.2.7 Conclusion ................................................................................................................ 53 3.3.1 Introduction .............................................................................................................. 53 3.3.2 Methodology ............................................................................................................ 54 3.3.3 Data analyses ............................................................................................................ 56 3.3.5 Discussion ................................................................................................................ 62 3.3.6 Conclusion ................................................................................................................ 67 CHAPTER FOUR .................................................................................................................. 68 4.0 Identification of Other Insect Species that may Populate the Rearing Substrates .... 68 4.1 Introduction .................................................................................................................... 68 4.2 Methodology ................................................................................................................... 69 4.2.1 Collection of other insect species attracted to the different rearing substrates ........ 69 4.2.2 Assessing the incidence of parasitism in a BSF production system ........................ 69 4.3 Data Analysis .................................................................................................................. 70 4.4 Results ............................................................................................................................ 70 4.5 Discussion ....................................................................................................................... 72 4.6 Conclusion ...................................................................................................................... 74 University of Ghana http://ugspace.ug.edu.gh ix CHAPTER FIVE ................................................................................................................... 76 5.0 Termites as supplementary protein sources for poultry in Four Regions of Ghana . 76 5.1 Introduction .................................................................................................................... 76 5.2 Methodology ................................................................................................................... 77 5.2.1 Study area ................................................................................................................. 77 5.2.2 Surveys to ascertain the use of termites in the four regions ..................................... 79 5.2.3 Identification of the termite species collected .......................................................... 80 5.3 Data Analysis .................................................................................................................. 80 5.4 Results ............................................................................................................................ 81 5.4.1 The use of termites as supplementary feed for poultry ............................................ 81 5.4.2 Identification of the termite species cited by respondents ....................................... 83 5.4.3 Termite collection methods ...................................................................................... 86 5.4.4 Relationship between collection method and region of study.................................. 89 5.4.5 Seasonal availability of termites .............................................................................. 91 5.4.6 Poisonous/ toxic termite species .............................................................................. 92 5.5. Discussion ...................................................................................................................... 92 5.5.1 The use of termites as supplementary feed for poultry ............................................ 92 5.5.2 Termite species identified in the survey ................................................................... 93 5.5.3 Termite collection methods and availability during the different seasons ............... 94 5.5.4 Poisonous/toxic termite species ............................................................................... 95 5.6 Conclusion ...................................................................................................................... 97 5.7 Assessment of indigenous termite collection methods ................................................... 97 5.7.1 Introduction .............................................................................................................. 97 5.7.2 Methodology ............................................................................................................ 98 5.8 Data Analysis ................................................................................................................ 100 5.9 Results .......................................................................................................................... 100 5.9.1 Macrotermes species .............................................................................................. 100 5.9.2 Odontotermes species ............................................................................................. 101 5.10 Discussion ................................................................................................................... 102 CHAPTER SIX .................................................................................................................... 106 6.0 General discussion ........................................................................................................ 106 6.1 General Conclusions ..................................................................................................... 107 University of Ghana http://ugspace.ug.edu.gh x 6.2 Recommendations ........................................................................................................ 109 REFERENCES ..................................................................................................................... 111 LIST OF APPENDICES ..................................................................................................... 153 University of Ghana http://ugspace.ug.edu.gh xi LIST OF TABLES Table 1. The nutritional composition of the various substrates was tested. ............................ 42 Table 2. Effect of the substrate on the weight of eggs: detailed results of the linear mixed effects model .............................................................................................................. 43 Table 3. Estimate of the weekly mean weight of eggs laid and confidence intervals (CI). ..... 43 Table 4. Effect of the substrate on the weight of prepupae: detailed results of the linear mixed effects model .............................................................................................................. 45 Table 5. Estimate of the means and confidence intervals (CI) of the weight of individual prepupae ..................................................................................................................... 45 Table 6. Effect of substrate on total prepupae yields, the total number of individuals surviving to prepupae, and the time for development from egg to prepupae stage…………...46 Table 7. The nutritional composition of black soldier fly prepupae reared on different substrates .................................................................................................................... 47 Table 8. Weekly harvest of black soldier fly prepupae (mean ± SE) during the dry and wet season ......................................................................................................................... 58 Table 9. Weekly harvest of black soldier fly per kilogram of substrate for the different treatments and climatic seasons considered .............................................................. 61 Table 10. Other insect species collected from the different substrates .................................... 71 Table 11. Percentage of adult emergence of BSF larvae from enclosed and exposed pupae .. 72 Table 12. The percentage of farmers from the four regions using termites in feeding their poultry (n = 1817) ...................................................................................................... 82 Table 13. Results of the binomial linear regression showing the factors affecting the use of termites ...................................................................................................................... 83 Table 14. Percentage of farmers that used the eight termite genera to feed poultry in the four regions ........................................................................................................................ 85 Table 15. Percentage of farmers employing the different collection methods of harvesting termites ...................................................................................................................... 91 Table 16. Toxic species of termites and their symptoms of illness usually to chicks ............. 92 Table 17. Mean weight of Macrotermes species harvested using the different substrates .... 101 Table 18. Mean weight of Odontotermes species harvested using the different substrates .. 101 University of Ghana http://ugspace.ug.edu.gh xii LIST OF FIGURES Figure 1. Evolution curve of the weight of eggs in the function of the substrate and the week ................................................................................................................................... 44 Figure 2. Trends of the weekly harvest of black soldier fly larvae according to the treatment applied and climatic season considered. .................................................................... 59 Figure 3. Trends in the cumulative weekly harvest of black soldier fly prepupae according to the treatment applied and climatic season considered. .............................................. 60 Figure 4. Trends of the weekly harvest of black soldier fly prepupae per unit (kg) of substrate according to the treatment applied and climatic season considered. ......................... 61 Figure 5. Trends of the cumulative daily harvest of black soldier fly prepupae when different weights of substrates are added to bins in different seasons ..................................... 62 Figure 6. Principal component analysis results for the description of relationships between termite genera and regions. (A) correlation circle of termite genera. (B) projection of the regions in the first factorial plane formed by axes 1 and 2 defined by the termite genera ......................................................................................................................... 86 Figure 7. Principal component analysis for the description of the relationship between termite species collection method and regions. (A) correlation circle of collection methods. (B) projection of the regions in the first factorial plane formed by axis 1 and 2 defined by the collection method. .............................................................................. 90 Figure 8. The relationship between the four major termite species collected and seasonal availability ................................................................................................................. 91 University of Ghana http://ugspace.ug.edu.gh xiii LIST OF PLATE Plate 1: Life cycle of the black soldier fly, Hermetia illucens................................................. 23 Plate 2. Organic substrates tested: (A) roots and tubers; (B) pig manure; (C) pito mash; (D) fruit waste; (E) millet porridge mash (F) chicken manure ........................................ 36 Plate 3. (A) Bowl containing substrate with dried plantain leaves and oviposition cardboards; (B) flute of cardboard containing laid eggs; (C) flute opened to expose eggs .......... 37 Plate 4. (A) plastic bowls used in the incubation of eggs; (B) Incubated eggs in a screen house to prevent oviposition by other insects ...................................................................... 38 Plate 5. Experimental set up in a screen house to prevent alien fly oviposition ...................... 39 Plate 6. (A) BSF rearing bins with oviposition cardboards; (B) BSF rearing bins with the treatments arranged in a completely randomized design ........................................... 56 Plate 7: Map of Ghana showing the four regions where the study was conducted ................. 78 Plate 8. Termites species collected from the four regions ....................................................... 84 Plate 9. Collection methods used in harvesting termites: (A) method 1; (B) method 2; (C) method 3; (D) method 4; (E) method 5 ..................................................................... 87 Plate 10. Containers used in harvesting termites: (A, B, C) jerry can; (D, F) open top gourd; (E) earthen pot ........................................................................................................... 89 Plate 11. Pots filled with substrates for trapping termites in the field ..................................... 99 Plate 12. Harvesting of termites. (A) pots buried under Macrotermes species mound; (B) pot buriedon Odontotermes species nest ......................................................................... 99 Plate 13. Termites collected from the trapping. (A) Odontotermes species (B) Macrotermes species ...................................................................................................................... 100 University of Ghana http://ugspace.ug.edu.gh xiv LIST OF ABBREVIATIONS ARI Animal Research Institute BSF Black solider fly BSFL Black soldier fly larvae DM Dry matter FFA Fish for Africa FAO Food and Agricultural Organization GDP Gross Domestic product IFWA Insect as Feed in West Africa IPRI The International Food Policy Research Institute MoFAD Ministry of Fisheries and Aquaculture MOFA Ministry of Food and Agriculture MT Metric tons OECD Organization of Economic Co-operation and Development USDA United States Department of Agriculture University of Ghana http://ugspace.ug.edu.gh 1 CHAPTER ONE 1.0 General Introduction 1.1 Background to study The increase in world population is the utmost challenge facing world food production systems. Coupled with this, urbanization in developing countries will lead to a shift in food and diet patterns particularly toward livestock products (Delgado, 2003). A rise in purchase power due to increased incomes will accelerate the consumption of meat products (Steinfield et al., 2006). Hence, the demand for livestock products is expected to nearly double in Sub-Saharan Africa and Asia, from 200 kcal per person per day in 2000 to 400 kcal per person per day by 2050 (Thornton, 2010). Since the 1960s, global meat production has seen a tremendous increase, particularly for poultry meat (Asante-Addo and Weible, 2020). Chicken production increased by nearly a factor of 10, whereas beef production doubled in the same period (Landes et al., 2004; Thornton, 2010). With an estimated mean global consumption of 14.2 kg/capita in 2018 for poultry meat, its demand has surpassed pork as the favoured meat (OECD-FAO, 2019). Similarly, Ghana has experienced an increase in poultry meat consumption (Asante-Addo and Weible, 2020). Over the past decade, poultry meat consumption has become a very common addition to the Ghanaian diet, the bulk being imported poultry meat (Asante-Addo and Weible, 2020). However, Asante-Addo and Weible (2020) indicated that higher-income earning households tend to consume locally-produced birds more than lower-income earning households due to their palatability and health benefits. Poultry meat consumption increased from 1.7 to 6.1 kg per capita between the year 2000 to 2017, more than double the average for Sub-Saharan Africa (OECD-FAO, 2017). The average probability of consuming chicken in a Ghanaian household was found to be 13.5, 16.9, 25.9, 21.0, and 22.7% for occasional University of Ghana http://ugspace.ug.edu.gh 2 consumers, once a month, 2-3 times a month, once a week, and twice or more a week, respectively (Asante-Addo and Weible, 2020). Before the mid-1980s, domestic poultry production (both backyard and commercial poultry) was a very vibrant activity in the country. In the past, almost every home in Africa reared scavenging indigenous chicken for home consumption and sold it when surplus money was needed (Sonaiya, 1993). It plays a significant role in the agrarian economy of developing countries and serves as a source of livelihood for unemployed youth and women (Padhi, 2016). Concurrently, aquaculture is the fastest-growing animal-producing sector and provides half of the world’s fish for consumption (FAO, 2012a; FAO, 2016). About 8-9% of the protein consumed by humans is obtained from aquaculture (FAO, 2012a). Over the past thirty years, world aquaculture production has increased from 5 million to 65 million tons (World Bank, 2013). The aquaculture sector contributes about 3-5% of the GDP of Ghana and is a source of employment for the populace (Bank of Ghana, 2008). It represents about 11.3% of the total national fish production in the year 2016 and is projected to represent 15% in the next three to five years (MoFAD, 2016). The national production increased from over 32,512 MT per year in 2013 to 52,470.49 MT per year in 2016 and is expected to hit 72,000 tons is expected in the next 3 to 5 years (MoFAD, 2016). 1.2 Justification The indigenous poultry and aquaculture sector in Ghana have the potential to improve livelihoods, provide employment for the youth and women, and contribute to the economy. However, these two sectors still suffer from basic constraints such as high cost of feed, scarcity of feed, access to high-quality protein sources, and limited knowledge of the cost of investment (Amenyogbe et al., 2018). Generally, feed represents the highest cost in an animal production system, representing about 70% of the total cost of production (Omole et al., 2005). The two key sources of protein used University of Ghana http://ugspace.ug.edu.gh 3 in livestock feed, fishmeal and soybean meal are economically and ecologically unsustainable. Fishmeal is rapidly declining as a feed source because overexploitation of ocean resources has significantly reduced fish stocks (Naylor et al., 2000). Soybean meal, is a stable food resource for both human and animal production systems, instigating both competition and pressure on arable lands (Steinfeld et al., 2006). Indigenous poultry production, practised in almost every home suffers from qualitative and quantitative feed shortages (Dankwa, 2004; Pousga et al., 2007), leading to low live weight of birds, low egg production, and death and thus reduce family income. Similarly, smallholder fish farmers due to scarcity and high cost of feed often resort to feeding fish on low-quality feed such as rice bran, brewers waste, maize bran, and anchovies resulting in a slow growth rate and increased susceptibility of fish to diseases and eventual death. The failure of many fish farms and the sale of low-grade fish in Ghana have been attributed to low- quality fish feed and the high cost of feed ingredients (Kassam, 2014; Kaunda et al., 2010). A solution to developing sustainable household poultry farming and aquaculture systems is to use untapped local, easily available, and cheap protein sources. Insects, which are a natural food source for poultry and fish are one source. The protein in insects is comparable to that of fishmeal or soymeal and can also be produced cheaply (Heuzè and Tran, 2013). Insect larvae and pupae are a rich source of proteins (40-70% dry weight), other valuable nutrients such as iron, vitamins A and B, and other essential amino acids (DeFoliart, 1995; van Hius, 2013). The use of insects as a food and feed source is an old-age practice in Africa, Asia, and Latin America that is gaining popularity in recent times. Local farmers often feed insect larvae and termites to poultry, especially young birds (Pomalégni et al., 2016; Sankara et al., 2017). Furthermore, scavenging birds are occasionally found feeding on both insect larvae and adults on their own during their feed search. The use of insect protein to replace or supplement University of Ghana http://ugspace.ug.edu.gh 4 conventional protein sources is highly recommended by FAO as an alternative and feasible strategy to reduce food insecurity in the world (FAO, 2010). Many insects are potentially suitable as animal feed (van Huis et al., 2013), however, some of these such as caterpillars, grasshoppers, and mealworms require agricultural products which would otherwise be used for human consumption for their mass rearing. Fly larvae, on the other hand, can be produced cheaply on organic waste streams and simultaneously serve to manage organic waste produced by humans. Fly larvae are potential biodegrading agents capable of reducing a large amount of organic waste and converting them to body mass, which can be used to feed livestock and fish. The remaining digestate is also suitable manure for improving soil fertility. The most promising and commonly used fly larvae species for animal feed is the larvae of the black soldier fly, Hermetia illucens Linnaeus, 1758 (Diptera: Stratiomyidae). Black soldier fly larvae are preferred over house fly larvae because they are not known pests of any crop and are not vectors of diseases or a nuisance to humans (Kenis et al., 2018). Termites are also commonly used by farmers throughout West Africa (Chrysostome, 1997). Termites collected from chippings of termite mounds or by trapping from mounds are usually fed to chicks and guinea fowl keets to promote growth and increase egg production. A preliminary survey in 2015, showed a moderate use of fly larvae and termites as feed by smallholder poultry and fish farmers. About 90% percent of farmers (sample size 1960) surveyed in parts of Ghana used termites and 9% produced fly larvae (house fly) as feed for their poultry (unpublished report by CSIR-ARI). Furthermore, surveys in Benin and Burkina Faso showed that 5.7% and 15.6% of indigenous farmers produced fly larvae respectively, to feed their poultry (Pomalégni et al., 2016; Sanou et al., 2019). Other farmers also collect fly larvae from decomposing waste when they chance on it to feed their poultry. Similarly, Sankara et al. (2018) reported that 78% of farmers in Burkina Faso use termites as poultry feed. University of Ghana http://ugspace.ug.edu.gh 5 The full adoption of this novel approach is slow due to insufficient data on production techniques of fly larvae (Kensi et al., 2018), concerns about the nutritional quality of the resulting larvae when different substrates are used, and collection (termites) and production (fly larvae) of sufficient quantities to replace the animal protein component of fish and poultry feed. The project ‘Insects as Feed in West Africa (IFWA)’ was therefore initiated to develop appropriate methods for fly larvae and termites production and utilization in smallholder systems in West Africa, based on waste material. The focus of this study was to evaluate the insect rearing and collection methods used by smallholder farmers, which will be complemented by studies by other researchers on the economic, environmental safety and nutritional acceptability by poultry. 1.3 Broad Objective Evaluate the production of black soldier fly (BSF) larvae and termite collection techniques. 1.4 Specific Objectives 1. To assess the suitability of different substrate(s) as rearing media. 2. To evaluate the production of BSF larvae under natural oviposition in garden bins. 3. To collect indigenous knowledge on the use of termites as poultry feed. 4. To evaluate termite collection methods University of Ghana http://ugspace.ug.edu.gh 6 CHAPTER TWO 2.0 Literature Review 2.1 Edible Insects The consumption of insects by humans and use as livestock feed is an old-age practice commonly seen among the people of Africa, Asia, and Latin America. Perhaps, the earliest record of entomophagy is found in the old testament of the Holy Bible. “Of these, you may eat any kind of locust, katydid, cricket or grasshopper” (Leviticus 11: 22, New International version). John the Baptist was said to be a man whose meat was locusts and wild honey (Matthew 3:4). In Africa, entomophagy was reported as far back as 1685 by Simon van der Stel in his expedition to Namaland, south of Namibia (Waterhouse, 1924; Palmer and Pitman, 1972). Insect as food is known to supplement the dietary needs of about 2 billion people worldwide (Makkar et al., 2014). Many of the edible insects are collected from the forest/wild by local people and was consumed or sold to raise funds to supplement the family budget. Overall, about 1,900 species of insects are consumed by humans (van Huis, 2013). A hundred and four families from 14 orders of insects are consumed worldwide (Malaisse, 2005). A total of 470 species of edible insects are consumed in Africa alone (Kelemu et al., 2015). Out of this, 256 are eaten in the Central African Republic, 164 in Southern Africa, 100 in Eastern Africa, 91 in West Africa, and only 9 species in North Africa (Kelemu et al., 2015). Commonly consumed species are Orthopterans, Hymenopterans, Coleopterans, Isopterans, Homopterans and Heteropterans, and caterpillars of Lepidopterans. About 30% of total consumed insects in Africa are Lepidopteran, 29% Orthopteran, 19% Coleopteran, 7% Hymenopteran, and 15% belonging to Heteropteran, Homopteran, Isopteran, Dipteran, and Odonata (van Hius, 2005). Popular comestible lepidopteran families include Saturniidae, Notodontidae, and Sphingidae (Malaisse, 2005). In almost every part of Sub-Saharan Africa, the species Cirina forda University of Ghana http://ugspace.ug.edu.gh 7 (Westwood), Bunaea alcinoe (Stoll), and Anaphe panda are eaten (Kelemu et al., 2015). Orthopteran species widely consumed across the African continent include Schistocerca gregaria, Locusta migratoria migratorioides, Nomadacris septemfasciata, Locustana pardalina, and Anacridium melanorhodon melanorhodon (Kelemu et al., 2015). Alates, soldiers, queens, nymphs, and eggs of Macrotermes bellicosus, Macrotermes subhyalinus, Macrotermes falciger, and Macrotermes natalensis are of continent-wide importance in terms of consumption (van Hius, 2003). The honey produced and larvae of Apis mellifera Linnaeus and A. melliferra adansoni Latreille are the main species of Hymenoptera consumed across the African continent (Takeda, 1990; Muthali and Mughogho, 1992). Edible insects are an important protein source in the diet of many people in Africa. In Congo, more than 40% of the animal protein in the diets in some parts of the country is insect protein (Gomez et al., 1961). On average, the major source of protein is obtained from 300g of caterpillars consumed per week and 96 tonnes annually by households in Kinsasha (Vantomme et al., 2004). In the Central African Republic, the protein intake of 95% of the forest people is obtained from insects (FAO, 2004) and sometimes, serves as the sole source of essential proteins available to them (van Huis, 2013). In contrast to the popular belief that insects are eaten due to the scarcity of food or starvation, research confirms the contrary, that they are a delicacy for many people (DeFoliart, 1999; van Hius, 2003; Kelemu et al., 2015). In Uganda, during November when the tettigoniid Ruspolia differens, commonly called “nsenene” appears in large numbers, the sale of meat and fish dwindles as there is a preference for the insect (Mulissa, 1997). The season of “nsenene” is waited upon with much anticipation among these people. The edible moths, Anaphe venata Butler and C. forda Westwood are extensively marketed in Nigeria and purchased for about twice the price of beef (Agbidye and Nongo, 2009; Agbidye et al., 2009). In Western Kenya, the Luo people living along Lake Victoria, consume the black ant Carebara vidua for its University of Ghana http://ugspace.ug.edu.gh 8 nutritional and medicinal properties (Ayieko et al., 2012). Ghana is not left out, as larvae of the palm weevil, Rhynchophorus phoenicis known in the Akan language as “akokono” is a traditional delicacy enjoyed by the locals (Anankware, 2016). Indeed, insect protein is a valuable source of nutrients for many populations across the world and the African continent. They are readily available and a cheap source of protein that can satisfy the need and ease the problem of food insecurity in an ever-increasing population of the world. 2.2 The use of insects as Animal Feed Information on the traditional use of insects as animal feed is scarce; mostly restricted to anecdotal reports in general articles and reviews (Hein et al., 2005; Kenis et al., 2014) or technical notes and unpublished thesis (Farina et al., 1991; Chrysostome, 1997; Naidoo, 2000; Chrysostome, 2009; Diawara, et al., 2013,). In the available records, termite use has been the most frequently cited as a feed supplement given by local farmers to their poultry. Termites collected from chippings of mounds or trapped using containers are sometimes the only available protein at the disposal of indigenous farmers. In KwaZulu Natal, South Africa, rural farmers feed their local chickens with termites and ants (Naidoo, 2000). The extensive use of termites as supplementary feed by indigenous poultry farmers is reported in Benin and Burkina Faso (Chrysostome, 1997; Chrysostome et al., 2009; Diawara et al., 2013; Sankara et al., 2017). Moreover, Sankara et al. (2017) revealed that 78% of the respondents interviewed in parts of Burkina Faso used termites at least occasionally to feed their poultry. In another report of a study conducted in Benin, 5.7% of farmers interviewed produced and used fly larvae (Musca domestica) as supplementary feed for indigenous poultry (Pomalégni et al., 2016). University of Ghana http://ugspace.ug.edu.gh 9 However, in several recent studies, experimental works have demonstrated the successful replacement of the protein component of livestock feed (poultry, fish, and pig) with insect protein (Bondari and Sheppard, 1981; Téguia et al., 2002; Agunbiade et al., 2007; Stammer et al., 2014;). The larvae of the black soldier fly (H. illusecns) and house fly (M. domestica) dominate these studies (Mohammed et al., 2017; Zhou et al., 2017; Dabbou et al., 2018; Wallace et al., 2018; Belghit et al., 2019). Pupae of the silkworm Bombyx mori and larvae of mealworm Tenebrio molitor have been proven as an excellent replacement for the protein component of livestock feed (Bovera et al., 2015; Jin et al., 2016; Asimi et al., 2017; Sheikh et al., 2018). Newton et al. (1977) successfully reared black soldier fly larvae on dried swine manure. Bondari and Sheppard (1981) demonstrated the possibility of replacing the conventional protein with black soldier fly larvae to feed catfish and tilapia. A diet of 1:1 fish meal and maggot meal ratio resulted in high egg production in old layer (50 weeks old) hens (Agunbiade et al., 2007). Furthermore, Téguia et al. (2002), showed that the replacement of fish meal at different levels with maggot meal in the starter and grower-finisher diets for broiler resulted in significantly higher final weight gained than in the control diet containing exclusively fishmeal. 2.3 Nutritional value of edible insects Insects are consumed for both nutrients and medical properties. In general, insects are known to be rich in proteins, fats, fibre, vitamins, and minerals (van Hius, 2013). The nutritional composition of edible insects is highly variable between and within species, the metamorphic life stage consumed, and the diet of the insects (Rumpold and Schlüter, 2013). The major contribution of nutrients obtained from edible insects is protein, fat, and chitin as this is the major body composition of insects (Roos, 2018). However, insects in any metabolic stage are a good source of various micronutrients that a fully functional insect requires for its metabolism (Roos, 2018). University of Ghana http://ugspace.ug.edu.gh 10 The protein composition of various insect species generally ranges between 40-70% dry weight (Rumpold and Schlüter, 2013). On dry matter bases, the black soldier fly contains 35-50% (Henry et al., 2015; Shumo et al., 2019); house fly 50-76% (Hwangbo et al., 2009; Pretorius, 2011); mealworm 46 - 70% (Ravzanaadii et al., 2012; Zhao et al., 2016) and silkworm 50% (Mitsuhashi, 2010) protein. Insect protein is comparable in quality to meat and fish protein (Srivastava et al., 2009; Mitsuhashi, 2010; Schabel et al., 2010) and protein from Acheta domesticus is superior to soy protein (Finke et al., 1989). A 100g of caterpillar provided 75% of the daily amount of proteins required by humans (Agbidye et al., 2009). When compared to 1 chicken egg, 3 pupae of silkworm had similar nutrient content (Mitsuhashi, 2010). An overview of the amino acid profile of some common edible insects showed that generally, concerning daily human amino acid requirements, these insects have a high amount of phenylalanine and tyrosine except for methionine (Rumpold and Schlüter, 2013). Insect fat range from 5% to more than 30% (DeFoliart, 1991) and is higher in pupal and larval stages than in the adult stages (Chen et al., 2009). However, others such as the palm weevil, R. phoenicis, and maguey grub have a high-fat content of about 62.1% (Omotoso and Adedire, 2007) and 58.55% respectively based on dry matter (Melo et al., 2011). The fat content of as high as 77% (dry matter) has been reported in larvae of the butterfly Phasus triangularis (Ramos-Elorduy et al., 1997). Insects are not lacking in micronutrients. Generally, the majority of insects contain high amounts of potassium, calcium, iron, magnesium (Schabel, 2010), zinc (DeFoliart, 1992), and selenium (Finke, 2002). Termites are particularly very high in iron (Banjo et al., 2006). According to Schabel (2010), caterpillars generally provide the required minerals in abundance. A 100g of caterpillars on average supply 33.5% of the minimum daily required University of Ghana http://ugspace.ug.edu.gh 11 amount of iron needed (DeFoliart, 1992). Also, insects supply several vitamins; bee broods are rich in vitamins A and D, and caterpillars in Vitamins B1, B2, and B6 (Schabel, 2010). It is noteworthy that the nutritional values of some of the insects considered above are those of wild insects collected. However, as stated earlier the nutritive values of insects highly depend on the feed consumed by the insects and this is likely to vary when insects are reared in the laboratory or semi-farm conditions on organic waste streams. 2.4 Safety of insects as food and feed The safety of insects as a food and feed source is a priority study area for the total acceptance of entomophagy. Several authors have investigated the safety of insects reared for food and feed (Klunder et al., 2012; Charlton et al., 2015; Diener et al., 2015a; van der Fels-Klerx et al., 2016; Quaye et al., 2018; van der Fels-Klerx et al., 2018; Schrögel and Wätjen, 2019). Contaminants such as veterinary medicines, heavy metals, pesticides, mycotoxins, allergens, and dioxins, occurring mainly in the substrates used in their production or their environment are of concern (van der Fels-Klerx et al., 2018). The contaminant in insect meal varies depending on the substrate used in growing the insects. Insects reared on agricultural waste are likely to contain pesticide residues and mycotoxins, whereas, those grown on animal manure contain veterinary drugs accordingly (van der Fels- Klerx et al., 2018). The bioaccumulation of these chemicals is positively correlated with the type of chemical, the concentration in the substrate, the insect species, and the growth phase of the insect (van der Fels-Klerx et al., 2016). Copper and zinc are efficiently metabolised by insects as such no correlation has been found between substrate concentration and internal concentration in insects (Maryanski et al., 2002; Vijver et al., 2003). However, the concentration of cadmium, lead, mercury, and arsenic is reported to be positively correlated with the concentrations in the substrate and the internal concentrations in the insect (Zhang et al., 2009; Diener et al., 2015; van der Lee and Oonincx, 2016). University of Ghana http://ugspace.ug.edu.gh 12 Charlton et al. (2015), reported that contaminants such as veterinary medicine, heavy metals, pesticides, and mycotoxins were below the recommended maximum concentrations by WHO, European Commission and Codex for black soldier flies, house flies, blow fly, and blue bottle flies. Low levels of mercury were detected in house fly larvae meal produced on pig manure in Ghana. Similarly, no lead contamination was found in house fly larvae meal produced on several substrates (Nkegbe et al., 2018a). However, there were concerns with the concentration of cadmium in three housefly samples analyzed (Charlton et al., 2015). Bioaccumulation of cadmium was observed in BSF and arsenic in yellow mealworms (Diener et al., 2015; van der Fel-Klerx et al., 2018). There is no evidence of accumulation of various mycotoxins in distinct insect species when fed with high concentrations of mycotoxins (van Broekhoven et al., 2014; Sanabria et al., 2017; Sanabria et al., 2019; Schrögel and Wätjen, 2019;). It seems insects can metabolize mycotoxins, but further research is needed for confirmation and to identify the metabolites formed (van Broekhoven et al., 2017). Likewise, no bioaccumulation of veterinary drugs was found in H. illuscens larvae reared on animal manure (Lalander et al., 2016), but, Charleton et al. (2015), reported the accumulation of nicarbazin in M. domestica grown on poultry manure. Information on the accumulation of veterinary drugs is limiting. The investigation of the effect of BSF larvae meal on poultry meat revealed that there was no significant effect on blood and serum parameters except for phosphorus (Dabbou et al., 2018). Red blood cell and white blood cell counts, packed cell volume, monocytes, and basophils were not affected by the inclusion of house fly larvae meal in poultry feed at different inclusion levels (Nkegbe et al., 2018b). Likewise, the total cholesterol, uric acid, albumin levels of yolk, and triglycerides were not affected by the inclusion of house fly larvae meal (Nkegbe et al., 2018b). Furthermore, insects contain toxins or allergens which may cause allergic reactions or disease in some individuals (Broekman et al., 2016). The African silkworm Anaphe venata has University of Ghana http://ugspace.ug.edu.gh 13 been associated with the seasonal ataxic syndrome and unconsciousness in Nigerians due to the presence of the enzyme thiaminase which when ingested renders thiamine (Vit B1) inactive (Adamolekun, 1993; Adamolekun et al., 1997; Nishimune et al., 2000; Moyo et al., 2014). In addition, forest insects harvested in the wild may be unwholesome due to poisoning by insecticide treatment. The collection of the desert locust Schistocerca gregaria for consumption could be detrimental to health when aerial spraying during outbreaks is used as a means of control. Poor handling, processing, and storage of insect meals can predispose them to the infestation of microorganisms, rendering them unsafe for consumption. Klunder et al. (2012) reported that bacterial levels of dried and roasted domestic house cricket were higher than boiled crickets. Mujuru et al. (2014) showed that hot-ash roasting of G. belina retained higher levels of Escherichia coli and Staphylococcus aureus than boiled and open-pan roasted. Also, they observed that solar drying of boiled samples resulted in recontamination by moulds. Several spore-forming fungi, mycotoxigenic fungi, and insect pests have been identified on stored insect meals. Aspergillus sp, Penicillium sp, Fusarium sp, Dermestes maculatus, Sitophilus zeamais, Corcyra cephalonica, Tribolium confusum, Tribolium castaneum, Oryzaephilus surinamensis, Bracon hebetor, Anisopteromalus cavandrae, Stathmopoda species, and mites were collected on mopane caterpillar stored for five months (Mpuchane et al., 2000). Furthermore, due to their richness in nutrients and moisture, insects offer a favourable environment for many microbial organisms (Klunder et al., 2012). Pathogenic and non- pathogenic microflorae have been isolated from the gut and body walls of some edible insects. Pathogenic microflorae such as Staphylococcus aureus, Aspergillus tamarrii, and Bacillus cereus were isolated from the body wall and gut of the common housefly Musca domestica reared on fresh fish (Banjo et al., 2005). Freshly harvested palm grubs contained Escherichia coli, Staphylococcus sp., and Klebsiella aerogenes (Opare et al., 2012). University of Ghana http://ugspace.ug.edu.gh 14 The levels of Salmonella sp. and Klebsiella sp. recorded in palm weevil exceeded the recommended standard for meat consumption by the food administration manual of the ministry of health for New Zealand (Opare et al., 2012). There is, however, no evidence of insects harbouring pathogenic viruses, although they could as vectors in their transmission (van der Fel-Klerx et al., 2018). Nonetheless, proper treatment before consumption has been reported to be effective in eliminating most microbial especially Enterobacteriaceae, but to a lesser extent spore-forming bacterium (Klunder et al., 2012). 2.5 Farming Edible Insects Although the exploitation of insects has gone on for thousands of years, only three insect species (honeybee, silkworm, and cochineal) have been fully domesticated. The majority of edible insects are obtained through collection or harvesting from nature. However, in recent years, with the exigency to find alternate sources of protein to curb food insecurity, the domestication of edible insects has become a necessity. Also, wild collections are seasonal, and their availability is threatened by increased deforestation, agricultural intensification, and environmental pollution due to insecticidal spraying. Moreover, increased demand has led to the overexploitation of wild resources. Semi-domestication and farming of edible insects are essential for sustainability, continuity, and reduced cost if insects are to become a component of stable diets or as livestock feed ingredients. There is a need for a regular supply of large quantities and high-standard qualities of insects to be produced for use as food and feed. Insect farming is a rising economic venture. A 14% increase in the livestock feed market between 2011 and 2015 (van Huis and Oonincx, 2017) created a potential opportunity for insects as a resource for this industry. The most common species of insects farmed are Gryllodes sigillatus, Gryllus bimaculatus, Acheta domestius, Tenebrio molitor, Zophobas University of Ghana http://ugspace.ug.edu.gh 15 morio, Alphitobius diaperinus, Locusta migratoria, Pachnoda marginata peregrina, Blaptica dubia, Rhynchophorus ferrugineus, Hermetia illuscens, and Musca domestica (Durst and Hanboonsong, 2015). Semi-domestication of insects is ongoing in Vietnam, Laos, and Thailand on a community scale (Dicke et al., 2019). Farmers, for example, enhance weaver ant populations by providing stringing ‘ant highways’ from vines to assist movement from tree to tree and also providing household food scraps for ants to enhance growth and nest formation (van Mele and Cuc, 2007; Hanboonsong and Durst, 2014). In recent times, active insect farming is a lucrative venture in Thailand with over 20,000 local people engaged in small-to-medium scale enterprises (Durst and Hanboonsong, 2014). In the USA, the cricket farming industry is a multimillion-dollar business, mostly producing insects for pet and zoo animals (Weissman et al., 2012). About 50 million crickets are produced weekly for pet and zoo animals (Weissman et al., 2012). Africa known to be among the top continents consuming insects is not left out in insect farming. In recent years, reliance on wild collections in Africa is being discouraged, as such, several institutions have developed production technologies for breeding insects such as crickets, palm weevil grubs, black soldier fly larvae, and house fly larvae. The Jaramogi Oginga Odinga University of Science and Technology has developed protocols for large-scale production of the house cricket, Gryllus bimaculatus, and black soldier fly larvae. The rearing technique for palm weevil grubs was developed and disseminated to some rural women in Ghana to empower them economically and improve nutrition (Parker et al., 2018). In Bamako-Mali, farming of house fly larvae has been going on for about a decade at the Institut d’Economie Rurale, Centre Régional de Recherche Agricole de Sotuba (Kone et al., 1998; Kone et al., 2017). The various organic waste streams that support house fly larvae growth and seasonal variations in harvest have been established. University of Ghana http://ugspace.ug.edu.gh 16 Furthermore, research on black soldier fly larvae and house fly larvae production has been conducted at the Biotechnology and Nuclear Agriculture Research Institute (BNARI) (Ewusie et al., 2019) and Animal Research Institute of the Council for Scientific and Industrial Research, respectively. The focus of their research work has been to develop techniques, assess the safety, and improve upon existing techniques for farming insects to help inform farmers on choices to make. Indigenous farmers in Burkina Faso (Sanou et al., 2018) and the Northern Region of Ghana (personal communication with farmers), produce housefly larvae to feed their poultry. Rumen content from livestock is collected, and exposed to natural fly populations for oviposition. The larvae and prepupae developed on the rumen are fed to poultry. Irrespective of the scale of domesticating insects, the overall benefits are immense. Insect farming has the potential to improve food security, alleviate poverty, and improve the nutrition of poor communities. 2.6 Advantages of farming insect Farming insects offer some environmental benefits over commercial livestock production. When compared to livestock production, insect production requires less water and land resources, emits lower greenhouse gases, has high feed conversion efficiencies, and can transform low-value organic by-products into high-quality food or feed (van Hius and Oonincx, 2017). When compared to mealworm production, 1g of chicken protein required two to three times as much land and 50% more water (Oonincx and De Boer, 2012). Similarly, a gram of protein from beef required 8-14 times more land and 5 times more water than mealworm protein production. Direct quantification of greenhouse gas emissions of five insects, three of which are edible (mealworm, house crickets, migratory locust, flower beetle, and Blaptica dubia) showed a University of Ghana http://ugspace.ug.edu.gh 17 lower greenhouse gas emission compared to livestock emissions (Oonincx et al., 2010). Oonincx and De Boer (2012), again compared the CO2 emissions between broiler chicken and mealworm protein and showed that broiler chicken emits 32-167% more CO2 than mealworm. Likewise, in Thailand, greenhouse gas emissions from poultry production were 89% higher than from cricket production (Halloran et al., 2017). Land use, water use, and greenhouse gas emissions from livestock and insect production are mainly associated with feed production in these systems (van Hius and Oonincx, 2017). A major reason that puts insects ahead of other livestock as a sustainable source of animal protein is their high feed conversion efficiency (Premalatha et al., 2011; Looy et al., 2013). Black soldier flies convert about half of their diet into edible protein (Oonincx et al., 2015b) in contrast to poultry which converts 33% (Wilkinson, 2011). The Argentinean cockroach and mealworms utilise 51-88% and 22-45%, respectively of their feed into edible protein (Oonincx et al., 2015b). Furthermore, a number of these potential insect species can be grown on organic waste streams, accumulating low-value organic by-products into high-value proteins. An attribute particularly important due to the huge quantities of organic waste generated annually worldwide. The black soldier fly, for instance, is capable of utilizing a wide range of organic waste substances from kitchen waste, animal manure, agricultural by-products, and market waste (Newton et al., 2005; Oonincx et al., 2015a; Surendra et al., 2016). Moreover, it can sterilize the waste by killing bacteria such as Escherichia coli and Salmonella enterica (Erickson et al., 2004; Liu et al., 2008). Others, such as the oriental ground cricket and mealworms can be reared on agricultural by-products (Oonincx et al., 2015a; Megido et al., 2016; Miech et al., 2016). The substrate chosen, however, is based on a legislative framework and food and feed safety issues. University of Ghana http://ugspace.ug.edu.gh 18 2.7 Nutritional requirements of insects Nutrition is the chemical requirement by an organism for its growth, tissue maintenance, reproduction, and energy required for these functions (Douglas and Simpson, 2011). Whiles insects can synthesize these chemicals, most are obtained from the food ingested (Behmer, 2009; Douglas and Simpson, 2011). “Insects as a group feed on a remarkably diverse list of organic substances” (Waldbauer, 1968). There is a high specificity in the type of food utilized by a group or species of insects (Waldbauer,1968). Nonetheless, generally, all insects have the same qualitative nutritional needs (Fraenkel, 1953, 1959). However, nutritional requirements vary with age, sex, stage of development, and physiological stress of the insect (Nation, 2001). Insects share much in common with other animals in terms of qualitative nutritional requirements (Thompson and Simpson, 2009). The basic essential macronutrients, protein, carbohydrates, and lipids required by other animals are necessary for insects as well. For example, insects share the same ten essential amino acids needed by humans (Thompson and Simpson, 2009). Nonetheless, there are specificities to their nutritional requirements. Proteins are especially a limiting nutrient for insects since they are unable to synthesize the essential amino acids (Thompson and Simpson, 2009; Barragan-Fonseca et al., 2018). These amino acids act as enzymes for transport and storage and are used for structural purposes (Behmer, 2008). The essential acids are; arginine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, and valine, these are essential dietary requirements for insects (Thompson and Simpson, 2009). These essential amino acids are important in some physiological functions in insects. For example, tyrosine (produced naturally in the body from phenylalanine) is important for the production of phenolic and quinone metabolism, a critical component of the cross-linking of proteins during sclerotization (Thompson and Simpson, 2009). Another essential nutrient required is sterols, which function in membrane formation and production of ecdysone and other moulting hormones (Behmer, University of Ghana http://ugspace.ug.edu.gh 19 2009; Thompson and Simpson, 2009). Cholesterol is the dominant tissue sterol in insects as well as many other animals. Water-soluble B vitamins (biotin, folic acid, niacin, thiamine, riboflavin, and pentatonic acid) are also particularly important for insect survival. They are the principal precursors for coenzymes of intermediary metabolism. Phytophagous insects require dietary ascorbic acid for normal function but not for other insects utilizing other types of foods (Behmer, 2009). The notable beneficial fat-soluble vitamins required by insects are tocopherol (E) and retinol (A), they aid reproduction and vision respectively. Carbohydrates, on the other hand, are non-essential for insects, but needed for energy in the absence of dietary fat or proteins (Behmer, 2009; Simpson, 2009). The ability to digest carbohydrates depends on the digestive capabilities of the insect (Behmer, 2009). However, most insects can utilize sucrose but not all non-sugars such as cellulose and dextrin (Behmer, 2009). Some species of insects such as the screw-worm fly, Chrysomya, normally feed on live animal tissues and the wax moth, Galleria, survives on artificial diets containing no carbohydrates (Behmer, 2009). There are exceptions to the extent to which carbohydrate is needed by insects. The desert locust, Schsitocerca gregaria, for example, requires at least 20 % of digestible carbohydrates in an artificial diet for optimal growth (Behmer, 2009). Similarly, the flour beetle, Tenebrio molitor, develops normally when dietary carbohydrate content is about 70 % but poorly when it drops below 40 % (Behmer, 2009). For some insects such as adult lepidopterans, dipterans, and hymenopterans, carbohydrates are predominantly the main energy source for metabolism. Sucrose acts as a phagostimulant, stimulating feeding in some insects (Thompson and Simpson, 2009). Likewise, fatty acids are non-essential for many insects. Mosquitoes and some lepidopterans are exceptions to this, requiring polyunsaturated fatty acids, a lack of it resulting in a nutritional University of Ghana http://ugspace.ug.edu.gh 20 disease called “crumpled wings” syndrome (Thompson and Simpson, 2009). In crumpled wing syndrome, the wings of newly emerged adult insects fail to expand making it impossible for the insect to fly. The fatty acid required is arachidonic acid and α -linolenic acid for mosquitoes and lepidopterans respectively (Behmer, 2009). Insects require potassium, magnesium, and phosphate in greater proportions relative to sodium, calcium, and chloride (Thompson and Simpson, 2009). Zinc and manganese are also important, aiding in the hardening of the mandibular cuticle. Insects require L-ascorbic acid and vitamin C for growth and development. Ascorbic acid is required in relatively larger amounts than vitamin C for insects (Thompson and Simpson, 2009). Deficiency in ascorbic acid results in abnormalities in moulting. Lipogenic growth factors such as choline and inositol are necessary for the proper growth and development of many insects (Thompson and Simpson, 2009). It is worth noting that insects, however, regulate the uptake of the important nutrients needed to maximize their fitness and for proper functioning. It has been demonstrated that specific proportions and amount of food is ingested to meet the daily nutritional requirement (Behmer, 2009). 2.8 The black soldier fly The black soldier fly, Hermetia illucens (Linnaeus 1758), belongs to the order Diptera and the family Stratiomyidae. It is a ubiquitous insect found in tropical, subtropical regions, and warmer temperate regions of the world (Üstüner et al., 2003). The wasp-like-looking adult measure between 15- 20mm in length, is bluish-black with yellowish-white tarsi and has two lateral translucent spots on the second abdominal segment (Hall and Gerhardt, 2002). They lack a functional mouthpart, therefore do not feed but depend on fat reserves accumulated during the larval stage and require only water during their approximately 5 to 9 days of life on earth (Tomberlin et al., 2002; Newton et al., 2005). University of Ghana http://ugspace.ug.edu.gh 21 Furthermore, adults are not known to vector any disease nor do they bite or sting humans (Sheppard et al., 2002; Čičková et al., 2015). The adults are neither attracted to human habitats nor food crops (Furman et al., 1959). The female black soldier fly oviposits near larval food sources rather than directly on the food source (Booth and Sheppard, 1984) and therefore does not come into direct contact with the organic waste to pick up pathogens or transmit pathogens. The dull whitish larvae, when mature, measure about 20mm long. They are flattened dorsoventrally with a narrow head-bearing mouthpart (Hall and Gerhardt, 2002). Unlike, adults, larvae, are voracious detritivores, consuming large volumes of fresh organic waste streams and converting them to body mass (Hardouin and Mahoux, 2003; Diener et al., 2011; van Huis et al., 2013; Makkar et al., 2014). They consume a wide variety of fresh organic waste, from decaying fruits and vegetables, restaurant waste, animal manure, market waste, fish offals, distillers’ grains, and coffee pulp to human excreta (Diener et al., 2011; van Huis et al., 2013). They have the potential to reduce up to 50% of fresh organic waste material (van Hius et al., 2013; Makkar et al., 2014), consuming about 25-500 mg of fresh matter per larvae per day (Hardouin and Mahoux, 2003; Diener et al., 2011). Moreover, due to the rapidity of decomposition of organic waste by larvae, bacteria growth is suppressed and bad odour is minimized drastically (Diener, 2011). Black soldier fly larvae are also competitors of house fly larvae by having a repulsive effect on oviposition by house fly, thus reducing the population of houseflies where they are found (Furman et al. 1959). 2.9 The life cycle of black soldier flies, Hermetia illucens Adults become sexually mature two days after emergence. A male grabs a passing female in mid-air and they descend in copulation (Tomberlin and Sheppard, 2001). A gravid female lays between 400 to 800 eggs in dry crevices near decaying organic matter (Dortmans et al., 2017). The oval-shaped, creamy white eggs hatch into larvae in about 96 hours. The larvae undergo 6 University of Ghana http://ugspace.ug.edu.gh 22 larval instars, requiring about 14 days to complete the larval phase (Hall and Gerhardt, 2002). The sixth larval instar called prepupae crawls out of the growing medium in search of a dry suitable protected environment to pupate. The mouthpart transforms into a hook that enhances easy movement out and away from the wet waste and the larvae darken into dark brown or charcoal grey colour (Dortmans et al., 2017). Pupation takes approximately 14 days and the adult emerges (Hall and Gerhardt, 2002). During this phase, the pupa becomes stiff and immobile with the posterior end slightly curved. When ready to emerge, the fly breaks off what used to be the head section, crawls out, dries up, spreads its wings, and flies off (Dortmans et al., 2017). The adults live for about 9 days under favourable conditions, find a suitable mate, copulate, and the female lays eggs and dies. Important factors for adult survival are natural light and warm temperatures between 25-32oC (Dortmans et al., 2017). The entire life cycle from the egg phase to the adult lasts for approximately 40 days (Furman et al., 1959, May 1961, Tomberlin et al., 2002; Myers et al., 2008). University of Ghana http://ugspace.ug.edu.gh 23 Plate 1: Life cycle of the black soldier fly, Hermetia illucens 2.10 The effect of diet (substrate) on growth, development and nutrient composition of black soldiers fly flies Good knowledge of the methods for the mass production of black soldier fly larvae is necessary to promote the larvae as an alternate source of protein in livestock feed. Among the demands of mass rearing, the larval diet and nutrition are of great importance (Danieli et al., 2019). The black soldier fly larvae are highly polyphagous and able to feed on a wide range of organic waste streams. The nutritional composition of the organic waste stream determines the body composition and growth of the black soldier fly (Tomberlin et al., 2002; Barragán-Fonseca et al., 2018). University of Ghana http://ugspace.ug.edu.gh 24 The quality of food (substrate) is one of the major factors that influence the development of the black soldier fly (Barragán-Fonseca et al., 2018). Several studies have shown the relationship between the nutrient composition of the organic waste stream on the crude protein, crude fat, crude fibre, ash, dry matter, and weight of the larvae (Newton et al., 2005; St-Hilaire et al., 2007; Rachmawati et al., 2010; Li et al., 2011; Oonincx et al., 2015b; Barragan-Fonseca et al., 2018; Danieli et al., 2019; Shumo et al., 2019). A highly nutritious diet produces larvae and adults of high body mass and increases the fecundity of the females (Boggs and Freeman, 2005). Protein levels are especially important for survival and larval development. Oonincx et al. (2015b) reported the reduction of the development time of larvae from 37 to 21 days when a low protein diet was replaced with a high protein and fat diet. Larvae fed with swine manure were higher in protein content than larvae fed with cow manure (Newton et al., 2005; St-Hilaire et al., 2007). Danieli et al. (2019) reported that the crude protein of prepupae reared on three different substrates was significantly different from each other. Shumo et al. (2019) also gave an account of the effect of different substrates on the crude protein, ether extract, minerals, amino acid, crude fat, and vitamins of BSFL. The substrate used for the production of larvae affected all the aforementioned proximate factors except the vitamin content of BSF larvae. In the same way, larvae fed chicken manure had higher crude fat content when compared to those grown on pig manure and cattle manure (Li et al., 2011). The survival rate and duration of development have also been reported to be positively correlated with food quality (Newton et al., 2005; Gobbi et al., 2013; Oonincx et al., 2015b; Chia et al., 2018). Larval and pupal mortalities between 1-7 % were recorded when BSF was fed hen feed diet alone, but higher (60-80 %) when given a meat diet (Gobbi et al., 2013). Likewise, development time was faster on a higher-quality diet than on a lower-quality diet, University of Ghana http://ugspace.ug.edu.gh 25 sometimes taking twice as much time (Gobbi et al., 2013; Oonincx et al., 2015b; Chia et al., 2018). Furthermore, larvae, prepupae, pupae, and adult weights are significantly affected by food quality and quantity (Chia et al., 2018; Meneguz et al., 2018). Likewise, the dry matter content of the larvae is affected by the diet but generally ranges between 20 - 44 % for fresh larvae (Sheppard et al., 2008; Diener et al., 2009; Finke, 2013; Nguyen et al., 2015; Oonincx et al., 2015a;). However, the micronutrient concentration is to a lesser extent affected by the substrate used as feed (Barragan-Fonseca et al., 2017). 2.11 Substrates Suitable for rearing black soldier fly larvae Black soldier fly larvae are the best-known species for utilizing organic waste streams (van Hius and Oonincx, 2017). Generally, due to the polyphagous nature of the larvae, several organic waste streams can serve as good rearing media for BSF larvae. Of particular importance in choosing a substrate (organic waste stream) is the nutrient composition (Barragán-Fonseca et al., 2018), the moisture content (Cammack and Tomberlin, 2007), and the particle size (Palmer et al., 2019). Substrates rich in proteins promotes the growth and development of the larvae and are positively correlated with adult longevity and female fecundity (Cammock and Tomberlin, 2017; Oonincx et al., 2015b). Furthermore, the fat content and fatty acid profile of the resulting larvae are dependent on the substrate (Meneguz et al., 2018; Ewald et al., 2020). A moisture content of 70-80 % is optimum for BSF larvae (Myers et al., 2008; Cheng et al., 2017; Lalander et al., 2019), below and above this threshold, the growth and survival of the larvae are negatively affected. The moisture content of substrates can be enhanced when necessary by the addition of water or reduced by dewatering or the addition of bulking agent. To increase the surface area of the substrate to enable easy access to the available nutrients by larvae, the large particle-sized substrate should be shredded. The need for high dietary moisture University of Ghana http://ugspace.ug.edu.gh 26 is due to the morphology of the BSF larvae mouthpart (Kim et al., 2010; Purkayastha et al., 2017), high moisture makes scraping off food from feeding surfaces easier (Banks, 2014). For mass production of BSF larvae, several organic waste streams have been tested on their suitability as rearing media. Among these, the most tested are from markets (Rana et al., 2015; Barragan-Fonseca et al., 2018), animal farms (Sheppard et al., 1994; Newton et al., 2005; Oonincx et al., 2015a; Shumo et al., 2019), catering services (Driemeyer, 2016; Surendra et al., 2016; Shumo et al., 2019), food-processing by-products (Lardé, 1990; St-Hilarine et al., 2007; Manurung et al., 2016; Permana et al., 2018), and brewery distillers ( Webster et al., 2015; Bava et al., 2019; Chia et al., 2019; Shumo et al., 2019). Larvae effectively processed rotten fruit and vegetable from markets, pig manure, chicken manure, cow manure, food scraps from restaurants, spent coffee grain, coffee palp, and brewers’ grain. Other substrates tested are human faecal matter (Lalander et al., 2013; Banks et al., 2014) and chicken feed (Diener et al., 2009; Gobbi et al., 2013; Bava et al., 2019). 2.12 Termites Termites are eusocial insects widely distributed through the tropical and sub-tropical regions of the world (Eggleton, 2000). They are a highly ecologically successful species due to their sophisticated social organization with the unique ability to feed on recalcitrant plant matter such as wood (Khan and Ahmed, 2018). Termites make up about 10 % of total animal biomass and 95 % of soil insect biomass of tropical ecosystems (Jones and Eggleton, 2000). The populations of termites can reach enormous levels as much as 1000 individuals per square meter (Eggleton et al., 1996). Over 2,600 species belonging to 281 genera and nine families of termites have been described (Kambhampati and Eggleton, 2000; Engel et al., 2009). Africa alone owns 1000 species out of the 2,600 known species worldwide (Lewis, 2003). University of Ghana http://ugspace.ug.edu.gh 27 Termites have economic importance to man, they are either highly beneficial or highly deleterious. Cellulose being the primary food source for some termites, they seek this food source causing damage to vegetation, buildings, and other man-made wooden structures. Globally, on a per anum basis, the economic losses incurred in controlling termite pests are about $40 billion (Rust and Su, 2012). In Africa (including Ghana), accurate information on economic losses as a result of termite pests is not well documented (Akutse et al., 2011; Ugbomeh and Diboyesuku, 2019). However, due to the enormous damage caused, the perception of people about termites is mostly negative. Nonetheless, they play very vital roles in tropical ecosystems through plant decomposition, nitrogen, and carbon recycling (Holts and Le Page, 2000). Termites bring about soil formation and nutrient recycling through the consumption of plant necromass. About 50 to 100% of the leaf litter in tropical forests is decomposed by termites (Bignell and Eggleton, 2000; Brauman, 2000). Mounds and soils of termites are used in geochemical prospecting, plastering houses, and making bricks and pots (van Hius, 2017). Mound samples are a good geochemical sample media for mineral exploration (Affam and Arhin, 2005; Arhin et al., 2015) and have aided in gold exploration in parts of Northern Ghana (Arhin and Nude, 2010). The enzymes found in a termite's digestive system can aid in the production of biofuel from woody biomass (Khan and Amad, 2018). The current biomass conversion technology for fuel and chemicals can be improved by the use of the lignocellulolytic enzyme system in wood- feeding termites (BenGuerrero et al, 2015). Termites also serve as food for many indigenous folks in Africa, Asia, and South America. Swarming reproductive, soldiers and queens are collected and eaten fresh, cooked, or fried. They are also harvested and used as feed for poultry and as bait in fishing. The mushrooms that spring up from the mounds annually are a delicacy. The fungus garden, soldiers, and mounds University of Ghana http://ugspace.ug.edu.gh 28 are used in popular medicine. They are used in the treatment of bronchitis, asthma, whooping cough, tonsillitis, and sinusitis (Alves, 2009; Alves and Dias, 2010). 2.13 Termites as food and feed Termites are an economically and socially important source of protein consumed in many parts of the world for many generations. The alates, soldiers, and queens, are frequently enjoyed as a delicacy. Indigenous poultry farmers unable to afford conventional protein sources offer termites as an alternative for their poultry (Sankara et al., 2018). Forty-three species of termites are used as food or feed worldwide (Figueirêdo et al., 2015). The African continent is very popular with termite consumption. It has been reported in almost all parts of Africa (19 countries) except in Northern Africa (Fombong and Kinyuru, 2018). van Hius (2003) reports that about 14 species of termites from the subfamily Macrotermitinae are eaten in sub-Saharan Africa alone. The most consumed termites belong to the family Termitidae, representing about 87 % of the total edible termites (Fombong and Kinyuru, 2018). The species frequently recorded as human food and livestock feed is Macrotermes bellicosus (van Hius, 2003). Others M. subhyalinus, Nasutitermes macrocephalus, and Pseudacanthotermes spiniger are equally popular (van Hius, 2003; Fombong and Kinyuru, 2018). Other families commonly consumed are Hodotermitidae, Kalotermitidae, and Rhinotermitidae (Figueirêdo et al., 2015). In a study conducted by Anankware (2016), nine edible insects belonging to five orders were recorded from Ghana with termites representing 45.9 % of this number. The termites consumed in Ghana are predominantly Macrotermes bellicosus (Anankware, 2016). The sale of sun-dried termites is common in local markets in many East African towns and villages (Fambong and Kinyuru, 2018). Fried and boiled termites are consumed as snacks between main meals among the Baganda and Bantu-speaking people of Uganda. University of Ghana http://ugspace.ug.edu.gh 29 The use of termites as livestock feed is not well documented. Most of the literature is found in general reviews on entomophagy, technical reports, and unpublished thesis (Chrysostome, 1997; van Hius, 2003; Dao, 2016; Khan and Ahmed, 2018). A recent study by Sankara et al. (2018), reported the use of termites by 78 % of the total number of poultry farmers interviewed in 25 provinces in Burkina Faso. Sogbesan and Ugwumba (2008), demonstrated the possibility of replacement of the conventional fishmeal with termite protein in rearing Heterobranchus longifilis. A 50 % termite meal inclusion diet yielded the highest mean weight gain of 9.6 g/fish, the lowest feed conversion ratio (2.9), and the highest protein efficiency ratio of 0.8. The study showed the practicality of using termites as a possible replacement for conventional protein in fish diets. 2.14 Collection of termites Termite harvesting is mostly done seasonally during the rainy months. In Ghana, Macrotermes bellicosus is collected in June and July (Anakware, 2016); in East Cameroon, in March, April, and May (Muafor et al., 2014), and in Kenya, March to May and September to December raining seasons (Fombong and Kinyuru, 2018). Various trapping/collection methods are used in obtaining termites from their mounds or nests. In the countries where termites are eaten, sexual winged reproductives on nuptial flights are collected during the maiden rains. They are collected in the evenings by placing a basin filled with water right under a source of light. Termites attracted to the source of light, fall into the water-filled basin and get trapped (van Hius, 2003; Chung, 2008; Kinyuru et al., 2010). In the Democratic Republic of Congo, the emergence hole on the mound is covered with a basket turned upside down. Termites that cling to the bottom of the basket are detached every few minutes by shaking into a container (van Hius, 2003). In another method, an emergence hole is covered with a dome-shaped framework of sticks or elephant grass covered with banana University of Ghana http://ugspace.ug.edu.gh 30 leaves, while all other holes are blocked. An opening at one side of the dome-shaped structure has a light source that attracts flying termites into a receptacle under it. The women and children of the Central African Republic, push saliva-wet grass blades or tree barks into the open shaft of the mound. Smoke is blown into the opening, causing the soldiers to cling to the grass blades. The blades are pulled out and soldiers are stripped into a receptacle. The ground around the mounds of some species is continuously beaten or drummed to trigger them to emerge and soldiers collected. The queens are collected by digging up the entire mound and in the process potentially destroying the colony. In Togo and Benin, harvesting is done by making an opening in a termite mound and placing a fibrous and humidified substance in a calabash over the hole (Farina et al., 1991; Chrysostome, 2009). Termites collected by this method are used in feeding poultry. Some of the substances used are cow dung, maize cobs, maize stalks, and stalks of sorghum. The calabash is protected against excessive heat by covering it with branches and grasses. 2.15 Poisonous termites Chrysostome (1997), reported that some termites are not suitable for feeding poultry. The humus-feeding termites in the genus Noditermes were reported by farmers to be poisonous to poultry. All poultry fed with Noditermes died after one week in an experiment conducted by Chrysostome (1997), with the highest mortality recorded for guinea fowls (76.9 %). Reports on the toxicity of termites to livestock seem to be scarce. However, soldier castes are known to secrete toxic substances or have powerful mandibles that are used in defending the colony against intruders. Defence against termites is a well-known phenomenon. But in the past 25 years, very little work has been done (Šobotník et al., 2010), with the last exhaustive review published by Prestwich in 1984. The task of defence is primarily that of the soldiers and workers in University of Ghana http://ugspace.ug.edu.gh 31 soldierless colonies. The colony is defended by employing either mechanical and chemical defence systems or both (Diyana et al., 2018). The mechanical defence involves soldiers using their heavily sclerotized mandibles to bite or snap an intruder. Complementary to mechanical defence is the release of chemical secretions from the exocrine glands (Šobotník et al., 2010). The predominant compounds found in these secretions are terpenes (monoterpenes, diterpenes, and sesquiterpenes), aromatic compounds, quinones, and macrocyclic (Evan et al., 1977; Evans et al., 1979; Mill, 1983; Prestwich, 1984; Plasman et al., 1999). However, across the different species, colonies, and populations, there are extreme variations in the secretions both in quantity and quality (Nelson et al., 2001). These chemicals either act as a repellent, irritant, immobilizing agent, anti-healing, or toxins (Mill, 1983; Šobotník and Dahlsjö, 2017). The glandular secretions of soldiers of Armitermes spp. contain poisons that are applied topically onto the target (Mill, 1983). A GC/MS analysis separated four compounds from Macrotermes carbonarius, three of which were unidentified and one identified as lauric acid methyl esther (Diyana et al., 2018). The insecticidal activity of lauric acid has been demonstrated by Mohamed et al. (2013), where 100 % mortality within 24 hours was recorded for Aphis gossypii. A polycyclic diterpene, acting as an irritant and glue was extracted from the frontal gland of Trivervitermes and Nasutitermes soldiers (Laurent et al., 2005). The available information indicates that many of these secretions are principally directed against ant predators (Mill, 1983). Nonetheless, they are effective against vertebrate predators such as anteaters (Lubin and Montgomery, 1981), aphids (Mohamed et al., 2013), fungal pathogens, centipedes, and other insects (Šobotník and Dahlsjö, 2017). The mechanism involved in the lethality of toxic termites to poultry seems to be unknown or yet to be investigated. University of Ghana http://ugspace.ug.edu.gh 32 CHAPTER THREE 3.0 Evaluate black soldier fly larvae rearing systems 3.1 Introduction Larvae of the black soldier fly (BSF) are a readily available source of protein that can be utilised in animal feed to replace non-sustainable and expensive protein sources (Tomberline and van Huis, 2020; van Huis et al., 2020). In Africa, in particular, there is advocacy for poultry and fish farmers to include insects, particularly BSF, in their feed to improve the nutrition of their livestock and reduce production costs (Abro et al., 2020; Chia et al., 2019; Ssepuuya et al., 2017), even though there is no data yet on BSF adoption rates by farmers (Abro et al., 2020). Black soldier fly larvae can be produced cheaply on a wide range of organic waste materials. Several studies have tested the suitability of many organic waste streams on the growth, development, and proximate composition of BSF, with promising results (Banks et al., 2014; Čičkova et al., 2015; Danieli et al., 2019; Miranda et al., 2019). In most BSF production systems, adults are reared in cages to obtain eggs that are placed on the most suitable and available substrates (Caruso et al., 2014, Diener et al., 2009). In Ghana, BSF larvae production in the adult production system was tested on a small-scale production at Fish for Africa (FFA), Accra (Devic et al., 2014; Anankware, 2016). Eggs collected from caged adults were inoculated onto organic wastes in metallic troughs and larvae were manually removed from the waste after 14 days. Another study conducted on the production of BSF larvae also in Ghana between 2014-2015, by ENTO-PRISE an AgriTT Research Challenge Fund Project, used a bay system. The bay system utilises eggs collected from reared adult flies and inoculated into concrete bays to allow the development of larvae. One edge of the bay was inclined at an angel that led into a trench, where prepupae crawled out and were collected. However, egg production is a complicated technique that requires specific expertise. Therefore, small systems have been developed for individual farmers or hobby gardeners, consisting of University of Ghana http://ugspace.ug.edu.gh 33 exposing substrates to naturally occurring BSF females for laying eggs (Kenis et al., 2018). Koné (1998) developed a similar system in Mali to produce housefly larvae by the exposure of organic substrates to attract wild fly populations for oviposition. They used cement beds which served as substrates holding chambers to attract wild flies to oviposit and develop into larvae. On the fourth day after exposure for oviposition, larvae were separated from the waste by removal of the upper layer and sifting the rest through a colander to release the larvae. The system was later improved as described in Koné et al. (2017). In a simple system such as natural oviposition that relies on wild fly populations, it is crucial to select substrates that are suitable for stimulating oviposition and also meet the nutritional requirement for the development of the larvae. The “preference-performance principle” postulates that gravid female insects prefer to oviposit in substrates that maximize offspring fitness (Jaenike, 1978). Such behaviour is common among phytophagous species (Gripenberg et al., 2010), and it has also been observed in detritivorous flies (Baleba et al., 2019). In holometabolous insects such as BSF, where the juveniles are incapable of relocating after hatching and with no parental care, females should favour oviposition substrates that are most suitable for larval development. Egg-trapping efficiency is paramount to the effectiveness of a BSF larvae production system based on natural oviposition (Sripontan et al., 2017). Moreover, several studies (Newton et al., 2005; Li et al., 2011; Oonincx et al., 2015a; Barrangen-Fonseca et al., 2018) have indicated that the quality of BSF larvae meal is contingent on the nutritional composition of the rearing substrate. The substrate used also affects the survival rate and development duration of BSF larvae (Newton et al., 2005; Oonincx et al., 2015a). The literature on the natural oviposition system especially for BSF larvae is limiting. The only documented study was conducted by Nyakeri et al. (2016) in Bondo, Kenya, where the ability to attract BSF using an open system rearing bin was studied. However, YouTube and other online blogs (DipTerra.com, insectus.com, bsffarm.com, Protera.com) are flooded with a great University of Ghana http://ugspace.ug.edu.gh 34 variety of household-based BSF larvae treatment rectors developed and promoted by the enthusiastic hobbyist (Diener et al., 2015b). The motivation here is to make farmers self- sufficient by treating their farm waste while producing the protein component of feed for the farm animals. The information from the blogs seems to suggest that the production of BSF larvae under natural oviposition is an easy task, needing low-tech and low-cost solutions. This study, therefore, sought to evaluate the effect of organic wastes on BSF egg trapping efficiency, the development of larvae, and natural oviposition rearing using a bin system. The result of the study is expected to provide information on the selection of the most suitable substrate and methods for use by smallholder fish and poultry farmers. 3.2 Hypothesis 1: The substrates most attractive for black soldier fly (BSF) oviposition are also those that are most suitable for larval development. Objectives: i. To determine the substrate(s) that can maximize oviposition by female black soldier flies (BSF). ii. To determine the substrate(s) appropriate for enhanced larvae development 3.2.1 Methodology Substrates used Six organic waste substances (substrates) collected from markets, livestock farms, and local food processing industries were tested for their suitability as oviposition and larval rearing media. The substrates used were pito mash (waste from a locally brewed sorghum drink), millet porridge mash, pig manure, chicken manure, fruit waste, and waste from roots and tubers (Plate 2). The pito mash used in the study was obtained from a pito processer in Ashaiman and the millet porridge waste from Kisseman (both suburbs in Greater Accra Region). Fruit waste and roots and tubers were from the Madina market (the main foodstuff market in the La University of Ghana http://ugspace.ug.edu.gh 35 Nkwantanang district of the Greater Accra Region). Pig manure was obtained from Council for