Potential Utilization of Edible Insects in Extruded Rice Products to Address Malnutrition Issues in Developing Countries

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POTENTIAL UTILIZATION OF EDIBLE INSECTS

IN EXTRUDED RICE PRODUCTS

TO ADDRESS MALNUTRITION ISSUES IN DEVELOPING COUNTRIES

A Thesis

Presented to the

Faculty of

California State Polytechnic University, Pomona

In Partial Fulfillment

Of the Requirements for the Degree

Master of Science

Agriculture

Jaynie Tao

2016

SIGNATURE PAGE

THESIS: POTENTIAL UTILIZATION OF EDIBLE INSECTS IN EXTRUDED RICE PRODUCTS TO ADDRESS MALNUTRITION ISSUES IN DEVELOPING COUNTRIES

AUTHOR: Jaynie Tao

DATE SUBMITTED: Spring 2016

College of Agriculture

Dr. Bonny Burns-Whitmore Thesis Committee Chair College of Agriculture Human Nutrition and Food Science

Dr. Olive Yao Li College of Agriculture Human Nutrition and Food Science

Dr. Eileen M Cullen College of Agriculture Plant Science

Dr. Gabriel Davidov-Pardo College of Agriculture Human Nutrition and Food Science

ACKNOWLEDGEMENTS

Perhaps like a child left on her doorstep, I came to Dr. Olive Li for an advisor, except I was the one who did the knocking and insisted on staying. With the shadow of my past disappointments, I was determined to make a difference as a graduate student.

Although not completely familiar with whom she was, her innate willingness to see the success in others inspired me to seek her guidance. While I was intentionally searching for an advisor, I very luckily came across a mentor as well. With plenty of knowledge and experience to challenge me intellectually, I’ve learned that being a scientist is not something to take for granted and that research is truly a meaningful and fulfilling experience. And, with the brain of an engineer and a heart of an engine, Dr. Li has given me the opportunity to become more “rough and tough” which is a valuable lesson for someone who is prone to big-girl-tears. And so, I sincerely give thanks to my mentor, Dr.

Li, for her patience and leadership through my time as a Master’s student. Without her, I would not be where I will be tomorrow.

Though it was me who knocked, it was Dr. Bonny Burns-Whitmore who helped me find Dr. Li’s door. Dr. Burns-Whitmore has been there for me, and who has known me since the beginning. Without her initial guidance, I would still be searching for a place in the world. For that, I am grateful for all the help that she has given me. Without her, I would not be, quite literally, where I am today.

I also give thanks to Dr. Eileen Cullen and Dr. Gabriel Davidov-Pardo for their genuine interest and encouragement throughout this project. I never thought I’d work with insects nor that I would have fun doing so. I am grateful to have an entomologist,

iii such as Dr. Cullen, as a committee member who can share my newfound interest. And while sensory evaluation has always drawn my attention, it is the expertise and assistance from Dr. Davidov who has made this research the historic success that it is. It was also his small piece of unexpected wisdom that “if you don’t hate your project, then you aren’t putting enough effort into it” which has encouraged me to make this Master’s thesis all that it can be.

I’d also like to show my gratitude for all of the other Cal Poly Pomona Staff and

Faculty who have supported my research and helped with the recruitment of participants.

Their assistance certainly played a major role in the gathering of the sensory panel throughout the campus that my paper fliers simply couldn’t accomplish. I’d also like to give a special thanks to Dr. Jack Fong, PolyCentic, and the Poly Post. Dr. Jack Fong, a

Cal Poly Pomona sociology professor, not only shared his enthusiasm for my research with me and my thesis committee but also towards his entire class as he invited them all to join the tasting sessions. He also provided an insightful interview, along with Dr. Li and I, for an article later published in PolyCentric, the official news source for Cal Poly

Pomona. The article was written by Dan Lee and contained photographs from the university photographer, Tom Zasadzinski, who wonderfully captured the experience.

The study was also published in the Poly Post, the student newspaper for Cal Poly

Pomona, with the story being written by Alicia Balderrama. With their help to spread the word about entomophagy to the public, perhaps the future of edible insects will be here to sooner than we think. With that, I am also thankful for all of the sensory participants who participated in my study. Their many encouraging comments made the data a pleasure to analyze and I hope the experience was meaningful and special to them.

And, of course, I am immensely grateful for all the help from my research assistant and peer, Yuguang Zheng. He has provided me with much support for the data collection of this project and I appreciate all the long hours he has spent with me in the lab. Not only has he given me help with research but, as the other food science graduate student, he has made me feel less like a lonely island. I’d also like to thank my other research assistants: Haley Smolinski, Lisa Mai, Megan Telebrico, Ali Hasan, and Spencer

Jones for their work in the development of the insect rice; Jonathan Guo, Helene Mecate,

Jamie Lam, Kristine Frohnauer, Philip Toledo, and Casey Knox for their assistance in sensory evaluations.

A last few thanks are for my family. First, is for my mom, who has been my hero since the dinosaurs. Her unconditional support has kept my feet on the ground and my head on my shoulders. Without her, I would not be who I am today. Second, is for my

Aunt Ingrid, who would have been proud to see what I’ve accomplished at her alma mater. I miss her very much and I hope she is chuckling at the idea of me eating insects.

And lastly, for my life-long partner and best friend, Thomas Nguyen, who has washed more than his share of dishes and laundry, cleaned the litter box, and taken out the trash for me these past three years. I hope he enjoys the next three years as I repay him with mouthful of complaints but a heart full of appreciation for all that he does for me. But jokes aside, he has been my loyal companion through this rollercoaster journey. Without him, I would not be able to accomplish all that I have today.

ABSTRACT

Although unconventional in the United States, entomophagy, or the practice of consuming insects, can provide a nutritious relief to the many malnourished people in developing countries. Edible insects are part of numerous traditional diets found in over

113 countries, including those in Asia, Africa, and South America. Currently, there are two billion people consuming over 2,000 recorded edible insects. Many of these worldwide insects contain amounts of protein, fat, vitamins, and minerals comparable to commonly eaten livestock.

Accordingly, the main objective was to investigate the feasibility of incorporating an edible insect ingredient in an extruded brown rice product. Four formulations were developed, including cricket or locust flours at either 10% or 15% addition levels. A second objective was to determine the physicochemical and nutritional differences between the novel insect rice products and market brown rice. Overall, the insect rice was found to be darker in color, rougher in surface morphology, and softer and more adhesive in texture than market brown rice. Nutritionally, the insect rice offered an excellent source of protein and 10-12% of the daily value of fat which are both at greater quantities than typical rice products. The products were also suggested to contain considerable amounts of dietary fiber and iron, although these quantities require further investigation.

To determine consumer acceptance, the third objective of the study, sensory evaluations involving a 120 untrained panel was conducted. Results suggested cricket formulations were well accepted and worth continuing research and development, while

15% locust formulation was the least preferred of the four formulations.

TABLE OF CONTENTS

Signature Page ...... ii Acknowledgements ...... iii Abstract ...... vi List of Tables ...... x List of Figures ...... xii Chapter 1 – Introduction ...... 1 Food Insecurity ...... 3 Nutrient Content of Edible Insects ...... 3 Entomophagy – More than Survival ...... 6 Nutritional Variability of Edible Insects ...... 7 Environmental and Economic Impact ...... 9 Getting Over the “Yuck” Factor ...... 11 Edible Insect Fortification in Rice Flour ...... 12 Extrusion Technology ...... 13 Chapter 2 – Research Objectives and Approach ...... 16 Research Objectives ...... 16 Research Hypothesis ...... 17 Objective 2 ...... 17 Objective 3 ...... 18 Research Approach ...... 19 Chapter 3 – Experimental Methodology...... 22 Materials ...... 22 Methods...... 23 Formulation Development ...... 23 Extrusion Process for Insect Rice Production ...... 24 Analyses of Physicochemical Properties ...... 25 Nutritional Analysis ...... 27 Total Dietary Fiber Analysis ...... 27 Iron Analysis ...... 28 Protein and Fat Content ...... 29 Optimal Cooking Standardization ...... 29

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Sensory Evaluation ...... 31 Participant Recruitment ...... 31 Questionnaire ...... 32 Samples ...... 35 Location and Schedule ...... 36 Sensory Preparation Flow ...... 37 Research Compliance and Subject Confidentiality ...... 38 Statistical Analysis ...... 39 Chapter 4 – Results and Discussion ...... 40 Formulation Optimization ...... 40 Physicochemical Properties of Uncooked Insect Rice...... 42 Color and Shape ...... 42 Texture ...... 46 Water Activity...... 49 Moisture Content ...... 51 Shelf Life Study ...... 53 Nutrition Content ...... 55 Fiber ...... 55 Iron ...... 58 Protein...... 60 Fat ...... 62 Standardization of Optimal Cooking Conditions for Insect Rice ...... 64 Sensory Evaluation ...... 68 Demographics ...... 68 Perception of Entomophagy ...... 69 Color ...... 76 Aroma ...... 79 Flavor ...... 81 Mouthfeel ...... 85 Overall Liking ...... 88 Chapter 5 - Conclusion ...... 91 Objective 1 ...... 91 Objective 2 ...... 91 Objective 3 ...... 93

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Chapter 6 – Limitations and Recommended Future Studies ...... 95 Limitations ...... 95 Formulation and Extrusion Optimization ...... 95 Fiber and Iron Analysis ...... 96 Sensory Evaluation ...... 96 Recommended Future Studies ...... 97 Formulation Improvement ...... 97 Culinary Exploration ...... 98 Nutrition Analysis ...... 99 Extrusion Standardization ...... 99 Shelf Life Studies ...... 100 References ...... 101 Appendix A ...... 106 Appendix B ...... 110 Appendix C ...... 111 Appendix D ...... 113 Appendix E ...... 171 Appendix F ...... 173

LIST OF TABLES

Table 3.1 - Insect rice formulation ...... 24 Table 4.1 - Rates and percent yields from insect rice extrusion ...... 40 Table 4.2 - Insect rice formulations ...... 43 Table 4.3 - 60x magnification view of insect rice formulations and control ...... 46 Table 4.4 – Texture measurements of rice products ...... 48 Table 4.5 - Water activity of rice products ...... 50 Table 4.6 - Moisture content for rice products ...... 51 Table 4.7 - Changes in water activity (aw) after 3 months for insect rice products ...... 53 Table 4.8 – Percent recovered residue of insect and rice products ...... 55 Table 4.9 - Iron content of insect and rice products ...... 58 Table 4.10 - Protein content of insect and rice products ...... 60 Table 4.11 - Fat content of insect and rice products ...... 62 Table 4.12 - Cooked insect rice products of different rice to water ratios ...... 66 Table 4.13 - Mean scores for overall liking of insect rice formulations ...... 88

Table A.1 - Insect Flours ...... 106 Table A.2 - L*a*b* values for Next Millennium Farms cricket flour ...... 106 Table A.3 - L*a*b* values for Thailand Unique locust flour ...... 106 Table A.4 - Water activity for Next Millennium Farms cricket flour ...... 106 Table A.5 - Water activity for Thailand Unique locust flour ...... 107 Table A.6 - Moisture content for insect flours ...... 107 Table A.7- Percent recovered residue of insect flours* ...... 107 Table C.1 - Percent recovered residue for insect flour and their rice products ...... 112 Table D.1 - Free response regarding participants' experience with entomophagy and their trends ...... 115 Table D.2 - Comments on the color of 10CR and their trends ...... 121 Table D.3 - Comments on the color of 15CR and their trends ...... 123 Table D.4 - Comments on the color of 10LR and their trends ...... 126 Table D.5 - Comments on the color of 15LR and their trends ...... 128 Table D.6 - Comments on the aroma of 10CR and their trends ...... 131 Table D.7 - Comments on the aroma of 15CR and their trends ...... 133 Table D.8 - Comments on the aroma of 10LR and their trends ...... 136 Table D.9 - Comments on the aroma of 15LR and their trends ...... 138 Table D.10 - Comments on the flavor of 10CR and their trends ...... 141 Table D.11 - Comments on the flavor of 15CR and their trends ...... 143 Table D.12 - Comments on the flavor of 10LR and their trends ...... 146 Table D.13 - Comments on the flavor of 15LR and their trends ...... 148 Table D.14 - Comments on the mouthfeel of 10CR and their trends ...... 151 Table D.15 - Comments on the mouthfeel of 15CR and their trends ...... 153 Table D.16 - Comments on the mouthfeel of 10LR and their trends ...... 155 Table D.17 - Comments on the mouthfeel of 15LR and their trends ...... 158 Table D.18 - Other comments about 10CR and their trends...... 160 Table D.19 - Other comments about 15CR and their trends...... 162

Table D.20 - Other comments about 10LR and their trends ...... 164 Table D.21 - Other comments about 15LR and their trends ...... 166 Table D.22 - Any other comments provided by the sensory panel and their trends ...... 168

LIST OF FIGURES

Figure 3.1 - Presentation of samples on a tray ...... 35 Figure 4.1 - L*a*b* values for insect rice products...... 43 Figure 4.2 - Microscopic view of insect rice formulations and control ...... 45 Figure 4.3 – Texture analysis of 15LR, in quintuplicate ...... 47 Figure 4.4 - Age of sensory panel ...... 68 Figure 4.5 - Ethnicity of sensory panel ...... 68 Figure 4.6 - Gender of sensory panel ...... 68 Figure 4.7 - Education level of sensory panel...... 68 Figure 4.8 - Familiarity with entomophagy ...... 70 Figure 4.9 - Future willingness to consume edible insects ...... 70 Figure 4.10 - Experience with entomophagy ...... 70 Figure 4.11 - Future willingness to purchase edible insect products ...... 70 Figure 4.12 - Perception of edible insects in the diet ...... 74 Figure 4.13 - Color intensity of insect rice formulations ...... 77 Figure 4.14 - Aroma intensity of insect rice formulations ...... 80 Figure 4.15 - Flavor intensity of insect rice formulations ...... 82 Figure 4.16 - Mouthfeel intensity of insect rice formulations ...... 86 Figure 4.17 - Overall liking of insect rice formulations ...... 89

Figure A.1 - Next Millennium Farm cricket flour certificate of analysis ...... 108 Figure A.2 - Locust flour protein certificate of analysis provided by Thailand Unique 109 Figure B.1 - Acid digestion of insect and insect rice products ...... 110 Figure B.2 - Steaming method for determining optimal cooking conditions ...... 110 Figure C.3 - Texture analysis of 10CR, in quintuplicate ...... 111 Figure C.4 - Texture analysis of 15CR, in quintuplicate ...... 111 Figure C.5 - Texture analysis of 10LR, in quintuplicate ...... 112 Figure C.6 - Texture analysis of MBR, in quintuplicate...... 112 Figure D.1 - Sensory evaluation room set up ...... 113 Figure D.2 - Entomophagy questions from the sensory evaluation study ...... 114 Figure E.1 - Poly Post publication of the edible insect sensory evaluation study ...... 171 Figure E.2 - Page 1 of the PolyCentric publication of the sensory evaluation study ..... 172

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CHAPTER 1

INTRODUCTION

It is no secret that most of the hungry people of the world live in developing countries. Although unconventional in the United States, entomophagy, or the practice of consuming insects, can provide a nutritious relief to the malnourished population. Indeed, edible insects have already become a part of numerous traditional diets found in over 113 countries, including those in Asia and Africa where the majority of hunger resides

(Rumpold and Schluter 2013). With 2 billion people already taking part in entomophagy, the concept of an edible insect product would certainly be familiar and accepted.

Currently, there are more than 1900 species of edible insects documented in the literature

(Jongema 2014). Many of these insects worldwide contain considerable amounts of protein, fat, vitamins, and minerals, which is comparable to even commonly eaten livestock, such as beef and chicken (Ramos-Elorduy and others 1997, Womeni and others

2009, Xiaoming and others 2010, Sirimungkararat and others 2010).

While entomophagy can be a means of survival for developing countries, edible insects can also be an alternative food source for developed countries, such as the United

States. According to the Food and Agriculture Organization (FAO) of the United Nations

(2009), as the global population is projected to reach over 9 billion in 2050, the availability of food will become a serious issue. As edible insects can serve as a nutrient- dense source of food, they offer economic and environmental advantages as well. Insects have a higher feed conversion efficiency and fecundity, thus reducing the overall food production cost. Furthermore, farming edible insects is substantially less taxing to the

1 environment as it requires no or minimal land clearing and produces less greenhouse gases than other livestock (Rumpold and Schluter 2013, Oonincx and others 2010).

However, the “yuck” or “disgust” factor remains an obstacle towards achieving successful product development. Whole insects especially, whether due to nature or nurture, can be uneasy to accept as a safe and appealing food source.

Thus, the objective of this study is to investigate the feasibility of the obscure incorporation and fortification of a major food staple, such as rice, by utilizing edible insect flour as a value-adding ingredient. By removing the original form of the insect and incorporating its flour into a food matrix that is widely consumed around the world, the aim is to provide an innovative solution for food insecurity, especially in developing countries. Furthermore, the hidden form of the edible insect is designed to reduce the

“yuck” factor and increase the acceptance of entomophagy. Rice flour has been specifically chosen to be the vehicle based on the consideration that the nutrients carried by the edible insect will supplement the nutritional composition of rice. The two main ingredients will be combined in different ratios to form a dough mass, which can be processed through an economically feasible, cold-forming extrusion technology. Thus, through this production method, a final rice product containing optimal addition level of insect flours will be developed. Subsequent objectives will include the measurements of its physicochemical properties, nutrient profile, and consumer acceptability via sensory evaluations.

Food Insecurity

While not as widespread in the United States, food insecurity remains a serious and prevailing issue for much of the world. There is an estimated 805 million people who are without enough food to maintain a healthy and active lifestyle (WFP 2015b).

Developing countries, especially those in South Asia and Sub-Saharan Africa, have the highest rates of hunger. While Asia contains the largest population of hungry people, about two-thirds of the total, Africa has the highest prevalence (WFP 2015b).

Malnourishment occurring in early childhood and if untreated, results in life- threatening consequences. Chronic malnutrition will lead to irreversible stunting and has been reported to be the most important risk factor for illness and death from diseases

(WFP 2015a, Muller and Krawinkel 2005). Approximately 150 million children do not consume the required amounts of energy or nutrients for growth and development, with more than half (52%) of these children residing in South Asia and almost a quarter (21%) living in Sub-Saharan Africa (UNICEF 2001).

Nutrient Content of Edible Insects

Interestingly, although perhaps not surprisingly, entomophagy is especially practiced in developing nations. Countries in South Asia, where more than 40% of the population is chronically malnourished, have already long accepted entomophagy as part of their culture. For instance, countries such as India and Lao People’s Democratic

Republic have identified 24 and 21 edible insect species, respectively. Collectively,

Africa has been reported to have some 246 species of edible insects. Countries within this continent that exhibit more severe rates of chronic malnutrition, such as Madagascar or

Zambia, possess 22 to 33 species (Ramos-Elorduy and others 1997). These insects are consumed for their comparable levels of energy and nutrients, although the levels may vary depending on species type, metamorphic stage, habitat, and diet. For instance, a study conducted in Thailand demonstrated that 100 grams of insects (fresh weight) had comparable, if not more, calories than that of equal weights of commonly eaten livestock, excluding pork (Sirimungkararat and others 2010). Ramos-Elorduy and others (1997) reported that of 78 insect species found in Mexico, the caloric content ranged from 293-

762 kJ per 100 grams of dry matter. Specifically, migratory locusts (Locusta migratoria), the species of which are currently plaguing Madagascar, were discovered to have calories varying between 598-816 kJ per 100 grams of fresh weight in a study conducted in the

Netherlands (Oonincx and van der Poel 2011). Moreover, besides the caloric value, these animals can provide crucial macronutrients as well.

Likewise to the evidence supporting the caloric value of edible insects, it also suggested that insects to be a substantial source of protein. Certain insects found in China were reported with higher protein contents than those in most plants and commercial meat, fowl, and eggs (Xiaoming and others 2010). In the study, 11 orders of insects were analyzed with results of protein contents ranging from a low 13% to a high 77% (dry weight basis). Other researchers reported a range of 37-54% protein content in eight insects found in Thailand (Raksakantong and others 2010). In spite of these variations in protein content, edible insects overall demonstrate to be an exceptionally good source of protein. While this macronutrient is often difficult to obtain in developing countries and so is lacking from their daily diets, edible insects are relatively more available than other meats, demonstrating yet again its worth as a solution for reducing global food insecurity.

Furthermore, while malnutrition is not limited to protein deficiencies but includes the overall lack of caloric intake, populations within developing countries could also benefit from energy dense foods provided by edible insects (DeFoliart 1992). Energy intake can be greatly enhanced, to an extent, with higher levels of fat, like those potentially found in edible insects. In a study lead by Womeni and others (2009), six insects from Cameroon of sub-Saharan Africa were analyzed to determine their lipid contents and essential fatty acid profiles. Results showed that not only were these insects an excellent source of fat, ranging from 9.12 to 67.25% (dry weight basis), but also that they were rich in polyunsaturated fatty acids. These types of fats, along with monounsaturated fats, have been widely accepted to offer more nutritional benefits and should be used to replace the intake of saturated fatty acids (Yang and others 2012, Enos and others 2014). Furthermore, the polyunsaturated fats included essential fatty acids, such as linoleic (omega-6) and linolenic (omega-3) acids (Womeni and others 2009).

Comparable findings were demonstrated in Thailand as well, varying between 0.34 to

23.98% of total fat content depending on the species of insects (Raksakantong and others

2010). Thusly, edible insects have the potential to be an exceptional source for energy and macronutrients.

Micronutrients, on the other hand, are just as essential for a healthy life. A major concern for developing countries is iron intake as its deficiency is the world’s most common and widespread nutritional disorder (WHO 2014). Many of the insects safe for consumption contain abundant levels of iron that often exceed other commonly eaten animals. For instance, edible insects such as the popular palm weevils (Rhynchophorus phoenicis) or mopane caterpillars (Imbrasia belina), both species found in Africa, can

5 provide 12 and 31 mg of iron per 100 grams of weight, respectively (Banjo and others

2006). Chicken and beef, on the other hand, provide only 1.2 and 3 mg of iron, respectively (Sirimungkararat and others 2010). Zinc, another mineral important for growth and development can be generally found in most insects. For example, it has been reported that the aforementioned palm weevil larvae contains 26.5 mg per 100 grams

(Bukkens 2005).

Data for dietary fiber found within edible insects is largely inconclusive, especially in regards to fiber for human consumption. While, acid detergent fiber and neutral detergent fiber are the common indicators for fiber levels specific for animal feed

(Nowak and others 2014), crude fiber is nutritionally meaningless as it refers to the remaining fiber post acid and alkali treatments. In food analysis, soluble and insoluble fibers, and their sum as total dietary fiber, are used for product formulation development and nutritional labeling. However, there currently is little information about these values in edible insects. Thus, this study will also aim to establish the values of total dietary fiber in the edible insect raw materials and in the products that are developed utilizing these raw materials.

Entomophagy – More than Survival

Thus, the overall nutritional value of edible insects presents them to be a remarkable alternative for alleviating the food insecurity in developing countries, with the exception of dietary fiber. Although it could be said that the underprivileged do turn to edible insects for sustenance since other forms of livestock are either unavailable or expensive, the tradition of consuming edible insects is not always a tactic for survival.

Banjo and others (2006) reported edible insects being included in a planned diet throughout the year and not solely as an emergency food supply. A collective group of people in South Africa called Pedis, for instance, value insect meals, such as those derived from caterpillars, even more so than beef as the latter sales decrease during caterpillar harvesting seasons (Womeni and others 2009). Palm weevils, especially the larvae form of Rhynchophorus phoenicis, have been a part of a traditional African diet for centuries and are, like the locusts there, prepared by frying (DeFoliart 1993, Banjo and others 2006). Besides Africa, this insect has also been appreciated across other tropical regions including Latin America and Asia (DeFoliart 1993). In Mexico, investigators reported exceptional consumer acceptance for their maize flour tortillas supplemented with ground yellow mealworm larvae (Tenebrio molito) (Aguilar-Miranda and others

2002). The practice of eating insects can also be found in the East. Lethocerus indicus, or giant water bug, is a particularly popular insect consumed throughout several Asian countries such as Thailand, Laos, Vietnam, and Cambodia (Kiatbenjakul and others

2015). Its odor provides a flavor profile that is essential for consumer acceptance of food products such as chili pastes or fish sauces. As entomophagy is practiced around the world, a novel edible insect product has certain potential for being accepted and enjoyed.

Nutritional Variability of Edible Insects

However, it is important to note that while promising, edible insects are still highly variable between/within species and metamorphic stages. When comparing the amount of energy found between edible insects in previously cited studies, these values varied by hundreds of calories per gram (Ramos-Elorduy and others 1997, Oonincx and

7 van der Poel 2011). Protein content had differences as great as 64% while fat had variations as high as 44% (Xiaoming and others 2010, Raksakantong and others 2010,

Womeni and others 2009). It could be easily discerned from these values that the benefits of each edible insect species can vary significantly. In addition, analogous to shellfish or prawns, the type of habitat and diet of these insects can also change their flavor and even their nutritive value (Klasing and others 2000, Ramos-Elorduy and others 2002, Finke

2003, Raksakantong and others 2010, Oonincx and van der Poel 2011, Nowak and others

2014). As grasshoppers (Zonocerus variegatus) in Nigeria were fed with bran, which contains more fatty acids than that of corn, these insects demonstrated almost doubled protein levels (Ademolu and others 2010). In yet another study, migratory locusts were fed with three different diets, consisting either solely of grass, of grass and wheat bran, or of grass, wheat bran, and carrots (Oonincx and van der Poel 2011). The investigators found the wheat bran diets reduced the protein content and increased the fat content of the locusts while the addition of carrots further enhanced the fat content and provided greater levels of β-carotene. Hence, although entomophagy has been revealed with great potential to be an extraordinary resource for relieving global food insecurity, it is essential to obtain complete nutritional profiles in order to determine the actual nourishment available in the finished product. As this study reveals some of the nutritional content of the insects used, specifically crickets and locusts, this data will further supply knowledge to the edible insect database.

Environmental and Economic Impact

While developing countries are currently faced with the burden of food insecurity, the FAO (2009) predicts the global population will increase to 9 billion by 2050. As the world population proliferates, the demand for food and feed will escalate along with it, subsequently requiring food production to increase by 70% (FAO 2009). With the current state of the environment, and as it continues to degrade, there is a call to conserve natural resources such as land and water. Evidently, rearing insects requires remarkably less land than farming other categories of livestock. Oonincx and de Boer (2012) discovered that to produce 1 kg of edible protein, mealworms (Tenebrio molitor) required only 10% of the land that is needed for beef production. While this difference may not be as dramatic when compared with pork or chicken production, mealworm farming still requires 29-

50% less arable land. Due to their obvious smaller size, edible insects also have the potential to be farmed vertically, thereby requiring no additional land clearing to advance production (van Huis and others 2013). Greenhouse gases, including carbon dioxide, methane, and nitrous oxide, are also produced at a lesser rate by insects such as crickets

(Acheta domesticus) and mealworms (Tenebrio molitor). When compared with pigs and cattle, the difference is by a factor of about 100 (Oonincx and others 2010). Furthermore, the manure produced from common livestock also contaminates the surface and groundwater while releasing ammonia and acidifying the land (van Huis and others

2013). Whereas pigs have demonstrated to produce ammonia at a rate of 10 times or greater than crickets and mealworms, the quantity produced by cattle is undoubtedly even less favorable (Oonincx and others 2010).

As land becomes scarce, the water supply is equally threatened. The FAO (2012) predicts that by 2025, two-thirds of the world will be under stress from water shortages.

When evaluated against the quantity required to produce 1 kg of grain protein, 100 times more water is required to produce the same weight of animal protein, especially as water is necessary for forage and feed production (Chapagain and Hoekstra 2003). While assessments for the quantity of water required for farming edible insect is currently unavailable, the results are likely in favorable directions, in parallel with that of greenhouse gas and ammonia emissions.

In addition to the heavy usage of land and water, farming livestock also requires feed, which in turn requires further land clearing. Alternatively, insects are cold-blooded and so only require feed for energy and for warmth. Studies have shown that to produce 1 kg of livestock weight, at least 2.5 kg of feed is required for poultry meat, 5 kg for pork, and 10 kg for beef (Smil 2002). Edible insects, such as crickets (Acheta domesticus), require only 1.7 kg to produce the same weight (Collavo and others 2005). In addition, only a percentage of this livestock weight is edible, consequently reducing the actual production and availability of meat protein and other nutrients. While chicken and pork provide 55% of edible weight, beef only provides 40% (Nakagaki and DeFoliart 1991).

Crickets (Acheta domesticus) conversely offer 80% of its live weight for consumption, making its feed conversion efficiency exceptionally high. Furthermore, insects in general reproduce more rapidly and in greater quantities. For instance, the same aforementioned cricket species can lay 1,200-1,500 eggs within 30 days (Patton 1978). Insects also reach their adult stages much quicker than their livestock counterparts and so are capable of reproduction sooner.

Getting Over the “Yuck” Factor

However, despite the undeniable benefits for consuming and rearing edible insects, this does not deter from the “yuck” or “disgust” factor. A study conducted by

Rozin and others (1999) revealed that despite American students being willing to touch insects with their hands, many declined touching those insects to their lips. The concept of food neophobia, the fear of trying new or novel food products, has been suggested as the reason for this rejection (Megido and others 2014). This concept is explained by

Rozin and Fallon (1980) to be a dismissal due to distaste for the organoleptic qualities, fear of danger to the body, or disgust stemming from a prior impression of what the product is or of its origin. While either of these three explanations or the combination thereof may be understandable, perhaps the neophobia of entomophagy could be lifted.

Lobster, once considered “junk” food in the 17th and 18th centuries, was established to be cruel and unusual punishment for feeding to servants and prisoners more than twice a week (Crowley 2015, Greenlaw 2002). Today, lobster is regarded as a fine dining food, depicted as a pleasure enjoyed by consumers with high incomes. Another interesting example is the history of the ever popular sushi dish, the California roll. When first introduced in Los Angeles, Ichiro Mashita, the sushi chef of Tokyo Kaikan, innovatively substituted the toro (fatty tuna) with avocados as it had a similar texture but was more familiar to Americans. He also placed the nori inside the rice as this too was an unusual ingredient (Crowley 2015). Presently, California rolls have become an icon of sushi and are enjoyed all across the globe. Whatever the cause of the prior neophobia, lobster and the California roll have had their status momentously and positively changed. Edible insects have the same potential. Megido and others (2014) suggest that the promotion of

11 knowledge and acceptance of edible insects will begin with the understanding of the relationship between insects and shellfish, as they are simply arthropods of the land and sea, respectively. Correspondingly, increasing the frequency of positive exposures and tasting trials would also be effective.

Edible Insect Fortification in Rice Flour

What is also proposed for increasing acceptability is the form of the edible insect ingredient. While traditional cooking methods such as roasting or frying are frequently used to prepare tastier or more palatable dishes, the insects often remain whole, especially in tropical regions (van Huis and others 2013). Instead, edible insects can be made into granular or paste forms that may result in improved acceptability. While in a paste or powder form, they can be incorporated into other foods. For instance, in

Thailand and other South Asian countries, ground giant water bugs (Lethocerus indicus) are an essential ingredient in chili paste and fish sauces (Kiatbenjakul and others 2015).

Whereas the chili paste is used as an ingredient in dishes, the current study suggests the fortification of a main staple food could be more effective as the ultimate vehicle for delivering valuable nutrients offered by edible insects.

Rice is a staple food for more than half of the world’s population, especially in developing countries (IRIN 2010, USDA 2012). It provides 20% of world’s dietary energy and supports the livelihood of more than 1 billion people (FAO 2004, IRIN 2010).

It is regularly consumed and composes a great portion of the diets in both developing and developed countries. The overall percent of dietary energy supplied by rice for Asia,

Africa, and South America has been reported to be about 30%, 10%, and 10%,

12 respectively (FAO 2004). Specifically, developing countries such as Bangladesh, Laos

People’s Democratic Republic, and Indonesia have more than half of their dietary energy supplied by rice. Other than energy (363/100 grams), rice is also a good source of thiamin, riboflavin, niacin, as well as amino acids such as glutamic and aspartic acid

(FAO 2004). While rice lacks lysine, edible insects such as caterpillars (Gynanisa maja), grasshoppers (Ruspolia differens), and winged termites (Macrotermes falciger) can complement this essential amino acid (Siulapwa and others 2014).

To obtain rice, the grain is milled to remove the outer husk and bran layer. During this process, broken rice is produced and can be used to create rice flour. For the current study, the objective is to determine the feasibility of combining an edible insect flour ingredient with rice flour for the fortification of a staple food. Successful results will first, extend the yield of rice harvest and create an additional economic prospect for this milling byproduct (Qian and Zhang 2013), and second, improve the acceptability of a nutrient dense ingredient such as edible insects for general consumers.

Extrusion Technology

To create this fortified rice product, extrusion, a method that is growing in popularity especially in developing countries, will be employed. This is a continuous process that combines cooking, mixing, shaping, and forming (Fellows 2009). The process begins by the raw materials, typically in granular form, being fed into the barrel where water is added to convert the dry ingredients to a semi-moist dough mass that is processed further. As the screw within the barrel kneads the mixture, it becomes plasticized with a desirable rheological property suitable for extrusion. The screw pushes

13 the dough mass out of the die holes where it expands and cools rapidly, forming the shape set by the type of die plate used. This type of technology can generate a remarkable number of products, ranging from cereals, pasta, soup and beverage bases, hot dogs and sausages, processed cheese, and chewing gum (Fellows 2009).

Extruders are comprised of either one screw, called a single-screw, or two screws, called a twin-screw. They are further classified as either cold-forming extruders or cooking extruders. The current study aims to utilize the low-cost, single-screw, cold- forming extrusion process, where the temperature of the food mixture does not increase over 70°C, leaving the food uncooked and undistorted (Fellows 2009). The advantage of utilizing single-screw cold-forming extrusion is in its comparably lower capital, operating, and maintenance costs to its twin-screw counterpart. Also, this type of extruders generally has fewer procedural parameters to manipulate and so require less skilled labor as its operation is continuous and automatic. Even so, the processing costs are lower while providing higher productivity rates than other cooking and forming techniques (Fellows 2009). Resulting products are then dried and therefore remain shelf stable. Lastly, the process produces no effluents (Fellows 2009).

Action towards promoting the application of extrusion technology has already been set forth. ExtruAfrica, an initiative of the Center of Excellence in Advanced

Manufacturing at the North-West University in South Africa, encourages the extrusion method through conferences and training workshops. The first seminar was first held in

2011 and continues on (ExtruAfrica 2015). Filli and others (2014) suggest extrusion technology to be a key to adding value to agricultural commodities to reduce food insecurity in developing countries. Staple cereal grains, including rice, have yielded

14 positive results. In fact, several investigators have already made considerable progress in creating products that incorporate indigenous materials through extrusion applications

(Filli and others 2014) while increasing nutrient density (Obatolu 2002). Moreover, utilization of extrusion technology will also offer potential employment and an improved livelihood for local citizens.

CHAPTER 2

RESEARCH OBJECTIVES AND APPROACH

Research Objectives

Whether from a perspective of developing or developed countries, the entire global population is in need of alternative food sources. People living in developing nations, including those countries in Sub-Saharan Africa or South Asia, are often found to be dangerously malnourished. With beef and chicken being difficult to come by, they have been enjoying edible insects as a dietary resource for generations. This is because edible insects actually contain several nutritional benefits. They offer plenty of calories, protein, fat, vitamins, and minerals, depending on their species, metamorphic stage, and diet.

Although still widely unconventional, developed countries have also begun considering entomophagy as a means to support our ever growing populations. In fact, as aforementioned, the FAO projects the world population to approach 9 billion by 2050. As the numbers swell, natural resources such as land and water are being rapidly degraded.

Edible insects, with their high feed conversion efficiency and fecundity, as well as their minimal space for rearing, certainly presents as an advantageous solution for present and future food insecurity.

Even so, the “yuck” or “disgust” factor is not so easily eliminated from today’s cultures. While nutritious, perhaps the unappealing nature of entomophagy is in the consumption of the whole insect. With that, the obscure incorporation of this food source through the utilization of insect flours is considered. To further enhance the acceptability

16 of edible insects, a worldwide staple such as rice flour and the simple, low-cost processing of extrusion will be used in combination to develop a novel product, which can offer as a solution for food insecurity.

Hence, the objectives of this current study are three-fold:

1. To investigate the feasibility of incorporating edible insect flours ingredients

into an extruded rice product, leading to a novel food product.

2. To assess the physicochemical and nutritional properties of the insect rice

product using the optimized formulations from the extrusion process and to

compare these insect products between each other and against market brown

rice as a control

3. To determine consumer acceptability of the insect rice products for potential

implementation in developing countries

Research Hypotheses

Objective 2

Null Hypothesis 1 (H01): there is no discernible difference between the extruded insect rice products and the control, market brown rice, in terms of physical characteristics.

Alternate Hypothesis 1 (H A1): differences between the extruded insect rice products and the control, market brown rice, will be discernible through the measurements of physical characteristics.

Null Hypothesis 2 (H02): there are no discernible differences between the insect rice formulations, developed using either cricket or locust flours, in terms of physical characteristics.

Alternative Hypothesis 2 (HA2): differences between the insect rice formulations, developed using either cricket or locust flours, will be discernible in terms of physical characteristics.

Null Hypothesis 3 (H03): there are no discernible differences between the 10% and 15% addition levels of the same insect flour within the extruded rice product in terms of physical characteristics.

Alternative Hypothesis 3 (HA3): differences between 10% and 15% addition levels of the same insect flour within the extruded rice product will be discernible in terms of physical characteristics.

Objective 3

Null Hypothesis 4 (H04): there are no discernible differences between the insect rice formulations, developed using either cricket or locust flours, in terms of appearance, aroma, flavor, mouthfeel, and overall liking, as determined by a sensory panel.

Alternative Hypothesis 4 (H04): differences between the insect rice formulations, developed using either cricket or locust flours, will be discernible by a sensory panel in terms of appearance, aroma, flavor, mouthfeel, and overall liking.

Null Hypothesis 5 (H05): there are no discernible differences between the 10% and 15% addition levels of the same insect flour within the extruded rice product in terms of appearance, aroma, flavor mouthfeel and overall liking, as determined by a sensory panel.

Alternative Hypothesis 5 (HA5): differences between 10% and 15% addition levels of the same insect flour within the extruded rice product will be discernible by a sensory panel in terms of appearance, aroma, flavor, mouthfeel, and overall liking.

Research Approach

Brown rice flour will be the major bulk ingredient for the production of the novel insect rice products. In contrast to white rice flour, its brown rice counterpart is more nutritious. Brown rice flour offers an additional 18% protein, 48% dietary fiber, 82% iron, and 67% zinc (National Nutrient Database for Standard Reference, Release 27,

USDA). In general, white rice is also inferior in its supply of vitamins and minerals when compared to brown rice. Utilizing a brown rice ingredient will also provide some relief to the intrinsic dark color provided by the edible insects as consumers will associate a brown rice product with a darker appearance.

Edible insect flour will be derived from cricket and locust of which are both enjoyed by many populations around the world. Crickets are commonly consumed in

Thailand where there are 20,000 registered cricket farmers (Dunkel 2015, van Huis and others 2013) as well as other Asian countries such as Lao People’s Democratic Republic and Cambodia (van Huis and others 2013). They are also making their way into the

United States market as they are appearing in commercially produced products such as energy bars from Chapul (Colorado) and “Chirps” (cricket chips) from Six Foods

(Boston, MA). Nutritionally, per 100 grams, crickets can approximately offer 125 kcals,

15 g protein, 6.3 g fat, 41 mg iron, and 75 mg calcium as well as other benefits

(Sirimungkararat and others 2010). Crowley (2015) stated that crickets provide 15% more iron than spinach, 2 times more protein than beef, and equal quantity of vitamin B12 as salmon. Locusts, while currently not as available as crickets in the market, are potentially superior in nutrient density. They can offer 598-816 kcal, 13-28 g protein, and

8-20 mg iron per 100 grams (Oonincx and van der Poel 2011, van Huis and others 2013).

Inclusion of locust may also provide a solution to present or future infestations, as these insects often occur in swarms. Despite being viewed as pests, locusts are popular as fried dishes in countries such as Thailand and Madagascar.

Development began at a 5% (dry weight basis) addition level of insect flour in the extruded rice formulation and was then increased to 10% and 15%. Concerns for the increasing addition levels of edible insect ingredients included an intensified aroma and taste, which although may be familiar in other dishes, is not currently associated with rice. Regardless, the protein fortification of insect flour to a product such as brown rice was certainly advantageous. For instance, by referencing the certificate of analysis

(COA) provided by the vendor of the cricket flour (Appendix A.1 and A.2), we predicted that a 15% insect flour formulation would theoretically increase the protein content by

8.5 – 9g of protein per 100 grams of insect rice on a dry basis. Together with the protein content originally in existence with brown rice flour (3 g protein per 40 g serving), 100 g of the insect rice would hypothetically provide an estimated 13.5 – 14.6g of protein or 26

– 29% of the daily value (DV) of protein.

Vegetable shortening will provide lubrication during the extrusion process by reducing the level of friction within the extruder barrel caused by the high level of starch in the mixture (Li 2009). Water will be utilized to hydrate the flour particles to aid in the formation of plasticized dough mass suitable for extrusion. While having the same granule size as brown rice, tapioca starch will be used as a thickening agent that provides an additional binding effect other than what is supplied by rice starch (Mishra and Rai

2006). Sodium alginate, a gum extracted from the cell walls of brown algae, will be included for its gelling and as well as thickening properties to enhance the structural

20 qualities of the rice dough. A calcium chloride solution will be sprayed on the surface of the extruded rice pieces which will result in a crosslinking reaction at the surface between the sodium alginate and the calcium salt. This will enable the starch granules from the extrusion formulation to become entrapped in a continuous network, or shell, at the surface of the extruded rice particles (Li 2009, Koh 2009).

In determining the success of the three aforementioned objectives, first, the measurement of the extrusion flow rate (as the weight of samples collected per unit of time) and the yield of samples with desirable shape and size will be measured to determine whether Objective 1 has been achieved. Second, a market brown rice purchased from a local supermarket will serve as the control when evaluating the physical properties of the insect rice products. Statistical analysis of these experimental measurements will determine the failure to reject or the rejection of the null hypothesis under Objective 2. Finally, a sensory evaluation, along with its respective statistical analyses, will be involved to determine the fulfillment of the Objective 3 and the status of its respective hypotheses.

CHAPTER 3

EXPERIMENTAL METHODOLOGY

Materials

Organic cricket flour (Gryllodes sigillatus) was obtained as “Protein 2050-C” from Next Millennium Farms, which later changed their company name to Entomo

Farms (Ontario, Canada). Locust flour (Locusta migratoria) was purchased from

Thailand Unique (Udon Thani, Thailand). Both sources offer products that contain 100% farm raised insects of which only the floured forms of these insects were employed in this study. Brown rice flour and tapioca starch were both acquired from Bob’s Red Mill®

(Milwaukie, OR) while the All-Vegetable Shortening was from Crisco® (Orrville, Ohio).

Sodium alginate and calcium chloride are both food grade materials typically found in product development labs and were purchased from VWR Chemicals (Radnor,

Pennsylvania). The insect rice products were cooked using a steaming method to be explained in the Optimal Cooking Standardization section.

Market brown rice (MBR), long whole grain (Signature Kitchen, Pleasanton, CA) was purchased from a local grocery store and used as a control to assess the physicochemical properties of the insect rice formulations for the hypotheses under

Objective 2. The brown rice was cooked in an ARC-926D Aroma rice cooker (San

Diego, CA) as per the instructions provided by the operation manual of the equipment.

Methods

Formulation Development

As the current research pioneers a pathway to fill the knowledge gap, a method of trial and error was required to create an optimized product. The initial rice formulation suitable for cold-forming extrusion was derived from a patented technology, Ultra Rice®

(Li 2009), which was originally developed for fortification of vitamin A and iron, as well as zinc and other B vitamins. The current study adopted its base formulation while excluding the fortifying agents of micronutrients and replacing white rice flour with brown rice flour, as well as incorporating tapioca starch to gain an extra binding effect.

Other formulation components, including sodium alginate, water, and shortening were kept at their original addition levels. A calcium chloride solution was sprayed on the surface of the extruded rice pieces before dehydrating the insect rice product. The incorporation of edible insect flours began with an addition level of 5% (dry weight basis) and then increased to 10% and 15%. Preliminary development of the insect rice at

5% addition levels presented no major difficulties observed from both the extrusion process and the physical and organoleptic properties. However, the incorporation of insect flours at10% and 15% resulted in an apparent difficulty to the extrusion operation and observable differences in particle shape, size, and organoleptic properties. Therefore, the current study focused on the development of optimized formulations incorporating higher addition levels (10% or 15%) of either cricket or locust flour. With that, a total of four formulations were developed: 10% cricket rice (10CR), 15% cricket (15CR), 10% locust (10LR), and 15% locust (15LR) as shown in Table 3.1 below.

Table 3.1 - Insect rice formulation Ingredient Percent (%) Brown rice flour 65.5 - 70.5 Insect flour 10 - 15 Tapioca starch 15 Sodium alginate 1.5 Vegetable shortening 3 Water 33 (db) Calcium chloride coating (20%, w/v) 10* *50 mL solution was sprayed per kg of extruded rice, leading to 10% calcium coating level

Extrusion Process for Insect Rice Production

Insect rice production began with sifting of the dry ingredients including the brown rice flour, insect flour, tapioca starch, and sodium alginate before blending them together. A countertop KitchenAid stand mixer, model K45SS (Benton Harbor, MI) was used to blend the dry materials until homogenous before liquefied vegetable shortening and water were slowly introduced. When all the ingredients were added, the dough was mixed for at least 5 minutes. The homogenized dough was then transferred to a La

Monferrina single-screw, cold-forming, pilot-scale P12 pasta extruder. A rice-shaped die plate #305 (Asti, Italy) and a face cutter with a variable speed controller were assembled together to obtain the rice-shaped pieces. The extrusion flow rate, as weight (g) of sample collected per unit time, and the total yield (g) per 1kg batch were recorded.

After extrusion, the insect rice was transferred to a rotating pan coater (12 inch diameter, Union Standard Equipment Company; Bronx, NY) powered by a Dayton DC gear motor 90VDC (Dayton, Texas). A calcium chloride solution at a concentration of

20% (w/v, 20g per 100mL) was prepared and approximately 50mL of this solution was sprayed on the 1kg batch of extruded insect rice as it underwent a continuous tumbling

24 mode within the coating pan. After the surface coating, the insect rice was then placed in an Excalibur NSF Commercial Food Dehydrator (Sacramento, CA) for at least 6 hours at

165°F. The dry weight (g) of the insect rice was recorded.

Next, the dried insect rice was screened to obtain a final product with desirable shape and size. This was a two-step process where the insect rice was first separated using a colander with a range of 3.00 – 4.60 mm holes. Larger pieces were removed from the top of the colander and bagged as “chunky” rice. The remaining insect rice was then screened once more using a sieve with a 1.56 x 2.10 mm mesh. The pieces remaining in the sieve were collected as the final product while the flakey pieces from the bottom of the sieve were bagged as “powderized.” The weight of both the rice-shaped final product and irregular shaped pieces (“chunky” and “powderized”) were used to calculate the yield of the final product using the following equation:

Yield (%) = weight of rice-shaped particles with desirable size total weight from extruded insect rice batch

Final product yield and extrusion flow rate were compared between the four formulations, allowing the evaluation of the extrusion performance for each formulation.

All insect rice products were stored in labeled, re-sealable, Ziploc® bags and placed in a cool, dark space, ready for analyses and sensory evaluation.

Analyses of Physicochemical Properties

Assessments of color was performed with the filling of a petri dish with a sample and a Konica Minolta CR-400 Chroma Meter (Ramsey, NJ). The values were reported as

L*a*b*. Water activity was measured with the employment of an AquaLab Pre (Pullman,

WA) and by loading just enough of the sample to cover the bottom of the sample cup.

Moisture content was determined using a Shimadzu MOC-120H moisture analyzer

(Kyoto, Japan) and approximately 5 g per sample. These analyses were performed using the intact insect rice and MBR (non-powderized) and in triplicate for all four insect rice samples at 0 months, with the exception of moisture content which was measured once for each sample. The latter had only a single measurement because, unlike water activity evaluations which took 5 – 10 minutes, the moisture analyzer required up to 35 minutes to complete one sample measurement. To evaluate the shelf stability, water activity was then measured again, in triplicate, after 3 months of storage for all four samples.

Texture analysis of cooked insect and market brown rice was conducted using a

Stable Micro Systems TA-XT Plus texture analyzer (Godalming, UK) equipped with the

Texture Expert Software package. The probe used was a 35mm cylinder probe (p/35) with a test speed of 0.5 mm/s and a post-test speed of 10.0 mm/s. The strain was set at

90% and the trigger force was automated at 3.0 g. Three pieces of cooked rice were used per trial with a total of five measurements performed for each product.

Qualitative examination of the surface morphology, in terms of shape and size, was performed using a Digital Blue QX7 microscope (Atlanta, GA) at both 10 and 60 magnification for all insect rice formulations with the market brown rice used as the control.

Nutritional Analysis

Total Dietary Fiber Analysis

An Ankom TDF Analyzer (Macedon, NY) was employed to estimate the total dietary fiber content of the insect rice formulations. This state of the art equipment automates the AOAC 991.43 procedures for following the enzymatic-gravimetric method in total dietary fiber (TDF) analysis of grain and cereal, fruit and vegetables, and processed food products. Procedures for the assessment followed the operation protocol provided along with the equipment. The process began with 0.5 g of insect rice sample being ground into a powder and then placed in the patented filter bags of the fiber analyzer. The samples were then digested via α-amylase (Bacillus licheniformis), protease (Bacillus licheniformis), and amyloglucosidase (A. niger). Programmed and mechanized movement of heated bars from the equipment mimicked human digestion without laborious manual shaking. The remaining samples after enzymatic digestion then underwent an acetone wash to remove all other sugars and were then left to air dry.

Finally, the bags were placed in an oven and were further dried for two hours.

Afterwards, the bags were weighed and the difference between the starting and final weights revealed the % recovered residue. This value included a portion of other constituents, such as undigested protein and ash, along with the TDF content. This recovered residue should undergo further analyses to determine the actual levels of ash and undigested protein, which would then be used to correct the initial values of weight difference for the actual TDF content. However, due to the limited capacity in the lab for ash and protein analyses, this recovered residue value was used as a rough estimation of the content of total dietary fiber.

Iron Analysis

A graphite furnace atomic absorption spectrometer (AAS) was utilized to analyze the iron content of the insect rice formulations. First, the samples were digested via concentrated hydrochloric acid and nitric acid at 300°C. About 1 g of each insect rice sample was ground into powder and then placed in a 500 mL Erlenmeyer flask, which was then added with 10 mL of each of the two concentrated acids and placed on a hot plate (Appendix B.1). The mixture was boiled until the majority of acids had been evaporated, or the yellow fume produced from the nitric acid oxidizing the organic matters in the sample had disappeared. Once removed from heat, the flask was cooled and then rinsed with deionized (DI) water three times with all of the assay solution being transferred into a 100 mL volumetric flask. The remaining volume was filled up with DI water. During the experiment, it was observed that the assay solutions contained white- colored precipitates, which may be related to undigested fat in the samples. Thus, a gravimetrical filtration was carried out prior to iron analysis using AAS. To remove the residue, the assay solutions were filtered using a Buchner funnel and flask where filter paper (pore size of 0.5 µm) and rubber tubing were employed in the apparatus assembly.

The filtered solution was placed back into a 100 mL volumetric flask.

Once digestion and filtration were completed, the samples were analyzed for their iron content using a Perkin Elmer AAnalyst 400 Atomic Absorption Spectrometer

(Waltham, MA). A calibration curve was created using standard iron reference solutions in a concentration series of 1 to 5 ppm, while 1 N hydrochloric acid was used as the blank.

Protein and Fat Content

Percent protein and fat analyses of the insect rice formulations were outsourced to

Certified Laboratories in Anaheim, CA, which adopted standard AOAC methods for protein and fat analyses. The vendor of the cricket flour, Next Millennium Farms

(Entomo Farms) was able to supply a complete certificate of analysis (COA) while

Thailand Unique, the supplier of the locust ingredient, was only able to provide its COA for protein (Appendix A.1 and A.2).

Optimal Cooking Standardization

In order to achieve consistent cooking properties of the insect rice products for the sensory evaluation study, the cooking preparation process was standardized. It was imperative to consider that while market brown rice is of the actual rice grains, the novel insect rice product is a reconstitution of floured ingredients, thus lacking the outer bran layer which provides a physical barrier for the inner starch. Instead, the crosslinking between the sodium alginate and calcium chloride coating offers a surface shield, or shell, to maintain the shape integrity of the insect rice throughout the cooking process.

While effective, the insect rice still required steaming in place of boiling. The proportion ratio of rice to water had to be determined as well.

Three ratios were tested, including 1:1, 1:1.5, and 1:2 (rice to water) on a weight to weight basis. The rice and water was gently mixed in polystyrene portion cups and then placed on the steaming basket accompanying with a stovetop pot (Appendix B.2). It was necessary to steam the insect rice immediately after combining with water to prevent any initial breakdown of the products. After the insect rice was steamed for 15 minutes, it

29 was then removed from heat. The appearance (color), aroma, texture, and taste were noted. This process was completed for each of the four insect rice formulations.

With the lack of an actual bran layer acting as a protective barrier, the major concern when preparing the insect rice for consumption is certainly in its shape integrity and texture. Accordingly, these were the attributes for which the optimal rice to water ratio was decided. The insect rice prepared with the most desirable mouthfeel was one that was neither too gummy nor too firm, with the former resulting from the addition of too much water or being overcooked, and the latter being from insufficient water or being undercooked. Observations on shape integrity and textural properties were used to guide the decision of an optimal rice-to-water cooking ratio, which was used for preparation of the four insect formulations in the sensory evaluation study.

Thus, based on the preparation method described above, 20 grams of each insect rice formulation mixed with 30 grams of water (1:1.5 ratio) was steamed in a 3.25 oz polystyrene cup. To efficiently and quickly mix the correct ratio of insect rice to water, several 20 gram aliquots of each insect rice formulation were prepared before beginning the sensory evaluation, along with test tubes containing 15 mL of water, where two test tubes or a total of 30 mL of water would be ready for mixing for each 20g sample. Two stovetop pots were employed, each with their own steaming basket and on a hot plate. As each steaming basket could hold one for each of the four insect rice formulations, leading to 8 cooked samples per batch, so a total of 16 test tubes were prepared. Once the insect rice was steamed for 15 minutes, they were removed from heat and divided into 8 two- ounce sample cups per 20 grams of insect rice. Thus, two pots yielded a total of 16 taste- portion samples for each formulation.

Sensory Evaluation

Participant Recruitment

To determine the acceptability of the insect rice product, a panel of 120 untrained participants was recruited via paper fliers posted across the Cal Poly Pomona campus and the University Village – the neighboring student residential area. Fliers were also advertised on the College of Agriculture and the College of Engineering digital displays.

Assistance in sending mass emails through listervs was received from the College of

Science, the College of Agriculture, the Office of Undergraduate Research, and the

Kellogg Honors College. The College of Environmental Design helped in forwarding the recruitment email to their faculty as well. While professors from various departments, including Plant Sciences, Biology, Kinesiology, and Sociology encouraged their students to participate, so did club officials from the Food Science Society and Agricultural

Biology Club to their student members. Recruitment continued for about 3 weeks.

Panelists were required to be: without shellfish allergies, consumers of brown rice, and between the ages of 18 – 65 years old. It was heavily stressed that those with shellfish allergies were unable to partake in the study as an allergy to shellfish was likely to produce a similar reaction to land insects (Mlcek and others 2014, van Huis and others

2013). It was also encouraged that the participants were either familiar with or enjoyed brown rice as this was the base of the products. The age range of the participants was determined as a means to exclude minors and those with a higher risk of compromised immune systems. Another prerequisite for participants was that they were willing to consume edible insects. Thus, subjects knew that the products to be tasted contained such an ingredient. While the “yuck” factor remains a major issue in entomophagy, those who

31 unknowingly consumed edible insect products could become seriously distraught after learning that this is what they’ve consumed. Hence, despite the potential bias that may occur from having this information, it was in the best interest of the subjects that they were informed of the products containing edible insects.

Qualified and interested individuals were asked to RSVP on a Google form found on the recruitment flier and emails. This was a short online form that asked a total of three questions: 1) “Do you have any shellfish allergies?” 2) “Do you eat brown rice?” and 3) “When are you available to conduct sensory evaluation on edible insect products?”

The last question allowed the subject to choose one of three available tasting sessions.

However, if any persons were to select “yes” for the shellfish question or “no” to the brown rice question, they would be prompted to the following message instead,

“Thank you! We appreciate your willingness to participate in this study!

Unfortunately, you are not eligible for this study. Please have a nice day!”

The purpose of these RSVPs was to again exclude those who do not qualify and to estimate the number of participants for each tasting session. Nonetheless, walk-ins were still welcomed.

Questionnaire

Designing of the questionnaire was completed through the sensory evaluation computer software, RedJade, developed by Tragon Corporation (Redwood Shores, CA).

It incorporated the Informed Consent Form, the evaluation questionnaires, and two other surveys.

Each questionnaire began with a Welcome page, which again stated that the product that the panelist was about to consume contained edible insects and that any persons with a shellfish allergies were asked not to eat this product. They were also requested to taste their samples from left to right and to rinse with water in between. At the end of the Welcome page, panelists were asked to input a participant number, given to them by a research assistant. This number is required in order for the questionnaire to begin.

After keying in their participant number, the panelist was presented with the

Informed Consent Form (ICF). This is a critical document that conveys the overall sensory evaluation objective and procedures, the benefits and risks of the evaluation, the bill of rights of the subject, and also provides contact information for any questions or concerns. While reading the entire ICF is dependent upon each participant, the program at least mandated the participant to scroll through the ICF before the “I Agree” button became available. Only when the subject accepted the ICF were they brought the insect rice samples.

Evaluation of the edible insect rice samples involved the four finalized formulations: 10CR, 15CR, 10LR, and 15LR. Organoleptic attributes and the overall liking were asked for each product. The samples were randomly served. The attributes of concern comprised of appearance, aroma, taste, and mouthfeel, primarily because the introduction of the raw insect material into the rice formulation led to some apparent alteration to each of these characteristics. For instance, both the cricket and the locust flour created a rice product that was darker in color, fishier in aroma and taste, and grainier in texture, which will be discussed in Chapter 4. To assess the degree of these

33 changes, each of the mentioned characteristics was rated on a Just About Right (JAR)

Scale. This particular scale was selected as it is capable of not only determining the acceptance of a sample, but also indicates the intensity of the attribute as perceived by the consumers. As such, the appearance was rated between “much too dark” and “much too light”; the aroma between “much too fishy” and “much too odorless”; the flavor between

“much too earthy” and “much too bland”; and the mouthfeel between “much too grainy” and “much too smooth.” The overall liking, on the other hand, was scored on a 7-point hedonic scale. While a 9-point hedonic scale is perhaps the most traditional, it requires extensive training to the sensory panelists so that they can distinguish the subtle difference in between these many anchors. On the other hand, a 5-point scale may be simpler for most untrained panelists to follow but providing insufficient information for investigators to conduct sufficient assessment of the differences between panelists’ perceptions.

As mentioned, in tasting the insect rice, the panelists were asked to try the samples from left to right, starting with the left most sample. To begin the questions for the first sample, the participants entered the sample’s 3-digit code. After completing their evaluation of the first sample, the panelist moved on to the second sample. The questions were the same for each sample. Between samples, the participants were provided a chance to rinse with water as the questionnaire rerouted to the next set of questions, counting down 10 seconds.

When all four samples have been assessed, a survey of the panelists’ perception of entomophagy was presented. Questions in this survey pertained to their familiarity and experience with, as well as their willingness to consume edible insects (Appendix D.2). It

34 also inquired about their perception of edible insects in the diet (e.g. appetizer, main entrée, etc.) and their willingness to purchase edible insect products in the market. Lastly, the questionnaire ended with a demographic survey, which inquired about their age, gender, ethnicity, and current education level.

Samples

Samples of each insect rice formulation was presented in a 2 oz polystyrene cup, along with a small cup of drinking water for rinsing, a fork, and napkin on a food tray

(Figure 3.1). The amount of insect rice was just enough to cover the bottom of the cup.

To keep the samples warm, the samples cups were lidded until ready to be given to the participants.

Figure 3.1 - Presentation of samples on a tray

Location and Schedule

Three tasting sessions were offered. The evaluations took place at the Human

Nutrition and Food Science labs in building 7 of the Cal Poly Pomona campus and were open from 5:00pm to 8:00pm on Tuesdays and Thursdays of week 3 and 4 of the 2016 academic spring quarter. This schedule was organized as a means to avoid interfering with class and club activities while still being held on days with a higher volume of classes.

A total of three areas were utilized for the entirety of the evaluation which included a Waiting Area, an Evaluation Room (Eval Room), and a Preparation Room

(Prep Room). The Waiting Area was the building 7 courtyard, furnished with patio tables and chairs of the Human Nutrition and Food Science (HNFS) Department, which allowed incoming participants to sit and wait for the next available booth if necessary. Subjects were told to meet at this outdoor area where two research assistants would be there to greet them.

In the Eval Room, a total of 12 booths were set up into three rows of four computers (Appendix D.1). Eight of these booths were composed on four silver-top metal benches, each housing two laptops and were isolated by a 3-walled makeshift booth.

These laptops, along with their chargers, were rented from the College of Agriculture.

The remaining four booths consisted of desktop computers with actual built-in booths.

Each booth was activated with the RedJade project. Two research assistants were assigned for this room to help monitor the evaluations. Near the exit was a bowl of chocolate with a sign that thanked the panelists and invited them to take a chocolate upon leaving. The chocolate included an assortment of fun sized Hershey’s and Mars brand

36 chocolates. The Eval Room took place in the sensory laboratory of the HNFS

Department, in building 7, room 117.

Samples were prepared in room 113, a neighboring classroom and kitchen adjoined by a small storage room. This Prep Room was designated for cooking, preparing, and arranging of the samples.

Sensory Preparation Flow

As participants arrived at the Waiting Area, research assistants were there to welcome them. Each subject was asked whether they had a shellfish allergy and if they were comfortable with consuming edible insects. Although these questions were already stated as requirements on recruitment fliers and emails, and also asked for on the Google

Form, this further ensured the safety of all participants. Besides this, walk-ins would not have RSVP-ed, leaving this as the best opportunity to confirm their qualifications. Next, when a booth became available, the research assistant ushered the participant to the booth and provided them a participant number that the subject then entered at the bottom of the

Welcome page. When a participant number is used, the research assistant crossed off the number on their list of four digit numbers, starting from 1001.

After inputting their participant number, each subject was asked to agree to the

ICF. Meanwhile, the Prep Room assistants would receive the presentation order for that subject’s samples from RedJade. Only after the panelist had accepted the ICF would the

Eval Room assistant have notified the Prep Room assistants that the participant was ready for their samples. At this time, the samples would be brought out by Prep Room

37 assistants. When the samples were delivered, the assistant explained the following to the participant:

“You’ll be tasting the samples from left to right, starting with [this] sample. After

the first set of questions, move on to the second sample. If you have any questions,

please feel free to ask [Eval Room research assistant].”

In order for the Prep Room to work efficiently, especially with high volumes of incoming participants, three research assistants along with the investigator were required.

Two of the research assistants divided the cooked insect rice into the 2 oz sample cups.

The other research assistant arranged the trays with drinking water, forks, napkins, and the cooked samples, in the order presented by the RedJade program. The investigator helped with the sample setup, mixed the uncooked insect rice with water, and refilled the test tubes with 15 mL of water in preparation for the next batch of samples.

When the panelists completed their evaluation, the Eval Room assistant thanked them for their participation, acquired their tray, and threw away any remaining materials.

Departing subjects were allowed to take a chocolate as they exited the Evaluation Room.

Research Compliance and Subject Confidentiality

Approval to conduct research with human subjects was obtained from the

Institutional Review Board of Cal Poly Pomona, IRB protocol # IRB-16-45. A total of 7 research assistants were involved in the sensory evaluation study. Each assistant obtained training from the Collaborative Institutional Training Initiative Program (Miami, FL) and were qualified to work with human subjects.

This study was established as a low-risk study. While the panelists were isolated to conduct their tasting session to provide privacy and to limit biases resulting from mutual influence or exchange, the subjects were still able to see each other upon entering and departing from the Eval Room. Even so, their personal information, such as their first or last names, was not collected at any stage of the study and the data collected was highly improbable to be linked to any individual subject. No sensitive information such as medical records or school transcripts was requested and those identifiers that were asked in the demographics survey were not enough to identify any one panelist.

Furthermore, the participant numbers were given by only the research assistants. Without the investigator, the sole person with access to the data, being involved in providing participant numbers, the link between the subjects’ data and their participant numbers is untraceable.

Statistical Analysis

For the water activity, color, and texture data, an analysis of variance or ANOVA with a post-hoc t-test was conducted on the average values to determine significance.

This was accomplished using the Data Analysis application in Microsoft Excel.

Statistical analysis for the sensory data was done automatically through the

RedJade computer program. This included a chi-square test for the scores on four sensory attributes when utilizing JAR scales and an ANOVA test with a Tukey post-hoc test for the hedonic scale questions (i.e. the overall liking).

CHAPTER 4

RESULTS AND DISCUSSION

Formulation Optimization

Extrusion of insect rice formulation was carried out using the single-screw, cold- forming extruder. Table 4.1 displays the extrusion flow rates and yields for all insect rice formulations.

Table 4.1 - Rates and percent yields from insect rice extrusion Mass flow rate (g/sec) Extrusion total yield (%, w/w) Desirable shape yield (%, w/w)

10CR 4.69 84.25 56.00 15CR 3.43 76.22 58.40 10LR 1.11 74.42 61.04 15LR 1.99 76.87 61.37

Between the cricket and locust formulations, the cricket rice products presented greater extrusion flow rates of 3.43 and 4.69 g/sec than those of locust formulations. This was certainly observed as the physical extrusion of the locust rice dough was noticeably slower than the cricket products. When comparing the % extrusion yield – the weight of samples collected after extrusion versus the initial weight of dough mixture, it is suggested that an improved total yield was also correlated with an increase in extrusion flow rate. This is seen as the higher extrusion flow rates for 10CR and 15CR formulations also resulted in relatively greater total yields of 84.25 and 76.22%, respectively, compared to the 10LR and 15LR formulations. Overall, it is suggested the incorporation of cricket flour, regardless of the addition levels, did not cause too much difficulty to the extrusion process. The incorporation of locust flour, on the other hand, caused some

40 unfavorable effect not only on the extrusion process, but also on the quality of the extruded rice products.

While the moisture content offers negligible variability between locust and cricket formulations (Table 4.6), perhaps the difference in extrusion flow rates rather lies in the composition of these two edible insects. For instance, as required in starch-based extrusion formulations, vegetable shortening is typically added at a pre-determined, optimized level to ensure consistent extrudability by reducing friction or providing lubrication between the dough mass and the extruder barrel/screw set. Therefore, the addition of cricket flour, with its slightly higher fat content (Table 4.11), may have caused some enhancement to the functionality of the lubricating oil. On the other hand, the suggested greater quantities of fiber in locust rice products (Table 4.8) as discussed later, may have instigated some negative effect to the extrusion process. In combination, these qualities consequently resulted in a faster mass flow rate for cricket rice formulations when compared to locust rice products. However, as extrusion is a very complicated process, influenced by a wide range of operation parameters, such as feed rate, barrel temperature and pressure, and more importantly, the composition of dough mixture (Fellows 2009), multiple extrusion runs should be conducted for each insect formulations. Doing so would further confirm these conclusions.

Nonetheless, all four formulations achieved a similar yield of final rice particles with desirable shapes and size, all within a range of 56% to 61% (w/w), which means about one third of the raw materials blended in the dough mass was lost during extrusion, dehydration, and/or size screening. This is a normal occurrence for small scale, 1 kg batch productions as it was necessary to shut down the laboratory extrusion equipment

41 between the productions of each insect formulation. In doing so, a thorough cleaning before processing the next formulation could be performed to avoid any potential contamination between formulations. However, this did cause some wastage at the beginning of each extrusion run as proper shear stress and barrel temperature were required to allow the dough mass to become compressed and subsequently pushed out with desirable consistency and then cut into properly-sized rice-shaped pieces (~5.90mm x 1.98mm x 1.70mm). On the other hand, continuous production utilizing large batches, such as with industrial extruders, can improve this production yield to over 95%.

Even so, the extrusion of rice flour formulations containing either cricket or locust flour up to a 15% addition level was feasible. The visual representations of those products can be found in Table 4.2.

Physicochemical Properties of Uncooked Insect Rice

Color and Shape

Although visual observation could qualitatively differentiate the color variation between some of the insect rice products, a colorimeter (Konica Minolta, Ramsey, NJ), was employed to objectively quantify the color of the insect rice formulations while using market brown rice as the control. The L*a*b* values obtained from this device provides a three-dimensional coordinate for all perceivable colors, where L* = lightness to darkness on a scale of 100 to 0, a* = red-to-green values, and b* = yellow-to-blue values.

Specifically, L* = 0 denotes the darkest black and L* = 100 as the brightest white, while when a* and b* = 0 provides a true neutral grey. On the other hand, positive a* values represent red colors and negative a* values represent green colors. For b* values, positive

42 numbers characterize yellow colors and negative numbers as blue colors. The L*a*b* values can be found in Figure 4.1 and can be referenced to the images of these rice products as shown in Table 4.2.

Table 4.2 - Insect rice formulations 10CR 15CR 10LR 15LR MBR

70 d a 60 a b c 50

20 d b b 10 a a a a b c d 0 10CR 15CR 10LR 15LR MBR L* values 57.30 56.95 55.84 53.37 64.47 a* values 4.89 4.85 5.69 5.49 4.04 b* values 7.98 8.54 11.31 10.80 15.25 Figure 4.1 - L*a*b* values for insect rice products Color of the insect and market rice products are displayed as L*a*b* values where L* = lightness to darkness on a scale of 100 to 0, a* = red-to-green values, and b* = yellow-to-blue values Different lower case letters distinguish a significant difference for that color parameter and the error bars represent the standard deviation.

Between cricket and locust rice formulations, there was a notable difference in color shades, in terms of both a* and b* values. Cricket rice products, at both 10% and

15% additions, were generally greyer in color as their a* and b* values were significantly

(p < 0.05) lower than those of locust formulations. Subjectively, this was also easily discerned through visual observations. These differences could be attributed to the color of the starting insect ingredients since the cricket flour was greyer than the locust flour as shown in Appendix A.1.

In comparing the two addition levels, 10% versus 15%, statistical analyses showed different directions. For example, for the two cricket formulations, there were no significant differences (p > 0.05) in all color attributes between the two addition levels.

This complemented what was perceived via visual inspection as both 10CR and 15CR were both grey in color and with little distinguishability between the two formulations.

On the other hand, locust rice products became noticeably darker with significantly lower

L* value (p < 0.05) while lessening in red shade (significantly smaller a* value) when increasing the addition from 10% to 15% in the rice formulation. The change in darkness was an anticipated result as the red-brownish locust flour (Appendix A.1) was expected to be a major source of color in the final product. On the other hand, greater red values were predicted to follow an increase in locust flour levels. In spite of objective results from statistical analysis disclosing a significant difference between the two locust formulations in L* and a* values, there was little perceivable differences between them by visual observation. Overall, the quantity of locust flour in the rice formulation seems to present a stronger influence than cricket flour on the color of the final product.

However, it should be noted that these differences in color were observed for the uncooked rice. After cooking, it was witnessed that the color differences between the four insect rice products became noticeably enhanced. Sensory evaluation results also paralleled this finding, which will be discussed in a later section.

Contrasting the insect rice products with the control, all formulations exhibited darker shades (lower L* values) and were redder (greater a* values) and less yellow

(smaller b* values), regardless of insect source. Again, this was supported by visual observations and was attributed to the insect ingredients as they are the chief supplier of the dark color in the brown rice flour-based formulations while other ingredients within the formulation, such as brown rice flour, tapioca starch, and sodium alginate, present with a white color.

To examine the shape, size, and surface morphology of the extruded insect rice, a digital microscope (Digital Blue, Atlanta, GA) was employed at both 10x and 60x magnifications. Figure 4.2 illustrates the shape dimension and surface properties of rice particles as a result from extrusion.

Figure 4.2 - Microscopic view of insect rice formulations and control At 10x, from left to right: 10CR, 15CR, 10LR, 15LR, and MBR

In figure above, the frontal view of the rice particles is the smooth surface that was pressed alongside the die plate while the left and right edges are where the extruded strand was cut. The right most rice particle is an actual brown rice kernel. In contrast to

45 the coarse and sometimes flat edges of the extruded rice, the entire surface of the actual kernel is smooth, polished, and curved.

Table 4.3 - 60x magnification view of insect rice formulations and control 10CR 15CR 10LR 15LR MBR

Differences between MBR and extruded rice products can further be seen in Table

4.3. Presented here are the cut edges of the extruded rice at 60x magnification, where the surfaces can be seen as flakey, uneven, and rough. The cracked spots on the surface indicates that the incorporation of relatively high levels (10% and 15%) of insect flours into the rice flour matrix actually reduced the starch binding, impacting on the consistency of the product. MBR shows, as in Figure 4.2 and Table 4.3, an entirely smooth and polished layer. While these variations may be an obvious outcome it is important to note that the fine layer of bran with the brown rice kernel serves as a protective structure for the starch inside. Extruded rice products were homogenously composed and instead relied on chemical structures (i.e. alginate-calcium crosslinking at the surface) to maintain structural integrity.

Texture

In determining the texture of the insect rice products while utilizing market brown rice as the reference, a Stable Micro Systems TA-XT plus texture analyzer (Godalming,

UK) was employed. In particular, the hardness and adhesiveness were of concern.

Presented below in Figure 4.3 is the texture analysis profile of LR15, in quintuplicate, used as an example to illustrate these texture attributes. The texture analysis profiles for all other insect rice samples are presented in Appendix C.1 – C.4. In the figure, hardness is the peak force (kg) and measures the amount of strength required to bite down on the product. Adhesiveness is represented as the pull-down force (kg) with a negative value and reflects the sticking effect of a product when it adheres to the teeth during mastication. Together, these two characteristics can deliver some implication on how difficult a product is to chew. For instance, the greater the hardness value, the more energy is required to bite through the rice. A product that is very adhesive, on the other hand, tends to attach to the “teeth” while chewing. The texture values for all rice products are summarized in Table 4.4.

Figure 4.3 – Texture analysis of 15LR, in quintuplicate

Table 4.4 – Texture measurements of rice products Rice product Hardness (kg) Adhesiveness (kg) 10CR 0.675 ± 0.090a -0.009 ± 0.001a 15CR 0.621 ± 0.040a -0.009 ± 0.001a 10LR 0.669 ± 0.066a -0.007 ± 0.000a 15LR 0.662 ± 0.116a -0.008 ± 0.001a MBR 1.048 ± 0.033b 0.000 ± 0.000b Different lower case letters distinguish a significant difference in texture

While most of the insect formulations, with the exception of 10LR, demonstrated to be different amongst each other, all formulations were revealed to be significantly softer (lesser hardness values) and much more adhesive (greater negative adhesive values) than the control MBR (p < 0.05). The softness of the insect rice is due to its lack of a complete bran structure that, conversely, is present and intact in regular brown rice kernels. Bran contains >20% of insoluble fiber of which is made up of equal fractions of cellulose and hemicellulose (Qi and others 2015). These components make this outer layer a sturdy exterior, protecting the inner starch from being leached during high stress conditions such as heat. For the insect rice, however, the product is instead held together by the physical compression during extrusion and chemical bonding at the surface through a loose alginate-calcium crosslinking network. Although these chemical bonds cannot be observed by the human eye, they are certainly productive in maintaining the shape integrity of the product through cooking. However, as displayed in the microscopic images seen in Table 4.3, the polished and smooth layer of the MBR bran can readily repel water, whereas the rough and uneven surface of the insect rice is easily penetrated.

This will ultimately result in some starch being leached out upon cooking. The gelatinization of starch leaching to the surface with surrounding water during cooking is the cause for the observed adhesiveness. The act of gelatinization, or the uptake of water

48 by amylose and the subsequent breaking of their crystalline structure when heated, creates a starch paste between the starch and water mixture which is sticky to touch

(BeMiller and Huber 2008). This paste was observed to be coated around the insect rice particles as well as pooled at the bottom of the container during the cooking optimization process. It was also noted that upon cooling, the adhesiveness of the insect rice products increased. As the temperature reached room temperature, the starch molecules began to reform their crystalline structure, following a process known as retrogradation. With this recrystallization, the insect rice became increasingly gummy. Both starch gelatinization and retrogradation are also typically observed when cooking market rice, albeit the adhesiveness is significantly more pronounced in the insect rice formulations.

There were no differences between 10% and 15% formulations or between cricket and locust rice products. Of the exception that was aforementioned, 10LR had demonstrated to be statistically less sticky than the other three formulations (p < 0.05).

Even so, the actual difference in adhesiveness was a mere 0.001-0.002 kg which is negligible.

Water Activity

A major indicator for a food’s shelf life is in its water activity (aw) as this pertains to the amount of free water available for microbial growth. Foods of higher water activity values, especially when above 0.6, often lead to faster spoilage and shorter shelf life due to the survival and proliferation of molds, yeasts, and bacteria.

Table 4.5 - Water activity of rice products

Rice product aw 10CR 0.160 ± 0.001a 15CR 0.160 ± 0.000a 10LR 0.171 ± 0.002b 15LR 0.180 ± 0.001c MBR 0.461 ± 0.001d

Different lower case letters distinguish a significant difference in aw

While the literature on the water activity of insect products is currently unavailable, the water activity of MBR in Table 4.5 was comparable with the value of

0.491 that was reported by Schmidt in the food chemistry laboratory in the University of

Illinois (Schmit and Fontana 2007).

All insect formulations showed water activities lower than the critical point of 0.6

(Table 4.5). Moreover, the insect rice formulations all exhibited aw levels that were significantly lower than that of the market brown rice (p < 0.05), thus indicating a longer shelf life. As the water activity values of the cricket and locust flours were respectively found to be 0.264 and 0.244 (Appendix A.4 and A.5), which are greater than the final insect rice products, these improved values lie within the processing of the insect products, including extrusion and dehydration. Specifically, the extruded rice was dehydrated for at least 6 hours under 165oF, which not only removes free water for microbial growth but also destroys existing microorganisms to some extent.

Contrasting between formulations shows a significantly different level of water activity (p < 0.05), with the exception of 10CR and 15CR which had similar values (p >

0.05). First, cricket formulations were established as shelf-stable products with a lower level of water activity, yet the addition level of cricket flour does not seem to present as a factor. On the other hand, while both locust formulations were shown with significantly

50 greater aw values, the higher incorporation level of locust flour further increased the amount of free, available water. In investigating the moisture content of the two types of insect flours, locust flour was found to have more moisture than cricket flour (Appendix

A.6). This suggests the locust flour itself likely to be a main cause of higher water activity values in locust rice formulations compared to their cricket counterparts.

Nonetheless, 10LR and 15LR had water activity values only greater by 0.011 and

0.020, respectively, compared to their cricket equivalents and the actual impact to shelf life is considered to be minimal. Furthermore, the water activity values for all insect rice formulations demonstrated to be well below the critical 0.6 limit, to the credit of proper dehydration after the extrusion process. Thus, the insect rice products demonstrate to be capable of averting microbial growth and spoilage.

Moisture Content

While water activity is the measure of the amount of free water available for microbial growth, moisture content is the amount of water within the food, including the free and bound forms of water. Table 4.6 shows the moisture content of the insect rice products and the control market brown rice.

Table 4.6 - Moisture content for rice products Rice product Moisture (%) 10CR 8.09 15CR 10.24 10LR 8.51 15LR 9.71 MBR 8.16

Just as in evaluating the water activity of the insect rice products, there is currently no information available in the literature on the moisture content of such products. The laboratory measurement of the control MBR in Table 4.6 is comparable to the value of 7.0% reported by Schmidt (Schmit and Fontana 2007). However, there seems to be some disparity between the two values, which could be a result from the type of rice species, time of analysis, or other confounding variables. This could also be due to the variance in analytical methods as the current study utilized the Shimadzu MOC-120H moisture analyzer (Kyoto, Japan) while Schmidt employed a vacuum oven.

With that, it is considered that the Shimadzu may have provided some overestimation of the moisture content of the insect rice formulations. Even so, as shown in Table 4.6, the 10% insect formulations (both 10CR and 10LR) had similar values to the control. Conversely, as the incorporation of the raw insect materials was increased to

15%, the residual moisture content of the final products was also raised by 2.15% and

1.2% in 15CR and 15LR formulations, respectively. This implies, yet again, that the residual moisture level in the finished products was associated with the addition level of the raw insect flours.

Based on the water activity measurements of the cricket and locust products, it was anticipated that 10CR and 15CR have lower moisture contents than their locust counterparts. However, this was not the case as 15CR demonstrates to have the highest moisture contents, even greater than MBR. It should be considered, however, that only one measurement was conducted for moisture content instead of triplicates, which was performed for water activity. If replicated measurements were completed for each sample, then the mean values for moisture content may show in a narrower range.

Nonetheless, while moisture content can provide some suggestion in regards to food stability, water activity remains the chief indicator of shelf life.

Shelf Life Study

With water activity being an essential indicator towards the spoilage of food products, these values were analyzed at 0 months and after a 3 month period (Table 4.7).

Table 4.7 - Changes in water activity (aw) after 3 months for insect rice products Rice product 0 Months 3 Months 10CR 0.160 ± 0.001a 0.219 ± 0.002a 15CR 0.160 ± 0.000a 0.236 ± 0.002b 10LR 0.171 ± 0.002b 0.369 ± 0.009c 15LR 0.180 ± 0.001c 0.310 ± 0.002d Different lower case letters distinguish a significant difference within that month of time

At month 3, each insect formulation presented with an increase in water activity.

Between the cricket and the locust products, the latter displays a greater amplification as

10LR increased by 115.79% and 15LR increased by 72.22%. Cricket formulations increased by 36.88% (10CR) and 47.50% (15CR). This was an expected result as the water activity of locust products at 0 months was greater than their cricket equivalents.

As it was established previously that the residual moisture content left in dehydrated products was primarily derived from the insect flours, it was predicted that the higher insect formulation would have a higher water activity over time. However, this was not demonstrated for the two locust samples as the greatest increase in water activity was in the 10LR instead of 15LR. With the discrepancy between locust rice products being only

0.059, perhaps this was a results of some calibration error in the instrument or other confounding variables.

Even with this inconsistency, the results further establish that the water activity is indeed greater in locust rice products than in cricket rice formulations. It also reveals that after 3 month storage, the insect rice products would have more free water available for microbial growth, which is a concern and needs further monitoring over a longer storage period. Despite this increase the water activity values of all four formulations were still well below the critical limit of 0.6 after 3 months, which suggests they should be still considered as safe for consumption upon proper cooking.

In general, as reported in the literature, brown rice is considered to have a shorter shelf life than white rice. This is perhaps due to the richer and more balanced nutrient profiles in brown rice versus white rice, which makes it more prone to fat oxidation and microbial proliferation overtime. The further grinding process to convert the grain kernels to the flour makes the rice flour more vulnerable to oxidation, which may halve the original shelf life as an intact grain. The Oldways Whole Grain Council (2013) recommends brown rice kernels to be stored for up to 6 month in the pantry while the flour derived from the whole brown rice grain to be kept only for 3 months. Together with the reference information, the shelf life of the insect rice products is estimated to last over 3 but not beyond 6 months. The water activity measurements, along with an oxidation study, of the four formulations after 6 months storage should confirm this statement.

Nutrition Content

Fiber

To assess the total dietary fiber content of the insect rice formulations, an Ankom

Dietary Fiber Analyzer (Macedon, NY) was utilized. The results are reported below in

Table 4.8 as % recovered residue that contains undigested protein and ash in addition to the total dietary fiber (TDF) content in the insect rice formulations. As explained in the methodology section, due to the limited laboratory capacity on protein and ash analyses, this % recovered residue value was used as a rough estimation of TDF content. The

USDA National Nutrient Database for Standard Reference, Release 28 (2015) was used to obtain the TDF value for MBR.

Table 4.8 – Percent recovered residue of insect and rice products Rice product % Recovered residue 10CR 12.35 ± 0.92a 15CR 12.20 ± 3.68a 10LR 14.25 ± 1.91a 15LR 21.75 ± 6.15a MBR* 3.60 Same lower case letters distinguish no significant difference in values *MBR value of 3.60 is the total dietary fiber value as obtained from the USDA National Nutrient Database for Standard Reference, Release 28 (2015)

Locust rice products were revealed to contain greater quantities of recovered residue. While this could be considered owing to the dietary fiber provided by the locust raw material, this may be unfolded as the current measurements over-estimated the TDF content without correction from other residual components. A particular concern is with the residual protein remaining undigested after protease digestion, as protein is shown to be available in relatively high amounts within both the raw insect flours (Appendix A.1

55 and A.2) and in the insect rice formulations (Table 4.10). This would suggest that the % recovered residue may contain less fiber and more of other constituents, especially protein. However, by associating the dietary fiber content of the cricket flour provided by the vendor (Next Millennium Farms) in (Appendix A.1) with the actual lab measurement using the Ankom dietary fiber analyzer (Appendix C.1), the overestimation of the latter could be roughly determined. As the COA from the vendor details the total dietary fiber content of the cricket flour as 7.7% and the current study reports the same material with

36.40% recovered residue, it is suggested that the values in Table 4.8 for 10CR and 15CR might be an overestimated by up to 78.85%. While this could be used for some approximation of the actual TDF content in the rice samples, it still does not factor the

TDF provided from the brown rice flour, the bulk ingredient of the insect rice products.

Nonetheless, taking into account that the protein, which is likely to be the other major constituent within the recovered residue, is not excessively diverse between the insect rice formulations (Table 4.10), and that the other major source of dietary fiber is the brown rice flour, which is constant within the four formulations, some comparisons of

TDF content between the insect rice products could be made. Thus, it is proposed that locust rice formulations do contain a superior level of TDF than their cricket counterparts. It is known that as opposed to providing typical insoluble or soluble dietary fibers, such as cellulose or pectin, edible insects are more likely to contain chitin (van

Huis and others 2013). This is an insoluble fiber, similar to cellulose, but instead contains amino group substitutes. This non-digestible carbohydrate is known to be found in the exoskeleton both the Insecta and Crustacea class of the animal kingdom. Considering the exoskeleton of locust is perhaps heavily composed with chitin, this implies the fiber

56 content that could be detected by the instrument was primarily derived from this compound. While chitin may provide some graininess to the texture of the rice, which is later discussed in the Sensory Evaluation section, the ability to interact with dietary lipids has led to some suggestion of chitin being included as a functional fiber (Gropper and

Smith 2013). In fact, chitin is already being utilized in Japan to fortify cereals to enhance the levels of fiber and calcium (Williams 2010).

While the brown rice flour does differ in quantity between 10% and 15% insect rice formulations, evaluations between 10CR and 15CR, as well as 10LR and 15LR remain inconclusive as it is uncertain whether the differences are caused by the insect or the brown rice flour. Furthermore, inconsistencies may have been caused by some procedural noise as samples containing 10% or more of fat may cause interference in the

Ankom dietary fiber assessment. With Table 4.11 displaying the fat content of 15CR to be near this value, this may be the reason for the greater value of recovered residue, since some lipid residue may be included without proper pre-defatting procedure. The recovered residue value is also inappropriate for contrasting with the theoretical dietary fiber level of MBR that is derived from the USDA, instead of an actual measurement with the Ankom instrument.

Even so, the results do reveal exciting evidence for the dietary fiber contents in cricket and locust products, which has not been reported in the literature. Previous research conducted for the fiber determination of edible insects has been mainly on the crude fiber content, which is irrelevant for human nutrition. However, the current study does demonstrate some results supporting locust to contain greater quantities of TDF than crickets.

Iron

A graphite furnace atomic absorption spectrometer (AAS) (Perkin Elmer,

Waltham, MA) was employed for the analysis of both the iron content of raw insect flours and the insect rice products. The values are shown in Table 4.9 below. It should be noted, however, that the iron content of MBR was collected from the USDA National

Nutrient Database for Standard Reference, Release 28 (2015).

Table 4.9 - Iron content of insect and rice products Product Iron (mg/ 100g) Coefficient of Variance (CV, %) Cricket flour 6.80 ± 2.55a 37% 10CR 6.68 ± 3.65a 55% 15CR 4.09 ± 1.80a 44% Locust flour 8.05 ± 0.50a 6% 10LR 4.42 ± 0.62a 14% 15LR 2.87 ± 0.17a 6% MBR* 1.29 n/a Same lower case letters distinguish no significant difference in values *MBR value of 1.290 is the iron value as obtained from the USDA National Nutrient Database for Standard Reference, Release 28 (2015)

Based on the literature information, the locust rice products were anticipated to be a better source of iron than the cricket formulations. For instance, Oonincx and van der

Poel (2011) found that, locusts of the same species of those used in the current study

(Locusta migratoria) have 15.10 mg of iron per 100g weight. In a separate study, crickets, Acheta domesticus, were reported to have 6.27-11.23mg / 100g (Rumpold and

Schluter 2013). When at first examining the iron content for the raw materials (i.e. cricket and locust flours) (Table 4.9), the laboratory measurements of 8.05 mg/100 g for locust flour and 6.80 mg/100 g for cricket flour showed to be similar to these literature findings, especially locust flour containing slightly higher iron levels than the raw cricket material.

The cricket value was also close to that of the COA provided by Next Millennium Farms

58 as well (Appendix A.1). Yet, when examining the iron contents measured for insect rice formulations, it was not easy to generate a clear conclusion. While greater iron levels were expected to be observed in formulations with increased additions of insect flours, this was not the case in the current study. The inconsistency is mainly related to the iron analysis protocol, where the wet ashing process using concentrated HCl and HNO3 should completely digest the organic matters in the samples. This digestion subsequently converts all minerals in the food samples into an elemental form that can be detected by

AAS. However, during the current lab operation, a lipid-like precipitate was observed to be present in most of the wet assay solutions, suggesting the acid digestion was not properly completed. Furthermore, the ensuing filtration step in an effort to remove this fat-like layer may have also caused some loss of iron as it may be entrapped in these deposits. This ultimately led to lower values of iron content in the wet assay solutions for all samples.

Each sample was prepared with two assay solutions, or measured with duplicates.

The standard deviation, or the coefficient of variance (CV, %) from the duplicates was significantly high for most of the samples, especially for samples of cricket flour and the two rice formulations made of it, ranging from 37% to 55%. This variation between different assay solutions even for the same sample confirmed that there might be some interference using the current iron analysis protocol, which certainly requires further standardization in order to produce repeatable and reproducible results.

What can be successfully drawn from the GF-AAS iron analysis is that this essential mineral is indeed found within these insect rice products, and is presented with

59 higher values when compared to the control MBR, which might be owed to the incorporation of insect flours.

Protein

An analytical lab located in Anaheim, CA called Certified Laboratories facilitated the protein and fat analyses of each of the four insect rice products. The results are displayed below in Table 4.10 for protein and Table 4.11 for fat, with the exception of the values for MBR. The MBR values were obtained from the USDA National Nutrient

Database for Standard Reference, Release 28 (2015).

Table 4.10 - Protein content of insect and rice products Product Protein (%, w/w) 10CR 12.72% 15CR 15.56% 10LR 14.06% 15LR 15.70% MBR* 7.54% *MBR % protein was obtained from the USDA National Nutrient Database for Standard Reference, Release 28 (2015)

60 which not only fills in some of the gap within this field of research but also offers support in the utilization of edible insects in the diet.

As established by the US Food and Drug Administration (FDA), the daily value

(DV) is a term that provides consumers a reference intake level for macro- and micronutrients. Based on a 2,000 calorie diet for adults and children of four or more years of age, the DV for protein is 50g (FDA 2015). While this value was created to ease consumer confusion and is utilized as a standard for the food industry, the 2010 Dietary

Guidelines for Americans, published by the US Department of Agriculture (USDA) recommends adults males, 19 – 55+ years old, to consume 56 g of protein and adult females of the same age range to consume 46 g of protein (USDA 2010). According to above results, the insect rice products provide 12.7 to 15.7 gram of protein per 100 g insect rice (Table 4.10); thus, with a standard serving size of ½ cup of rice, or ~80g, this would provide 10.18 – 12.56 g of protein. Based on the DV value of 50 g established by the FDA, just one serving of the insect rice would provide 20 – 25% of the daily value for this macronutrient, confirming the cricket and locust rice products to be excellent sources of protein.

Referring to the available information of protein content provided by the vendors

(Appendix A.1 and A.2), where the cricket flour and locust flour have a protein content of 59.46% and 54.67%, respectively (dry weight basis), it was predicted that the cricket rice formulations would offer a superior quantity of protein than their locust counterparts.

However, the results as displayed in Table 4.10 shows this was not the case as 10LR and

15LR showed greater amounts of protein, although perhaps not significantly dissimilar in value. Protein levels were reported to be within 64.38 – 70.75% for crickets (Acheta

61 domesticus) (Rumpold and Schluter 2013) and 55.5 – 64.9% for locust (Locusta migratoria) (Oonincx and van der Poel 2011). Although the crickets in the study conducted by Rumpold and Schluter (2013) have higher quantities of protein than the

Locusta migratoria, the crickets used in the current study are of a different species,

Gryllodes sigillatus. It is important to consider that there may be variability in nutritional content from edible insects as they differ between and within species and are also influenced by type of diet and environment. Furthermore, the method of analysis could also provide some disparities in the results. Thus, with these research findings solely as a reference, it seems plausible that the cricket and locust offer comparative protein content in the rice products developed in this study.

Fat

Below in Table 4.11 are the representative values for the % fat in the insect rice and MBR products. While the lipid assessment of the four insect rice formulations was outsourced to the aforementioned analytical lab, the MBR fat content was derived from the USDA National Nutrient Database for Standard Reference, Release 28 (2015).

Table 4.11 - Fat content of insect and rice products Product Fat (%) 10CR 7.76% 15CR 9.73% 10LR 7.44% 15LR 7.97% MBR* 3.20% *MBR % fat was obtained from the USDA National Nutrient Database for Standard Reference, Release 28 (2015)

(Acheta domesticus) had 18.55 – 22.80% fat (Rumpold and Schluter 2013) while locust

(Locusta migratoria) had 17.9 – 29.6% fat (Oonincx and van der Poel 2011). Thus, the ranges found in the current and previous research support crickets as the potentially more impressive resource for lipids, although perhaps only marginally.

According to the FDA (2015), the daily value for fat is 65g for a 2,000 calorie diet. In another perspective, the percent of calories obtained from fat should be in a range of 20 – 35% (or 400 - 700 calories for a 2,000 calories diet) for both males and females who are 19 – 55+ years old (USDA 2010). From the fat analysis results as displayed in

Table 4.11, the insect formulations provided 10-12% DV for fat (6 grams in 10% formulations and up to 7.7 grams in 15% formulations) when one serving of ½ cup or approximately 80g of such rice products were consumed, which did not exceed the

USDA recommended intake level. While in the United States, the USDA recommends

Americans reduce their calories from solid fats, this is not of concern for those in developing countries, for whom these insect rice products are currently aimed towards.

Protein-energy malnutrition is the most critical nutritional issues faced by populations in Asia, Latin America, and Africa, especially in children (Latham 1997).

Perhaps of the most infamous and severe cases of the malnutrition emergencies are marasmus and kwashiorkor. Here, marasmus is simplified to be caused by the overall

63 deficiency of food, while kwashiorkor is caused by lack of energy and protein as the diet is primarily composed of carbohydrates. The former results in the dangerous wasting of the body as fat and protein stores are utilized to create energy to survive. People inflicted with kwashiorkor, on the other hand, results in serious edema along with wasting. As the many populations within developing countries are already practicing entomophagy, the novel insect rice products can naturally offer a supplement to the plain rice staple within their diet. As previously discussed, the insect rice can provide not only 150 – 200% times more protein, but overall increase the caloric intake for these starved people. In comparison to typical brown rice, the insect rice products can provide up to 47 more of much needed calories per serving of 80 grams. The insect rice is also highly likely to provide other added advantages, such as increased consumption of iron and fiber, although the exact quantities are still to be determined.

Standardization of Optimal Cooking Conditions for Insect Rice

Without the outer shell of bran as seen in regular brown rice, the insect rice would not be able to withstand a complete immersion cooking method as typically employed by an automated rice cooker. Instead, the extruded rice containing edible insect flours requires a steaming method (Appendix B.2). To determine the optimum rice to water ratio when cooking the insect rice formulations, a series of 1:1, 1:1.5, and 1:2 ratios were assessed after which the appearance, aroma, taste, and texture were evaluated. In all formulations, increasing water levels exhibited lighter color in the cooked rice (Table

4.12) which suggests an increase in water uptake led to a reduced color intensity. Each formulation also demonstrated to become more slippery at the particle surface, mainly

64 due to over-gelatinization of starch. This can be seen by the greater amount of glean in samples prepared by a 1:2 rice-to-water ratio in Table 4.12. This is a common result from cooking regular rice, especially when boiling with greater amounts of water. The slipperier and stickier surface of the rice samples prepared by 1:2 ratio was not only considered to be unappealing, it also was perceived as overcooked as the core of the particle was overly mushy when it was cut through. Similarly, the 1:1 ratio caused the cooked rice to have a dry and grainy texture, as well as to have a harder core when chewing, suggesting this ratio was not sufficient to cook the rice sample properly. This was particularly the case when cooking locust formulations using low water levels as the resultant rice demonstrated distinctly grainy and even crunchy qualities. This was further evident in 15LR. Thus, it was determined that the 1:1.5 ratio was the best condition for cooking the insect rice formulations when using a steaming method.

Table 4.12 - Cooked insect rice products of different rice to water ratios 1:1 1:1.5 1:2

10CR

15CR

10LR

15LR

To prepare for the sensory evaluation study, small 20 gram batches of the insect rice was steamed using this 1:1.5 ratio, which would subsequently yield approximately 8 tasting-portion samples per batch. This technique allowed the rice samples to be prepared

66 with consistent results for tasting sessions while also making it convenient for the volume of expected participants.

After cooking the insect rice samples under the optimized conditions, preliminary sensory evaluations were performed. First, all formulations seemed to have become brighter in their color post steaming. This was especially noticed in the locust rice samples as the cooked rice had become a more vibrant red-brown version than its uncooked grey-brown form. During the cooking, water uptake by the rice perhaps allowed the color to be better reflected off the surface. With that, the color differences between each insect product had seemed to intensify as well. For instance, whereas the greyness was nearly undistinguishable between uncooked cricket rice formulations, the

10CR had become noticeably paler than 15CR after cooking. This occurred between the locust formulations as well as 10LR became remarkably lighter colored than 15LR after steaming. This can be explained by the fact that during cooking, the water was absorbed by the rice particles, followed by starch gelatinization and significant particle size swelling. This expansion consequently diluted the color intensity to some extent. Also, once the starch gelatinized, it changed from its crystalline form to an amorphous state of amylose, which also enhanced the transparency of individual rice particles, resulting in a paler appearance. With less starch being present in 15% formulations, this would explain the darker colors observed in 15CR and 15LR. In assessing the aroma and flavor of the cooked insect rice, both cricket and locust formulations had an earthy aroma and taste.

However, the cricket rice had slightly more umami flavor, reminiscent of oyster sauce in

Asian cuisine, while the locust rice presented with a more briny “shrimpy” flavor.

Although this may be peculiar discovery for some, crickets and locust (land insects) and

67 shrimp (shellfish) are actually both arthropods, which are invertebrates from the

Arthropoda phylum of the animal kingdom (Kaae and Kaae 2000). Being separated only by class, and thus being so closely related, explains the likeness in aroma and taste.

Sensory Evaluation

Demographics

Over the span of 3 days, a total of 120 untrained panelists were recruited from the

Cal Poly Pomona campus for this study. To profile the sensory panel, a demographics survey was included in the questionnaire and the age, gender, ethnicity, and the highest level of education were asked (Figure 4.4 – 4.7).

3% 1% 2%

55+ years old Male 18% 45% 45 - 54 years old Female 35 - 44 years old 55% 25 - 34 years old 76% 18 - 24 years old

Figure 4.4 - Age of sensory panel Figure 4.6 - Gender of sensory panel

3% 3% 3% 8% 5% PhD 17% 7% Other Master's Black 13% Latino Graduate Asian 4th year 30% 38% White 28% 3rd year 45% 2nd year 1st year

Figure 4.5 - Ethnicity of sensory panel Figure 4.7 - Education level of sensory panel

With recruitment taking place on the Cal Poly Pomona campus, it was no surprise that the majority of the panel were in their early 20’s, about 76% (Figure 4.4). Only 6% of the panel were 35 year old or above. It was also found that the ratio of men to women panelists was almost 50:50, with only 10% more females to males (Figure 4.6). In a previous study conducted in Belgium, a similar ratio was found in their sensory evaluation with mealworms (Tenebrio molitor) and crickets (Acheta domesticus) (Megido and others 2014). These similarities may suggest that either entomophobia or lack thereof holds no discrimination against gender, although this may require further analysis of the data to confirm this proposition. In terms of ethnicity, there were a greater number of

Latinos and Asians, together making up almost 70% of the sample population (Figure

4.5), which may simply be reflective of the campus population.

An issue, found only after surveys were conducted, was in the options available for the education level question. There was no option for subjects to identify themselves as persons who have graduated from their bachelors but were not pursuing any further education at the time. For instance, there were no choices for completing a 2 or 4 year degree while currently in the workplace and not continuing any education. There was also no option for those who were still in school but past their 4th year of college. With that, there may be some inflation within the data for the groups stating to be within their 1st -

4th year in college (Figure 4.7).

Perception of Entomophagy

Several inquiries pertaining to the concept of entomophagy were asked in a separate survey within the questionnaire. These questions and their responses are

69 displayed in Figure 4.8 – 4.11 below, with the exception of the free answer responses pertaining to the panelists’ experience with consuming edible insects, if any. These responses are presented and analyzed in Appendix D.1 – D.22.

18%

No 42% No Yes Yes 58% 82%

Figure 4.8 - Familiarity with entomophagy Figure 4.10 - Experience with entomophagy

Definitely would not eat Definitely would not buy Probably would not eat Probably would not buy 3% 3% Maybe/ maybe not 5% Maybe/ maybe not Probably would eat Probably would buy 15% Definitely would buy Definitely would eat 13% 34% 22%

38% 29% 38%

Figure 4.9 - Future willingness to consume Figure 4.11 - Future willingness to purchase edible insects edible insect products

As mentioned, two of the largest ethnicity groups within the sample population were Latinos and Asians. In Mexico, “escamoles,” or black ant larva, are fairly popular dishes and can cost $25 or more per plate (Aguilar-Miranda and others 2002, Premalatha and others 2011). “Ahuahutle” is a famed Mexican caviar that is composed of some seven different species of Hemiptera (true bugs) (van Huis and others 2013, Aguilar-

Miranda and others 2002, DeFoliart 1992). A perhaps an even more famous food, especially in Oaxaca, Mexico, is “chapulines,” or grasshoppers, and is consumed as street food (van Huis and others 2013). This particular dish can be found in Mexican restaurants here in the United States. While entomophagy is unquestionably strong within

Latin America, countries in Asia also share this tradition. The giant water bug, as aforementioned is a prominent edible insect used in multiple dishes for its unique flavor in Thailand and other South Asian countries (Kiatbenjakul and others 2015,

Sirimungkararat and others 2010, van Huis and others 2013). While silkworms are farmed for silk production, their pupae are also prized delicacies in China, Japan,

Thailand, and Vietnam (Sirimungkararat and others 2010, van Huis and others 2013). In

Korea, rice-field grasshoppers are consumed as side dishes or snacks (van Huis and others 2013). Considering that about 70% of the participants were self-reported to have cultural or ethnical background from countries where entomophagy is such a widely practice custom, it seemed appropriate that at least this many participants were familiar with the practice of consuming edible insects (Figure 4.8). Furthermore, as the said countries are all regarded as developing nations, the data collected from the sensory evaluation study could be somewhat extrapolated to provide indications of consumer acceptance of such insect products in those countries within Latin American and Asia, although domestic Latin Americans and Asian Americans may practice unique dietary habits that are more or less different to the customs found in their mother countries. Thus, despite the present investigation being unable to reach those currently living in countries such as Mexico or Laos People’s Democratic Republic, the results could still offer some

71 implications for such products being implemented in developing countries in the future, although these generalizations may be fairly limited.

Together, the 82% of panelists being familiar with entomophagy and the 73% of subjects declaring a high probability of consuming edible insects in the future (Figure

4.9) point towards a positive trend. In a separate study, Tuorila and others (1994) found a positive relationship between pleasurable first experiences with a novel food and the likelihood of future consumption. As more than 50% of the free-response comments included a statement of their enjoyment and/or support of the current study and its products (Appendix D.22), this certainly presents a progression towards the future applications of edible insects in the market, even in developed nations. Although, it was required that participants be willing to consume edible insects to taste the samples, it cannot be completely assumed that the participants would have chosen to eat edible insects after this experience. This is clear within the 6% who reported that they probably would not or definitely would not consume edible insects in the future. This unwillingness could imply that there was some other incentive for their participation in this particular project. This could include some potential extra credit offered by other professors or even the peer pressure from friends and family.

Even so, the majority of the subjects were willing to consume edible insects in the future. While 73% reported positively in this inquiry, only 42% were able to say they’ve consumed edible insects before their involvement in the current study (Figure 4.10). With more participants (58%) never having previously consumed edible insects, this is certainly a positive outcome of the study. It demonstrates the eagerness of a population to at least try edible insects. Even more dramatic are the findings from the Belgium study by

Megido and others (2014). In this particular survey, the researchers found that although

46.6% of the participants reported some negative attitude towards entomophagy, an even greater number of respondents, 77%, were willing to consume edible insects. The novelty, and perhaps curiosity, of entomophagy may be even more powerful than the

"yuck" factor. Progress is being made towards the acceptance of entomophagy, much like sushi and lobster in the United States.

On the other hand, when taking into account the willingness to purchase edible insect products in the future, there seems to be some disparity. Although people stated that they would probably and definitely consume edible insects in the future, the results from Figure 4.11 show that they may not actually purchase these insect products. It is proposed that this is somewhat correlated with the fact that the majority of the panel are college students with limited finances, where money is only spent on items that are absolutely essential or worth the expense. As mentioned in the free response portions of the questionnaire (Appendix D), knowing of the nutritional and environmental benefits of consuming and farming edible insects would likely provide enough information for these consumers to make a decision towards purchasing edible insect products in the future.

An inquiry pertaining to the opinion of how edible insects should be placed in the diet was also included in the survey. Participants were asked to select as many choices as they felt that described their opinions.

3% 2%

5% Snack 22% Appetizer 13% Side dish Main entree (hidden form) Dish topping/ Garnish 17% 19% Main entree (whole insect) Dessert Not Applicable 19%

Figure 4.12 - Perception of edible insects in the diet Perceptions of edible insects in the diet are displayed by the percentage of the sensory panel who selected those choices. This particular survey question had multiple selections and participants were asked to select all that applied.

When considering the marketing of edible insect products, the results from Figure

4.12 suggest that edible insects are most often preferred to be consumed as a snack

(22%), appetizer (19%), and side dish (19%). Edible insects being perceived most as a snack could be related to their small or, in general that snacking is on the rise. In fact, in

2014, the number of in-between snacks increased to 2.8 with more than half of adults consuming three or more snacks per day (Wyatt 2014). In a previous study, while

“snack” was not included as an option in the survey, edible insects were primarily distinguished as an appetizer. Investigators suggest this also be due to the small size and original form of the food (Megido and others 2014). With edible insects being perceived as fourth as a main entrée (17%) in the current research, this implies that this source of protein is primarily viewed as only supplementary and not yet as a main source of meat,

74 such as those from beef or chicken. Edible insects were even less identified as a main entrée if it were left in its whole form and as a dessert. While people generally do not consume their meat as whole animals, this is no surprise to be the same for insects. Even with shellfish, such as with shrimp, many consumers may prefer to have the head removed before consuming. Moreover, as edible insects are not particularly sweet, it is unlikely that edible insects would be perceived as dessert unless people were familiar with the chocolate covered insects or insect lollipops already available on the market.

Even so, comparing these results with the response pertaining to the panels’ personal accounts with entomophagy (Appendix D.1), it was also derived that a few previous experiences with edible insect candies were not positive ones. With these negative experiences, it seemed appropriate that the “dessert” option was not as highly selected even with the familiarity of edible insects as candies.

As mentioned, personal accounts of entomophagy were included as a free response question in the survey and these answers can be found in the Appendix D.1. A total of 50 people provided their own encounters with edible insects. Of these experiences, there were an unanticipated number of accounts where the insect was intact.

These foods were mostly roasted, fried, or seasoned products and were implied to be consumed as snacks, appetizers, or side dishes. About 20% of the experiences were either in Mexico or in Mexican restaurants and involved grasshoppers. However, an overwhelming majority of the experiences was with crickets (86%) and was followed by worms (i.e. such as mealworms, maggots, or silkworms) being the second most consumed insect (32%) (Appendix D.1). While some enjoyed their experiences, several panelists stated that consuming whole insects was a bit off-putting but suggested that if

75 the flavor was suitable, they would be more likely to enjoy the product. This comment about flavor being able to better allow the acceptance of a product was also reflected in the actual sensory evaluation portion of the questionnaire.

There were a total of five attributes considered by the panelists on the insect rice formulations, including: color, aroma, taste, texture, and overall liking. The first four were based on a five-point Just About Right (JAR) scale while the latter was rated on a 7- point hedonic scale. Participants were also encouraged to leave free response comments about each attribute (Appendix D.2 – D.21).

Color

In scoring color, this characteristic could be scored as either “much too dark,”

“somewhat too dark,” “just about right,” “somewhat too light,” or “much too light.” With the color being derived from the insect flours added to the base matrix, it was predicted that higher-level insect formulations would have scores reflecting this observation.

Results from this evaluation can be seen in Figure 4.13 below.

100%

90% c 80%

70% Dark 60% a 50% b Just about right 40% Light 30%

20% c 10%

0% 10% Cricket 15% Cricket 10% Locust 15% Locust

Figure 4.13 - Color intensity of insect rice formulations Responses are grouped by the four insect rice formulations and displayed as percentage of participants who provided the color-coded scores, where dark responses includes both “much too dark” and “somewhat too dark” selections and light responses includes both “much too light” and “somewhat too light.” Different lower case letters distinguish a significant difference for only just about right scores

Of the four insect rice products, the color of 10LR was exceedingly accepted over the other three formulations (p < 0.05). This was also reflected in the comments as a greater number of subjects mentioned 10LR to look the most similar to market brown rice. It was also suggested that the slightly brown shade of 10LR allowed the panelists to correlate its color as if it were toasted or served with soy sauce, which seems to be a general preference among these consumers. Unfortunately, this may have misled some participants into expecting a product with some Maillard browning or monosodium glutamate flavors, which will be discussed further.

For the other three formulations, these products were deemed either somewhat too light (10CR) or somewhat too dark (15CR and 15LR). As anticipated, these observations

77 were matched with the objective evaluations conducted using the Konica Minolta colorimeter. With the cricket rice having a* and b* values being closer to 0, this suggests the product to be closer to grey hues than the locust rice, which was certainly reflected in the free response section. According to the majority of the comments regarding the appearance of the two cricket formulations, only these products were considered too grey in color, which was unappealing. Some panelists stated that the lightness of 10CR suggested the rice to look undercooked, moldly, or stale. 15CR received mix reactions of the formulation being both too pale and too dark. Although this grey color is familiar in other products, such as in the Japanese soba buckwheat noodles, perhaps this hue is not as appetizing in food as brighter colors such as yellow or red colors. Also, it could be assumed that the subjects would be comparing the insect rice formulations with that of market brown rice which is typically off-white, brown, or even yellow in color when cooked.

On the other hand, the most reoccurring comment concerning the color of 15LR was that it was too dark in color (Appendix D.5), with most panelists scoring it as either

“somewhat too dark” or “much too dark.” While some darkening in color can be the indicator of those delicious flavors from frying, searing, or baking, overshooting this desirable “golden brown” color can suggest the food to be burnt, dry, and overcooked, which was the case for 15LR. The L* values of the locust rice products from the colorimeter measurements matched these comments as the products showed to become significantly darker as the level of locust flour increased.

As the grey and brown colors were demonstrated to be derived from the insect flour, and also considered to be a negative attribute, this may prove to be an issue for the

78 insect formulations, particularly 10CR, 15CR, and 15LR. There are two suggested solutions to this challenge. The first is the removal of the color through bleaching the insect flour before incorporation into food formulations. However, as this may deteriorate the valuable nutrients within the raw material, this may be the lesser of the two options.

The second consideration is the introduction of a separate ingredient where its addition will not only mask the greyness, but also offer a color which matches flavor expectations.

One such ingredient is cocoa powder as its inclusion could provide a more pleasing brown color and a corresponding chocolate flavor. This tactic is already being utilized by other marketed insect products such as Chapul’s Chaco and Aztec Cricket Bars, Exo’s

Cocoa Nut Cricket Bars, and Jungle Bars. A preliminary extrusion trial was also conducted to incorporate 5% cocoa powder into 10CR formulation, leading to promising results in terms of color and aroma. As this newly developed chocolate cricket rice has yet to be cooked, the flavor and mouthfeel are still yet to be determined.

Aroma

With cricket and locust being similar to ocean shrimp, it was anticipated that the participants would observe a fishy aroma when smelling the insect rice products. Thus, the possible responses were as such: “much too odorless,” “somewhat too odorless,” “just about right,” “somewhat too fishy,” and “much too fishy.” The ratings for the aroma of the insect rice can be seen in the figure below.

100%

90%

80%

70% Fishy 60%

50% Just about right a 40% a Odorless 30% b 20% c 10%

0% 10% Cricket 15% Cricket 10% Locust 15% Locust

Figure 4.14 - Aroma intensity of insect rice formulations Responses are grouped by the four insect rice formulations and displayed as percentage of participants who provided the color-coded scores, where fishy responses includes both “much too fishy” and “somewhat too fishy” selections and odorless responses includes both “much too odorless” and “somewhat too odorless.” Different lower case letters distinguish a significant difference for only just about right scores

While all formulations were perceived with some aroma, both cricket formulations were deemed to have aromas that were statistically just about right (p <

0.05) where the products were neither too fishy nor too odorless. On the other hand, the two locust formulations were observed to be somewhat too fishy.

In interpreting the comments provided by the panelists, it was implied that both grainy or wheat like aromas and lack of an odor coordinated with what was expected for market brown rice. With that, most of these observations were made in the cricket formulations and so were reflected in their subsequent scoring of being just about right in aroma. An interesting result was that it seemed that participants perceived the 10CR as more fishy in aroma than 15CR, despite the latter having more insect flour, the source of

80 the smell. However, this is only a 4% difference in the “much too fishy” category and is considered statistically insignificant by the RedJade program. Even so, perhaps this unexpected result could be account for by the color. With 10CR being observed as being paler in color, it would be appropriate to assume this product to be more odorless, as can be seen in the “somewhat too odorless” category. With that expectation and yet observing a fishy aroma, the participant would score the product to be fishier in aroma.

Locust formulations, on the other hand, showed a significant difference in its fishy aromas. Although both were considered to be somewhat too fishy, there were four times as many subjects who rated 15LR as much too fishy. In fact, there were specific comments regarding this product having smelled like crickets sold in pet stores.

Furthermore, a fascinating finding was that some panelists were able to match 10LR and

15LR to each other, stating that the aromas were somehow comparable. Thus, the odor originating from locust formulations has been determined to be much more pronounced than that from cricket products, with the smell being much more obvious when increasing the level of locust flour.

Flavor

A main concern for the flavor of the insect rice products was in its earthiness.

Figure 4.15 displays the percentage of scores for the following responses: “much too bland,” “somewhat too bland,” “just about right,” “somewhat too earthy,” and “much too earthy.”

100%

90%

80%

70% Earthy 60% a 50% a Just about right 40% b 30% Bland c 20%

10%

0% 10% Cricket 15% Cricket 10% Locust 15% Locust

Figure 4.15 - Flavor intensity of insect rice formulations Responses are grouped by the four insect rice formulations and displayed as percentage of participants who provided the color-coded scores, where earthy responses includes both “much too earthy” and “somewhat too earthy” selections and bland responses includes both “much too bland” and “somewhat too bland.” Different lower case letters distinguish a significant difference for only just about right scores

From the comments, lower insect formulations, such as 10CR and 10LR, were observed to be more parallel to market brown rice flavors. Although some suggested that this meant the insect rice tasted somewhat bland, they also mentioned that this was not necessarily a negative attribute as rice is generally considered a bland product when served alone.

Even so, flavor remained an important component to the acceptability of the insect products as several participants mentioned that a suitable flavor could positively

82 sway the overall opinion of a product (Appendix D). Multiple criticisms made by the panel recommended that the acceptability of the products would be greatly improved if served alongside other foods or seasoned, such as with salt or soy sauce. As aforementioned, a study was conducted by Megido and others (2014) involving sensory evaluation on mealworms (Tenebrio molitor) and crickets (Acheta domesticus), where these insects were prepared in a variety of ways including: baked, boiled, a crushed mix of both insects, mealworms flavored with vanilla, and mealworms with chocolate. It was found that the most accepted form of preparation was with chocolate, followed by flavored with paprika, and then baked. While each mealworm was crispy in texture, the investigators advocate the flavor to be the factor in preference (Megido and others 2014).

By being a rice product, these insect formulations do have the advantage of being versatile in their formulation and preparation. Indeed, one of the research assistants took it upon himself to create a kimchi fried insect rice dish which was very much well received by the other investigators. While this sensory evaluation reveals some introductory indication of how this product would be fairly accepted in a developed country, such as the United States, the insect rice has the potential of being further developed to include flavors custom to developing countries. Another potential ingredient aforementioned is the introduction of cocoa powder. Relating the color observed in

10LR, which was with a toasted brown hue, many participants expected it to be much more flavorful than actually experienced. In utilizing cocoa powder, and presenting the succeeding outcome as a chocolate product, this could reduce the misleading nature of the brown color derived from the locust flour. Besides some color matching/masking, the cocoa powder may provide some improved flavoring as a chocolate ingredient. The

83 possibility for the research and development of this insect rice into a marketable product is seemingly boundless.

Yet, it is essential to recognize that earthiness was certainly an undesirable attribute in the insect formulations. It was found to be more apparent in not only formulations with greater quantities of insect flour, but especially in locust products.

15LR was determined to be the earthiest in flavor as it was scored by nearly four times more subjects as “much too earthy” than its 10% counterpart. The sensory panel also made several remarks on all the insect rice as having some fishy flavors, which were also especially found within the locust rice formulations and particularly in 15LR. These were anticipated comments as this was the aroma of concern. Thus, those who perceived the fishy aroma would be likely observe the same quality in the flavor of the insect rice. By recognizing that land insects, such as cricket and locust, and shellfish, such as shrimp, are both arthropods, it was anticipated that they would have comparable flavors.

At this point, it is necessary to note that although the cooked rice samples were kept warm at the best capabilities of the investigators during the tasting sessions, it was difficult to maintain a consistently desirable temperature without proper equipment. This change in serving temperature would have undoubtedly had some effect on the overall experience of the panelists when taste-testing the product, particularly when grading the aroma and the flavor of the insect rice. When heated, the aroma and flavors are much better sensed as the volatile chemicals are able to travel and stimulate the olfactory senses, both orthonasally and retronasally. Although reading through the free responses did show that various serving temperatures did have some influence on the evaluation, it was not too drastic due to the fact that most of the samples were served at an appropriate

84 temperature. Regardless of changes in the temperature, the earthy and fishy flavors from the insect rice formulations were certainly present, particularly with stronger profiles in

15LR. As 20% of all the comments made on this product had indicated their distaste

(Appendix D.13), the intensity of those flavors in 15LR is determined to be unacceptable to the consumers.

Other observations obtained from the free response portion of the evaluation also showed that many people experienced a sort of aftertaste in all formulations. While some were unable to identify the flavor, others determined the aftertaste to be either bitter or mineral-like. Again, these were more heavily observed in 15LR while least discerned in

10CR. Thus, considering that both the earthiness and the aftertaste were the most prominent in 15LR, it is suggested that the observed bitterness is correlated to the earthy flavor, or more specifically, to the calcium chloride coating. Another remark, especially in 15CR, was some identification of saltiness that is likely to be derived from the sodium chloride that was the byproduct from the crosslinking reaction between sodium alginate and calcium chloride at the surface of extruded rice particles.

Mouthfeel

Graininess was the primary concern for the texture of the insect rice formulations.

Thus, the question was anchored as “much too smooth,” “somewhat too smooth,” “just about right,” “somewhat too grainy,” and “much too grainy.” The responses from the participants about this aspect of the rice products can be found in Figure 4.16 below.

100%

90%

80%

70% Grainy 60% a

50% Just about right a 40% Smooth 30%

20% b 10% d

0% 10% Cricket 15% Cricket 10% Locust 15% Locust

Figure 4.16 - Mouthfeel intensity of insect rice formulations Responses are grouped by the four insect rice formulations and displayed as percentage of participants who provided the color-coded scores, where grainy responses includes both “much too grainy” and “somewhat too grainy” selections and smooth responses includes both “much too smooth” and “somewhat too smooth.” Different lower case letters distinguish a significant difference for only just about right scores

Once again, the cricket formulations were rated statistically as just about right (p

< 0.05) whereas locust formulations were observed to have mouthfeels that were somewhat too grainy. In fact, 10CR and 15CR had between 1.8 and 3.4 times more people who considered the texture as just about right than their locust counterparts. While the graininess was more evident in locust formulations, it was even more apparent in

15LR. Thus, the source of the perceived graininess is derived from the locust flour itself.

In addition to the term grainy, the words “gritty,” “crunchy,” and “sandy” were used to describe the locust rice formulations in the free-response comments from the panelists.

Although it appeared that there were more participants who rated 10LR as somewhat too grainy than 15LR, which is an unexpected result, the overall distribution of subjects still

86 established 15LR as the grainier of the two locust products. While 15LR is received 2.5 times more scores of “much too grainy” compared to 10LR, it received 11 times more ratings when compared with 15CR. In studying the free response portion of this evaluation, there were an overwhelming number of comments pertaining to the gritty texture of 15LR (Appendix D.17). For example, 66% and 69% of the comments on 10LR and 15LR, respectively, were concerned with how grainy the products were.

Unquestionably, this attribute was perhaps the most unappealing characteristic experienced by the panelists. Considering that locust rice products were determined to contain greater quantities of fiber and that the principal component is likely to be chitin, this insoluble fiber is highly suggested to be the culprit behind the unpleasant texture. In contrast, a sensory evaluation study on the supplementation of maize flour tortillas with ground mealworms (Tenebrio molitor) was conducted in Mexico. Since these particular species and stage of insects are without the tough exterior that insects have within the

Orthoptera order, such as crickets and locusts, perhaps this is partly the reason for the positive results in that study, including an enhanced mouthfeel and improved rolling of tacos in the fortified tortillas (Aguilar-Miranda and others 2002). Still, preferences for rice texture varied among the participants as there were some subjects who expressed their appreciation for the crunchiness of the insect rice products. Furthermore, in the aforementioned Belgium study, the investigators found the crispy preparations of the insects were significantly favored over the boiled versions (Megido and others 2014).

Even so, those who identified 15LR as much too grainy remains to be the overwhelming majority.

In contrast with these sensory evaluation results, the objective texture analysis revealed that all of the insect rice formulations were actually softer than MBR. While locust rice formulations were rarely considered soft in the sensory evaluations comments, perhaps it is the grittiness/crunchiness that prevented the rice from being perceived as soft.

Overall Liking

To determine the overall acceptance of each of the four insect rice formulations, a

7-point hedonic scale was employed. The results from this evaluation are represented in both Table 4.13 and Figure 4.17.

Table 4.13 - Mean scores for overall liking of insect rice formulations Insect rice Mean score 10CR 4.76 ± 1.49a 15CR 4.45 ± 1.56a 10LR 3.97 ± 1.47b 15LR 2.92 ± 1.48c Highest score for overall liking was a 7 with the lowest score being 0. Different lower case letters distinguish a significant difference in overall liking of products

100%

90%

80% Dislike extremely 70% Dislike moderately 60% Dislike slightly

50% Neither like nor dislike Like slightly 40% Like moderately 30% Like extremely 20%

10%

0% 10% Cricket 15% Cricket 10% Locust 15% Locust

Figure 4.17 - Overall liking of insect rice formulations Responses are grouped by the four insect rice formulations and displayed as percentage of participants who provided the color-coded scores.

By and large, Table 4.13 reveals no significant difference between how 10CR and

15CR were preferred (p > 0.05). Specifically, the cumulating response from the “like slightly,” “like moderately,” “like extremely” were about 60% for 10CR and over 50% for 15CR (Figure 4.17). Each of these products was scored between being neutrally enjoyed and liked slightly. Locust products with 10% addition levels were nearly neutrally accepted as its mean value was 3.97 out of 7. The formulation with the highest locust content, however, was significantly the least liked product, as it showed a mean score of 2.92 out of 7, denoting an acceptance between dislike slightly and dislike moderately.

While both cricket formulations were evaluated to have aroma, flavor, and mouthfeel that were just about right, the color may have reduced their final mean scores for acceptance. As aforementioned, the color was reported to be too gray and somewhat

89 too light (10CR) or somewhat too dark (15CR) to be appealing. This suggests that the color is a major factor in the acceptance of the insect rice products. This is an appropriate response as color is certainly the primary attribute when evaluating food (Giusti and

Wrolstad 2003).

Next, it can be supposed that the overwhelming graininess of 15LR played a chief role in the acceptance of this particular product as it received the lowest mean score, thus being the least liked. In addition to the negative reaction to the texture of 15LR, all of the other attributes, including color, aroma, and flavor, also was rated rather poorly. The color was considered somewhat too dark, the aroma as somewhat too fishy, and the flavor as much too earthy.

Interestingly, 10LR was reported to be more accepted than the 15% locust product. Despite it having received scores for individual attributes almost parallel to

15LR, where it was perceived to be somewhat too fishy in odor, somewhat too earthy in taste, and somewhat too grainy in texture, this product had the best rating for color.

Again, this attribute demonstrates itself as a powerful means to increase the acceptability of a product. Although the color of 10LR was undeniably darker than that of the other cricket formulations, this color was associated with pleasant and desirable flavors.

Furthermore, since this attribute was the first to be inquired, there was no bias prior to some negative experience encountered later for the other characteristics. Of course, 10LR was not rated with a higher score in overall liking as the subjects became disappointed in the aroma, flavor, and mouthfeel of this formulation.

CHAPTER 5

CONCLUSIONS

Objective 1

A total of four insect products, each with the same brown rice flour base, were developed using cricket or locust flours at either 10% or 15% addition levels. The formulations were determined to be feasible for extrusion, with only about one third of the raw material being lost during the multiple-step process, which is a standard occurrence for small scale, 1kg batch productions. By successfully producing an optimized formulation where an edible insect flour ingredient was incorporated into an extruded rice product, the Objective 1of the current research was achieved.

Objective 2

Extrusion of the insect rice formulations yielded cricket products that were grey in color while the locust formulations were darker and browner in appearance. Both these grey and brown colors were darker and certainly unique to market brown rice, which was used as a reference.

Also in contrast with market brown rice, these insect rice products lacked an outer bran layer, typical in whole grain rice kernels. The absence of this protective barrier subsequently resulted in a product that presented with a rougher surface morphology and yet still a softer and more adhesive texture after cooking. These qualities also consequently required a steam cooking technique, as opposed to a full water submersion boiling method that is typically employed in automated rice cookers.

In evaluating the stability of the product, it was found that each insect product had a shelf life of at least three months. Further studies to determine whether this shelf life could be extended to six months should be considered, especially since the water activity, although significantly lower than the critical 0.6 aw level, increased at a relatively rapid rate over three month storage.

By and large, physical properties of the insect rice products were found to be darker in color, rougher in surface morphology, softer and more adhesive in texture, and more shelf stable than market brown rice. These results led to the rejection of H01 null hypothesis and the failure to reject the alternative hypothesis HA1 as the insect rice products show to have physical characteristics which are distinguishable from market brown rice. In comparing between insect formulations, only color was determined to be significantly different. For instance, it was found that the cricket rice was greyer in color while the locust products had browner hue. With that, the H02 null hypothesis was rejected and the HA2 alternative hypothesis was failed to be rejected. When contrasting between 10% and 15% formulations, H03 was rejected for locust formulations as increasing the insect flour led to products with a darker shade.

Nutritionally, the insect rice formulations showed much promise as a solution to global food insecurity. With 10 – 12.5g per serving (1/2 cup or 80 g) and 20 – 25% of the daily value, the cricket and locust rice products were confirmed to be excellent sources of protein. Furthermore, the insect products demonstrated to offer 150 - 200% more protein than market brown rice. Future studies in identifying the amino acid profile would yield even more valuable information concerning the nutritional benefits of edible insects.

Fat content of the insect rice products were revealed to provide 10-12% of the daily value. This created an energy dense food that is necessary for developing countries.

Products such as the insect rice could help to alleviate the hunger being experienced in countries such as Sub-Saharan Africa and South Asia, where rice a staple food, as they continue to undergo serious protein-energy malnutrition diseases.

Another nutrient of global concern is iron as it is proclaimed to be the number one nutrient deficiency in the world. Although the iron analysis of the insect rice formulations requires further investigation, the results suggest that this micronutrient to be present and in theoretically superior levels than market brown rice. Total dietary fiber is also determined to be available, with locust formulations are proposed to contain greater quantities, of which is composed chiefly of chitin.

Overall, the measurements of physicochemical and nutritional properties confirmed the insect rice products were quite different from the control – market brown rice. The statistical analyses suggested the rejection of the null hypotheses H01 – H03, thus the alternative hypotheses HA1 – HA3 was failed to be rejected (p < 0.05). Arriving at these conclusions fulfills Objective 2 of this study.

Objective 3

It is further indicated that perhaps the higher level of chitin led to a displeasing texture during sensory evaluations. In addition to being perceived as too grainy, the locust formulations at 15% addition levels were also determined to be excessively dark in color, fishy in aroma, and earthy in flavor, resulting in it being the least liked formulation. On the other hand, both cricket rice products have demonstrated as the best

93 formulations worth continuing research and development. Thus, with such significant differences found between cricket and locust rice formulations, the H04 null hypothesis could not be accepted, hence, the HA4 alternate hypothesis was established at p < 0.05.

Between the two cricket rice products, the major issue is perhaps in its appearance as the sensory panel considered the products to be either too light or too dark in appearance. Despite this distinction between color, 10% and 15% cricket rice products were equally enjoyed. Locust products, however, were overall liked at different levels. As aforementioned, 15% locust rice was the least liked product. On the other hand, its 10% counterpart, while having been reported with similar negative scores for aroma, flavor and mouthfeel, this product offered a toasty brown color that was the most accepted of the four formulations. This appreciation, in effect, allowed 10% locust formulations to be significantly better received than 15LR. Therefore, while H05 null hypothesis fails to be eliminated for the cricket rice formulations, it is still rejected for the two locust products.

Overall, both cricket rice products and the 10% locust rice formulations were scored between neutrally enjoyed and liked slightly, demonstrating positive results towards consumer acceptance. With this establishment Objective 3 has been successfully completed.

CHAPTER 6

LIMITATIONS AND RECOMMENDED FUTURE STUDIES

Limitations

Formulation and Extrusion Optimization

All four formulations achieved similar yield of final rice particles, yet about one third of the raw material was still lost during extrusion, dehydration, and/or size screening. While this may be a normal occurrence for small scale, 1 kg batch productions, continuous production utilizing large batches (i.e. industrial extruders), can improve this production yield to over 95%.

Furthermore, as the insect raw materials were obtained internationally from

Canada (cricket flours) and Thailand (locust flour), the cost and the availability presented some restraint for the current study. With limited supply, the restrictions on the production of the insect rice products and on the freedom to conduct more thorough research were unavoidable. While the careful planning and organization still led to significant findings, this study could certainly be improved with industrial equipment and local availability of insect ingredients.

With a positive outlook for the future adoption of entomophagy in developed nations, the increasing need and interest for the continued development of fortified food products with edible insects will perhaps offer some third party facilitation for this field of research, specifically in terms of financial support. The lift of monetary burdens will allow for more extensive research in the utilization of this alternative food source which

95 ultimately can give some relief to the malnourished populations within developing countries.

Fiber and Iron Analysis

As the laboratory was limited in its abilities to perform further ash and protein analysis after the enzymatic-gravimetric analysis for the total dietary fiber, the resulting value of recovered residue only provided a rough estimation for the fiber contents. In comparing with the certificate of analysis provided by the cricket flour vendor, Next

Millennium Farms, it is certain that the values obtained from the Ankom Dietary Fiber

Analyzer were an overestimation. Further assessment of the ash and protein in the residue is necessary to determine the actual quantity of total dietary fiber.

A relatively high fat content was also found in the insect rice products and may have caused interference in the iron analysis. An additional defatting step may be required to remove this particular constituent. The current procedure for iron analysis using AAS should be standardized to generate more reliable data.

Sensory Evaluation

While a market brown rice was excluded from the tasting sessions due to the product being obviously differentiable from the insect rice products, this did pose a limitation to the study. Without a reference product, the level of acceptability of the insect rice formulation in comparison with a control was unable to be determined. With concerns of panelists experiencing sensory fatigue with multiple products and/or a time bias from a lengthy questionnaire, it was decided that the four insect formulations would

96 first be compared between each other. The product with the best results from this initial study would then be selected to be further optimized in its formulation and compared with a market brown rice reference in a second round sensory evaluation in a future study.

Also, since the people living in developing countries could not be directly reached, convenience sampling was utilized to recruit subjects for the sensory evaluation study. While the high percentage (~70%) of participants are of Latino and Asian descent, the results from this study offers only a suggestion for how the insect rice formulations would be received in those developing countries. Moreover, while only a handful of the participants were of African ethnicity, these results would poorly reflect the perception of this major population.

Even so, the sensory evaluations do demonstrate a positive outlook for the acceptance in entomophagy in developed nations. While countries such as the United

States can be powerful in setting global trends, perhaps their positive reception of edible insects will provide a much needed advancement towards utilizing this alternative food source as a means for alleviating present and future food insecurities.

Recommended Future Studies

Formulation Improvement

Cricket formulations were reported to be accepted between neutrally enjoyed and liked slightly with the color strongly suggested to being the least pleasurable attribute observed in these products. With this respect, further studies incorporating cocoa powder is considered. This ingredient would not only mask the displeasing grey and pale colors

97 of the cricket rice products, but it may also lend to an improved appreciation for the overall flavor and mouthfeel as chocolate is particularly known to provide. In addition, the cocoa powder may be able to present other enhanced functionality during extrusion processing.

Another potential modification would be the inclusion of a preservative. While the insect rice is predicted to have a shelf life between 3 to 6 months, a preservative such as calcium propanoate, sodium benzoate, or butylated hydroxyanisole that are commonly used in wheat flour-based products, could be added into the formulation to improve the overall stability of the product.

Culinary Exploration

With flavor being a major determinant for the overall liking of the product, further studies integrating additional preparation/serving methods are highly recommended. For instance, the current insect rice products could be sautéed with soy sauce and other vegetables, such as with Asian style fried rice. The product could also be presented with other steamed vegetables which could improve the perceived color of the rice products.

From the entomophagy survey, it was conveyed that edible insects are primarily viewed as snack by the sample population. With this in mind, an extruded rice product containing edible insects can instead be shaped into a different form, such as a puffed chip similar to the popular Frito Lays Cheetos or as rice crackers similar to those marketed by Quakers.

Nutrition Analysis

Protein assessment confirmed the major nutritional advantageous of incorporating edible insects, such as cricket and locust, into the diet. Next, analyzing the amino acid profile is suggested to further understand the quality of the protein content. Perhaps this could be an interesting direction for those within the field of sports nutrition and looking for an alternative to whey or soy protein isolates.

Similarly, the lipid content of the insect rice formulation could be further profiled.

While providing 10-12% of the DV of fat, the type of fatty acids present should be examined. Some studies have shown that the oils of some edible insects were found to be high in polyunsaturated fats, including the essential linoleic (omega-6) and linolenic

(omega-3 or omega-6) fatty acids (Womeni and others 2009). Crickets, especially, were reported to contain 62.39% polyunsaturated fatty acids, with 45.63% being linoleic acid and 16.19% α-linolenic acid (omega-3). By offering important polyunsaturated fats, such as those that are essential, this could further promote the incorporation of edible insects into an everyday diet.

Extrusion Standardization

While offering several advantages, such as having relatively low capital, maintenance, and processing costs and being fairly straightforward in its operation, there are multiple factors that can affect product quality during extrusion. Besides the variations in the raw materials itself, the composition of the dough mixture (e.g. the type of starch and its particle size, macronutrient, and moisture content) can certainly influence the extrusion process. Besides the formulation composition, extrusion

99 conditions, such as temperature and feed rate, can also affect the characteristics of the final product (Fellows 2009). With several variables providing various functionalities in this technology, it is essential to standardize the extrusion process. Thus, to truly comprehend the effect of the insect flours on the extrusion production of these novel rice products, multiple trials using the same formulations and conditions should be performed to validate the extrusion process with acceptable, reproducible, and repeatable results.

Shelf Life Studies

As there were some inconsistencies between the 10% and 15% formulations, it is recommended that the shelf life study be repeated. The study could also be extended to six months, as well as include an oxidation study, to determine the stability during this extended length of time. If the formulation were to be modified with the inclusion of cocoa powder, a shelf life study should unquestionably be conducted as the higher lipids within this raw ingredient may be prone to oxidation.

Another factor to incorporate into this particular assessment is in the packaging of the potential marketed product. For instance, previous studies have researched various storage methods of brown rice. Sharp and Timme (1986) revealed that refrigerated temperatures (3°C) and in hermetically sealed conditions, such as polyethylene film bags, provided improved results in terms of sensory and fatty acid contents. With this knowledge, the insect rice products could be stored in polyethylene bags in both refrigerated and room temperatures and observed for a 6 month period. In the study, the color and texture can also be measured to determine whether time or storage environments will influence these attributes in addition to water activity.

100

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APPENDIX A

SPECIFICATIONS OF RAW INSECT FLOUR

Table A.1 - Insect Flours cricket flour (Next Millennium Farms) locust flour (Thailand Unique)

Table A.2 - L*a*b* values for Next Millennium Farms cricket flour trial # L* value a* value b* value trial 1 44.50 5.40 9.16 trial 2 44.24 5.29 8.95 trial 3 44.23 5.3 8.75 average 44.32 5.33 8.95

Table A.3 - L*a*b* values for Thailand Unique locust flour trial # L* value a* value b* value trial 1 47.89 7.58 14.17 trial 2 47.63 7.46 13.98 trial 3 47.52 7.53 14.01 Average 47.68 7.52 14.05

Table A.4 - Water activity for Next Millennium Farms cricket flour

trial # aw temperature °C trial 1 0.272 24.9 trial 2 0.257 25.0 trial 3 0.264 25.0 average 0.264 25.0

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Table A.5 - Water activity for Thailand Unique locust flour trial # aw temperature °C trial 1 0.247 25.1 trial 2 0.250 25.0 trial 3 0.235 25.0 average 0.244 25.0

Table A.6 - Moisture content for insect flours insect flour moisture (%) NMF cricket 3.76 locust flour 5.00

Table A.7- Percent recovered residue of insect flours* insect flour % recovered residue cricket flour 36.40 locust flour 54.40 *Percent recovered residue was obtained from the Ankom Dietary Fiber Analyzer and provides a rough estimation of the total dietary fiber content

107

Figure A.1 - Next Millennium Farm cricket flour certificate of analysis 108

Figure A.2 - Locust flour protein certificate of analysis provided by Thailand Unique

109

APPENDIX B

EXPERIMENTAL SETUP

Figure B.1 - Acid digestion of insect and insect rice products In preparation for graphite furnace atomic absorption spectrometry iron analysis, acid digestion of the insect flour and their rice products was performed. 500mL Erlenmeyer flasks containing 1g of either insect flour or ground insect rice product and 10mL of both concentrated hydrochloric acid and nitric acid were boiled at 300°C on hot plates

Figure B.2 - Steaming method for determining optimal cooking conditions A steaming basket accompanying a stovetop pot was used to steam three inset rice-to-water ratios in order to determine to optimal water additions levels for cooking the insect rice products. Water was first brought to a boil using a hot plate. Once boiling, the insect rice, mixed with water in a 3.25 oz polystyrene cups, was place in the steaming basket within the pot and covered. The rice was steamed for 15 minutes before being removed from heat.

110

APPENDIX C

ORIGINAL EXPERIMENTAL DATA

Figure C.3 - Texture analysis of 10CR, in quintuplicate

Figure C.4 - Texture analysis of 15CR, in quintuplicate

111

Figure C.5 - Texture analysis of 10LR, in quintuplicate

Figure C.6 - Texture analysis of MBR, in quintuplicate

Table C.1 - Percent recovered residue for insect flour and their rice products trial # 10CR 15CR 10LR 15LR cricket flour locust flour trial 1 11.70 9.60 12.90 26.10 n/a n/a trial 2 13.00 14.80 15.60 17.40 36.40 54.4 average 12.35 12.20 14.25 21.75 36.40 54.4

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APPENDIX D

SENSORY EVALUATION SETUP AND ORIGINAL DATA

Figure D.1 - Sensory evaluation room set up Twelve booths, each equipped with a laptop activated with the RedJade questionnaire and a 3-walled booth, was prepared for the sensory evaluation study. Photo credit: Tom Zasadzinski, Cal Poly Pomona photographer

113

Figure D.2 - Entomophagy questions from the sensory evaluation study Questions pertaining to entomophagy were inquired in a separate survey in the RedJade questionnaire. There was a total of 5 questions. An additional free response question would appear if participants answered “yes” to the question “Have you ever consumed edible insects before today?”

114

- - 1 1 1 1 1 1 1 1 1 1 1 + + 2 2 1 1 Other Other 1 1 1 1 1 1 1 1 1 1 Worms Worms Grass- hopper hopper 1 1 1 1 1 1 1 1 1 1 1 1 1 1 Cricket Cricket 1 1 Mexico Mexico 1 1 2 2 Hidden Hidden Dessert Dessert late late Choco- 1 1 Candy Candy 2 2 2 2 2 2 2 1 1 fried/ fried/ Roasted/ Roasted/ seasoned seasoned 1 1 2 2 2 2 2 insect insect Whole Whole worms. worms. Table D.1 - Free response regarding participants' experience with entomophagy and their trends mealworm, spider mealworm, as you'd like. like. you'd as previously tasted the edible insect insect the tasted edible previously e hav Please describe your experience with with experience your describe Please edible insects, including what type of type what including insects, edible insect was tasted. Share as much detail detail as much Share was tasted. insect rice before. It was locust and cricket. cricket. and locust was It before. rice and roasted crickets, live and Roasted mealworms seasoned unpleasant had an mealworms Roasted had somewhat but (crunchy) mouthfeel (toasted) flavor okay to eat tolerable were crickets Roasted I've eaten dried crickets and silk and silk crickets dried eaten I've tasting. quite earthy were They I have tried cricket flour but in the form the form in but flour tried cricket have I with seasoned was It chip. a puffed of and with familiar are that people flavors liked. can easily be Crickets, locust, locust, Crickets, I I again never crickets - Live corn those and crickets eaten have I thing a fun it thought was things. I worm on was I ate them when I to do so a taco ate them in I Mexico. in vacation and seasoned cooked fully were they and eating tell was I really couldn't so I eating of concept think the I insects. been have cultures many great, is insects already. so doing Silk worms where fried I believe, they they believe, fried I where Silk worms had an and odd salty crispy, were were fried they where Crickets aftertaste. the than experience better salted, lightly silkworms worms I crunchy the crunchy, were they had the crickets tasty while tried were them to taste a strange much of too I have eaten live insects, fried insects, insects, fried insects, live eaten have I tried a I insects. containing products and made with candy a was that product but nasty plain just was that flour cricket poor to a due just be could that formulation. 1 1 2 9 9 7 7 8 8 3 3 4 4 5 6 6

115 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 Worms Worms Other + - Grass- hopper hopper 1 1 1 1 1 1 1 1 1 1 1 1 1 Dessert Dessert Hidden Mexico Cricket late late Choco- 2 2 1 Candy Candy 2 2 fried/ fried/ Roasted/ Roasted/ seasoned seasoned 2 2 2 2 2 1 1 1 insect insect Whole Whole with with come from a from come a lot experience experience as you'd like. like. you'd as Ant candy and i and candy Ant eats silkworms. silkworms. eats Please describe your experience with with experience your describe Please edible insects, including what type of type what including insects, edible tried a cricket and maggot in an in an maggot and tried a cricket insect was tasted. Share as much detail detail as much Share was tasted. insect I have tried have I that culture considerable have I eaten have I edible. I tried the exo flour chips made of cricket cricket of made chips flour tried the exo I Their exo). is the brand believe (I flour at flavor to no fishy/insecty little product seasoning to the due be but may his all the flavor to mask added with chili powder so the flavor was of was the flavor so powder chili with which mealworms, eaten have i and chili. part one candy; in the no uniformity was crumbly was the next while sweet was bitter. and probably buy them if I could. them if I buy probably cooked were which ants, eaten have I nutty of sort crunchy, and roasted were real with candies have they Mexico In in for them worms or scorpions so insects out dried are These decoration. displeasing but the flavor was more to more was flavor the but displeasing liking. my taste They before. crickets had I fried they but chips normal like good, pretty were crickets the because scary look my around i see ones the than bigger house. and mealworms, crickets, eaten have I Agriculture class. A professor brought in brought professor A class. Agriculture roasted been had insects which the whole had also a I seasoning. with and flavored bar. I insect an bit of a to try change that felt I insects, roasted the didn't mind the with flavor better had a they distinct a noticed the bars I In seasoning. liking. to my not was which aftertaste, bit a was the insects of texture The I

There pleasant. not was them consuming would and liked them I dubia roaches. 10 15 16 17 13 14 11 12

116 1 1 1 1 1 1 1 1 1 Worms Worms Other + - 1 1 Grass- hopper hopper 2 2 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 Dessert Dessert Hidden Mexico Cricket 1 1 late late Choco- 1 1 Candy Candy fried/ fried/ Roasted/ Roasted/ seasoned seasoned 2 2 1 1 1 1 1 1 2 2 2 2 2 2 1 1 1 1 insect insect Whole Whole I I a brownies brownies mealworms; mealworms; original original the mountains. the mountains. grasshoppers are are grasshoppers can find in some in some can find crickets and and crickets bars. add much flavor but just but flavor much add as you'd like. like. you'd as Please describe your experience with with experience your describe Please edible insects, including what type of type what including insects, edible insect was tasted. Share as much detail detail as much Share was tasted. insect I participated in a Science Day event at event Day a Science in participated I out andhelped ago years a few Mt. SAC of out section insects edible inthe meal tried were I insects The curiosity. were crickets. They believe and I worms tried both but I people most for fried worms meal The uncooked. and cooked the legs can feel in I as texture had more that me bother didn't it but tongue on my much. and cookies powder Cricket to identical taste was where product I ate crickets in Mexico when I a was I when Mexico in ate crickets I tried haven't and them didn't like kid. I idea of eating insects and not necessarily necessarily not and insects eating idea of the taste and cream sour and maggots tried bbq I tasty neither were which crickets onion protein had cricket also have gross. I nor favored. strongly too were bars which Seasoned dried dried Seasoned powder cricket In Mexican culture, culture, Mexican In you snack common high in towns small have tried an insect bar and enjoyed it. enjoyed bar and insect an tried have they if insects edible more try would I have eaten edible insect products products insect edible eaten have I crickets tried candied have I before. I ate a chocolate covered cricket. It was It cricket. covered ate a chocolate I could feel I as experience interesting an in my insect the whole much pretty bit past I it, once chewing was I as mouth leg cricket Had a layer. chocolate that a while. for teeth my between in stuck didn’t Overall texture. added

the to due be could it But them since. 1 salt and lemon with Crickets

I experience. the and enjoyed before

21 22 19 23 20 24 25 26 18

117 1 1 1 1 1 1 1 1 1 1 Worms Worms Other + - 1 1 1 1 Grass- hopper hopper 1 1 1 2 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 Dessert Dessert Hidden Mexico Cricket 1 1 late late Choco- 1 1 Candy Candy fried/ fried/ Roasted/ Roasted/ seasoned seasoned 1 1 1 1 1 1 1 1 1 1 1 1 1 1 insect insect Whole Whole to the the to 1 and maggots and maggots as you'd like. like. you'd as Please describe your experience with with experience your describe Please edible insects, including what type of type what including insects, edible insect was tasted. Share as much detail detail as much Share was tasted. insect consume insects in their original form. form. original in their insects consume bar not finish did bar; nutrition chapul although grainy, too was texture because sweet slightly was the flavor Chocolate. Chocolate. cricket being covered in sugar. in sugar. covered being cricket I have eaten crickets before and it was andwas it before crickets eaten have I I had previously consumed a processed processed consumed a hadpreviously I a from replacement) (meal bar protein utilized that market foods natural popular main as the powder protein cricket an had product The ingredient. protein entirely but not was flavor earthy the purchase might only I unpleasant. recipe their changed they if again product enough strong curiosity found my and I another purchase to me to compel or am not vegan I bar. flavored protein sources alternative but vegetarian weird am certainly I interest. my peak will that I see don't for so I texture about I love eating fried grasshoppers, grasshoppers, eating love fried I of One Mexico. Oaxaca, from preferably also was and as such, tasted the samples liked the most. I the one Fried Crickets and Crickets with with Crickets and Crickets Fried were more available. I have an have available. I more were palette. adventurous I’ve had crickets, scorpion hadscorpion crickets, I’ve all i liked them before Teriyaki flavored crickets. They were were They crickets. flavored Teriyaki sauce. teriyaki with whole roasted and crispy. Crunchy I ate some small grasshoppers in an in an grasshoppers small ate some I grasshoppers The restaurant. Oaxacan with and seasoned and roasted dried are

due be this may tasteless almost

crickets small like really I

bars Chapul

30 28 31 29 32 33 27 34 35 36

118 1 1 1 1 1 1 1 1 1 1 Worms Worms Other + - 1 1 1 1 Grass- hopper hopper 1 1 1 1 1 1 1 1 1 1 Dessert Dessert Hidden Mexico Cricket late late Choco- 1 1 Candy Candy fried/ fried/ Roasted/ Roasted/ seasoned seasoned 1 1 1 1 1 1 1 1 1 insect insect Whole Whole it. it. i was was i liked liked when when bugs is not a not is bugs 1 1 as you'd like. like. you'd as little girl. It was spicy and I and I spicy was girl. It little ate grasshoppers in Mexico Mexico in ate grasshoppers Please describe your experience with with experience your describe Please edible insects, including what type of type what including insects, edible insect was tasted. Share as much detail detail as much Share was tasted. insect Edible insect food products are very very are products food insect Edible no and taste, my pungent for too likely professional of the amount matter the to spit am tempted I salesmanship, products ate insect I back out. mouthful project this and expos, food aat couple athis very is say, would I and in2015, insects The product. refined well-made, think I and crickets, locusts, had I were problem of taste. taste. of problem I've had crickets, roasted with ranch with roasted hadcrickets, I've They can't remember. I grasshopper... I and smell offensive very a have often such with to eat food like don't characteristics. aggressive They before. grasshoppers ate edible I they and salt seasoned with heavily were taste liked the actually I crunchy. were somewhat. restaurant in Downey, CA. Texture of Texture CA. in Downey, restaurant the like but didn't good was crickets taste. earthy Tried cricket tacos at a Mexican Mexican aat tacos cricket Tried I I a lemon or lime. They were okay. The okay. were They lime. or lemon have would as I good as wasn't texture are they when that understand liked. I were these crisper, a little are they fresh flavor The crisp crunchy. or not very the picking that to say have okay. I was body its by or a leg up by grasshopper large crickets, of the size were (They at hard a little was it eating and crickets) first. interesting, didn't love/hate it. It was was it. It didn't love/hate interesting, I tried fried crickets once, I was was once, I crickets tried fried I

eating ok, Was dressing.

in lollipops. crickets consumes have I insect. of the type know do not I was It the experience. about indifferent 40 37 41 39 38 42 43 44

119

1 1 1 1 13

1 1 1 17

1 1 16 Worms Worms Other + - 1 1 Grass- hopper hopper 8 1 1 2 2 2 1 1 1 4 1 1 1 43

1 1 1 1 1 1 10

13 1 1 1 Dessert Dessert Hidden Mexico Cricket 1 1 late late Choco- Candy Candy 8 4 2 1 1 34 fried/ fried/ Roasted/ Roasted/ seasoned seasoned

2 2 1 1 1 3 2 1 1 2 2 2 41 insect insect 82.0% 82.0% 68.0% 16.0% 8.0% 4.0% 26.0% 20.0% 86.0% 16.0% 32.0% 24.% 34% 26% Whole Whole as you'd like. like. you'd as Please describe your experience with with experience your describe Please edible insects, including what type of type what including insects, edible insect was tasted. Share as much detail detail as much Share was tasted. insect I've tried dehydrated crickets and and crickets tried dehydrated I've both were They mealworms. dehydrated a of pieces hard brown to the similar flavorless. but relatively kernel, popcorn crunchy. were They I had salt and vinegar crickets before, before, crickets vinegar had and I salt in a had crickets I good. Also, were they pleasant, not was that bar once, chocolate was it that due fact to the likely but most crickets. and chocolate, chili, insects. The cooked insects were good were insects cooked The insects. different that taste didn't it because I have eaten fried crickets and and crickets fried eaten have I crickets and grubs, cooked grasshoppers, tried chocolate also have I in lollipops. small of sort some ants or had that except for maybe an earthy taste to them. tastethem. to an earthy maybe for except didn’t strange, very were lollipops The good was chocolate the and them; like chocolate. the tasted just i because chili and lime with ate grasshoppers I once. restaurant in Mexican a powder in their was appeal their Most of aspect Every uniqueness. and exoticness but unpleasant mildly them was eating of to due part in large was this that suspect I old. too being the food I tasted a brownie made with insect insect with made tastedbrownie a I made a it powder believe was I powder. very tasted brownie The crickets. from The mint. and chocolate of strongly liking. my for grainy too was texture on my visits to Mexico I have tasted tasted have Mexico I to visits on my were that crickets and Grasshoppers paste a tamarind coated with more of a snack. of more

45 46 47 48 49 50

120

Table D.2 - Comments on the color of 10CR and their trends

Too pale/light Like rice Grey Liked color Comments about the color of 10CR

1 1 i personally prefer a darker color

2 1 This one looks a little gray.

3 1 ..looks sort of unappetizing because of how light it is.

4 looks milky brown

5 1 A bit too gray, brown is more appealing.

6 1 looks a bit undercooked

7 1 The brown rice I consume usually doesn't have this dark tinge. It reminds me of old oatmeal.

8 1 The color of the product is light but slightly dull, with a gray tint.

9 1 It appears marginally more pale than what I would consider desirable. It is a muddled brown, with sporadic dots.

10 1 Something about the light color makes it look uncooked.

121 11 this is very light for what i think is a brown rice.

12 1 the color was too light

13 1 It seemed similar to brown rice which made it comforting and familiar to eat.

14 1 lighter grey color with a tinge of yellow.

15 1 looks like brown rice

16 1 The light color makes it look very mushy and unappealing.

17 it looks fine

18 1 similar to brown rice

i rated it as somewhat to light due to the uniformity of the color. there were grains of rice that looked lighter than others. 19 1 overall it was a nice color. 20 has the lightest brown color

21 1 I like the color

22 Looks like a cinnamon cereal.

23 1 The colour is more of a brown rice

24 purple rice color

25 1 Very pale coloration, almost looks like maggots.

26 1 The color of it was really close to looking like actual rice not too dark or light.

27 light brown/tan, sort of like sand

28 1 Looks like normal brown rice, not too dark, not too light.

29 1 It looks as if the food might be going bad

Too pale/light Like rice Grey Liked color Comments about the color of 10CR

30 1 It is too light and almost seems to be cooked with milk like in Spanish cuisine.

31 1 i think it's too grey

32 The color doesn't deter me from eating the samples.

33 1 The color is unappealing

34 1 greyish looking

35 1 normal looking brown rice

36 1 lighter grey and tinted white

37 Lightest of the colors

38 1 light gray in color, I perceive this to be slightly unappetizing. I associate gray with mold.

The sample was dark enough to notice that it was not generic white rice, and much too light to be distinguished as 39 brown rice. 40 1 compared to other products, this sample is lighter and has a faded color; not as much pigmentation

similar color to sample 693 except lighter, has that grey undertone. If I didn't see samples 825 and 965 it probably 41 1 wouldn't give me an impression of being stale.

122 42 I don't really have a color preference? This is a weird question.

43 1 looks too light, like, raw/undercooked

44 1 also more on the gray side not appealing to the eye

45 1 this product seems a bit too light, if the comparison is supposed to be to brown rice...

46 1 The pale tan look is just unappealing. I prefer the darker colors, as it has more character.

47 1 It is very light, almost looks like it hasn't been cooked. The appearance is the least attractive of the four samples.

48 really light

49 the color looks similar to oatmeal

50 1 The lightness of it is a little offputting

51 1 too grey.

52 1 The color was not very appetizing.

53 1 appeared very dull colored with a tinge of gray. Not as appealing to the eye as the other samples.

54 Not to dark nor light.

55 Too light to be brown rice but that doesnt really matter.

56 1 Resembles brown rice quite well. A little bit darker, but I wouldn't say, 'somewhat too dark' though.

57 1 The color of this sample reminds me of brown rice and looks like a nice sand color.

58 1 looked to light

59 1 By comparing to regular brown rice, this sample is grey.

60 Looks somewhat 'synthetic' as if the product was put into a pasta maker, then chopped into rice shaped pieces

Too pale/light Like rice Grey Liked color Comments about the color of 10CR

61 It may be a good thing because people will assume it is just regular rice, not insect rice.

62 1 The color gives off an undercooked/bland look.

63 the color is the lightest of all the other rice samples

64 Not terribly pleasing.

65 1 The color of this rice had me very hesitant to try the product.

66 1 this one has a pale gray color to it

67 Good consistent color

sum 28 9 8 2 % 41.79% 13.43% 11.94% 2.99%

Table D.3 - Comments on the color of 15CR and their trends

Too gray Grey Dark Like rice Comments about the color of 15CR or pale or dull 1

23 1 1 Too muted, not very appealing

2 1 Looks a little gray.

3 1 A little darker than the first, but I actually like the dark tinge better. Somewhat appetizing.

4 looks earthy brown

5 1 Much too gray.

6 A bit lighter than brown rice.

7 1 Not sure what I am comparing it to. It is kind of grey.

8 1 It doesn't appeal to me

Without being denoted as a cooked dish, this rice appears like a muddy semi-dark brown that looks like the 9 1 food that an animal may consume. The brown color is more grey compared to its brown counterparts. Definitely the other samples look more 10 1 1 appealing. 11 N/a

12 noticed that it is speckled.

13 1 1 looks like a greyish dark

14 1 dark brown

The color is okay but if it went any lighter, it would not appeal to me. The lighter color makes it look 15 somewhat mushy. 16 it looks fine to me

17 1 slightly too dark

Too gray Grey Dark Like rice Comments about the color of 15CR or pale or dull 18 nice brown color

19 reminds me of a purple rice

20 1 The color reminds me of chocolate milk. It's not too dark but only slightly more dark than normal.

21 1 A very pale, dull color

22 1 The colour is right

23 darker than the sample 249. Very healthy looking

24 1 1 It has an odd grey coloration to it that does not make it look too appetizing.

25 1 The color of it was good though a little dark.

26 1 brownish grey color

27 1 Has the right color, looks like normal brown rice.

28 Knowing that I was going to eat rice I was expecting somewhat white or brown color.

29 Not as good as the first two, the color seems slightly off.

30 1 kind of grey 124 31 Looks like microwaveable quinoa.

32 light in some parts and dark in some parts

33 1 it seem to be an okay normal color

34 1 some colors of grey and brown

35 There is a color variance in his sample. the more moist spots are lighter while the drier spots are darker.

Cool gray-brown color. Color seems evenly distributed. The cool color does not make it seem so appetizing. 36 1 1 I associate gray with mold. 37 1 1 The color was dark and dull, visually unappealing.

38 I know brown rice to be somewhat more dark

39 1 slightly darker than other tested products

40 1 1 has a grey undertone which some may find displeasing- may appear as stale.

41 No preference.

42 looks too light on one side and too dark on other, like no one flipped the frying pan

43 1 looks more on the gray side than the previous two samples

44 the product is unevenly colored. some areas are darker than others

45 The rice is more appealing with the brown woody color, over the bland beige color.

46 1 1 Has a greyish color to it that isn't very appealing.

47 1 dark

48 has more of a light purple brown

Too gray Grey Dark Like rice Comments about the color of 15CR or pale or dull I wouldn't say the color is too dark, but it does look a little weird. I can't really explain it, but it just looks a 49 1 little weird to me. 50 nice brown

51 1 It is kind of a brownish grey.

52 1 1 a little on the gray side, but not so unappealing. Would look better with a rich brown color.

53 Some grains are slightly darker

54 Does not discourage me but it does not necessarily appeal to me.

It looks like B. cereus invaded this rice, and is the product of a mutated spoonful of rice with its grey, slimey 55 (moist), appearance. Looks like it came straight from Oliver Twist. 56 reminds me of a beach sand color mixed with rice.

57 1 looks like a brown rice

58 It is a light brown

59 By comparing to normal brown rice, this sample is too light.

it's almost just about right, it's a little creamy looking as if brown rice was boiled in milk (but less than the 60 125 first sample) 61 1 The color seems just right, not too light, but a nice crisp tan.

62 the color of this rice looks too light, but it looks similar to maybe a 'whole grain' type of food.

63 1 1 The color looks a little too gray which I don't like.

64 1 1 It is a dark gray color.

65 1 The color is a little too dark. It looks like it is overcooked.

sum 13 16 12 5

% 20.00% 24.62% 18.46% 7.69%

Table D.4 - Comments on the color of 10LR and their trends

Like rice Burnt/toast Soy sauce Like color Comments about the color of the 10LR

1 Inconsistent

2 Nothing about the color is unsettling

3 1 The brown reminds me of brown rice!

4 1 a light color that looks like undercooked brown rice

5 1 It's a nice brown color, similar to that of whole grain rice. I like it.

6 1 it looks a bit overcooked at some parts and others under

7 Splashes of darker color

8 This color appeals to me more than 249 and 693.

9 It looks like cinnamon cereal.

10 1 Has a nice brown color that can be easily associated with brown rice.

The rice appears to be a hue of brown that does not seem too dark nor too pale, serving to be optimal to 11 1 my personal preference. 12 1 The product looks like a toasty brown rice. 126 13 1 the products color is appealing to the eye.

14 1 similar to brown rice. normal colored food that I would consume.

15 1 light brown yellow, slightly burned in appearance

16 darker than normal brown rice but not super dark

The color is just right. The color makes me want to eat it. It reminds me of the precooked packaged 17 1 1 rice I buy from the store. 18 1 i like the way this one looks the best

looks somewhat like oatmeal, which makes this a more familiar food and more comfortable to 19 1 consume 20 1 a little dark but looks like a burnt brown rice

21 1 The colour is right to me

22 golden dark brown color

23 The color seems inconsistent throughout the product. But, the color itself does not seem out of place.

24 The color was good nothing too dark.

25 1 it's a brown color, like the crust of bread

26 1 Has great color for brown rice. Not too light, not too dark.

27 1 Looks like actual rice

It actually looks like a Chinese dish of rice cooked in a clay pot with sweet soy sauce drizzled. It 28 1 personally reminds me of home. Good color. 29 1 It seems about the same color I would see in regular brown rice but a little lighter

Like rice Burnt/toast Soy sauce Like color Comments about the color of the 10LR

I am comparing to typical brown rice color. However, the darker color does not bother me. It reminds 30 me of wild rice. 31 the color is a right blend of light and color. it is a golden brown almost

32 1 has the correct color of brown rice, but its grain shape is too lengthy to be brown rice.

I am judging the color of this product in comparison to the other ones presented in front of me and a bit 33 in comparison to the usual brown rice that I consume. 34 1 Warm brown color, seems vaguely appetizing. Reminds me of soy sauce mixed into white rice.

This sample had the perfect coloring, light brown with just a hint of orangeness to make it visually 35 1 palatable. 36 good color darker then the first

37 preferred brown color compared to all of the other samples

38 a pleasing color- I would assume that it is an preexisting grain unless notified otherwise

39 No preference for color.

40 light brown

41 1 perfect color.

127 42 1 I find the color to be pleasant.

Darker color is more appealing than the pale brown, however the rice does not have entirely consistent 43 color, which is slightly offputting. May be a cooking issue, though. It is verging on being too light, but there is a brownish color that looks like it has been cooked. I 44 associate that darker color with better flavor. 45 light

46 1 looks good

47 1 I like the color of this one. It's not too dark and not too light. It's perfect.

48 1 looks like toast or brown sugar

49 It looks like a golden brown.

50 I would like it if it was a little darker like product 965

51 Very light when compared to the other samples and normal brown rice.

52 A bit light for brown rice but it doesnt matter to me. Its not discouraging.

53 Looks slightly ashen, in comparison to brown rice.

54 1 looks like brown rice

55 1 looks like brown rice

56 N/A

57 light brown

58 By comparing to normal brown rice, it is light and grey.

59 The color isn't completely uniform, but mostly a darker shade.

Like rice Burnt/toast Soy sauce Like color Comments about the color of the 10LR

60 1 looks like fried brown rice with seasoning

61 1 Looks like brown rice.

62 light brown texture

63 1 the color looks almost similar to brown rice, as if there is cinnamon cooked with it

64 1 The color of the product is a good representation of typical brown rice.

65 it has a decent color to it

66 The color seems pretty consistent and not too dark.

67 1 normal brown

sum 15 7 3 11

% 22.39% 10.45% 4.48% 16.42%

Table D.5 - Comments on the color of 15LR and their trends 128 Burnt/ Chocolate Brown Too dark Dark Orange Like rice Comments about the color of 15LR

cooked 1 A little bit inconsistent but vibrant color

2 Nothing out of the ordinary.

3 1 1 reminds me of...feces - the color that is. And burnt brown rice.

4 1 1 dark, but vibrant in its brown color

5 The color is not uniform, there are some black 'pepper-like' specks.

6 1 looks somewhat overcooked

7 Also somewhat uneven.

8 pleasant color

9 1 Little more brown compared to actual brown rice

The rice has a dark appearance, however it looks like a cooked-dark appearance so it 10 1 1 is not necessarily bad. It looks dark for any conventional rice dish, but in terms of

food in general it does not look bad. 11 1 Looks like old chocolate rice crispies, but looks well cooked.

12 1 1 a bit too dark, however it does seem appealing.

This was noticably the darkest sample of the 4 presented to me. I felt that because it 13 1 was darker it did not look as appealing. 14 1 slightly darker than brown rice

15 1 1 very very dark, looks like burnt rice

Burnt/ Chocolate Brown Too dark Dark Orange Like rice Comments about the color of 15LR cooked The color is slightly too dark by just a little bit. It looks as if it might have too much 16 1 1 of the product on it. 17 1 1 too dark in my opinion

18 1 orange-ish

19 1 1 although it does not taste like it, the dark color gives the impression of being burnt

20 1 1 a little dark does not look that appetizing

21 1 Reminds me of a chocolate flavored cereal.

22 The colour was fine

23 1 milk chocolate color

Looks a little too dark than what I have come to associate with brown rice. Some 24 1 1 color inconsistency as well. 25 1 1 The color of it was very dark similar to overcooked rice.

26 1 dull brown

129 27 1 1 It was very dark, almost looked burnt.

28 1 1 For rice it is a bit too dark

29 It looks very interesting and also appetizing. Almost as if it was seasoned with herbs.

30 1 It look doesn't look very good.

This looks like the darkest sample but doesn't deter me from trying/ eating; it looks 31 1 1 like its been fried if I had to say anything. 32 1 darker complexion

33 1 very dark looks like too much insects are in this

34 looks like the rice might have soy sauce flavoring. misleading.

35 1 1 Darkest/most brown of the products presented to me. Not too dark.

36 Seems a bit grainy in color. Uniform color throughout product

The rice product appears very similar to a brown rice or whole grain product in color. 37 1 I would not think anything was unusual about this product. 38 1 darkest out of all the samples

darker than sample 825 but it is still a pleasant, warm shade. Gives it an earthier, 39 1 more 'natural' impression. 40 No opinion.

41 1 looks to dark to pass for rice, but might just be the normal color for whatever it is

42 1 the color is dark, yet you cannot notice there is something in the rice

43 1 it looks just like brown rice, nothing abnormal about the color.

44 1 it's a bit darker than what I expected.

Burnt/ Chocolate Brown Too dark Dark Orange Like rice Comments about the color of 15LR cooked 45 1 Overall color is good, however about 30% of the grains have a darker, dried out look.

I'm not sure how good the lighting is in this room. It might look different in sunlight, 46 but I think that the product looks appetizing. 47 light

48 1 medium brown

49 1 1 It looks a little too dark

50 nice color

51 1 It is dark and reminds me of brown sugar.

52 My favorite coloring out of all the products.

53 1 Fairly dark when compared to brown rice.

54 About the color of brown rice

Looks like dog food! :( Or it looks like I was trying to fry rice, and I burnt it... andd then I scraped it off the 55 1 side of the wok to eat it because I have no more food... or rice... in my fridge to live 130 off the rest of the week's budget. I could compare it to the purple rice again, but it... is... poop.... colored.

looks like wet dirt or the sand that is on the shores of a beach that has pieces of sand 56 left in it. 57 1 it looked like a color similar to brown rice

58 1 1 By comparing to brown rice, it is too dark.

59 One of the more tanner colors, but it is appealing to me

60 1 makes me think of over fried rice and soy sauce

Although this one has a darker tint to it, the toasted look makes it look more 61 1 1 appetizing. the color looks similar to the first rice sample. looks almost like brown rice but 62 thinner The product looks a little too orange, especially when compared to brown rice. This 63 1 is slightly unappetizing. 64 1 1 this one has a somewhat dark brown color to it

Compared to the brown rice I prepare, this is slightly too dark and does not look like 65 1 1 consistent coloring 66 Color makes it look like brown rice

sum 3 7 11 11 30 2 3

% 4.55% 10.61% 16.67% 16.67% 45.45% 3.03% 4.55%

Table D.6 - Comments on the aroma of 10CR and their trends Rice/ Dog/pet Odorless/ grainy/ Fishy Playdoh/ clay Earthy Dislike Comments about the aroma of 10CR food faint wheaty 1 1 Odorless, no preconceptions

2 1 Almost no smell to this one. Does not bug me.

All the products smell about the same...this one had just as much intensity as the 3 second test. a strong smell that resembles what brown rice would smell like. i did not notice any 4 1 fishy smell 5 1 Does not smell fishy at all, smells more like wheat or some other type of grain.

6 ok

7 1 Very faint scent, but I prefer that.

8 1 Smells like cooked rice.

9 1 It is not a fishy smell, it has a smell that reminds me of dog food.

The fragrance does not seem far from the typical scent that I associate with rice, 10 1

131 indicating that it does seem unpleasant but rather potentially enjoyable. The aroma is somewhat that of fresh baked bread. It reminds me more of wheat 11 1 bread. 12 1 1 the product's aroma was too fishy upon smelling.

13 1 smells like wet dog food.

14 1 kind of resembles the smell of play doh

15 1 light smell of fish.

16 1 It has a very earthy smell but it is not a bad thing.

17 1 smells like dog food

18 1 I didn't smell any type of fish in this rice mostly a earthy smell

19 1 has a slight starch smell

20 The aroma was right for me

21 1 this one has the rice smell

22 1 Was not able to smell much from the product.

23 1 It had no off putting smell. It does have a very floury scent to it.

24 1 1 it smells like rice but a little earthy

25 1 Smells like normal brown rice.

26 1 The smell is apparent but not too much

27 1 The smell is inviting and smells almost like brown rice. Very good

28 1 also smells like play dough

Rice/ Dog/pet Odorless/ grainy/ Fishy Playdoh/ clay Earthy Dislike Comments about the aroma of 10CR food faint wheaty 29 It smells fine, I wouldn't have even known insects are in it.

Initial impression reminds me of the smell of dog food (kibble) but only if I had to 30 1 relate it to something. 31 1 Reminded me of dog food smell

32 1 smells like shellfish

33 1 smells mild not too strong

34 1 Kind of smells like dog/cat food.

35 Hay like aroma.

36 1 There is barely a fishy scent unless I stick my nose into the sample cup.

37 1 The product had a weird smell (reminiscent of dog food), similar to another sample.

38 1 faint and nice

39 1 slight odor, not noticeable

40 1 Smell is less potent than the other three samples.

132 41 It doesn't smell bad?

42 1 barely fishy scent, just how i would prefer it. 825 and 965 were to fishy for me

43 1 smells like wheat

44 1 not really any smell

45 1 It doesn't smell fishy at all; I only can smell a wheaty aroma

This has the same earthy type of smell as the last one. There is an aroma, but it isn't 46 1 off-putting. 47 good

48 1 smells grainy and flavorless

49 It smells a little like a taco shell. I like it.

50 1 very slight smell, not strong, hard to tell what it is I am smelling.

51 1 I could not detect an aroma.

52 1 Smell reminded me almost of pet food. I liked it though.

53 1 Very faint smell.

54 1 1 Smells like wet dog food. Kind of unappealing

I had to put it pretty close to my nose, so I think I like the odor as well. It's quite 55 1 faint. 56 1 The aroma of the product reminds me of whole grain cereal, smells like a lot of flour.

57 1 smelled of dog food

58 1 doesnt really have an aroma to it

Rice/ Dog/pet Odorless/ grainy/ Fishy Playdoh/ clay Earthy Dislike Comments about the aroma of 10CR food faint wheaty 59 1 It smells a little bit like white rice and flour.

60 1 Smells like dog food, but it's fine.

61 1 smells really grainy - like whole grain, with very subtle hints of savory

62 1 It kind of smells like pet food.

63 1 Strong fishy smell

64 1 this rice smells like fish, just fish

65 1 Smells somewhat like clay

66 1 Out of all the rice samples, this one had the most fishy odor.

67 1 it did not seem like it had an odor to it

68 1 Just a little smell of fish flakes

69 Smells bland.

sum 12 17 16 9 3 4 2

133 % 17.39% 24.64% 23.19% 13.04% 4.35% 5.80% 2.90%

Table D.7 - Comments on the aroma of 15CR and their trends Grain/ Fishy Earthy Liked Dislike Pet food Comments about the aroma of 15CR wheat 1 No strong odor

2 The smell reminds me of some type of plastic.

It's a little fishy I guess is the smell? I mean, I would say that it's not bad, but it's 3 1 1 something I'd rather not smell for my meal. 4 1 not a strong smell, but does smell earthy, like brown rice

5 1 Too fishy, like those fish food flakes.

6 1 The smell is pretty good

7 1 Pleasantly grainy.

8 Smells similar to 249

9 1 There is a slight off smell, that isn't necessarily fishy but is a little off putting.

Despite the color, this product has the scent of a conventional rice dish; the aroma is 10 1 1 mildly desirable. 11 1 The aroma is that of flour.

12 1 the smell is appealing, and balanced.

Grain/ Fishy Earthy Liked Dislike Pet food Comments about the aroma of 15CR wheat 13 1 smells earthy and I do not notice any fishy smells that deter me

14 1 1 smells like a combination of cereals and dog food

15 1 smells pretty bad

16 1 This aroma is just right. I smell brown rice but with a slightly earthy scent.

17 need to be close to notice the smell

18 To me it smelled a little sweet

19 1 odor is not bad smells starchy

20 1 Smells somewhat like pasta

21 1 The aroma was good for me

22 1 does not have a strong smell. Just like rice

23 A mellow aroma not pungent or repulsive.

24 Didn't have a scent that was out of the ordinary.

25 there's a smell but i can't distinguish what it smells like

134 26 1 Smells like normal brown rice, nothing distinguishable about it.

27 The smell doesn't remind me of fish but it is a bit strong.

28 1 A nice inviting aroma.

29 1 smells earthy rather than fishy.

30 1 1 I really liked the smell of this one it smelled like my rice at home

31 1 smell of shellfish but not too strong

32 1 it smelled less fishy and it was earthy

33 1 smells like brown rice

34 More of a Hay aroma.

35 1 Slightly seafood-like. Not as overwhelming as I expected.

36 1 The sample had a strong mineral scent and fishy scent.

37 1 smells gain like

38 odor not too noticeable

39 smell is more potent than samples 825 and 965.

40 It smells ok.

41 Looks similar to brown rice.

42 1 subdued odor. wouldn't even say it smelled fishy... just sort of grainy/earthy

43 1 smells like rice

44 1 smells pretty good i really liked the smell

but it smells like there are are there like smells it but locust) in this product. this in locust) when eating the product, product, the eating when cooking rice with fish water; it smells like rice a bit but bit a has a rice like it smells it water; fish with rice cooking Comments about the aroma of 15CR of 15CR about the aroma Comments to to it it it comes in more amount or denser, it might be much too fishy. It fishy. It too much be it denser, might amount or in more it comes is bad. If isIf bad. pet food. pet food.

it smells wheaty; I don't detect any shellfish aroma any don't detect I wheaty; it smells but not fishy smell strong No discernible scent, other than a cooked starch aroma. starch a cooked than other scent, discernible No isn't offensive. but it is a smell, There good chip.tortilla a like a little smells all. atIt not fishy It's this one. of the smell like really I food. not great maybe but food like smells bland, unoffensive, store. fish a of reminds me smells The much smell not Could noticeable It's bad. isn't aroma The be might insects (crickets?... 'fewer' just reminds me of grains and flours like the others the like flours and grains of me just reminds bad smelling not smell The like smell smells earthy, but not fishy. also nutty also not butfishy. earthy, smells It has an earthy smell, pleasant grain smell. smell. pleasant grain smell, has an earthy It similar smells the rice to go with smell fishy Smells like a grain to me to me a like grain Smells one strong a not is it but aroma an did have It flakes fish of food slight smell Very 5 5 1 1 1 pet food. like smells it again, Once discouragement. towards me moves Definetly food. dog wet like smells It 1 1 offood dog smells 1 1 1 1 1 5 5 Dislike Dislike food Pet 1 1 8 8 Liked Liked

1 1 1 1 1 1 1 1 1 17 wheat wheat Grain/ Grain/ 1 1 1 1 Fishy Fishy Earthy

1 1

1 % 13.64% 12.12% 25.76% 12.12% 7.58% 7.58% 45 46 49 50 47 48 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 sum sum 9 8

135

Table D.8 - Comments on the aroma of 10LR and their trends

Earthy Fishy Mild Pet food Rice Disliked Liked Comments about the aroma of 10LR

1 1 no strong odor

2 Somewhat smells like plastic.

3 1 ...it still reminds me of brown rice..!

4 1 1 a bit earthy, but not overwhelming or distasteful

5 1 It doesn't smell fishy to me, but the aroma is not very appealing.

6 1 smells a little bit like fish food

7 1 The aroma makes me not want to taste it.

I wouldn't call it fishy but it definitely smells funny. A cross between shrimp and pet 8 1 1 store fish food. 9 1 Smells like rodent food

The product has a somewhat earthy scent to it, such to the point that I would be able 10 1 to distinguish the fragrance from a typical rice dish. 11 1 There is a definite strong smell. Reminds me of turtle foods, that are flakes.

136 12 1 1 it smells like cat food, and not an appealing smell

13 I didn't smell any essence of fishiness.

14 1 smells like dog food

15 1 smells way too strong

16 1 1 It smells just like rice. It has a somewhat earthy smell.

17 1 smells like animal food again to me

18 the smell is noticeable without being too close to the food but it is not overpowering

19 1 I can smell a little bit of fish in this sample

20 has a smell cant pin point

There is a distinctness to the aroma like a food product I've smelled before, somewhat 21 like a pasta. 22 1 I could taste the fishy aroma

23 no smell compare to others

24 1 Has a rather noticeable and pungent aroma. Not repulsive, but off-setting.

25 1 Didn't smell bad at all had a rice smell to it.

26 it has a slight sandy smell

27 Smells fine, nothing distinguishable about the smell.

28 1 Because it has the appearance of rice the smell is too strong for me

29 1 Slightly too fishy but not unappetizing.

Earthy Fishy Mild Pet food Rice Disliked Liked Comments about the aroma of 10LR

30 smells kind of like play dough

31 It smells okay.

32 1 1 There are hints of 'fishy-ness' but not as strong as other samples

33 1 Smells about the same as regular brown rice with a bit of a strong smell at the end

34 The aroma is similar to wild rice, come to think of it.

35 smell was bearable, not too strong

36 1 smells very fishy and smells like cardboard

37 1 mild smell

38 1 1 Still has fishy odor, but not overpowering or overwhelming

This product smells like dog food. I can finally properly describe it as a strong mineral 39 1 scent. 40 1 never thought of the smell to resemble fish but it does

41 For some reason smells like plastic to me. Like elementary school cafeteria rice.

42 1 Smells too strong imo. 137 43 1 doesn't smell appetizing

44 1 smells like rice

45 stronger smell than previous sample

46 1 1 The aroma is definitely a mixture of wheat and shellfish, but it's not overpowering

I wouldn't describe it as fishy, perhaps musty, but it has a cooked starch smell, which 47 1 is appropriate. 48 1 1 Again, an earthy smell to the product, but not overly off-putting. Not too strong.

49 good

50 1 grainy scent

51 1 1 Smells a little fishy, not too much though.

52 unsure about what i am smelling.

53 It smelled like fiber cereal.

54 1 1 Also smelled like pet food and was a little enjoyable.

55 Can only smell if it is very close to my nose.

56 1 1 Smells like wet dog food. Somewhat unappealing

For insect rice, not too bad. It's rather pleasant (in comparison to other insect 57 1 1 products). Not too noticeable. 58 1 smells of dog food

59 1 I did not get a lot of smell on this sample.

desirable desirable a not not

is is

that that

and 15LR smells like. smells

me.

food food appealing. first petsmart, petsmart,

fish fish aroma the is not is dettered dettered the the It It what what or or smelled. to to

me me crickets at at crickets food. food. wet of of have have

I I similar similar

obvious then then obvious turtle turtle when when smell smell bothered bothered smell.

from this sample was again, very pet-food aromas. Like pet-food Like aromas. again, very was this sample from like like smell fishy, fishy, more more

the the Comments about Comments food food anything anything food food

bit too too aroma aroma Comments about the aroma of 10LR of 10LR about the aroma Comments strong. food. dog dog 825 fish fish bit the the tank. to to a little little cat cat almost as if you cooked rice with fish water. it still has a similar similar a has still it water. fish with rice cooked if you as almost a fishy smell, it honestly smelled like pretty strong green tea to me me tea to green strong pretty like smelled it honestly smell, a fishy offputting offputting smells just smells of of than reminds of of reminds isa fish fish about about little little a bit bit smell smell fishy, fishy,

aroma is a isaroma unlike isaroma smell smell has a smells like like smells somewhat somewhat Nothing Nothing a smells the extremely smells aroma This strong It It not even It's wouldn't I it, but mind do nor I smell, the hate don't it. I kept on sniffing as I plate. on my minutes than five more it to smell want a The like It's fishy Less smell The smell. The it smells a bit smells rice cooked Has a distinct odor but it does not smell bad. not smell does it but odor distinct aHas got I aroma of hint the slight pellet store feed animal an The aroma of the rice did not smell of anything fishy. fishy. of anything smell not did the rice of aroma The it did not have an odor to it to an odor have not didit There is a slight odor, but not too bad. bad. too not but isodor, a slight There pet store a bit a like little smells

arthy E 3 3 1 1 it like but I fishy a little smell does savory, smelling up something picking 4.48% 4.48%

ice R 9 9

1 nsect I 1 1 1 1 9 9

1 825

Table D.9 - Comments on the aroma of 15LR and their trends 1 1 1 1 11

1 1 trong S food Pet Rice Disliked Liked ild 1 1 1 1

food 1 1 et et P

1 1 1 1 14

1 1 1 islike D

1 1 1 1 ishy F Earthy Earthy Fishy M

% 5.97% 20.90% 17.91% 16.42% 13.43% 13.43% 7 8 9 2 3 4 5 6 1 60 63 64 61 62 65 66 67 13 14 15 10 11 12 sum sum 4

138

Fishy Dislike Pet food Strong 825 Insect Rice Earthy Comments about the aroma of 15LR

16 1 the smell is noticeable and somewhat lingering

17 1 1 The smell of fish hits you pretty hard

18 not bad cant pin point the smell.

19 1 Smells a bit stronger than the previous sample, 825.

20 The aroma was somewhat odorless

21 1 smell like rice

22 1 1 1 Very pungent fishy smell. Very off-putting.

23 The aroma was fine but had a hay scent to it.

24 1 it smells like fish

25 musty, grainy smell

26 1 Smelled a little bit fishy, but it was not enough to cause dislike or disgust.

27 1 The smell is a bit strong

28 1 It does seem too fishy but not enough to be unappetizing.

139 29 1 It smells like the fish flakes you would feed your goldfish.

30 1 Smells like fish food but I'm open minded.

31 1 It smelled more earthy than fishy to me.

32 1 the smell is a bit too strong

33 1 1 smells very fishy and earthy!

34 1 im scared to taste the product.

35 1 Strong aroma.

Smells like dried fish. Strong odor, but not smell-able unless I place it 2-3 inches 36 1 1 under nose. The product had a scent reminiscent of dog food. I can't describe it any other 37 1 way. 38 1 same as 825 for me.

39 1 Doesn't smell very good.

40 1 not far off from sample 825...

41 1 it smells like fried grasshoppers

42 had the smell of freshwater plants you find in ponds or lakes. not a terrible smell.

43 1 There's definitely a shellfish-y smell to the product

I don't know if I would have identified it as fishy, but there is a definite smell to 44 1 1 it that is strong. 45 okay

46 1 very earthy/ grass

smells like like dog smells unpleasant. could be. (It be. (It could Comments about the aroma of 15LR of 15LR about the aroma Comments resembles the smell of dried insects, specifically the crickets at the crickets driedspecifically of insects, the smell resembles

smells just fine smells ok fishy. a little smelled It pet food like smells nose my from away it inches 2 to smell able was I unappealing Somewhat food. dog wet like Smells it as bad as not although, too, food dog like Smells asVery well). my eyes to close if were I even food smelled like dog food dog food like smelled product. the of a little can smell I definitely petsmart unappealing bit and a strong is very on this sample the smell disagreeable not but pet food a bit like smells had first sample the like aroma strong a have not did this one flakes food fish of me reminds smell The yikes barn a like farm smells 3 3 1 1 1 1 4 3 1 1 11 1 1 1 1 1 8 1 1 1 1 1 1 1 1 10

17 Fishy Fishy Dislike food Pet Strong 825 Insect Rice Earthy 1

1 % 27.42% 16.13% 12.90% 17.74% 6.45% 4.84% 1.61% 4.84% 48 49 50 51 52 53 47 54 55 56 57 58 59 60 61 62 sum sum

140

Table D.10 - Comments on the flavor of 10CR and their trends

Bland Aftertaste Like Rice Tortilla Bitter Comments about the flavor of 10CR

1 1 Bland but can be seasoned when used as part of meals

2 1 Taste like wild rice. Slight aftertaste nothing bad.

I mean...I'm used to eating quite unflavorful foods so this didn't exactly taste bland to me 3 1 1 but it probably is bland for most people..not much flavor in this, not much earthiness as the

last ones, and the aftertaste is almost as undesirable and bleh as the first test!! :( 4 1 1 a weak starchy taste that was kind of dull

5 1 I enjoyed the flavor.

6 1 It had practically no taste.

7 1 1 No flavor while eating it but it does have an aftertaste that is somewhat strong.

The flavor does not lean too strongly toward a good flavor, however, it does not taste very 8 1 different from brown rice. If served warm, I believe that the flavor profile and retention

would be stronger in a more desirable way. There is a very slight bitter after taste. Although the bitter after taste goes away after it is 9 1 1 completely swallowed.

141 10 1 the product lacked flavor

I enjoyed the flavor initially and it was my most favorite. But it did have a slight aftertaste,

11 1 similar to 693 which deterred me a little bit and was not to my liking. 12 tastes a little like dog food.

13 1 flavor is pretty bland, it tastes less strong than the smell

I was not able to tell that there is something added to the rice. The flavor is somewhat 14 earthy but it is good. 15 1 no flavor

it tastes a couple of bites for the flavor to kick in. the first few are a bit too bland but once i 16 1 1 recognized the flavor is was enjoyable 17 1 I didn't taste anything when trying this rice.

18 1 Tastes good reminds me of rice

19 1 The flavour was similar to the brown rice that I am used to

20 1 taste really good.

21 1 1 Good flavor, tastes just like rice.

22 Does have an earthy taste to it but it isn't bad.

23 1 there isn't much flavor

24 1 weird aftertaste

25 1 Was fine while chewing, but when swallowing and aftertaste were somewhat earthy.

26 1 1 Leaves a slightly bitter? taste on tongue

27 1 A very nice taste that allows the sweetness to come out. Very pleasant.

Bland Aftertaste Like Rice Tortilla Bitter Comments about the flavor of 10CR

28 1 Just tastes like cold rice, but is edible.

29 1 bland, no season

30 1 taste very mild and like brown rice not bad flavor

31 1 taste like actual rice

32 This earthy flavor leaves no bitter aftertaste.

33 1 The product had a mild flavor but had a strong earthy aftertaste that was mildly unpleasant.

34 1 1 i believe this is the best yet, a slight aftertaste of gain as expected

35 1 tastes good

36 1 Not as much flavor as the previous samples. I don't taste much at all actually.

37 1 It has a really weird aftertaste, don't like it that much.

38 More fishy than earthy

39 nondescript taste

40 tastes like wheat bread

142 41 1 couldnt make out any taste on this sample

it almost tastes like plain, white corn tortillas. it's not necessarily bad, but I feel that it 42 1 1 could be better seasoned 43 Flavor is appealing, almost has a mushroomy undertone.

44 1 This one is also bland, but in a good way. There is a flavor but it isn't very strong.

45 good

46 its not bad. i would eat it

47 1 1 Again, a little bland, but I like this one. It tastes a little bit like a tortilla chip.

48 could use stronger flavor

49 1 There was not too much flavor in this product.

50 1 very bland. Didn't really enjoy nor dislike it.

51 1 Tasted like corn tortilla flour.

52 1 Bland but not too bad

53 1 Earthy aftertaste

The initial taste of the product was too bland, but as I chewed a little more into the product, it had a bit of an aftertaste... the cricket. If I ate this with a hearty meal or a lot of sauce, I might not recognize it. I really like the initial flavor though-- bland is a good thing for 54 1 1 rice. Maybe adding salt could work? But in terms of rice as a staple food, I could see myself eating this as part of my diet. As an American, probably once a month. However, eating a 4th bite, it seems to be pretty inconsistent to me. 55 1 1 The flavor of the product is very bland , reminds me of plain cooked rice

I I

good. not be be would would I I would would

flavor of a bug (which bug a(which of flavor plain oatmeal, and and oatmeal, plain consistency. consistency. 15CR components it it components like like of of

that it has a has it that tastes like a a like tastes other other bread bread flavor with with

my first two bites were kind of...enjoyable. bites were two first my

the the very very in in A A

right in aspects of earthiness. There was a slight a slight was There earthiness. of aspects in right and became more noticeable with each bite. each with noticeable more andbecame good.

off rice mixed mixed it it not bland. but product. product. like like like like not not bad, just about about just the own own feel feel and and more more Not Not i

Comments about Comments

Comments about the flavor of 10CR 10CR flavor aboutof the Comments it's it's

but but little little rice. rice. on on

rice. strong strong a as well bad like like not

flavor flavor was not exactly favorable favorable not exactly was brown brown the product was was the product flavor flavor

taste taste Aftertaste to to tasted tasted isgood bland. the the flavor, flavor, strong strong the close close flavor flavor flavor was a was was too too was enjoyed enjoyed like like mind eating this again. this mind eating not does this It it of flavor The that aftertaste I displeasing displeasing The aftertaste the and first, the to compared earthy as really at hit me first flavor mean.....the I more seems aftertaste, an of bitter too Not either. first to the compared bad too isn't all first,than so the way?) (in earthy an flavorful I The Very that it sense in the earthy was the rice of flavor The the thought induces in it insects has is that knowing flavor) a good associate with particularly don't to ischewiness a good There i really enjoyed the taste to it in my mouth in my it to the taste enjoyed i really flavor much too Not There wasn't really any flavor, just a hint of grainy taste. grainy of hint just a flavor, any really wasn't There no flavor all, at anything like taste doesnt the rice taste. thought would it initially I as notbad as was rice this of the taste Taste more like real rice, which is bland. bland. is which rice, real like more Taste It taste like flour from white rice and grain. and rice white from flour taste like It first bite), (especially hintssavory of good some nutty, of kind earthy' 'savory the initial of instead 'dirt earthy' islittle a but later tasted very earthy earthy very tasted

1 itter 2 2 B

1 1 arthy E 3 3

- Comments on the flavor 15CRof and their trends alty S 11

1 1 1 1 1 1 13

Table D. eeds salt N 1 1 7 7

ice 1 1 1 R

13 1 land B

1 1 1 26 ftertaste Bland Bland Aftertaste Like Rice Tortilla Bitter A 1 1 1 1

7 8 9 2 3 4 5 6 1 % 40.63% 20.31% 10.94% 20.31% 4.69% 3.13% 64 61 62 63 60 58 59 57 11 12 13 10 56 sum sum

143

Aftertaste Bland Rice Needs salt Salty Earthy Bitter Comments about the flavor of 15CR

14 1 taste very plain

15 1 1 tastes like dirt

The taste was okay at first but I continued to chew it became too bland. At first, it 16 1 1 reminded me of the basmati rice that my roommate prepares Pakistani style but the taste

soon disappeared and I felt as if I needed more flavor. 17 tastes better than the last one i just had

18 1 aftertaste not offensive either

19 1 although not bad, it leaves somewhat of an after-taste

When taking the first couple of bites it didn't give any flavor but then it gave a strong 20 1 1 1 earth after taste 21 Initial taste and texture was good then BAM some weird funk hits you

22 I liked it

23 1 a little bit salty and oily

24 1 Good initial flavor, somewhat of an aftertaste.

25 1 The flavor at the moment was good but it did leave a weird after taste.

144 26 it taste pretty good, there isn't a strong earthy taste but not bland either

27 Tasted fine, nothing bad about it.

28 1 The flavor is a bit too earthy for me

29 Taste is nice and inviting. No strange after taste.

30 Although I've said the first two samples were just right, sample 693 tasted the best.

31 1 Didnt really taste anything

32 1 not much of a taste

33 1 it was very mild

Initial taste was good but then the earthiness was tasted in the back of my mouth. A bit 34 1 between somewhat earthy and just right, more towards the just right. 35 1 1 Strongly earthy to the point of being bitter.

The flavor was fishy but most pleasant compared to the other sample due to its lack of 36 1 overpowering after taste. 37 1 1 very mild 'brown rice earthy taste', not sweet

38 1 flavor has an earthy taste but it's enjoyable

Earthier than the previous two samples, but a different kind of earthy which I find more 39 1 pleasant. Reminds me of barley tea. 40 1 Pretty tasteless, but I like that about it.

pretty good, if I could add sauce I wouldn't think twice about eating it. honestly I'm pretty 41 hungry so if I had to pick one cup to finish it would be this one 42 1 1 1 tastes like salt was added, but has a bitter after-taste

evenly, tiny bit of fishiness fishiness bit tiny of evenly, Comments about the flavor of 15CR 15CR flavor aboutof the Comments in terms of taste. A very tiny aftertaste occurs, but nothing to to nothing but occurs, aftertaste tiny very A taste. of in terms a little salt, but the flavor is much better than the first sample. Bland, sample. the first than better is much flavor the but salt, a little the flavor was really good on this sample sample good on this really was the flavor of me reminds product that the flavors the three are and oaty grainy, earthy, mothball. a licked i like taste to started of but sort first, at pleasant was Flavor metallic somewhat or sour slightly turns after flavor flavor, initial Good okay This could also use use also could This way. in a butgood bland. somewhat but flavor still heavy It's a little bland. I would have to flavor it with something to eat it. to eat something it with to flavor have would I bland. a little It's needs salt salt needs sample the best far By about. complain chew. as you but fades inbeginning the flavor strong Very all at used insects there was tell not could it. butdont like i why tell cant I the flavor. dont like I in Although, the flavor. to a 'kick' there's and pungent Not this one. like really actually I had flavor but not overpowering not butoverpowering had flavor good actually. very tortillas, flour of me reminds glavor The bad to eat seem doesnt it but rice regular real it isn't cantell you good. pretty tastes the next mouthfull, I tasted more of an off-flavor (could be a different part of its body, body, itsof part different a be (could off-flavor an of tasted more I mouthfull, next the 'minerally'. slightly tasted more It little a had), which was I bite previous to the compared flavor minerally the bittery, of longer lingers taste the but bite, next the in better the for synergistically work may it eating, be may I food to other addition In described. flavor. a have flavor really doesn`t flour. takes like It distributed everything. of most. Hints the this like one I satisfying really was savoriness the but though. bland. did seem It corn tortillas of me Reminds is good. flavor This bad but not flavor unusual different a ait's like 1 1 to me. bitter tasted It 6 6 1 1 1 days. a few for out sitting been has product the like taste, and bitter issalty like the flavor 8.96% 8.96%

1 1 1 1 12 1 1 3 3 1 1 1 1 2 2 5 1 1 1 1 1 1 1 1 1 14

12 Aftertaste Aftertaste Bland Rice salt Needs Salty Earthy Bitter

% 17.91% 20.90% 7.46% 2.99% 4.48% 17.91% 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 59 60 63 64 65 58 61 62 66 67 sum sum

145

Table D.12 - Comments on the flavor of 10LR and their trends

Aftertaste Bitter Earthy Dislike Rice Bland Comments about the flavor of 10LR

1 1 strong aftertaste

A little bit of flavor could be added. I feel like there is a weird aftertaste that i cannot 2 1 describe. It's a light earthy taste at first, just reminds me of brown rice, but then the after taste is kind of.....undelightful. Not the best after taste in the world. It doesn't exactly taste fishy per say, 3 1 1 1 1 but it just tastes a little...bitter. BUT the taste doesn't linger too long. Initial taste is similar to brown rice. 4 1 a taste would expect from a type of rice

5 1 1 It was too earthy for me.

6 Some spots were worse than others.

7 1 1 Earthy tones as well as a bitterness to the flavor.

8 1 The flavor is reminiscent of undercooked brown rice, with a very grain-based flavor. 146 9 1 1 There is a grainy texture, and slight bitter after taste.

10 1 it had a bitter taste

The flavor seemed to be more pundgent than #693. The aftertaste was not as noticeable but 11 1 that could be because overall the flavor was more intense. 12 feels slightly hard, kind of like when rice isn't quite cooked

13 1 tastes like dirt

This sample tastes just like rice. If I didn't know what I am sampling I would assume it is 14 1 some sort of brown rice. I can definitely see myself eating this in the future. 15 1 not my favorite taste, kind of strong

16 1 there's an initial hint of flavor that is pleasant but then it becomes a bit bland

17 1 I don't taste much in this sample. There is a flavor but i cant tell what it is.

18 1 weird tastes bland and earthy ,

19 1 It tastes like plain rice.

20 The flaour was right for me

21 1 bland taste. Nothing surprisingly.

22 Flavor was quite pleasant. No lingering aftertaste. Nothing to complain about.

It tasted good nothing out of the ordinary. Tasted like regular rice, though did have a grainy 23 1 feel to it at times. 24 1 Had a mild earthy flavor, was noticeable.

25 1 It has a strong aftertaste

Aftertaste Bitter Earthy Dislike Rice Bland Comments about the flavor of 10LR

There is no taste, however I'm not sure if that is a bad thing as far as rice goes. The sweet 26 1 taste of rice is still present. 27 Does not taste any different than sample 249

28 1 Tastes like cold rice.

29 1 It has a strong after taste

I like the flavor, but compared to brown rice it is somewhat too earthy if the consumer is 30 1 expecting typical brown rice. 31 just the right amount of crunch and softness

32 the smell is overpowering and it taste a little like cardboard. smells a little bit like brown rice

33 1 it was really dry. like eating dirt, which is similar to eating earth.

I wanted to rate this between somewhat too earthy and between just about right. Slightly 34 1 1 bitter, but tolerable. 35 1 This product has a mineral-like earthy taste that was mildly pleasant.

36 1 1 decent flavor, however leaves an earthy aftertaste

37 1 Has a nice earthy taste, somewhat salty, overall pleasant.

147 38 Not bad actually. Its pretty tasteless, but thats actually sort of a relief.

39 1 Too bitter

40 needs sweet and sour sauce

41 1 tastes a little bitter

42 not bad at all, kind of bland.

43 This product is very, very bland.. It is almost tasteless.

44 1 Flavor is decent, it has somewhat of a 'wheat pasta' type flavor.

45 1 I don't know how to describe the flavor, it almost has a stale quality to it. Very unappealing.

46 good

47 full flavor

I don't like the flavor of this one. I selected 'earthy' but I'm not sure if that's how I would 48 1 1 describe it. It tastes a little fishy, as well as earthy. 49 1 flavor could use some intensity

50 1 It tasted like grains to me.

It was good that this sample actually had favor, but it did feel a little earthy. I prefer an 51 1 earthy flavor to a bland flavor. 52 I could taste the insects, but it was not strong.

53 It wasnt too bland or too earthy. It was alright. Not so discouraging

Tastes like flavored rice. If I didn't know it was insect rice, it would taste like the purple rice 54 or... low-quality, long-grained rice. But not bad.

Aftertaste Bitter Earthy Dislike Rice Bland Comments about the flavor of 10LR

it reminds me of very dry bread that has been left out for a few day, overall it taste okay and 55 wouldn't mind eating it. the taste also has a very strong herb taste as well. 56 1 tastes like dirt

57 1 The taste is not significant.

58 1 Not too much flavor, but that is okay since rice is somewhat of a side dish.

59 1 pretty earthy, also kind of resembles below al dente cooked noodles

60 1 A little bit earthy but it taste just right.

61 taste like cereal

62 1 There was not any flavor what so ever. It just tasted like I was eating plain powder.

the flavor is not at all that much of anything, almost like brown rice. it doesn't taste like how 63 1 it smells 64 1 The rice did not have much taste.

65 1 it was really dry and bland

66 1 This has a pretty good flavor. Reminds me of the brown rice I prepare at home.

148 sum 8 7 17 5 11 9

% 12.12% 10.61% 25.76% 7.58% 16.67% 13.64%

Table D.13 - Comments on the flavor of 15LR and their trends

Earthy Dislike Fishy Aftertaste Bitter Rice Bland Comments about the flavor of 15LR

This one is really too earthy. A bit difficult to eat and I cannot get over the fish food 1 1 1 1 smell. Oh goodness it isn't even too earthy it tastes like bitter and too strong of green tea 2 1 1 1 and then the aftertaste is just

BLARGH I want it out of my mouth!! bleh that was not so great. 3 1 had no noticeable flavor, and is rather boring

4 1 Too earthy, similar to what wood/dirt.

5 1 had a strange aftertaste

The flavor has veered away from what might taste like rice and has transformed 6 1 into something earthy, wheaty, and insect-like. The initial taste is earthy, and very mushy. There is a definite bitter taste as I 7 1 1 swallow. 8 It was strong but when added to other ingredients I could see it being fine.

9 1 1 it was bland, however it did have a salty after-taste.

10 1 The flavor was significantly more intense. It was too strong for my liking.

Earthy Dislike Fishy Aftertaste Bitter Rice Bland Comments about the flavor of 15LR

11 1 taste more bland than dog food.

12 tastes decent

When I first put it in my mouth, I can taste the smell. It felt as if I was eating turtle 13 food flavored rice. 14 1 i strongly dislike it

15 1 very earthy

16 the product has a nice taste that stays throughout, no bad after-taste and not bland

17 1 It has a strong fish taste to it

18 tastes a bit gritty

19 It was somehow rough

20 a little bit hard

21 1 Dense flavor, not exactly repulsive but is off-setting.

The after taste of the sample was very off. It was fine during but after it had a stale 22 1 taste to it.

149 23 1 it taste earthy but not too strong

24 1 somewhat strong aftertaste

25 1 Had a slight earthy taste.

26 1 1 Flavor way too strong. Leaves a bitter after taste

27 1 It is a nice flavor but too earthy. It is still appetizing.

28 The taste is a lot more stronger than the previous two samples.

29 1 odd aftertaste.

30 1 I taste the 'earthy-ness' at the end.

31 1 Very strong after taste

32 1 somewhat earthy taste

33 1 very unpleasant

34 i think there is some cricket antennae in this one.

35 1 Very earthy.

36 1 Tolerable, but strong aftertaste.

37 1 1 The product had a mild earthy flavor, not 'too earthy'. It was not unlikable.

38 1 more pungent then the others i would even say bold leaves an interesting aftertaste

39 1 tastes too earthy, not comparable to typical rice

40 similar to sample 825. pleasant, more wheat like this time.

The grainy-ness of this one overrides any appreciation of its flavor I might be able 41 to extract.

that I'm scared to go in public to talk to talk public in to scared go I'm that of slight bitter after taste that sticks to your your to sticks that taste after bitter slight of for me. for aftertaste aftertaste it could use salt. There is a flavor to it that isn't bad, but bad, isn't to that it a flavor is There salt. use could it Comments about the flavor of 15LR 15LR flavor aboutof the Comments The prior samples has an appealing woodiness, but this is is this but woodiness, appealing has an samples prior The this tastes almost like dirt without the texture of it, taste as unappealing unappealing as it, taste of texture the without dirt like this almost tastes great either. either. great product tastes like like tastes product the flavor of of the flavor as it smells smell the of me reminds It taste. my for earthy too is a flavor little The I did not enjoy the flavor at all, it had a fishy taste to it. taste to a fishy had atit all, the flavor didnot enjoy I okay it was Slightly bitter after flavor, not displeasing or overwhelming overwhelming or displeasing not flavor, bitter after Slightly The isn't taste bad hay to taste smell/ similar earthy. very flavor flour nice strong. too was flavor The aftertaste. earthy very protein tasted like It a lingering There's out. back it Spat pungent too little A with. to people me reminds the product of the flavor dry, somewhat is and mouth flour. with blend herb taste like I tiny savory bit of a verryyyyy bitterness, bit of tiny Definitely unpleasant. unpleasant. Definitely terrible. pretty yuck, not good at all all at not good yuck, grashoppers fried tastes like bad. its not but overall earthiness a or hint get somewhat i could bad flavor a not into 'fishy' evolves it then but is'earthy', experience I first flavor The Too fishy Too 6 6 1 1 oatmeal plain kind like eating of to it. It's flavor any isn't really There 1 much too discourage Doesnt bland. too or earthy too Not 1 bread bland like tasted 9.09% 9.09% 0 0 6 6 1 1 1 1

1 1 1 1 1 14 4 4 1 1 1 1

1 1 1 1 1 1 1 1 1 1 1 1 14

20 Earthy Earthy Dislike Fishy Aftertaste Bitter Rice Bland

1 1 1 1 1

1 1

% 30.30% 21.21% 6.06% 21.21% 9.09% 0.00% 42 64 65 66 49 51 52 53 54 55 56 57 58 60 61 62 63 48 50 59 44 45 46 47 43 sum sum

150

Table D.14 - Comments on the mouthfeel of 10CR and their trends Soft/ Too soft Grainy Liked Rice Comments about the mouthfeel of 10CR mushy 1 Not grainy or gritty

2 1 Does have a slight grainy feel that does not bother me but may discomfort others.

If you mean by smooth in the rate the mouthfeel as mushy, i would have to say a little too smooth. 3 1 1 but either way, this was fairly similar to the last products. this one has been the mushiest by far

though. 4 1 1 I enjoy soft rice so this was perfect.

5 1 Good.

The consistency of the rice did not feel different from typical brown rice with the minor exception 6 1 that it was marginally chalky in terms of mouthfeel. The product was chewy and was easy to swallow. The graininess was very similar to what wheat 7 1 bread tastes like. 8 the product was too smooth for me.

The product had a nice mouthfeel. It was gummy but I liked that there was not a noticeable 9 graininess to the product. 151 10 firm but not too firm.

11 1 very soft texture

12 1 i can feel the grain texture in my mouth as i chew the rice

I expected this sample to be too smooth but it was just right. It was not grainy and it did not leave 13 any residue in my teeth. 14 texture is super thick

15 enough bite, somewhat slimy

16 1 a little grainy but not bad

17 1 feels like a long grain rice

18 1 I enjoyed to mouthfeel

19 I like the flavor and it taste good

20 Good consistent texture throughout the product. Not grainy at all.

21 1 The feel of it was perfect it had a softness to it but was a little hard at times so it was well balanced.

22 1 1 it's soft but there are a crunchy bits

23 1 1 Was very soft, a little too soft for rice. Somewhat mushy.

24 1 1 The consistency is great and closely resembles rice

25 1 A very nice texture and it feels fibrous which is a plus.

26 1 1 Was a bit to mushy

27 1 soft, easy to chew

Soft/ Too soft Grainy Liked Rice Comments about the mouthfeel of 10CR mushy 28 1 it was soft and tasted like plain rice

29 1 Feels gummy, like rice does.

30 1 1 Slightly too soft.

31 Not too dry or wet/sticky. No sensation of sand-particles in rice.

32 1 The mouthfeel was great!

33 product easily dissolves

34 1 The mouthfeel and texture is smoother, like sample 693. a little grainier than 693 though.

35 Texture is fine.

36 unappealing

37 1 felt ok to eat just like regular rice

38 I don't detect many grain 'particles' for this sample.

39 1 1 1 Texture is inconsistent, however it's generally pleasant, similar to rice, if slightly softer.

40 I honestly don't find too much difference in mouthfeel and textures of the products so far.

152 41 1 grain

42 not grainy at all. easy to eat

43 Not too grainy and just smooth enough. It's perfect.

44 1 1 too soft

45 The mouth feel was not too grainy or smooth.

46 1 a little similar to actual rice.

47 1 I liked how smooth it was

I like the texture. It was similar to white rice, but not exactly brown rice. (i like white rice more 48 1 1 than brown rice texture) 49 1 Good texture, not grainy

There was a grainy aftertaste, but it seemed to linger the shortest time as compared to the other 50 1 1 ones. It feels like brown rice too, where it has just enough tension when biting into the rice. 51 1 grainy feel

52 By comparing to rice, this sample tastes like flour.

53 Seemed moister than the first sample.

there were some small chunks that were rigid, personally do not mind it but I believe it might lead 54 1 other people to think that this 'rice' wasn't washed enough 55 1 1 1 This one had much less grit, but wasn't too soft either. The texture was enjoyable.

56 1 it feels a bit like overcooked rice

57 1 1 1 Rice falls apart too easily. I would prefer it to be less grainy but more chewy.

this with with optimal optimal - teeth teeth was palitable palitable was it it sub my my But the the

on on rice. rice. with with

residue residue product. brown brown the 15CR

to to grainy grainy of

feel collaborated collaborated the

get get

feeling. throughout throughout mouth mouth when when compared compared not mouthfeel dry. mouth

side side gritty gritty did the the mushy mushy too too

I I your your firm. is a isa enjoyable enjoyable

not not soft soft soft consistency consistency in in

a side. be be but or or gummy gummy there there to to

it's fineit's part part Good Good have have mushy, mushy,

mushy mushy after after rice moment moment smooth smooth rice. did too too appealing gooey gooey 249 most most hard. hard. the it but like like Comments about the mouthfeel of 10CR of 10CR about the mouthfeel Comments too too

Comments about Comments first first texture. texture. like like not and on on the and and too too very very product. me of farro. of me little bit of the the bit of little felt felt

felt felt

the the for for

it not not rice! rice! 965 965 texture texture little little rice rice

however however is not is shaped shaped hard. from the from was great. was rice too too that was a was brown brown smooth smooth was good was great was sample sample

smooth smooth firm but firm grainier grainier of of noodle noodle not not grainy grainy was fine fine was a feel feel a texture texture bites where bites where is a isa than than complaints on on complaints normal normal texture texture it soft soft grainy grainy,

no no texture, texture, were were texture texture mouthfeel mouthfeel mouthfeel product seemed on a on product seemed too too I felt that it reminded it thatfelt reminded I overall overall feels very very feels feels like like feels grainy grainy have have it felt really good., really it felt there there It The The smoother Good The slightly slightly It me Reminds a Not initially chewy overly was rice The it profile retains. flavor like Tastes the The and slightly like feels mouth The sample. I

1 1 rice to brown close Very texture. Good 10 ice 1 1 1 Rice Rice R 16.95% 16.95%

1 1 1 1 1 1 1 - Comments on the ofmouthfeel 15CR trendsand their 13 ike L 15

Table D.

1 1 12 islike D

6 6 1 1 1 1 oft/ S mushy Too soft Too soft Grainy Liked

1 1 1 1 1 1 15 rainy/ Soft/ Soft/ gritty/ mushy mushy G crunchy

8 9 2 3 4 5 6 7 1 % 25.42% 10.17% 20.34% 22.03% 58 59 19 20 21 13 14 15 16 17 18 10 11 12 sum sum

153

Grainy/ Soft/ gritty/ Dislike Like Rice Comments about the mouthfeel of 15CR mushy crunchy 22 1 soft and not crunchy

23 1 Could feel small grains in it while chewing, and they remained in my mouth after swallowing.

24 1 I have the urge to drink water after tasting

25 1 A bit too grainy but it does have the sense of consuming higher fibrous food.

26 1 chewing makes it feel like mush or sticky

27 no taste but not crunchy at all

28 1 soft and well balanced

29 1 Can feel very small sand-like particles while chewing.

30 1 The mouth feel was great.

31 1 1 soft and paste like brown rice

32 1 lingering grainy texture

33 1 Very similar to normal Rice.

154 34 1 Pretty smooth, but I like that.

35 1 a little grainy

36 no complaints at all. this is the best sample so far

37 1 1 I enjoy the grainy/sandy texture of the product

38 1 1 Overall decent, but somehow has too much crunch, while also being too soft.

Again, the texture is not as chewy as what I normally associate with brown rice. The texture is 39 1 pretty good. 40 dry

41 1 grainy but is also a little too smooth

42 1 It's smooth and not at all grainy. It has the perfect feel.

43 1 too soft, falls apart to quickly

44 1 It was too smooth and I did not like the mouth feel.

45 1 1 Felt quite similar to rice, which I really liked.

46 Very chewy for me.

47 Too wet and smooth. however, i doesnt really bother me.

It's good except for the 'calcium'aftertaste, where it is grainy and grits with the teeth. However, this 48 1 goes away much faster than does 825; I like this mouthfeel better. 49 1 grainy feel

50 1 mushy and cold soft

51 It does not feel like rice, but it feel like flour.

mouth. knowing knowing though though

my my 965

it's something it's something inside inside

especially especially it it that

sample sample

me, me,

the teeth required. to to 10LR my my remnants of remnants of

recipe recipe similar similar feel feel I I me from eating it again. it eating from me I did not mind. There is some some is There didnot mind. I than between between taste taste

desirable.

graininess deterred deterred graininess it mouthfeel very very parts The The carbs such as brown rice. brown as such carbs the the overly moist so I would say say would moist so I overly

less water water less swallowed, swallowed, resistance resistance or or for for as I chewed it, chewed as I grainy grainy would not stop not stop would with with was not not was samples but samples as wholesome. grainy grainy chewing bits this this few few

that a

this product. thisproduct. feel feel completely completely other other

cooked cooked of of

rice rice rice

part. while while get get the only only Comments about Comments didnt didnt rice rice provides a provides all all this this like. it it not with with to to just just most most it to to Comments about the mouthfeel of 15CR of 15CR about the mouthfeel Comments of of some grainy grainy some swallow my teeth but but teeth my texture texture and the complaints. mouthfeel mouthfeel of insects. of to to out would would mixed mixed I I for for similar similar perfect perfect No No

seems to to seems crunch crunch hard hard over in over did feel did feel as chewing, chewing, mushy and not hard and dry, dry, and hard not and mushy crunchy crunchy grainy, grainy, rice rice sand y, y, grainy grainy - grainy grainy was a was isa slight slight product as soft as soft most most was too too was little little I did not like the the didnot like I is made this that like feels the like feels I Although left residue Sawdust alright. Feels not too It's eat in substitution could probably that I after Not a There There The almost this is it

the mouthfeel of the product was much too smooth. there was absolutely no crunch at allat no crunch absolutely was there smooth. too much was the product of the mouthfeel rice. overcooked slightly of reminiscent a islittle just chew The he feel was just right. Not too grainy, but not mushy either. not butmushy grainy, too justNot was right. feel he really small amounts of grittiness, feels a little dehydrated though dehydrated a little feels grittiness, of small amounts really not crispy or overly moist overly or crispy not

1 oft S 7 7 1 1 rice regular like almost feels Rice Rice

ice 1 R

- Comments on the ofmouthfeel 10LR and their trends 10

16 iked L Table D. 4 4

Dislike Dislike Like

1 1 1 1 islike D

1 1 12

Soft/ Soft/ mushy mushy 1 runchy C

18 1 1 1 1 1 1 1 1 1 1 1 gritty/ gritty/ rainy/ Grainy/ Grainy/ crunchy crunchy gritty/ G crunchy

1 1

9 3 4 5 6 7 8 1 2 % 31.58% 21.05% 7.02% 17.54% 12.28% 52 57 56 55 54 53 14 15 16 10 11 12 13 sum sum

155

Grainy/ gritty/ Crunchy Dislike Liked Rice Soft Comments about the mouthfeel of 10LR

crunchy 17 1 I do not like the texture of this one

18 1 1 i felt the mouthfeel was enjoyable, more on the smoother and softer side than grainy

19 1 I can feel some grain with this sample but not enough to be off putting.

20 1 i feel the grains as i chew after a big bite

21 1 The mouthfeel was a little grainy

22 1 1 the texture was not so good. I can feel it when I was eating it. Feels like eating little rocks

23 1 Slightly on the grainy side, but not too off-putting or noticeable.

24 1 1 Felt pretty close to regular rice but it did have a grainy feel at some points.

25 1 1 too much crunch

Grains were distinguished, could feel them while chewing and remained around my mouth 26 1 after swallowing. 27 It probably wouldn't be so bad combined with other food

28 1 Too grainy as if it's very high in fiber.

156 29 kind of falls apart in your mouth

30 Texture feels a bit thicker than the first sample.

31 1 The texture was good didnt feel to mushy or hard

32 wasn't too hard to chew but you could feel that seasoning was added to it

33 1 1 1 it felt soft but a little grainy i felt a little crunch

34 not as grainy as 693, but still quite unnatural to what i am use to.

35 1 somewhat grainy (not too grainy, just grainy.)

36 Rice grains stick together. Not overly dry or wet/sticky. No sensation of sand-like particles.

37 1 1 The product was a tad grainy to be confidently enjoyable.

38 1 1 slightly grainy, comparable to brown rice

Falls apart within quickly once within mouth. Grainy chunks, but may be pleasant to those 39 1 who like grainier textures. 40 Definitely chewier than the last one.

41 not a problem

42 1 less grainy than previous sample

43 1 1 I like the grainy texture - it's not like I have a mouthful of sand, but it's a nice feel.

Rice has an inconsistent texture. Some pieces have a good 'ricey' feel, others have a strange 44 1 1 crunch, like slightly undercooked pasta. 45 1 This product did have a less favorable texture to it. Almost like a powdery finish.

46 good

the the that accidentally were not cooked, it leaves a leaves it not cooked, were accidentally that this one, and I didn't like that. It almost feels like feels almost It that. didn't like and I one, this taste to it at all, but it is a bit soft and almost textured like like textured almost and bit soft a is but it all, at it taste to Comments about the mouthfeel of 10LR of 10LR about the mouthfeel Comments i think of pieces of rice grains grains rice of pieces think of i rough feeling in your mouth. in your feeling rough does not have a grainy a grainy have not does rice, not brown rice rice brown notrice, definitely feel the graininess in in the graininess feel definitely sand in your mouth. mouth. in your sand a little too grainy but would still enjoy it. eating enjoy still but would grainy too a little could I getting needs texture texture needs grainy. too It was Even after rinsing my mouth, there are some particulates lingering on my teeth and and teeth my on lingering particulates some are there mouth, my rinsing after Even present. is mineral other some or calcium eating like I'm Feels mouth. my of sides A bit similar to brown rice. rice. to brown bit A similar I liked the crunch it had. it liked the crunch I a bit grainy , grainy a bit and crunchy grainy very texture sample. in the texture a little are There flour. takes like It noodles cooked dente al below like feels also grit in there, some Just a hint of grainy/crunch. grainy/crunch. of hint aJust smooth enjoyable. feel the that made texture of hint grainy a slight was There the rice regular grainy. felt rice The it had some parts that were crunchy were that parts hadit some inthe chew. grittyness some had still but mushy, too not was This 1 1 6 1 1 9.38% 9.38% 1 1 1 1 5 5 1 1 1 1 1 1 6 1 1 1 1 9 9 1 1 1 1 1 1 1 1 8 8 Crunchy Crunchy Dislike Liked Rice Soft

1 1 42 gritty/ gritty/ Grainy/ Grainy/ crunchy crunchy

1 1 1 1 1 1 1 1 1 1 1 1 1 % 65.63% 12.50% 14.06% 9.38% 7.81% 47 48 49 50 51 53 52 54 55 56 57 58 59 60 61 62 63 64 sum sum

157

Table D.17 - Comments on the mouthfeel of 15LR and their trends Grainy/ gritty Sandy Dislike Like Rice Soft Comments about the mouthfeel of the 15LR

crunchy 1 1 Grainy, gritty

2 1 Uncomfortable feeling when chewing.

3 1 grainy AND some crunches in there. oof

4 1 a good texture, not too grainy and not too gooey

5 1 1 A bit too grainy for me, but maybe it's because I like softer rice.

6 Kinda squishy

There is a high level of crunchiness and graininess that made swallowing the rice an 7 1 arduous task (in the sense that it was not something I genuinely wanted to do). 8 I felt the product dissolved rather quickly, didnt get much time to chew.

9 1 the texture was about right.

I did not enjoy the mouthfeel, I felt that it was too grainy and there were small pieces of the 10 1 1 rice left in my mouth which was not to my liking and not pleasant. 158 11 falls apart in mouth, no chewing necessary. very smooth

12 1 1 taste like there is some a few tiny rocks mixed in the product

13 1 1 feels like sand mixed with rice

14 1 It felt grainy as soon as I put it in my mouth and it left far too much residue in my teeth.

15 1 1 it has nice texture that is more on the grainy side than smooth but still pleasant

16 1 Really grainy and somewhat hard

17 1 the texture is uneasy doesnt remind me of rice

18 1 Mouthfeel was grainy

19 not too smooth

20 1 Has a rather coarse texture throughout. Inconsistent texture.

21 1 It was a little grainy but not too much.

22 1 there a bits of crunch

Very grainy, almost felt like there was sand in it. Grains were noticeable while chewing 23 1 1 and after swallowing the grains remained in my mouth and throat. 24 It wasn't too grainy but still have the urge to drink afterwards

It is too grainy for cooked rice. It is also quite mushy but that does depend on the ratio of 25 1 water and rice. 26 1 it was a bit crunchy

27 1 1 i did not like this one at all it was too grainy

28 1 some of the grainy pieces would be lodged in between my teeth for days.

Grainy/ gritty Sandy Dislike Like Rice Soft Comments about the mouthfeel of the 15LR

crunchy The mouthfeel of this product reminds me of insects. I think I heard a crunch when I bit 29 1 into it. 30 1 1 1 I can feel small sand-like particles on the grains of rice. I don't really enjoy this.

31 1 The grainy quality was difficult to chew, and got stuck in my molars.

32 1 i am not as use to having a grainy rice unless i know I did not cook it properly

33 1 1 once dissolved within the mouth, it has a fine, sandy texture

34 1 Grainy is a good word to describe this.

35 1 it was grainy, but grossly smooth in places too. bad contrast

36 1 1 feels like having sand in your food.

37 1 1 there are larger chunks of particles in the product - I'm not sure if I find this pleasant.

38 1 1 Texture is actually pretty appealing, with a few pieces being too crunchy.

39 A bit too chewy

40 Very smooth for a brown rice.

159 41 okay

42 1 extremely grainy and not firm enough

43 Doesn't feel grainy. I'd say it feels just right

44 needs to be more chewy

45 1 This product was very grainy.

46 didn't really feel like actual rice.

47 1 1 I liked the crunchy sensation.

If i were eating actual brown rice i would be okay with crunchy bits but knowing that its 48 1 1 based from insects it grosses me out! Lingering, gritty particles on my teeth, although, not as long-lasting as Sample #825. I would not think it to be rice at all. For objective tasting, I tried it again and it had a very 'hard' feel to it, as if I added soil or rocks to it. However, it lasts longer than #693, making 49 1 1 it a pretty unpleasant mouthfeel. The 'calcium' feel, feels like the 'calcium' was ground up more fine, with an additional, 'magnesium'in a different rate of grinding. Almost like having sand and salt to eat with my burnt rice. 50 1 1 remind me of pieces of sand in my mouth , the texture was bit rough and grainy

51 1 texture of white rice

52 There is more texture into this sample.

53 felt really powdery in my mouth at the end

pretty dry feeling, gritty, makes me think these were rice grains that fell off the crop onto 54 1 the ground, and then picked up and cooked

it it soft

how how which which This This under under of of higher higher way way rice. rice. has has the the it in brown brown somewhat somewhat balls because because balls plain plain served served knowing knowing

looked looked :( into into But

when when and rice. rice. diet. was eating was right, I still like this product this product like still I right, other qualities especially color. especially qualities other I I if

as as

was lighter lighter was into my my into or white white or felt

I I lacks desirability desirability lacks 10CR

product product somewhere

bland.

simply simply This This it it taste appealing then i can look past the past can look i then this sample, this sample, and aroma as just about just about as and aroma but but foods or or foods was quite was great great this sample. very very Comments about the mouthfeel of the 15LR of the 15LR about the mouthfeel Comments It It liked liked a products. products. comments about comments

whole whole product product a finish finish

have have mouthfeel, mouthfeel, ther ther other other tastes good good tastes really really at it was not was much.

I I O to to talking. talking. the incorporate small amounts of it of amounts small incorporate of how much moisture and softness (sad) it had it (sad) softness and much moisture how of like mush....or things soft enough to be molded molded be to enough soft things mush....or like the far too too hate hate to to does it it does so so taste taste

about color. color. going sample sample texture texture a taste taste nor nor don't the the color color

I I the the but but strange tasteful tasteful the the I would prefer if the grains held together more and were more chewy. more and were more together held if the grains prefer would I either not smooth was it but grainy too not it was personal personal but but smooth smooth having having be be taste taste displeasing displeasing mind mind most most the the am probably am probably

after after I I just just bad bad the the and and somewhat somewhat a a attention didn't 1 1 thoughparts in some isit soft the rice. chew you when texture agrainy bit of can feel you 1 crunch. bit of a with mushy or cooked over a little seemed It 2 2 I I its its nothing nothing side. side. my my a might might

have have but

et et and taste better than than better andtaste paired with veggies. with paired g smell smell that that aftertaste aftertaste aftertaste aftertaste this product product this choose this product base on taste/texture/mouthfeel over brown brown over on taste/texture/mouthfeel base product this choose

is sustainable may incline me to me incline may issustainable this as as this bland bland did not rate the other qualities, color, color, qualities, other the not rate did the

it. great great but but smell smell doesn't doesn't

odd odd really really buy buy of of eat eat aftertaste. it it for for it, it, good, good, an an 1 1

didn't didn't left left would would could could enjoyed liked mean mean i pretty i although if it flavor. the with agree i because moderately It I an Has I I just don't I because again this eat wouldn't I because it seemed overcooked how and isit it I I acceptable, Overall served. been has comparing dislike was only My watery. and more cooked, n/a not would I and protein except kind its the like I be would - Other comments about 10CR and their trends 18

5 5 1 1 utrition Table D. N

8 8 food

1 1 other ith ith W 8 8 Sandy Sandy Dislike Like Rice Soft

1 1 1 olor C

40 gritty gritty Grainy/ Grainy/ crunchy crunchy

1 1 1 1 6 7 8 9 1 2 3 4 5 16 17 18 10 11 12 13 14 15 % 68.97% 13.79% 13.79% 8.62% 1.72% 3.45% 55 56 57 58 sum sum

160

Color With other food Nutrition Other comments about 10CR

19 Flavor of this one if somewhat more bitter than the previous one (code 693)

20 I seemed like my regular brown rice meal

21 1 Other than the odd coloration, the flavor was spot on and the aroma, or lack thereof, was pleasant.

22 The overall feeling of this sample was good.

23 Overall it was satisfactory, but did not have quite the right texture, therefore I slightly disliked it.

24 1 It wouldn't be so bad if combined with other foods

The color is not a problem, it simply feels as if it belongs in Spanish cuisine. The taste and aroma is very good with no 25 strange after taste. 26 very bland overall, very little insect product in it

27 it was a good product very mild and not as earthy

28 tastes pretty normal like other types of rice.

29 1 May pair well with other foods but it is only moderately good alone.

I would purchase this product if it was nutritionally sound and affordable. I would also flavor this product to mask the 30 1 mildly unpleasant earthy flavor in a realistic kitchen setting. The mouthfeel skewed my overall liking of this product.

161 31 The taste was stronger than the aroma, I enjoyed that about this batch

32 A little too bland in my opinion.

33 1 its overall look (color/texture) were very off-putting as I tried to select a small bite

34 none

The product was interesting - I can't say I hated it, but it wasn't my favorite. This product may be popular with people who 35 aren't fond of brown rice. 36 1 I would like it better with soy sauce, or something else to provide more flavor, but this is an acceptable product.

37 good

I could definitely eat this for a meal, maybe with some kind of sauce or seasoning. I don't like the color too much, but I 38 1 1 really like the taste. 39 not sure I would re purchase this product if bought in store

40 1 This product is something I would eat with another food.

41 I would eat it if it was the only thing around. I didn't feel any aversion, but I wouldnt buy it.

42 Loved everything especially the flavor and texture.

43 1 Needs some herbs, salt, pepper, spices, or something! Texture was good. Smeel was unappealing - make it less apparent.

This is a good product! I did get different reactions when eating this rice, but overall, I would buy this if I were on a 44 budget. 45 the product was just ok

46 taste like bread

47 This sample taste as close as white rice, but the texture is not the same as rice.

48 leaves a little muddy aftertaste that lingers

o d

sauce, sauce, hot hot woul i

it.

mouthfeel. eat eat spices and spices and not it's something something it's satisfying. less appetizing. less texture texture loads of of loads would would I I very very the the smell, smell, look look with with

with with up up

choice choice

fishy fishy it a was not not was it. a first first

to to products, products, and at at

grainy. serve serve given given feel feel if earthy/grainy taste taste brown brown 15CR soft soft sweetness added to it. On it's own, it's a little to a it's little it's own, to it. On added sweetness could could mouthfeel and the taste falling more on the neutral neutral on the more taste falling andthe mouthfeel but necessarily necessarily slightly slightly I I

surprised after after

more more not not was excellent. was very very and was more more rice gets very very gets the the it it the the okay. weird weird taste, taste, to to as as tofu tofu a taste taste then then texture texture a are are and and

protein protein the left left but fishy fishy

feel feel It It comments about comments was pleasantly pleasantly was - want. with with I I No No eating disliked, disliked,

compared compared I I me. me. weird

aftertaste aftertaste tolerate tolerate me. me. aroma and and aroma say. say. to to mouth Other Other of of were were Other comments about 10CR about 10CR comments Other buds to to grey grey promising, promising, thing the can and

weird weird you you The The taste if offered offered aftertaste. is kind kind is only only deterred deterred the look look if if as as my my was able able was overall, overall, I I starts off off starts that color. color. than than for for

It It which which but again again was the was far far almost almost lingering lingering color other other a aspect aspect so so one one salty salty was the the was tastings, tastings, flavor, flavor, product. the the taste, taste, bit bit just one one this this best best a for for this this of of was lacking. lacking. was initial initial eat eat more more this sample sample this the little little disliked a a to to complaints complaints I I is horrendous.texture horrendous.texture is be be overall, overall, was the the was few few is is except except eating much much no no some some

flavor flavor 4 4 want want first first thing

might might taste taste use use

it the the have have it

product product the myself myself aftertaste aftertaste

of of graininess of of graininess overall overall only aftertaste first first buy buy was good good was was okay. was could could wouldnt wouldnt

love love see see That After I the Good It The The The all I is the first than better mean....it was I one. the first to eat over choose definitely It loaded was this rice that apparent itbecame Unless to ever enough appealing be to rice the do no find I The it The I doesnt none the of because recipes breakfast used with being this sample can see I some with cereal hot a as prepared great be this would like feel side. I bland. this one I'd best I The after taste is quite strong quite strong is taste after The it. i liked bread. or oatmeal like kind of to me, good tasted this just tasted.it that way the and mouth in my felt the way like I chew. a has good sample This - Other comments about 15CR and their trends 19

1 1 1 1 1 3 3 ftertaste Table D. A

8 8 1 1 egative N

6 6 1 1 1 1 1 ositive Color Color food other With Nutrition P

9 3 4 5 6 7 8 1 2 49 50 51 19 20 21 22 13 14 15 16 17 18 10 11 12 sum sum

162

Positive Negative Aftertaste Other comments about 15CR

23 1 I wouldn't eat this if I had a choice

24 A nice product over all. It does not seem out of the ordinary.

25 taste wasn't too strong to dislike

26 it was mild therefor it was okay but i didnt like the idea of insects in it, i might have it again

27 the grainy texture to the sample is the main reason for dislike extremely

28 No crunch in this product.

29 1 I would purchase this product, if it was cost effective and nutritionally sound.

30 as it was moist when i tippet the cup it look like their was a live insect inside as the product slid down the container.

31 The mouthfeel won me over the other two products. It's more between like moderately and like extremely.

32 1 I could see myself eating a lot of this one.

33 none

34 The product definitely isn't bland - it tastes like it was seasoned already.

35 Flavor killed it for me.

163 36 1 Except for the color, this is a nice product. I could eat this.

37 na

I like the color, odor, and mouthfeel of this product. However, the taste is lacking; there isn't much flavor to it. My 38 favorite was 249 because it tasted kind of like a tortilla chip. I thought this would taste similar because it smelled similar,

but the taste didn't really measure up. 39 1 not offensive but not great, would not repurchase if in store

40 1 I would not eat this product.

Out of all the products, it was the most similar to rice. This one was my favorite, but still had a small undesirable aftertaste 41 1 that could probably be eliminated with addition of flavors/spices etc. 42 1 Would not care to eat again.

It needs some herbs, salt, pepper, spice, or something! Also, it reminds me of rice but the texture needs to be more solid 43 like rice. Also, the smell needs to be less apparent. I like the mouthfeel and flavor of this rice, but the appearance is a little off-putting, yet, I think it was extruded very well 44 in resembling rice. However, when I chew it, it is a little bit more gummy. This is probably not too big of an issue though. 45 the product was not the best one but it just makes me think of what dirt would taste like.

46 taste was alright

47 1 It actually was good

48 The texture and smell is worse than the taste.

I think this one compared to the others is just right like goldilocks and the three bears story in the essence of 'not too hot' 49 and 'not too cold' but except with aroma, flavor, and mouthfeel. The color could use some work though - it's a little too

light the saying taste like chicken can hold true with this one. the overall feeling in my mouth was okay but the taste, it tasted 50 1 like cold chicken and i was not quite the fan.

yes! yes! a

to.

or or had was like like was

it it

had - that that

flavor flavor horrible a face face than than unless I I unless the bitter/fishy. putting. offer offer - in in other other

was not was

not not it it again again me me but me overly overly it it was off was did did but for for feel feel it it eat eat nor nor to to mouthfeel.

flavor flavor it texture texture bland bland grainy grainy to to want want however however and and decent decent

10LR unpleasant unpleasant that that

not dislike appealing came came and feel. to to too too it it than than aroma, aroma, daily. is is this rice, rice, this was neither neither was eat anymore..... eat taste just kept punching punching kept just taste flavor flavor eat eat strongly strongly when when mouth mouth other other I I taste taste enough enough I I that it it

somewhat somewhat flavor, flavor, it after after I can I the the

to to is is good good rice rice and and and bad. consume consume the comments about comments to to like like flavor flavor 825. rice rice Not Not much much not overall overall strong strong

the the best best surely surely Other Other liked liked similar similar trip. did I I have have too too but the enjoyable, enjoyable, is is item, texture texture item, grainy Other comments about 15CR about 15CR comments Other would would very very be be had texture, texture, than than I I -

didnt didnt is is times, times, graininess, disliked disliked graininess, to to complaints towards it, towards complaints

It It little little it it camping camping at dinner dinner

after a more more this one this one not and and reason. me. on on of of looks toasty.I toasty.I looks

much. being being since since 693 693 had It It grainy grainy taste taste weird weird this sample this sample i appeared appeared to to normal normal

side side some some every every flavor flavor this sample. sample. this

rest, rest, little little recommend this product to my friends and family. The smell was just right but as I continued continued as I but just right smell was The family. and friends to my product this recommend due right. right. for for smell a sample, sample, on on the the this

product product the this as a as this be be purchase the the this product product this this as a as this somewhat somewhat to to first first isjust the the of of eat eat uneven uneven can can then it was more of....oh gosh I don't think think don't I of....oh gosh more was it then eat eat eat eat not back....the more I ate it, the more bitter the bitter the more ate it, the I more back....the enjoy enjoy the the the

distinguishable distinguishable opinion fan fan moderately, a color color not a my my would would would would would take it liked like it, it, like I I Compared Like Texture satisfying very It was taste. pleasant Had I I I i despite Did Seemed not available foods were other If which excited mouthfeel The Although I not wouldn't I surprised. pleasantly was I but taste distinct a have to it expected I sample. first this with am impressed I definitely would to me allowing smell slight the overpowered it of taste good The food. turtle of a whiff me it, gave it to smell the product. enjoy In boring indifferent I'm I

This had a good taste, texture and the smell was very slight. slight. very was smell and the texture taste, good a had This - Other comments about 10LR and their trends

1 1 1 1 1 1 1 1 20 egative N 6 6 Table D. Aftertaste Aftertaste

1 1 1 1 1 1 ositive P 7 7 Negative Negative

1 1 1 1 10 olor good good olor Positive Positive C but disappointed but

4 5 6 7 8 9 1 2 3 51 20 21 22 14 15 16 17 18 19 10 11 12 13 sum sum

164 Other comments about 10LR about 10LR comments Other of the product. Color was very nice but the consumption of the sample sample the of consumption the nice but very was Color the product. of or something! Texture was alright. Smell was less apparent compared to to compared apparent less was Smell alright. was Texture something! or best color of the four, but I didn't like the taste at all. It tasted too fishy and fishy too tasted all. at It didn't the taste but I of like the four, color best really hungry, but this is my least favorite of the bunch. The texture and the flavor are are flavor the and texture The the bunch. of favorite least my is this hungry, but really I were were I all. all. texture was wonderful, but the flavor wasn't as good. However, the blandness is closer to brown to brown closer is blandness the However, good. as wasn't the flavor but wonderful, was texture this one. It was the was It one. this like fish pellets. But I like how closely it resembles brown rice. I predict it to be my favorite out of out of favorite my be to it predict brown I rice. it resembles closely how like But I pellets. fish like it was too grainy. too it was like like and

First impression by looking at it, thought it would be just right. Once tasting it, it was below expectations below it, was it tasting just be Once itright. would it, thought at looking by impression First bad not texture. a just taste, a of much Not smelling it. intentionally not if strong istoo not smell The too. texture the good and was flavor The wish it was not so soft not so was it wish product. eat this would I well. rather to taste it found but I aftertaste, aHas strange color. and liked the crunch I again. eat it would I salt, pepper, herbs, spice, some Needs unappealing bit still samples other a lot Looks the four. good not very it was bland. too is taste the but is good, smell The crispy. or fried slightly product seems The The color is a bit off-putting. is a off-putting. bit color The once to try okay was but distasteful too it wasn't tastedit like smelled it and it like didnt bad i pretty it was noodles. soba of the taste has of kind the product. of the grainyness didnot enjoy I the gains of some with crunchy bit more a seemed this one na didn't I earthy, It did remind me about the insect nature nature insect the about me did remind It appetite. decreased slightly have may good. pretty are bugsthese go, As none product's This rice. could if eat this I at appetizing not Has a good color color aHas good diet in my this include i would

1 1 1 1 1 11 Negative Negative

1 1 1 1 1 1 1 1 11 Positive Positive 5 5 1 1 1 1 1 Color good good Color but disappointed disappointed but

48 49 50 38 39 40 41 42 43 44 45 46 47 25 26 27 28 29 30 31 35 32 33 34 23 24 36 37 sum sum

165

Table D.21 - Other comments about 15LR and their trends

Positive Negative Grainy Earthy Other comments about 15LR

1 Tolerable but I could not imagine myself eating this product after a few bites

2 This one felt like if fish flakes were sprinkled into my wet rice crispies.

3 1 Nope, would not eat again period.

4 despite the smell, the smell and texture is not offputting, and somewhat welcoming, if it had more flavor

5 1 Was not awful, I would never purchase it though.

6 Too strong a taste and smell.

I do not mind the notion of eating insects with my rice, however if I were to partake in such a practice I would 7 like the impact on the flavor and texture of the rice to be much more subtle than it is in this product. 8 The only thing i disliked was the bitter after taste.

9 n/a

166 10 disliked 965 the most compared to 693 and 825

11 different from most foods that I have tried.

12 1 1 too grainy and earthy

I cannot see myself eating this in the future. So far, it is my least favorite. This is also the only sample I did 13 1 not want to finish. 14 1 it's horrible, what insect is this?

15 1 1 earthy and grainy

This one was the worst one for me the smell and the taste were really off putting and if it was offered to me 16 1 again i would definitely not eat it again I don't think i would pay a premium to try but if it had a higher protein to carb content which im assuming so i 17 would pay an equal price point as a standard bag of rice but the flavor is offsetting same with texture so i

would need a sauce to cover it. 18 It has a slight after taste.

19 too bitter

20 1 Like it slightly

Weird and inconsistent texture combined with off-setting taste and smell does not make this an appetizing 21 product. 22 1 1 The graininess and aftertaste of the sample made me dislike it.

It was much too grainy, as I felt like I had grains stuck in my throat and mouth even after washing down with 23 1 1 water. Would not eat this again. 24 1 Wouldn't eat if I had a choice

25 None.

Positive Negative Grainy Earthy Other comments about 15LR 26 I would like it a lot more if the smell wasn't so bad and if the color wasn't so dark. 27 1 would not try this sample again 28 1 1 too grainy and pungent and earthy did not like 29 1 1 1 i dont like grainy rice. i dont like earthy rice. no sam i am, i dont like this rice. I did not unlike this product and would consider purchasing it based on nutritional requirements, nutritional 30 value, and cost effectiveness. 31 Not really a fan of the mouthfeel. 32 Bad texture, but at least it doesn't have an aftertaste. 33 1 yuck, wouldn't eat that even with soy sauce 34 no other comments just wish i knew what i was eating. 35 The product was okay, but the flavor and mouthfeel was a little bit off In conjunction with other foods (vegetables, etc) I could eat a full serving of this, but I don't know that I would 36 eat very much on its own. 37 na

167 38 1 1 it was both too grainy and earthy. needs to be balanced 39 I would like it more if there was some kind of flavor to it. Other than that, it was fine. 40 it was ok. would make a good cracker. 41 1 I would not eat this product. 42 1 It would be nice to eliminate the earthy aftertaste. 43 I only liked the crunchy feel, not so much the color or taste Most discouraging sample. Needs some spice, herbs, salt, pepper or something! The smell is unappealing. The 44 1 texture is mentaly unsurpassable. 45 1 I hate the product. Please, no more :( 46 the smell made me dislike the product 47 1 disliked this one 48 1 I like it more than other sample, but I would not purchase it. 49 I liked the first product the most. 50 the taste and smell alone makes this rice unappealing to me 51 it is a little bitter/tart leaves a numbness/tingle to mu tongue. I think the smell was too much influence on the taste for me. I would have preferred it a bit more if a little 52 more firm. sum 1 16 7 6

Table D.22 - Any other comments provided by the sensory panel and their trends Enjoyed/ All other comments With food support 1 Support sustainability of eating insects for protein. Hard to overcome perception of eating insects.

2 I would love to know the protein levels in these insects.

3 It was pretty fun :) thanks for the try guys! (get it, try guys. if not search them up on youtube) :-) 1

4 Hello,

5 I have a strong sense of smell and taste and can be quite picky.

6 Probably would be better to eat with something, other than it was tasty and not much different compared to brown rice. 1

Overall I prefer the darker colored rice which had a purple/black tinge even though its appearance did not appeal to me. I thought I was going to like the brown 7 colored rice more because it resembled regular brown rice more. However, its fishy aroma and grainy flavor did made me feel like I was actually eating bugs.

The darker colored rice did not have a fishy aroma and tasted like regular brown rice. I would consume the dark rice again. 8 INSECTS YAY! XD 1

9 I really loved the flavor of sample 249 and then 825. Sample 965 had bitter notes.

10 Hi

I would have never imagined to overtly try insects, but was pleasantly surprised. on the uniqueness. I think color, odor, and texture to be the most important. I 168 11 1 definitely see a combination of the products to be the ideal.

12 This was a lot of fun :) :) :) :) 1

13 Thank you for the experience! 1

i had a lot of fun participating. 14 1 thank you I think that in the questionaire to include an 'other' in the gender category. Also, for the ethnicity section, it should be titled 'Race' instead. For education level it 15 was difficultn to accurately answer because I spent 3 years in community college and am in my second year at CPP. So technically I am a 5th year in college. 16 This was a very interesting survey. Considering this is the first time I ever tried bugs 1

17 I enjoyed this taste test and would be interested in purchasing the first and last sample. 1

18 this was an interesting experience maybe add some sauce to it so it can taste better 1

i feel that 'hiding' the insects by blending them into flour is a great idea. usually the idea of eating insects in uncomfortable for people because they see the whole 19 1 insect. this seems to be a great way to increase people's confidence in consuming edible insects and trying more familiar forms. This was a interesting survey and would be fun to participate in another just like this one. As for the rice none really blew me away as being delicious but some 20 1 had an interesting taste while one was just horrible. 21 Cool product cant wait to see it on the shelves. 1

22 I am excited about this concept of incorporating edible insects into diets as a way to have better nutrition. 1

The last sample, code 825, had the best flavor and look overall. However I do not think that is necessarily the issue with the product. I feel the texture of the 'rice' is too soft and could benefit from minimal cooking. Cooking the rice to an al dente style would definitely help with the overall taste of the product. There is a 23 1 definitely aftertaste from the products that may be an issue later for consumers. I think as long as it is well masked with other seasonings/foods it should be fine. Thanks! 24 let me know any time that you have edible insect for food 1

25 I have no clue which one is which, overall felt like all the products could be served at some capacity, maybe with some refinement here and there.

Enjoyed/ All other comments With food support The concept of having it look like rice was phenomenal due to most people being able to relate and ave a comparison to judge it on. Couple of them had a good 26 1 feel and taste but some of them had a bad after taste. 27 Not as crunchy as I thought it would be.

28 Great experience. Thanks! 1

29 No comments. Good luck and thank you for the invitation of trying out the samples.

30 Good luck Jaynie :)

31 Great idea. 1

I like that there are studies like this, I'm very fond of nature and the environment and believe that insects could serve as a protein substitute now and if there is a 32 1 food shortage; or be used in countries where there already are food shortages. 33 Overall the taste testing was well and all samples were mimiced rice well

34 Really love this idea it would be a great way to introduce it into peoples diet without people worrying about the gross factor that steers away most people . 1

35 I am an entomologist

36 great experience and interesting taste presented to me 1

37 i liked the sample 249. i hope that has some insects in it. because i like the idea of eating insects. they are such a pest. might as well use them for something. 1

169 38 I have completed my bachelors but I am not a graduate student. I do not have a master's or PhD

39 I enjoyed the products 1

40 Nice job one of the formulations is pretty good. Best of luck with your research. 1

Thanks for letting me be a part of this study! I do hope Jaynie (sorry if I misspelled your name) receives the information necessary to feed the malnourished in 41 1 other countries or the alt protein enthusiasts like myself :) 42 I think all the choices would be great accompanied with beans, veggies and or meat. 1

43 great insect-rice products overall 1

44 If I were to rank these from best to worst, it would be 693, 825, 965, 249. Hope that helps?

45 i wish i knew what each sample consisted of

I hope that more research like this would be done! The protein content, as well as the fact that these products would be 'gluten-free', make it a priceless topic of 46 1 studies 47 I think as rice, this product paired with other food would be extremely enjoyable! 1

48 na

49 249 was my favorite all around

50 I believe in helping create a sustainable future and I believe eating insects will be part of it. 1

51 Intersting experience. Definetly would not do it again unless the taste and smell get better. I can get past the texture if it smells and tastes good. 1

52 This would have been good for DSDC if it fit the theme this year!!!

53 the smell of dog food was the most unpleasant part of the experience.

54 Good luck Janie!

55 This was really good and adventurist hope it helps you guys out! And i actually liked them it was good. 1

Enjoyed/ All other comments With food support It would be more easier for me if the questions define what I should compare the sample with. When the questions are asking (too light, too dark) I did not know 56 what the standard of light and dark, therefore, I compare it with brown rice. I do not know if there is intention in not giving standard to this survey, but somehow

it is hard for me to answer without any comparison. I would be open to incorporating insects into my diet if they were accessible in markets, had nutrition contents included, and perhaps recipes to follow. 57 1 Alternative food choices to meats are of interest to me from an environmental standpoint. 58 10/10 would definitely participate in a similar experiment in the future 1

59 this is a great study and another way to look outside the box for sources of nutrients 1

60 I am interested to know which type of insects I have sampled. It would have been interesting to know which type of insect I sampled after each questionaire.

i would eat insects, i would! 61 1 Tha Some of the feeling of these samples reminded me of the non rice pieces in things like rice-aroni. If there was a flavor enhancement like that, I dont think some 62 people would be able to tell the difference. sum 33 4 % 53.23% 6.45%

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APPENDIX E

MEDIA PUBLICATIONS OF SENSORY EVALUATION STUDY

Figure E.1 - Poly Post publication of the edible insect sensory evaluation study Poly Post is the student newspaper of Cal Poly Pomona. The above article, titled “CPP community consumes bugs for research” was written by Alicia Balderama and published on April 19th, 2015. Available at: http://www.thepolypost.com/lifestyle/cpp-community-consumes-bugs-for- research/article_08f3c4d2-04f8-11e6-838c-d7c26def05f6.html

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Figure E.2 - Page 1 of the PolyCentric publication of the sensory evaluation study PolyCentric is the official news source for Cal Poly Pomona. The above article, titled “On the Menu of the Future: Insects?”, was written by Dan Lee and posted on May 19th, 2016. Available at: http://polycentric.cpp.edu/2016/05/on-the-menu-of-the-future-insects/

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APPENDIX F

JOURNAL ARTICLE MANUSCRIPT READY FOR SUBMISSION

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POTENTIAL UTILIZATION OF EDIBLE INSECTS IN EXTRUDED RICE

PRODUCTS TO ADDRESS MALNUTRITION ISSUES IN DEVELOPING

COUNTRIES

Jaynie Tao1, Gabriel Davidov-Pardo1, Bonny Burns-Whitmore1, Eileen Cullen2, Olive Yao Li1* 1Department of Human Nutrition and Food Science, College of Agriculture, California State Polytechnic University, Pomona 2Department of Plant Sciences, College of Agriculture, California State Polytechnic University, Pomona

*Corresponding Author: Tel: (909) 869-3021; E-mail: [email protected]

ABSTRACT: Although unconventional in the United States, entomophagy, or the practice of consuming insects, can provide a nutritious relief to the many malnourished people in developing countries. Edible insects are part of numerous traditional diets found in over 113 countries, including those in Asia, Africa, and Latin America. Currently, there are two billion people consuming over 2,000 recorded edible insects. Many of these worldwide insects contain considerable amounts of protein, fat, vitamins, and minerals that are comparable to commonly eaten livestock. Accordingly, the primary objective was to investigate the feasibility of incorporating an edible insect ingredient in an extruded brown rice product. Four formulations were developed, including cricket or locust flours at either 10% or 15% addition levels. A second objective was to determine the physicochemical and nutritional differences between the novel insect rice products and market brown rice. Overall, the insect rice was found to be darker in color, rougher in surface morphology, and softer and more adhesive in texture than market brown rice. Nutritionally, the insect rice offered an excellent source of protein and 10-12% of the daily value of fat, which are both at greater quantities than typical brown rice. The products were also suggested to contain considerable amounts of dietary fiber and iron, although these quantities require further investigation. Also part of the objective of this study, was the determination of consumer acceptability, which was achieved through sensory evaluations involving a 120 untrained panel. Results suggested cricket formulations at 10% and 15% levels were equal in acceptance and both were worth continuing optimization, while 15% locust formulations were the least liked of the four formulations.

Introduction

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the concept of an edible insect product would certainly be familiar and accepted. Currently, there are more than 1900 species of edible insects documented in the literature (Jongema 2014). Many of these insects worldwide contain considerable amounts of protein, fat, vitamins, and minerals, which is comparable to even commonly eaten livestock, such as beef and chicken (Ramos-Elorduy and others 1997, Womeni and others 2009, Xiaoming and others 2010, Sirimungkararat and others 2010). While entomophagy can be a means of survival for developing countries, edible insects can also be an alternative food source for developed countries, such as the United States. According to the Food and Agriculture Organization (FAO) of the United Nations (2009), as the global population is projected to reach over 9 billion in 2050, the availability of food will become a serious issue. As edible insects can serve as a nutrient- dense source of food, they offer economic and environmental advantages as well. Insects have a higher feed conversion efficiency and fecundity, thus reducing the overall food production cost. Furthermore, farming edible insects is substantially less taxing to the environment as it requires no or minimal land clearing and produces less greenhouse gases than other livestock (Rumpold and Schluter 2013, Oonincx and others 2010). However, the “yuck” or “disgust” factor remains an obstacle towards achieving successful product development. Whole insects especially, whether due to nature or nurture, can be uneasy to accept as a safe and appealing food source. Thus, the objective of this study is to investigate the feasibility of the obscure incorporation and fortification of a major food staple, such as rice, by utilizing edible insect flour as a value-adding ingredient. By removing the original form of the insect and incorporating its flour into a food matrix that is widely consumed around the world, the aim is to provide an innovative solution for food insecurity, especially in developing countries. Furthermore, the hidden form of the edible insect is designed to reduce the “yuck” factor and increase the acceptance of entomophagy. Rice flour has been specifically chosen to be the vehicle based on the consideration that the nutrients carried by the edible insect will supplement the nutritional composition of rice. The two main ingredients will be combined in different ratios to form a dough mass, which can be processed through an economically feasible, cold-forming extrusion technology. Thus, through this production method, a final rice product containing optimal addition level of insect flours will be developed. Subsequent objectives will include the measurements of its physicochemical properties, nutrient profile, and consumer acceptability via sensory evaluations.

Materials and Methods

Formulation The initial rice formulation suitable for cold-forming extrusion was derived from a patented technology, Ultra Rice® (Li 2009) which was originally developed for fortification of vitamin A and iron, as well as zinc and other B vitamins. The current study adopted its base formulation while excluding the fortifying agents of micronutrients and replacing white rice flour with brown rice flour, as well as incorporating tapioca starch to gain an extra binding effect. Organic cricket flour (Gryllodes sigillatus) was obtained as “Protein 2050-C” from Next Millennium Farms, which later changed their company name to Entomo

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Farms (Ontario, Canada). Locust flour (Locusta migratoria) was purchased from Thailand Unique (Udon Thani, Thailand). Both sources offer products that contain 100% farm raised insects. A total of four formulations were developed: 10% cricket rice (10CR), 15% cricket (15CR), 10% locust (10LR), and 15% locust (15LR) (Table 1).

Table 1 – Insect rice formulation Ingredient Percent (%) Brown rice flour 65.5 - 70.5 Insect flour 10 - 15 Tapioca starch 15 Sodium alginate 1.5 Vegetable shortening 3 water 33 (db) calcium chloride coating (20%, w/v) 10* *50 mL solution was sprayed per kg of extruded rice, leading to 10% calcium coating level

Brown rice flour and tapioca starch were both acquired from Bob’s Red Mill® (Milwaukie, OR) while the All-Vegetable Shortening was from Crisco® (Orrville, Ohio). Sodium alginate and calcium chloride are both food grade materials typically found in product development labs and were purchased from VWR Chemicals (Radnor, Pennsylvania). Market brown rice (MBR), long whole grain (Signature Kitchen, Pleasanton, CA) was purchased from a local grocery store and used as a control.

Extrusion of Insect Rice Formulations Insect rice production began with sifting of the dry ingredients including the brown rice flour, insect flour, tapioca starch, and sodium alginate. A countertop stand mixer was used to blend the dry materials until homogenous before liquefied vegetable shortening and water were slowly introduced. When all the ingredients were added, the dough was mixed for at least 5 minutes. The homogenized rice dough with edible insect flour was extruded via a La Monferrina single-screw, cold-forming, pilot-scale P12 pasta extruder and a rice-shaped die plate #305. The extrusion flow rate, as weight (g) of sample collected per unit time, and the total yield (g) per 1kg batch were recorded. A calcium chloride solution at a concentration of 20% (w/v, 20g per 100mL) was prepared and approximately 50mL of this solution was sprayed on the 1kg batch of extruded insect rice as it underwent a continuous tumbling mode within a rotating coating pan. After the surface coating, the insect rice was then placed in dehydrator for at least 6 hours at 165°F. The dry weight (g) of the insect rice was recorded. Next, the dried insect rice was screened to obtain a final product with desirable shape and size. The % yield was calculated as:

Yield (%) = weight of rice-shaped particles with desirable size total weight from extruded insect rice batch

All insect rice products were stored in labeled, re-closable, low-density polyethylene bags and placed in a cool, dark space.

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Physicochemical Analysis Assessments of color (Konica Minolta; Ramsey, NJ), water activity (AquaLab; Pullman, WA), and moisture content (Shimadzu; Kyoto, Japan) were performed in triplicate for all four insect rice samples at 0 months. To evaluate the shelf stability, water activity was then measured again after 3 months of storage for all four samples. Texture analysis of cooked insect rice was conducted using a Stable Micro Systems TA-XT Plus texture analyzer (Godalming, UK) equipped with the Texture Expert Software package. These measurements were performed five times for each formulation. Qualitative examination of the surface morphology, in terms of shape and size, was performed using a Digital Blue QX7 microscope (Atlanta, GA) at both 10 and 60 magnification for all insect rice formulations with the market brown rice used as the control.

Nutritional Analysis Total Dietary Fiber Analysis An Ankom TDF Analyzer (Macedon, NY) was employed to estimate the total dietary fiber content of the insect rice formulations. This state of the art equipment automates the AOAC 991.43 procedures for following the enzymatic-gravimetric method in total dietary fiber (TDF) analysis of grain and cereal, fruit and vegetables, and processed food products. Procedures for the assessment followed the operation protocol provided along with the equipment. Values obtained from the analysis were the % recovered residue, which included a portion of other constituents, such as undigested protein and ash, along with the TDF content. This recovered residue should undergo further analyses to determine the actual levels of ash and undigested protein, which would then be used to correct the initial values of weight difference for the actual TDF content. However, due to the limited capacity in the laboratory for ash and protein analyses, this recovered residue value was used as a rough estimation of the content of total dietary fiber.

Iron Analysis A graphite furnace atomic absorption spectrometer (AAS) was utilized to analyze the iron content of the insect rice formulations. First, the samples were digested via concentrated hydrochloric acid and nitric acid at 300°C. During this digestion, it was observed that the assay solutions for some samples contained white-colored precipitates, which may be related to undigested fat in the samples. Thus, a gravimetrical filtration was carried out prior to iron analysis using AAS. Once digestion and filtration were completed, the samples were analyzed for their iron content using a Perkin Elmer AAnalyst 400 Atomic Absorption Spectrometer (Waltham, MA). A calibration curve was created using standard iron reference solutions in a concentration series of 1 to 5 ppm, while 1 N hydrochloric acid was used as the blank.

Protein and Fat Content Percent protein and fat analyses of the insect rice formulations were outsourced to Certified Laboratories in Anaheim, CA, which adopted standard AOAC

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methods for protein and fat analyses. The vendor of the cricket flour, Next Millennium Farms (Entomo Farms) was able to supply a complete certificate of analysis (COA) while Thailand Unique, the supplier of the locust ingredient, was only able to provide its COA for protein.

Optimal Cooking Standardization In order to achieve consistent cooking properties of the insect rice products for the sensory evaluation study, the cooking preparation process was standardized. While market brown rice is of the actual rice grains, the novel insect rice product is a reconstitution of floured ingredients, thus lacking the outer bran layer which provides a physical barrier for the inner starch. Instead, the crosslinking between the sodium alginate and calcium chloride coating offers a surface shield, or shell, to maintain the shape integrity of the insect rice throughout the cooking process. While effective, the insect rice still required steaming in place of boiling. A rice-to-water ratio of 1:1.5 on a weight to weight basis was use to steam- cook the rice. It was necessary to steam the insect rice immediately after combining with water to prevent any initial breakdown of the products. After the insect rice was steamed for 15 minutes, it was then removed from heat.

Sensory Evaluation Participant Recruitment To determine the acceptability of the insect rice product, a panel of 120 untrained participants was recruited from the California State Polytechnic University, Pomona. Panelists were required to be: without shellfish allergies, consumers of brown rice, and between the ages of 18 – 65 years old. It was heavily stressed that those with shellfish allergies were unable to partake in the study as an allergy to shellfish was likely to produce a similar reaction to land insects. Another prerequisite for participants was that they were willing to consume edible insects. Thus, subjects knew that the products to be tasted contained such an ingredient. While the “yuck” factor remains a major issue in entomophagy, those who unknowingly consumed edible insect products could become seriously distraught after learning that this is what they’ve consumed. Hence, it was in the best interest of the subjects that they were informed of the products containing edible insects. Qualified and interested individuals were asked to RSVP on a Google form found on the recruitment flier and emails. The purpose of these RSVPs was to again exclude those who do not qualify. Nonetheless, walk-ins were still welcomed.

Questionnaire Designing the questionnaire was completed through the sensory evaluation computer software, RedJade, developed by Tragon Corporation (Redwood Shores, CA). It incorporated the Informed Consent Form, the evaluation questionnaires, and two other surveys. Only when the subject accepted the ICF were they brought the insect rice samples. Evaluation of the edible insect rice samples involved the four finalized formulations: 10CR, 15CR, 10LR, and 15LR. Organoleptic attributes and the overall liking were asked for each product. The attributes of concern comprised of appearance,

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aroma, taste, and mouthfeel, primarily because the introduction of the raw insect material into the rice formulation led to some apparent alteration to each of these characteristics when compared to MBR. To assess the degree of these changes, each of the mentioned characteristics were rated on a Just About Right (JAR) Scale. The overall liking, on the other hand, was scored on a 7-point hedonic scale. In tasting the insect rice, the panelists were asked to try the samples from left to right, starting with the left most sample. Between samples, the participants were provided a chance to rinse with water. When all four samples have been assessed, a survey of the panelists’ perception of entomophagy was presented. Questions in this survey pertained to their familiarity and experience with, as well as their willingness to consume edible insects. It also inquired about their perception of edible insects in the diet (e.g. appetizer, main entrée, etc.) and their willingness to purchase edible insect products in the market. Lastly, the questionnaire ended with a demographic survey, which inquired about their age, gender, ethnicity, and current education level.

Location and Sample Set Up A total of 12 booths were set up, each housing two laptops, activated with the RedJade project, and were isolated by a 3-walled booth. Two research assistants were assigned for this room to help monitor the evaluations. Near the exit was a bowl of chocolate with a sign that thanked the panelists and invited them to take a chocolate upon leaving. The evaluation took place in the sensory laboratory of the Human Nutrition and Food Science Department. Samples were prepared in a neighboring classroom and kitchen adjoined by a small storage room. This preparation room was designated for cooking, preparing, and arranging of the samples. The samples were prepared based on the method described above, with a 1:1.5 ratio of rice-to-water. These samples were presented in a 2 oz polystyrene cup, along with a small cup of drinking water for rinsing, a fork, and napkin on a food tray. The amount of insect rice was just enough to cover the bottom of the cup.

Research Compliance Approval to conduct research with human subjects was obtained from the Institutional Review Board of Cal Poly Pomona, IRB protocol # IRB-16-45. A total of 7 research assistants were involved in the sensory evaluation study. Each assistant obtained training from the Collaborative Institutional Training Initiative Program (Miami, FL) and were qualified to work with human subjects.

Statistical Analysis For the water activity, color, and texture data, an analysis of variance or ANOVA with a post-hoc t-test was conducted on the average values to determine significance. Statistical analysis for the sensory data was done automatically through the RedJade computer program. This included a chi-square test for the inquiries utilizing JAR scales and an ANOVA with a Tukey post-hoc test for the hedonic scale questions.

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Results and Discussion

Formulation Optimization Extrusion of insect rice formulation was carried out using the single-screw, cold-forming extruder. Table 2 displays the extrusion flow rates and yields for all insect rice formulations.

Table 2 – Rates and percent yields from insect rice extrusion Mass flow rate (g/sec) Extrusion total yield (%, w/w) Desirable shape yield (%, w/w)

10CR 4.69 84.25 56.00 15CR 3.43 76.22 58.40 10LR 1.11 74.42 61.04 15LR 1.99 76.87 61.37

Between the cricket and locust formulations, the cricket rice products presented with greater extrusion flow rates of 3.43 and 4.69 g/sec than those of locust formulations. The difference in extrusion flow rates lies in the nutritional profile of these two edible insects. For instance, the addition of cricket flour, with its slightly higher fat content (Table 10), may have caused some enhancement to the functionality of the lubricating oil. On the other hand, the suggested greater quantities of fiber in locust rice products (Table 7), as discussed later, may have instigated some negative effect to the extrusion process. In combination, these qualities consequently resulted in a faster mass flow rate for cricket rice formulations when compared to locust rice products. When contrasting the % extrusion yield – the weight of samples collected after extrusion versus the initial weight of dough mixture, it is suggested that an improved total yield was also correlated with an increase in extrusion flow rate. Overall, it is suggested the incorporation of cricket flour, regardless of the addition levels, did not cause too much difficulty to the extrusion process. The incorporation of locust flour, on the other hand, caused some unfavorable effect not only on the extrusion process, but also on the quality of the extruded rice products. All four formulations achieved a similar yield of final rice particles with desirable shapes and size, all within a range of 56% to 61% (w/w). Even with the employment of laboratory extrusion equipment, the extrusion of rice flour formulations containing either cricket or locust flour up to a 15% addition level was feasible. The visual representations of those products can be found in Table 3.

Physicochemical Properties of Uncooked Insect Rice Color and Shape A colorimeter was employed to objectively quantify the color of the insect rice formulations while using market brown rice as the control. The L*a*b* values can be found in Figure 1 and can be referenced to the images of these rice products as shown in Table 3.

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Table 3 – Insect rice formulations 10CR 15CR 10LR 15LR MBR

70 d a 60 a b c 50

20 d b b 10 a a a a b c d 0 10CR 15CR 10LR 15LR MBR L* values 57.30 56.95 55.84 53.37 64.47 a* values 4.89 4.85 5.69 5.49 4.04 b* values 7.98 8.54 11.31 10.80 15.25 Figure 1 - L*a*b* values for insect rice products Color of the insect and market rice products are displayed as L*a*b* values where L* = lightness to darkness on a scale of 100 to 0, a* = red-to-green values, and b* = yellow-to-blue values. Different lower case letters distinguish a significant difference for that color parameter and the error bars represent the standard deviation.

Between cricket and locust rice formulations, there was a notable difference in color shades, in terms of both a* and b* values. Cricket rice products, at both 10% and 15% additions, were generally greyer in color as their a* and b* values were significantly (p < 0.05) lower than those of locust formulations. These differences could be attributed to the color of the starting insect ingredients since the cricket flour was greyer than the locust flour. In comparing the two addition levels, 10% versus 15%, statistical analyses showed different directions. For example, for the two cricket formulations, there were no significant differences (p > 0.05) in all color attributes between the two addition levels. On the other hand, locust rice products became noticeably darker with significantly lower L* value (p < 0.05) while lessening in red shade (significantly smaller a* value) when increasing the addition from 10% to 15% in the rice formulation. Overall, the quantity of locust flour in the rice formulation seems to present a stronger influence than cricket flour on the color of the final product. Contrasting the insect rice products with the control, all formulations

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exhibited darker shades (lower L* values) and were redder (greater a* values) and less yellow (smaller b* values) (p < 0.05), regardless of insect source.

To examine the shape, size, and surface morphology of the extruded insect rice, a digital microscope was employed at 60x magnification. Differences between MBR and extruded rice products can be seen in Table 4.

Table 4 – 60x magnification view of insect rice formulations and control 10CR 15CR 10LR 15LR MBR

Presented here are the cut edges of the extruded rice at 60x magnification, where the surfaces can be seen as flakey, uneven, and rough. The cracked spots on the surface indicates that the incorporation of relatively high levels (10% and 15%) of insect flours into the rice flour matrix actually reduced the starch binding, impacting on the consistency of the product. MBR shows an entirely smooth and polished layer. While these variations may be an obvious outcome it is important to note that the fine layer of bran with the brown rice kernel serves as a protective structure for the starch inside. Extruded rice products were homogenously composed and instead relied on chemical structures (i.e. alginate-calcium crosslinking at the surface) to maintain structural integrity.

Texture In determining the texture of the insect rice products while utilizing market brown rice as the reference, a Stable Micro Systems TA-XT plus texture analyzer was employed. In particular, the hardness and adhesiveness were of concern. The texture values for all rice products are summarized in Table 5.

Table 5 – Texture of rice products Rice product Hardness (kg) Adhesiveness (kg) 10CR 0.675 ± 0.090a -0.009 ± 0.001a 15CR 0.621 ± 0.040a -0.009 ± 0.001a 10LR 0.669 ± 0.066a -0.007 ± 0.000a 15LR 0.662 ± 0.116a -0.008 ± 0.001a MBR 1.048 ± 0.033b 0.000 ± 0.000b Different lower case letters distinguish a significant difference in texture

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its lack of a complete bran structure that, conversely, is present and intact in regular brown rice kernels. Bran contains >20% of insoluble fiber of which is made up of equal fractions of cellulose and hemicellulose (Qi and others 2015). These components make this outer layer a sturdy exterior, protecting the inner starch from being leached during high stress conditions such as heat. For the insect rice, however, the product is instead held together by the physical compression during extrusion and chemical bonding at the surface through a loose alginate-calcium crosslinking network. Although these chemical bonds cannot be observed by the human eye, they are certainly productive in maintaining the shape integrity of the product through cooking. There were no differences between 10% and 15% formulations or between cricket and locust rice products. Of the exception that was aforementioned, 10LR had demonstrated to be statistically less sticky than the other three formulations (p < 0.05). Even so, the actual difference in adhesiveness was a mere 0.001-0.002 kg which is negligible.

Water Activity and Shelf Life A major indicator for a food’s shelf life is in its water activity (aw) as this pertains to the amount of free water available for microbial growth. Foods of higher water activity values, especially when above 0.6, often lead to faster spoilage and shorter shelf life due to the survival and proliferation of molds, yeasts, and bacteria.

Table 6 – Changes in water activity (aw) after 3 months for insect rice products Rice product 0 Months 3 Months 10CR 0.160 ± 0.001a 0.219 ± 0.002a 15CR 0.160 ± 0.000a 0.236 ± 0.002b 10LR 0.171 ± 0.002b 0.369 ± 0.009c 15LR 0.180 ± 0.001c 0.310 ± 0.002d Different lower case letters distinguish a significant difference within that month of time

All insect formulations showed water activities lower than the critical point of 0.6 (Table 6). Moreover, the insect rice formulations all exhibited aw levels that were significantly lower than that of the market brown rice (p < 0.05), thus indicating a longer shelf life. Contrasting between formulations shows a significantly different level of water activity (p < 0.05), with the exception of 10CR and 15CR which had similar values (p > 0.05). First, cricket formulations were established as shelf-stable products with a lower level of water activity, yet the addition level of cricket flour does not seem to present as a factor. On the other hand, while both locust formulations were shown with significantly greater aw values, the higher incorporation level of locust flour further increased the amount of free, available water. In investigating the moisture content of the two types of insect flours, locust flour was found to have more moisture than cricket flour. This suggests the locust flour itself likely to be a main cause of higher water activity values in locust rice formulations compared to their cricket counterparts. Nonetheless, 10LR and 15LR had water activity values only greater by 0.011 and 0.020, respectively, compared to their cricket equivalents and the actual impact to shelf life is considered to be minimal. Furthermore, the water activity values for all insect rice formulations demonstrated to be well below the critical 0.6 limit, to the credit of

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proper dehydration after the extrusion process. Thus, the insect rice products demonstrate to be capable of averting microbial growth and spoilage. At month 3, each insect formulation presented with an increase in water activity. Between the cricket and the locust products, the latter displays a greater amplification as 10LR increased by 115.79% and 15LR increased by 72.22%. Cricket formulations increased by 36.88% (10CR) and 47.50% (15CR). This was an expected result as the water activity of locust products at 0 months was greater than their cricket equivalents. Despite this increase the water activity values of all four formulations were still well below the critical limit of 0.6 after 3 months, which suggests they should be still considered as safe for consumption upon proper cooking. In general, as reported in the literature, brown rice is considered to have a shorter shelf life than white rice. This is perhaps due to the richer and more balanced nutrient profiles in brown rice versus white rice, which makes it more prone to fat oxidation and microbial proliferation overtime. The further grinding process to convert the grain kernels to the flour makes the rice flour more vulnerable to oxidation, which may halve the original shelf life as an intact grain. The Oldways Whole Grain Council (2013) recommends brown rice kernels to be stored for up to 6 month in the pantry while the flour derived from the whole brown rice grain to be kept only for 3 months. Together with the reference information, the results from the current formulation design without adding any food preservatives, the shelf life of the insect rice products is estimated to last over 3 but not beyond 6 months. The water activity measurements of the four formulations after 6 months storage should confirm this statement.

Nutrition Content Fiber To assess the total dietary fiber content of the insect rice formulations, an Ankom Dietary Fiber Analyzer was utilized. The results are reported below in Table 7 as % recovered residue that contains undigested protein and ash in addition to the total dietary fiber (TDF) content in the insect rice formulations. As explained in the methodology section, due to the limited laboratory capacity on protein and ash analyses, this % recovered residue value was used as a rough estimation of TDF content. The USDA National Nutrient Database for Standard Reference, Release 28 (2015) was used to obtain the TDF value for MBR.

Table 7 – Percent recovered residue of insect and rice products Rice product % Recovered residue 10CR 12.35 ± 0.92a 15CR 12.20 ± 3.68a 10LR 14.25 ± 1.91a 15LR 21.75 ± 6.15a MBR* 3.60 *MBR value of 3.60 is the total dietary fiber value as obtained from the USDA National Nutrient Database for Standard Reference, Release 28 (2015)

Despite the values in Table 7 being an estimation of the % TDF, comparison could still be made. While taking into account that all other ingredients within the 10CR and 10LR are constant, the only difference between these two formulations is the source

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of the insect. With that, an increase in estimated % TDF can be seen. As this is the same trend for 15CR and 15LR, it is proposed that locust rice formulations do contain a superior level of TDF than their cricket counterparts. It is known that as opposed to providing typical insoluble or soluble dietary fibers, such as cellulose or pectin, edible insects are more likely to contain chitin (van Huis and others 2013). This is an insoluble fiber, similar to cellulose, but instead contains amino group substitutes. This non- digestible carbohydrate is known to be found in the exoskeleton both the Insecta and Crustacea class of the animal kingdom. Considering the exoskeleton of locust is perhaps heavily composed with chitin, this implies the fiber content that could be detected by the instrument was primarily derived from this compound. While chitin may provide some graininess to the texture of the rice, the ability to interact with dietary lipids has led to some suggestion of chitin being included as a functional fiber (Gropper and Smith 2013). In fact, chitin is already being utilized in Japan to fortify cereals to enhance the levels of fiber and calcium (Williams 2010). Inconsistencies may have been caused by some procedural noise as samples containing 10% or more of fat may cause interference in the Ankom dietary fiber assessment. With Table 10 displaying the fat content of 15CR to be near this value, this may be the reason for the greater value of recovered residue, since some lipid residue may be included without proper pre-defatting procedure. The recovered residue value is also inappropriate for contrasting with the theoretical dietary fiber level of MBR that is derived from the USDA, instead of an actual measurement with the Ankom instrument. Even so, the results do reveal exciting evidence for the dietary fiber contents in cricket and locust products, which has not been reported in the literature. Previous research conducted for the fiber determination of edible insects has been mainly on the crude fiber content, which is irrelevant for human nutrition. However, the current study does demonstrate some results supporting locust to contain greater quantities of TDF than crickets.

Iron A graphite furnace atomic absorption spectrometer (AAS) was employed for the analysis of both the iron content of raw insect flours and the insect rice products. The values are shown in Table 8 below. It should be noted, however, that the iron content of MBR was collected from the USDA National Nutrient Database for Standard Reference, Release 28 (2015).

Table 8 – Iron content of insect and insect rice products Product Iron (mg/ 100g) Coefficient of Variance (CV, %) cricket flour 6.80 ± 2.55a 37% 10CR 6.68 ± 3.65a 55% 15CR 4.09 ± 1.80a 44% locust flour 8.05 ± 0.50a 6% 10LR 4.42 ± 0.62a 14% 15LR 2.87 ± 0.17a 6% MBR* 1.29 n/a *MBR value of 1.290 is the iron value as obtained from the USDA National Nutrient Database for Standard Reference, Release 28 (2015)

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Based on the literature information, the locust rice products were anticipated to be a better source of iron than the cricket formulations. For instance, Oonincx and van der Poel (2011) found that, locusts of the same species of those used in the current study (Locusta migratoria) have 15.10 mg of iron per 100g weight. In a separate study, crickets, Acheta domesticus, were reported to have 6.27-11.23mg / 100g (Rumpold and Schluter 2013). When at first examining the iron content for the raw materials (i.e. cricket and locust flours) (Table 8), the laboratory measurements of 8.05 mg/100 g for locust flour and 6.80 mg/100 g for cricket flour showed to be similar to these literature findings, especially locust flour containing slightly higher iron levels than the raw cricket material. The cricket value was also close to that of the COA provided by Next Millennium Farms as well. Yet, when examining the iron contents measured for insect rice formulations, it was not easy to generate a clear conclusion. While greater iron levels were expected to be observed in formulations with increased additions of insect flours, this was not the case in the current study. The inconsistency is mainly related to the iron analysis protocol, where the wet ashing process using concentrated HCl and HNO3 should completely digest the organic matters in the samples. This digestion subsequently converts all minerals in the food samples into an elemental form that can be detected by AAS. However, during the current laboratory operation, a lipid-like precipitate was observed to be present in most of the wet assay solutions, suggesting the acid digestion was not properly completed. Furthermore, the ensuing filtration step in an effort to remove this fat-like layer may have also caused some loss of iron as it may be entrapped in these deposits. This ultimately led to lower values of iron content in the wet assay solutions for all samples. Each sample was prepared with two assay solutions, or measured with duplicates. The standard deviation, or the coefficient of variance (CV, %) from the duplicates was significantly high for most of the samples, especially for samples of cricket flour and the two rice formulations made of it, ranging from 37% to 55%. This variation between different assay solutions even for the same sample confirmed that there might be some interference using the current iron analysis protocol, which certainly requires further standardization in order to produce repeatable and reproducible results. What can be successfully drawn from the GF-AAS iron analysis is that this essential mineral is indeed found within these insect rice products, and is presented with higher values when compared to the control MBR, which might be owed to the incorporation of insect flours.

Protein An analytical lab located in Anaheim, CA called Certified Laboratories facilitated the protein and fat analyses of each of the four insect rice products. The results are displayed below in Table 9 for protein and Table 10 for fat, with the exception of the values for MBR. The MBR values were obtained from the USDA National Nutrient Database for Standard Reference, Release 28 (2015).

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Table 9 – Protein content of insect and rice products Product Protein (%, w/w) 10CR 12.72% 15CR 15.56% 10LR 14.06% 15LR 15.70% MBR* 7.54% *MBR % protein was obtained from the USDA National Nutrient Database for Standard Reference, Release 28 (2015)

From the above table, it is recognized that the insect flour certainly demonstrated as a high source of protein. In comparison with the control, these insect rice products offer over 150 - 200% times more protein than MBR. While brown rice flour did provide a second major source of protein as a bulk ingredient, its reduced level in two 15% formulations still yielded products with raised levels of protein, thus attributing these increased levels to the incorporation of insect flours. The results from this side-by- side protein analysis of cricket and locust products provides critical nutritional information, which not only fills in some of the gap within this field of research but also offers support in the utilization of edible insects in the diet. As established by the US Food and Drug Administration (FDA), the daily value (DV) is a term that provides consumers a reference intake level for macro- and micronutrients. Based on a 2,000 calorie diet for adults and children of four or more years of age, the DV for protein is 50g (FDA 2015). While this value was created to ease consumer confusion and is utilized as a standard for the food industry, the 2010 Dietary Guidelines for Americans, published by the US Department of Agriculture (USDA) recommends adults males, 19 – 55+ years old, to consume 56 g of protein and adult females of the same age range to consume 46 g of protein (USDA 2010). According to above results, the insect rice products provide 12.7 to 15.7 gram of protein per 100 g insect rice (Table 9); thus, with a standard serving size of ½ cup of rice, or ~80g, this would provide 10.18 – 12.56 g of protein. Based on the DV value of 50 g established by the FDA, just one serving of the insect rice would provide 26 – 29% of the daily value for this macronutrient, confirming the cricket and locust rice products to be excellent sources of protein. Referring to the available information of protein content provided by the vendors, where the cricket flour and locust flour have a protein content of 59.46% and 54.67%, respectively (dry weight basis), it was predicted that the cricket rice formulations would offer a superior quantity of protein than their locust counterparts. However, the results as displayed in Table 9 shows this was not the case as 10LR and 15LR showed greater amounts of protein, although perhaps not significantly dissimilar in value. Protein levels were reported to be within 64.38 – 70.75% for crickets (Acheta domesticus) (Rumpold and Schluter 2013) and 55.5 – 64.9% for locust (Locusta migratoria) (Oonincx and van der Poel 2011). Although the crickets in the study conducted by Rumpold and Schluter (2013) have higher quantities of protein than the Locusta migratoria, the crickets used in the current study are of a different species, Gryllodes sigillatus. It is important to consider that there may be variability in nutritional content from edible insects as they differ between and within species and are also influenced by type of diet and environment. Furthermore, the method of analysis could

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also provide some disparities in the results. Thus, with these research findings solely as a reference, it seems plausible that the cricket and locust offer comparative protein content in the rice products developed in this study.

Fat Below in Table 10 are the representative values for the % fat in the insect rice and MBR products. While the lipid assessment of the four insect rice formulations was outsourced to the aforementioned analytical lab, the MBR fat content was derived from the USDA National Nutrient Database for Standard Reference, Release 28 (2015).

Table 10 – Fat content of insect and rice products Product Fat (%) 10CR 7.76% 15CR 9.73% 10LR 7.44% 15LR 7.97% MBR* 3.20% *MBR % fat was obtained from the USDA National Nutrient Database for Standard Reference, Release 28 (2015)

With all insect formulations providing more than 200% of fat than the MBR, edible insects unquestionably contain a significant supply of lipids. In the table above, both cricket and locust insect formulations demonstrate to provide about 7.5 to 8.0% of fat. However, the cricket formulations do exhibit higher percentages of fat than their locust equivalents, especially in 15CR. In previous studies, it was found that crickets (Acheta domesticus) had 18.55 – 22.80 % fat (Rumpold and Schluter 2013) while locust (Locusta migratoria) had 17.9 – 29.6% fat (Oonincx and van der Poel 2011). Thus, the ranges found in the current and previous research support crickets as the potentially more impressive resource for lipids, although perhaps only marginally. According to the FDA (2015), the daily value for fat is 65g for a 2,000 calorie diet. In another perspective, the percent of calories obtained from fat should be in a range of 20 – 35% (or 400 - 700 calories for a 2,000 calories diet) for both males and females who are 19 – 55+ years old (USDA 2010). From the fat analysis results as displayed in Table 10, the insect formulations provided 10-12% DV for fat (6 grams in 10% formulations and up to 7.7 grams in 15% formulations) when one serving of ½ cup or approximately 80g of such rice products were consumed, which did not exceed the USDA recommended intake level. While in the United States, the USDA recommends Americans reduce their calories from solid fats, this is not of concern for those in developing countries, for whom these insect rice products are currently aimed towards. Protein-energy malnutrition is the most critical nutritional issues faced by populations in Asia, Latin America, and Africa, especially in children (Latham 1997). Perhaps of the most infamous and severe cases of the malnutrition emergencies are marasmus and kwashiorkor. Here, marasmus is simplified to be caused by the overall deficiency of food, while kwashiorkor is caused by lack of energy and protein as the diet is primarily composed of carbohydrates. The former results in the dangerous wasting of the body as fat and protein stores are utilized to create energy to survive. People inflicted with kwashiorkor, on the other hand, results in serious edema along with wasting. As the

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many populations within developing countries are already practicing entomophagy, the novel insect rice products can naturally offer a supplement to the plain rice staple within their diet. As previously discussed, the insect rice can provide not only 150 – 200% times more protein, but overall increase the caloric intake for these starved people. In comparison to typical brown rice, the insect rice products can provide upwards to 47 more of much needed calories. The insect rice is also highly likely to provide other added advantages, such as increased consumption of iron and fiber, although the exact quantities are still to be determined.

Sensory Evaluation There were a total of five attributes considered by the panelists on the insect rice formulations, including: color, aroma, taste, texture, and overall liking. The first four were based on a five-point Just About Right (JAR) scale while the latter was rated on a 7- point hedonic scale. Participants were also encouraged to leave free response comments about each attribute.

Color In scoring color, this characteristic could be scored as either “much too dark,” “somewhat too dark,” “just about right,” “somewhat too light,” or “much too light.” With the color being derived from the insect flours added to the base matrix, it was predicted that higher-level insect formulations would have scores reflecting this observation. Results from this evaluation can be seen in Figure 2 below.

100% 90% c 80% 70% Dark 60% a 50% Just about right b 40% 30% Light 20% 10% c 0% 10% Cricket 15% Cricket 10% Locust 15% Locust

Figure 2 - Color intensity of insect rice formulations Responses are grouped by the four insect rice formulations and displayed as percentage of participants who provided the color-coded scores, where dark responses includes both “much too dark” and “somewhat too dark” selections and light responses includes both “much too light” and “somewhat too light.” Different lower case letters distinguish a significant difference for only just about right scores

Of the four insect rice products, the color of 10LR was exceedingly liked over the other three formulations (p < 0.05). This was also reflected in the comments as a

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greater number of subjects mentioned 10LR to look the most similar to market brown rice. It was also suggested that the slightly brown shade of 10LR allowed the panelists to correlate its color as if it were toasted or served with soy sauce, which seems to be a general preference among these consumers. For the other three formulations, these products were deemed either somewhat too light (10CR) or somewhat too dark (15CR and 15LR). As anticipated, these observations were matched with the objective evaluations conducted using the Konica Minolta colorimeter. With the cricket rice having a* and b* values being closer to 0, this suggests the product to be closer to grey hues than the locust rice, which was certainly reflected in the free response section. According to the majority of the comments regarding the appearance of the two cricket formulations, only these products were considered too grey in color, which was unappealing. On the other hand, the most reoccurring comment concerning the color of 15LR was that it was too dark in color, with most panelists scoring it as either “somewhat too dark” or “much too dark.” While some darkening in color can be the indicator of those delicious flavors from frying, searing, or baking, overshooting this desirable “golden brown” color can suggest the food to be burnt, dry, and overcooked, which was the case for 15LR. The L* values of the locust rice products from the colorimeter measurements matched these comments as the products showed to become significantly darker as the level of locust flour increased.

Aroma With cricket and locust being similar to ocean shrimp, it was anticipated that the participants would observe a fishy aroma when smelling the insect rice products. Thus, the possible responses were as such: “much too odorless,” “somewhat too odorless,” “just about right,” “somewhat too fishy,” and “much too fishy.” The ratings for the aroma of the insect rice can be seen in the figure below.

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100% 90% 80% 70% Fishy 60% 50% a Just about right 40% a 30% Odorless b 20% 10% c 0% 10% Cricket 15% Cricket 10% Locust 15% Locust

Figure 3 - Aroma intensity of insect rice formulations Responses are grouped by the four insect rice formulations and displayed as percentage of participants who provided the color-coded scores, where fishy responses includes both “much too fishy” and “somewhat too fishy” selections and odorless responses includes both “much too odorless” and “somewhat too odorless.” Different lower case letters distinguish a significant difference for only just about right scores

While all formulations were perceived with some aroma, both cricket formulations were deemed to have aromas that were statistically just about right (p < 0.05) where the products were neither too fishy nor too odorless. On the other hand, the two locust formulations were observed to be somewhat too fishy. In interpreting the comments provided by the panelists, it was implied that both grainy or wheat like aromas and lack of an odor coordinated with what was expected for market brown rice. With that, most of these observations were made in the cricket formulations and so were reflected in their subsequent scoring of being just about right in aroma. Locust formulations, on the other hand, showed a significant difference in its fishy aromas. Although both were considered to be somewhat too fishy, there were four times as many subjects who rated 15LR as much too fishy. In fact, there were specific comments regarding this product having smelled like crickets sold in pet stores. Furthermore, a fascinating finding was that some panelists were able to match 10LR and 15LR to each other, stating that the aromas were somehow comparable. Thus, the odor originating from locust formulations has been determined to be much more pronounced than that from cricket products, with the smell being much more obvious when increasing the level of locust flour.

Flavor A main concern for the flavor of the insect rice products was in its earthiness. Figure 4 displays the percentage of scores for the following responses: “much too bland,” “somewhat too bland,” “just about right,” “somewhat too earthy,” and “much too earthy.”

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100% 90% 80% 70% 60% a Earthy 50% a Just about right 40% b 30% Bland c 20% 10% 0% 10% Cricket 15% Cricket 10% Locust 15% Locust

Figure 4 - Flavor intensity of insect rice formulations Responses are grouped by the four insect rice formulations and displayed as percentage of participants who provided the color-coded scores, where earthy responses includes both “much too earthy” and “somewhat too earthy” selections and bland responses includes both “much too bland” and “somewhat too bland.” Different lower case letters distinguish a significant difference for only just about right scores

Similar to the aroma, both 10CR and 15CR were perceived to have flavors that were significantly considered as just about right (p < 0.05) while 10LR and 15LR were determined to be either somewhat too earthy or much too earthy. From the comments, lower insect formulations, such as 10CR and 10LR, were observed to be more parallel to market brown rice flavors. Although some suggested that this meant the insect rice tasted somewhat bland, they also mentioned that this was not necessarily a negative attribute as rice is generally considered a bland product when served alone. Even so, flavor remained an important component to the acceptability of the insect products as several participants mentioned that a suitable flavor could positively sway the overall opinion of a product. Multiple criticisms made by the panel recommended that the acceptability of the products would be greatly improved if served alongside other foods or seasoned, such as with salt or soy sauce. As aforementioned, a study was conducted by Megido and others (2014) involving sensory evaluation on mealworms (Tenebrio molitor) and crickets (Acheta domesticus), where these insects were prepared in a variety of ways including: baked, boiled, a crushed mix of both insects, mealworms flavored with vanilla, and mealworms with chocolate. It was found that the most accepted form of preparation was with chocolate, followed by flavored with paprika, and then baked. While each mealworm was crispy in texture, the investigators advocate the flavor to be the factor in preference (Megido and others 2014). Yet, it is essential to recognize that earthiness was certainly an undesirable attribute in the insect formulations. It was found to be more apparent in not only formulations with greater quantities of insect flour, but especially in locust products. 15LR was determined to be the earthiest in flavor as it was scored by nearly four times more subjects as “much too earthy” than its 10% counterpart. By recognizing that land

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insects, such as cricket and locust, and shellfish, such as shrimp, are both arthropods, it was anticipated that they would have comparable flavors.

Mouthfeel Graininess was the primary concern for the texture of the insect rice formulations. Thus, the question was anchored as “much too smooth,” “somewhat too smooth,” “just about right,” “somewhat too grainy,” and “much too grainy.” The responses from the participants about this aspect of the rice products can be found in Figure 5 below.

100% 90% 80% 70% Grainy 60% a 50% a Just about right 40% 30% Smooth 20% b 10% d 0% 10% Cricket 15% Cricket 10% Locust 15% Locust

Figure 5 - Mouthfeel intensity of insect rice formulations Responses are grouped by the four insect rice formulations and displayed as percentage of participants who provided the color-coded scores, where grainy responses includes both “much too grainy” and “somewhat too grainy” selections and smooth responses includes both “much too smooth” and “somewhat too smooth.” Different lower case letters distinguish a significant difference for only just about right scores

Once again, the cricket formulations were rated statistically as just about right (p < 0.05) whereas locust formulations were observed to have mouthfeels that were somewhat too grainy. In fact, 10CR and 15CR had between 1.8 and 3.4 times more people who considered the texture as just about right than their locust counterparts. While the graininess was more evident in locust formulations, it was even more apparent in 15LR. Thus, the source of the perceived graininess is derived from the locust flour itself. In addition to the term grainy, the words “gritty,” “crunchy,” and “sandy” were used to describe the locust rice formulations in the free-response comments from the panelists. In studying the free response portion of this evaluation, there were an overwhelming number of comments pertaining to the gritty texture of 15LR. Unquestionably, this attribute was perhaps the most unappealing characteristic experienced by the panelists. Considering that locust rice products were determined to contain greater quantities of fiber and that the principal component is likely to be chitin, this insoluble fiber is highly suggested to be the culprit behind the unpleasant texture.

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Overall Liking To determine the overall acceptance of each of the four insect rice formulations, a 7-point hedonic scale was employed. The results from this evaluation is represented in the Table 11.

Table 11 - Mean scores for overall liking of insect rice formulations Insect rice Mean score 10CR 4.76 ± 1.49a 15CR 4.45 ± 1.56a 10LR 3.97 ± 1.47b 15LR 2.92 ± 1.48c Highest score for overall liking was a 7 with the lowest score being 0. Different lower case letters distinguish a significant difference in overall liking of products

By and large, there was no significant difference between how 10CR, 15CR, and 10LR were accepted (p > 0.05). Each of these formulations were scored between being neutrally enjoyed and liked slightly (mean values of 4.7, 4.6, and 4.4 out of 7, respectively). The formulation with the highest locust content, 15LR, not surprisingly, was significantly less accepted as its average score (mean value of 3.6 out of 7) was between being disliked slightly and a neutral overall liking. While both cricket formulations were evaluated to have aroma, flavor, and mouthfeel that were just about right, the color was reported to be too gray and somewhat too light (10CR) or somewhat too dark (15CR) to be appealing. This suggests that the color is a major factor in the acceptance of the insect rice products. This is an appropriate response as color is certainly the primary attribute when evaluating food (Giusti and Wrolstad 2003). Next, it can be supposed that the overwhelming graininess of 15LR played a chief role in the acceptance of this particular product as it received the lowest mean score, thus being the least accepted. In addition to the negative reaction to the texture of 15LR, all of the other attributes, including color, aroma, and flavor, also was rated rather poorly. The color was considered somewhat too dark, the aroma as somewhat too fishy, and the flavor as much too earthy. Interestingly, 10LR was reported to be similarly liked as the cricket formulations. Despite it having received scores for individual attributes almost parallel to 15LR, where it was perceived to be somewhat too fishy in odor, somewhat too earthy in taste, and somewhat too grainy in texture, this product had the best rating for color. Again, this attribute demonstrates itself as a powerful means to increase the acceptability of a product. Although the color of 10LR was undeniably darker than that of the other cricket formulations, this color was associated with pleasant and desirable flavors.

Demographics and Perception of Entomophagy With recruitment taking place on the Cal Poly Pomona campus, it was no surprise that the majority of the panel were in their early 20’s, about 76% (Figure 7) and were between their 1st year to their 4th year in college (Figure 10). It was also found that the ratio of men to women panelists was almost 50:50, with only 10% more females to males (Figure 8). In terms of ethnicity, it seemed that there were a greater number of Latinos and Asians, which together made up almost 70% of the sample population (Figure 9). As

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many countries within Latin America and Asia are developing countries, the data collected from the sensory evaluation study could be somewhat extrapolated to those countries within Latin American and Asia, although Latin American and Asian American dietary habits can be certainly unique to the customs found in their mother countries. Thus, despite the current investigation being unable to reach those currently living in countries such as Mexico or Laos People’s Democratic Republic, the results still offer some indication of the potential acceptability and implementation in those countries in the future, although these generalizations may be fairly limited. Together, the 82% of panelists being familiar with entomophagy (Figure 11) and the 73% of subjects declaring a high probability of consuming edible insects in the future (Figure 13) point towards a positive trend. In a separate study, Tuorila and others (1994) found a positive relationship between pleasurable first experiences with a novel food and the likelihood of future consumption. As more than 50% of the free-response comments included a statement of their enjoyment and/or support of the current study and its products, this certainly presents a progression towards the future applications of edible insects in the market, even in developed nations. However, while 73% reported positively in their willingness to practice entomophagy in the future (Figure 13), only 42% were able to say they’ve consumed edible insects before their involvement in the current study (Figure 12). With more participants (58%) never having previously consumed edible insects, this is certainly a positive outcome of the study. It demonstrates the eagerness of a population to at least try edible insects. Even more dramatic are the findings from the Belgium study by Megido and others (2014). In this particular survey, the researchers found that although 46.6% of the participants reported some negative attitude towards entomophagy, an even greater number of respondents, 77%, were willing to consume edible insects. The novelty, and perhaps curiosity, of entomophagy may be even more powerful than the "yuck" factor. Progress is being made towards the acceptance of entomophagy, much like sushi and lobster in the United States (Crowley 2015, Greenlaw 2002). On the other hand, when taking into account the willingness to purchase edible insect products in the future, there seems to be some disparity. Although people stated that they would probably and definitely consume edible insects in the future, the results from Figure 14 show that they may not actually purchase these insect products. It is proposed that this is somewhat correlated with the fact that the majority of the panel are college students with limited finances, where money is only spent on items that are absolutely essential or worth the expense. As mentioned in the free response portions of the questionnaire, knowing of the nutritional and environmental benefits of consuming and farming edible insects would likely provide enough information for these consumers to make a decision towards purchasing edible insect products in the future. An inquiry pertaining to the opinion of how edible insects should be placed in the diet was also included in the survey. Participants were asked to select as many choices as they felt that described their opinions.

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1% 3% 2%

55+ years old Male 18% 45% 45 - 54 years old Female 35 - 44 years old 55% 25 - 34 years old 76% 18 - 24 years old

Figure 7 - Age of sensory panel Figure 8 - Gender of sensory panel

3% 3% 3% 7% 5% 8% Other PhD 17% Black 13% Master's Latino Graduate Asian 4th year 30% White 28% 3rd year 38% 45% 2nd year 1st year

Figure 9 – Ethnicity of sensory panel Figure 10 – Education level of sensory panel

18% No 42% No Yes Yes 58% 82%

Figure 11 – Familiarity with entomophagy Figure 12 – Experience with entomophagy

Definitely would not eat Definitely would not buy Probably would not eat Probably would not buy 3% 5% 3% Maybe/ maybe not Maybe/ maybe not Probably would eat Probably would buy 15% Definitely would eat 13% Definitely would buy 34% 22%

38% 29% 38%

Figure 13 – Future willingness to consume Figure 14 – Future willingness to purchase edible insects edible insect products

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3% 2%

5% Snack 22% Appetizer 13% Side dish Main entree (hidden form) Dish topping/ Garnish 17% 19% Main entree (whole insect) Dessert Not Applicable 19%

Figure 15 - Perception of edible insects in the diet Perceptions of edible insects in the diet are displayed by the percentage of the sensory panel who selected those choices. This particular survey question had multiple selections and participants were asked to select all that applied.

When considering the marketing of edible insect products, the results from Figure 15 suggest that edible insects are most often preferred to be consumed as a snack (22%), appetizer (19%), and side dish (19%). Edible insects being perceived most as a snack could be related to their small size or, in general that snacking is on the rise. In fact, in 2014, the number of in-between snacks increased to 2.8 with more than half of adults consuming three or more snacks per day (Wyatt 2014). In a previous study, while “snack” was not included as an option in the survey, edible insects were primarily distinguished as an appetizer. Investigators suggest this also be due to the small size and original form of the food (Megido and others 2014).

Conclusion

All objectives of the current study were achieved. First, a total of four insect products, each with the same brown rice flour base, were developed using cricket or locust flours at either 10% or 15% addition levels. The formulations were determined to be feasible for extrusion. By and large, physicochemical properties of the insect rice products were found to be darker in color, rougher in surface morphology, softer and more adhesive in texture, and more shelf stable than market brown rice. The insect rice formulations provided an excellent source of protein, 10-12% of the daily value of fat, and offered some iron and total dietary fiber. Both cricket rice products have demonstrated as the best formulations for continuing research and development. Between the two cricket rice products, the major issue is perhaps in its appearance as the sensory panel considered the products to be either too light or too dark in appearance. Locust products, however, were overall liked at different levels. While 15% locust rice was the least accepted product, its 10% counterpart, while having been reported with similar negative scores for aroma, flavor

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and mouthfeel, this product still offered a toasty brown color that was the most accepted of the four formulations. Overall, both cricket rice products and the 10% locust rice formulations were scored between neutrally enjoyed and liked slightly, demonstrating positive results towards consumer acceptance.

Acknowledgements President’s Research, Scholarship, and Creative Activities Program (RSCA) (2014- 2015).

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