CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to Singapore Provisional Patent Application

Serial No. 10202251212M, filed September 29, 2022, titled “FERMENTATION METHODS AND SYSTEMS OF MAKING INSECT FEED”, the contents of which are incorporated herein by reference.

BACKGROUND

[0002] Insect-derived materials such as proteins and oils are of increasing commercial interest. As a direct result, there is a growing need for high-quality insect feed materials.

[0003] Traditional insect feeds suffer from several significant drawbacks. First, they tend to harbor growths of undesired fungi and bacteria which leads to spoilage. Additionally, the source organic materials give off heat due to growth and metabolism of mesophilic aerobic bacteria. The heat is harmful to the insects, causing stress, dehydration, and potentially death. These factors lead growers to administer feed to insects multiple times in smaller batches, consuming more time and effort.

[0004] Despite considerable advances made to date, there still exists a need for insect feeds that are nutritious but do not spoil easily, cause excess heat, or require excess labor.

SUMMARY

[0005] Methods of preparing insect feeds are described. The methods of preparing insect feed can comprise: providing input organic material; providing a lactic acid fermentation system comprising a holding vessel configured to hold input organic material under anaerobic conditions or anoxic conditions; delivering the input organic material and lactic acid bacteria inoculant to the holding vessel to form a pre-fermented mixture; and fermenting the pre- fermented mixture in the holding vessel under anaerobic conditions or anoxic conditions for a time period to prepare a fermented mixture.

[0006] Methods of feeding insects are described. The methods can comprise providing a fermented mixture; and feeding the fermented mixture to the insects; wherein: the fermented mixture is prepared by a method comprising: providing a lactic acid fermentation system comprising: a holding vessel configured to hold input organic material under anaerobic conditions or anoxic conditions; delivering the input organic material and lactic acid bacteria inoculant to the holding vessel to prepare a pre-fermented mixture; and fermenting the prefermented mixture in the holding vessel under anaerobic conditions or anoxic conditions for a time period to prepare a fermented mixture.

[0007] Fermentation systems for the preparation of insect feeds are described. The systems can comprise at least one holding vessel configured to hold input organic material under anaerobic conditions or anoxic conditions; at least one input valve; at least one output valve; at least one pH measurement unit linked to the holding vessel; and at least one pump.

[0008] Insect feeds comprising a fermented mixture are described. The insect feeds can comprise a fermented mixture produced by the method comprising: providing a fermentation system comprising: a holding vessel configured to hold input organic material under anaerobic conditions or anoxic conditions; delivering the input organic material and lactic acid bacteria inoculant to the holding vessel to prepare a pre-fermented mixture; and fermenting the prefermented mixture in the holding vessel under anaerobic conditions or anoxic conditions for a time period to prepare a fermented mixture.

BRIEF DESCRIPTION OF THE DRAWINGS [0009] Fig. 1 shows a conical style grow-out silo system (100). The system can include an input screw conveyor (101), input valve (102), relief valve (103), temperature sensor (104), pH sensor (105), output valve (106), output screw conveyor (107), and conical silo (108).

[0010] Fig. 2 shows a live bottom rectangular fermentation silo system (200). The system can include a manhole (201), input valve (202), level sensor (203), dissolved oxygen sensor (204), breath valve (205), upper vessel part (206), screw auger (207), temperature sensor (208), pH sensor (209), sampling point (210), live bottom part (211), and output valve (212).

[0011] Fig. 3 shows a feed mixing and inoculation system (300) using ribbon blade mixers and spray-on inoculant. The system can include a material input gate (301), inoculant dosing nozzles (302), and a discharge gate (303).

[0012] Fig. 4 shows a feed dosing machine and black soldier fly larvae tray system

(400). The system can include a feed input hopper (401), feed dosing pump (402), growing tray (403), and weighing table (404).

[0013] Fig. 5 shows an inoculant storage and use system (500). The system can include an inoculant tank (501), valve (502), pump (503), and flow meter (504) connecting to mixer (505).

[0014] Fig. 6 shows a system process flow (600). Overall steps include loading (601), mixing (602), fermentation (603), and discharge (604). Loading (601) contacts raw material (605) and lactic acid bacteria inoculant (606). Mixing (602) is performed in a mixer (607).

Fermentation (603) is performed in a fermentation silo (608) under anaerobic or anoxic conditions (609). Discharge (604) includes delivering fermented feed (610) by feed dosing to larvae (611). [0015] Fig. 7 shows a system process flow (700). Raw materials are received (701) and subjected to a QA/QC inspection (702). Materials are either approved (703) or rejected (704). Rejected materials are returned to supplier (705) or destroyed. Approved materials are subjected to grinding (706), mixing (707), and a transferred to a fermentation silo (708) for fermentation (709). Fermented feed is dosed (710) onto larvae rearing trays.

[0016] Fig. 8 shows a graph of temperature evolution during black soldier fly larvae rearing with commercial (non-lactic acid) inoculant and in-house lactic acid inoculant. The x- axis is rearing days, and the y-axis is temperature in Celsius. The hexagon symbol is ambient environment temperature. The square symbol is an experiment with commercial inoculant #1. The diamond symbol is an experiment with commercial inoculant #2. The round symbol is an experiment with in-house lactic acid inoculant. The triangle symbol is an experiment with a mixture of commercial inoculant #1 and commercial inoculant #2.

[0017] Fig. 9 shows a graph of the daily development of black soldier fly larvae. The x-axis is days. The y-axis is individual larvae weight in grams. The triangle symbol is a control of fresh unfermented soy. The round symbol is using fermented soy.

DEFINITIONS

[0018] ‘Anaerobic” refers to a condition or status that is completely deficient of free oxygen, directly involved in microorganism metabolism.

[0019] ‘Anoxic” refers to an environment partially deficient of free oxygen, as it may include bound molecular oxygen.

[0020] The acronym “BSF” refers to black soldier fly (or flies).

[0021] ‘Lactic acid bacteria” refers to gram-positive bacteria which naturally produce lactic acid as a product of metabolism/fermentation of carbohydrates. [0022] ‘Lactic acid fermentation” refers to the anaerobic breakdown of sugars into cellular energy and lactic acid byproducts.

DETAILED DESCRIPTION

[0023] This disclosure is not limited to the particular systems, devices and methods described, as these may vary. The terminology used in the description is for the purpose of describing the particular versions or embodiments only and is not intended to limit the scope. [0024] The fermentation system and methods described here differ in fundamental ways from existing animal feed silage. One important difference is the intended outcome: animal feed silage is used to store feed for long periods of time, whereas insect feed fermentation is conducted primarily to control the temperature of insect rearing (as well as to control feed safety and improve digestibility). The organic acids produced during fermentation inhibit mesophilic bacteria, delaying the onset of heat production from the respiration of microorganisms in the insect feed, this allows insect farmers to optimize the larval rearing period and prevent overheating in the trays which can lead to larval mortality.

[0025] Fermentation is typically much shorter (about 3 days is typical) than livestock silage (which can take several months) and uses organic food waste and byproducts whereas livestock silage uses plant leaves and stems.

[0026] The design of equipment and storage tanks is also different from that of animal feed silage. Insect fermented feeds are typically high moisture and are typically moved by pumps or screw conveyors. Fermented insect feed material has density and flowability that makes it challenging or impossible to use livestock silos, silage bales or similar equipment for their production.

[0027] Methods of preparing insect feeds [0028] Various methods of preparing insect feeds are described. In one example, a method of preparing insect feed can comprise providing input organic material; providing a lactic acid fermentation system comprising a holding vessel configured to hold input organic material under anaerobic conditions or anoxic conditions ; delivering the input organic material and lactic acid bacteria inoculant to the holding vessel to form a pre-fermented mixture; and fermenting the pre-fermented mixture in the holding vessel under anaerobic conditions or anoxic conditions for a time period to prepare a fermented mixture

[0029] The input organic material can generally be any organic material. The organic material can be any organic material capable of being fermented by lactic acid bacteria. Many examples of organic material exist. For example, the organic material can include fruit, vegetable, meat, bread, dairy, or combinations thereof. In other examples, the organic material can include plant fibers, offcuts, food waste, spoiled food, or combinations thereof.

[0030] In some examples, the input organic material is not animal waste or does not include animal waste. For example, the input organic material is not or does not include urine, feces, or a combination thereof.

[0031] The input organic material can include water. The water can generally be present in any amount. For example, the input organic material can have a water content of about 10% w/w or more. In other examples, the input organic material can have a water content of about 20% w/w or more, about 30% w/w or more, about 40% w/w or more, about 50% w/w or more, or about 60% w/w or more. For example, the water content can be about 60% w/w, about 65% w/w, about 70% w/w, about 75% w/w, about 80% w/w, or ranges between any two of these values. In a specific example, the water content can be about 75% w/w. In a specific range example, the water content can be about 65% w/w to about 85% w/w. [0032] The holding vessel can optionally have one or more openings or valves to facilitate the addition or removal of material. For example, the holding vessel can include at least one input valve, at least one output valve, or at least one input valve and at least one output valve. In some examples, one opening or valve can be used for both addition and removal of material.

[0033] The input organic material and the lactic acid bacteria inoculant can be contacted or combined before delivery to the holding vessel or can be contacted or combined in the holding vessel. The input organic material can be added to the holding vessel before, after, or concurrently with the lactic acid bacteria inoculant.

[0034] The lactic acid bacteria inoculant can be a dry inoculant or a liquid inoculant.

The inoculant can contain one strain of lactic acid bacteria or two or more strains of lactic acid bacteria. For example, the inoculant can contain at least one homofermentive lactic acid bacteria and at least one heterofermentive lactic acid bacteria. The inoculant can generally have any concentration. For example, the concentration can be at least about 10 ⁵ CFU/mL, at least about 10 ⁶ CFU/mL, at least about 10 ⁷ CFU/mL, at least about 10 ⁸ CFU/mL, or ranges between any two of these values.

[0035] The fermenting can be performed at ambient temperature or at a higher or lower temperature. Performing the fermenting at ambient temperature has advantages including reducing energy consumption. Fermenting temperature is typically not controlled. Ambient temperature can vary by location and time of year. For example, the fermenting can be performed at a temperature of at least about 20 degrees C. Alternatively, the fermenting can be performed at a temperature of about 25 degrees C to about 45 degrees C. A narrower temperature range can be about 25 degrees C to about 35 degrees C. A specific example can include fermenting at a temperature of about 30 degrees C. Specific examples of temperatures include about 20 degrees C, about 25 degrees C, about 30 degrees C, about 35 degrees C, about 40 degrees C, about 45 degrees C, and ranges between any two of these values.

[0036] The time period can generally be any time period. For example, the time period can be at least about 1 day or at least 1 day. Ranges of time periods include about 1 day to about 30 days, about 1 day to about 10 days, or about 1 day to about 7 days. Specific examples of time periods include about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, about 14 days, and ranges between any two of these values. In a specific example, the time period is about 3 days. Longer time periods are not detrimental, but most or all of the fermentation would have been already completed.

[0037] Additionally or alternatively, fermenting can be performed until a certain value is reached. For example, the fermenting can be performed until the fermented mixture has a temperature equal to or less than ambient temperature. During fermentation, the temperature typically increases above ambient temperature. In another example, the fermenting can be performed until the fermented mixture has a pH of about 5 or less. During fermentation, the pH typically decreases due to production of lactic acid. In another example, the fermenting can be performed until the fermented mixture has a temperature equal to or less than ambient temperature, and the fermented mixture has a pH of about 5 or less.

[0038] Anaerobic conditions can be achieved by sparging the holding vessel with a gas

(such as nitrogen gas or carbon dioxide gas) until the dissolved oxygen concentration inside the holding vessel is zero. Alternatively, anaerobic conditions can be achieved by filling the holding vessel to about 80-90% capacity, and closing the holding vessel. [0039] The method can optionally further include purging the holding vessel with at least one gas before the fermenting step. For example, the gas can be nitrogen gas (N2), argon gas (Ar), carbon dioxide gas (CO2), or combinations thereof.

[0040] The method can optionally further include removing the fermented mixture from the holding tank through an opening or an output valve.

[0041] If the water content of the fermented mixture is lower than desired, the method can include adding water to the fermented mixture to reach a desired moisture level.

[0042] The method can optionally further include adding at least one salt to the input organic material or the pre-fermented mixture. The salt can be a metal salt. The salt can be sodium chloride, magnesium sulfate, potassium phosphate, sodium selenite, or combinations thereof. The salt can be any commonly used salt. The salt can be added at generally any concentration, such as at least about 0.1% w/w, or about 0.1% w/w to about 5% w/w. Specific concentrations include about 0.1% w/w, about 0.5% w/w, about 1% w/w, about 2% w/w, about 3% w/w, about 4% w/w, about 5% w/w, and ranges between any two of these values.

[0043] The method can optionally further include adding at least one organic acid to the input organic material or the pre-fermented mixture. The organic acid can generally be any organic acid, such as citric acid, propionic acid, acetic acid, lactic acid, formic acid, succinic acid, fumaric acid, malic acid, or combinations thereof. In some examples, the organic acid is citric acid. The addition of at least one organic acid is believed to enhance the feed fermentation process to produce feed with reduced pathogenic microbial burden and improves aerobic stability by inhibiting yeast, mold, and fungus proliferation on the feed upon release onto the feeding trays. The organic acid can be added at generally any concentration, such as at least about 0.1% w/w, or about 0.1% w/w to about 5% w/w. Specific concentrations include about 0.1% w/w, about 0.5% w/w, about 1% w/w, about 2% w/w, about 3% w/w, about 4% w/w, about 5% w/w, and ranges between any two of these values.

[0044] The method can optionally further include feeding the fermented mixture to insects such as black soldier flies or black soldier fly larvae.

[0045] Methods of feeding insects

[0046] Methods of feeding insects are also described. Methods can include providing a fermented mixture; and feeding the fermented mixture to the insects; wherein: the fermented mixture is prepared by a method comprising: providing a lactic acid fermentation system comprising: a holding vessel configured to hold input organic material under anaerobic conditions or anoxic conditions; delivering the input organic material and lactic acid bacteria inoculant to the holding vessel to prepare a pre-fermented mixture; and fermenting the prefermented mixture in the holding vessel under anaerobic conditions or anoxic conditions for a time period to prepare a fermented mixture. The fermented mixture and method for its preparation can be as described above.

[0047] The insects can generally be any insects. Specific examples include black soldier flies and black soldier fly larvae.

[0048] In some examples, the fermented mixture is maintained at a temperature equal to or less than about 40 degrees C for the first seven days during the feeding step, or the fermented mixture is maintained at a temperature equal to or less than about 35 degrees C for the first five days during the feeding step. In other examples, the fermented mixture is maintained at a temperature equal to or less than about 42 degrees C during the feeding step. This temperature control can be provided by, for example, inhibition of mesophilic bacteria by the fermented feed.

The controlled temperature can improve the health and viability of the insects. [0049] Fermentation systems

[0050] Also described are fermentation systems for preparing insect feed. The system can include a holding vessel configured to hold input organic material under anaerobic conditions or anoxic conditions. The system can also include at least one input valve; at least one output valve; at least one pH measurement unit linked to the holding vessel; and at least one pump. The input valve can be linked to the holding vessel. The output valve can be linked to the holding vessel. The pump can be linked to the holding vessel and configured to pump materials into, out of, or both into and out of the holding vessel.

[0051] In some examples such as in handling material types such as fermented mixtures with low pH values, system components can be made of acid resistant materials such as carbon steel, stainless steel or fiberglass with anti-rust paint. In certain examples, there can be two categories of silos to handle high solid and high viscous material namely the live bottom silo (straight vertical vessel from the top to the bottom) and conical silo (conical at the bottom with an angle ranging from 45 to 60 degrees) with an output opening of diameter 400 mm (DN 400) for the discharge material without any bridging.

[0052] The holding vessel can optionally have one or more openings or valves to facilitate the addition or removal of material. For example, the holding vessel can include at least one input valve, at least one output valve, or at least one input valve and at least one output valve. In some examples, one opening or valve can be used for both addition and removal of material. The input valve and the output valve can be the same or different.

[0053] The holding vessel can be made of various materials. For example, the holding vessel can be made of metal, plastic, or a combination thereof. Specific examples include stainless steel, carbon steel, fiberglass, or combinations thereof. Alternatively, the holding vessel can be made of cement or concrete.

[0054] The holding vessel can be of various shapes and sizes. For example, the holding vessel can be a tank, barrel, or drum. The holding vessel can generally have any volume.

[0055] The system can optionally further include a purging gas source. The purging gas source can be configured to deliver a gas to the holding vessel. The gas in some examples does not contain oxygen gas (O2). The gas can include, for example, nitrogen gas (N2), argon gas (Ar), carbon dioxide gas (CO2), or combinations thereof.

[0056] The system can further include one or more measurement units linked to the holding vessel. For example, at least one temperature measurement unit, at least one pH measurement unit, at least one oxygen measurement unit, and combinations thereof may be linked to the holding vessel. The measurement units may be placed inside the holding vessel or in communication with the holding vessel.

[0057] The system can further include temperature control units such as at least one heating unit, at least one cooling unit, or combinations thereof linked to the holding vessel. [0058] The system can further include at least one pump. The pump can be used to pump material into or out of the holding vessel. The system can further include at least one agitator to mix contents of the holding vessel.

[0059] While in use, the system can further include input organic material, lactic acid bacteria inoculant, or both in the holding vessel. After use, the system can further include fermented mixture in the holding vessel.

[0060] Insect feeds [0061] Also described are insect feeds. For example, insect feeds can comprise a fermented mixture produced by the method comprising: providing a fermentation system comprising: a holding vessel configured to hold input organic material under anaerobic conditions or anoxic conditions; delivering the input organic material and lactic acid bacteria inoculant to the holding vessel to prepare a pre-fermented mixture; and fermenting the prefermented mixture in the holding vessel under anaerobic conditions or anoxic conditions for a time period to prepare a fermented mixture.

[0062] The fermented mixture can be any of the fermented mixtures described above.

For example, it can have a pH of about 5 or less. The fermented mixture can have a water content of about 60% or more, such as about 70% or about 80%.

[0063] The fermented mixture and/or the insect feed can be stable at ambient temperature for at least about 30 days. Longer shelf-life times include at least about 60 days, at least about 90 days, at least about 120 days, at least about 180 days, and ranges between any two of these values.

EXAMPLES

[0064] Example 1 : Construction of a conical anaerobic fermentation system

[0065] A conical style grow-out silo system (100) is shown in Figure 1. Screw conveyors can be used for moving both for inputs (101) and outputs (107). An input valve (102) can be used to control the addition of input materials into the system, as well as controlling the rate or amounts of input materials. A relief valve (103) can be used to prevent overloading of the system, as well as to allow release of gas or other pressure buildups. Input materials can flow from the input valve (102) to the conical silo (108). Monitoring of the fermentation can be performed using sensors such as a temperature sensor (104) and a pH sensor (105). An output valve (106) can be kept in a closed configuration during fermentation, and placed in an open configuration after fermentation to allow output material to move to the output screw conveyor

(107).

[0066] Example 2: Construction of a rectangular anaerobic fermentation system

[0067] A live bottom rectangular anaerobic fermentation system (200) is shown in

Figure 2. The system is suitable to handle high-solid and highly viscous materials. The system can include a straight vertical upper vessel part (206) connected to a live bottom part (211). The live bottom part (211) can include multiple screw augers (207) to facilitate handling and continuous discharge of material. The system can further include an input valve (202) and output valve (212), which when placed in a closed configuration can make the system water-tight and air-tight to achieve a desired anaerobic fermentation of input materials. A manhole (201) can be included to facilitate cleaning and maintenance of the system. A breath valve (205) can be used to allow release of built-up gasses from fermentation and prevent excess pressure from forming. The breath valve (205) can also be opened during discharge to prevent a vacuum from forming.

[0068] Accessories can be used to monitor and control the quality of the fermentation process. Example accessories include a dissolved oxygen sensor (204), temperature sensor (208), pH sensor (209), and sampling point (210).

[0069] Example 3: System process flows

[0070] Figures 6 and 7 show system process flows.

[0071] Figure 6 shows overall steps of system process flow (600) including loading

(601), mixing (602), fermentation (603), and discharge (604). Loading contacts raw material (605) and lactic acid bacteria inoculant (606). The raw material (605) and lactic acid bacteria inoculant (606) can be contacted before addition to the mixer (607), or upon addition to the mixer (607). The raw material (605) and lactic acid bacteria inoculant (606) can be added to the mixer (607) separately or together.

[0072] Mixing is performed in a mixer (607). Fermentation is performed in a fermentation silo (608) under anaerobic or anoxic conditions (609). Discharge includes delivering fermented feed (610) by feed dosing to larvae (611).

[0073] Figure 7 shows a system process flow (700) that includes additional upstream steps. Raw materials are received (701) and subjected to a QA/QC inspection (702). Materials are either approved (703) or rejected (704). Rejected materials are returned to the supplier (705) or destroyed. Approved materials are subjected to grinding (706), mixing (707), and are transferred to a fermentation silo (708) for fermentation (709). Fermented feed is dosed (710) onto larvae rearing trays.

[0074] Example 4: Side-by-side comparison of anaerobic and aerobic fermentation

[0075] Fermentation alters the pH of the larvae feed from an initial pH of greater than pH 5 to a lower pH of pH 4.38. Low pH values such as pH 4.38 inhibit the growth of pathogenic bacteria, undesirable yeast, and mold, and thus reduce proteolysis and control dry matter loss during fermentation. Table 1 shows the comparison of fresh feed without fermentation and feed after fermentation.

[0076] Table 1: Comparison of the fresh feed without fermentation and feed after fermentation

[0077] This table shows surprising, meaningful, and significant increase in pupae yield and performance from using fermented feed.

[0078] Example 5 : Comparison of lactic acid inoculant against non-lactic acid inoculant

[0079] Lactic Acid Bacteria known as “LAB” is gram-positive, microaerophilic, generally non-spore forming, either rod-shaped (bacilli) or spherical (cocci) bacteria that share common metabolic and physiological characteristics. The main function of LAB is to ferment water soluble carbohydrates to organic substances as their major end product. LAB generally belongs to the taxonomic group of Lactobacillales (order) and also genus Bifidobacterium, which are most widely recognized and applied as probiotics in food, beverage, and feed development. The LABs’ order Lactobacillales comprises all the members of all genera under the taxonomical Family Lactobacillaceae and also including other genera under the same order Lactobacillales, namely genus Leuconostoc, Pediococcus, Streptococcus, Enterococcus, Weissella, Oenococcus, Sporolactobacillus , Vagococcus, Tetragenococcus, and Lactococcus. The genus Bifidobacterium (phylum Actinomycetota) comprises all bifidobacterial strains that produce lactic acid as the major product of carbohydrate metabolism. The majority of silage LAB can grow under mesophilic conditions, including different temperatures ranging from 20-50 °C, the optimum temperature for growth is between 25 and 40 °C. They are naturally present in various resources such as fermented foods (meat, yogurt, kimchi), forage crops (alfalfa, corn, rye, sorghum and triticale), dairy products, rumen juice, and infant feces. [0080] Lactic acid bacteria inoculant is used to improve the larvae feed quality and prevent spoilage of the feed by increased production of organic acids, such as lactic acid and rapid decrease in pH. The fermented larvae feed with lactic acid inoculant is associated with prevention of spoilage microorganism, stabilization of feed temperature, prevention of nutrient loss, and minimization of the dry matter loss. Temperature is an important abiotic factor which influences the black soldier fly larvae performance.

[0081] Organic waste fermentation can result in high butyric acid, high acetic acid, or high lactic acid depending on factors such as moisture, inoculants, and protein content. The ratio of these organic acids indicates the quality of fermented substrates. Table 2 shows the comparison between three different types of fermentation and their respective outcomes in terms of concentration of acid produced and larvae yield.

[0082] Table 2: Comparison between different fermentation modes of insect feed and their output for larvae yield

SLD. = not detected

[0083] High butyric acid fermentation exhibits significant activities of Clostridial microorganisms which is undetectable in well-fermented insect feeds. However, they reduce dry matter and crude protein content of the feed with increased ammonia, thereby deteriorating feed digestibility for insect larvae. Due to the increased butyric acid production, the population of LABs and lactic acids are inhibited in the insect feeds.

[0084] The acetic acid produced in the insect feed fermentation could be beneficial depending on the ratio of acetic acid and lactic acid. Apart from some LABs being heterofermentative wherein they ferment carbohydrates into lactic acid and acetic acids, there exist groups of acetic acid which produce microorganisms. Such microorganisms also increase acetic acid concentration by converting lactic acids. High acetic acid concentration will reduce butyric acid and Clostridial population and detrimentally affect larvae growth performance. [0085] In the high lactic acid fermentation mode, the presence of LAB inoculants and the fermentation system assist in enriching lactic acid production in the insect feeds. If the ratio of lactic acid to acetic acid is high such as 3: 1, this will significantly reduce Clostridia population and butyric acid production. The LAB inoculants also reduce other competitive microbial populations including Clostridia and acetic acid producers. In addition, the LABs are tolerant to higher concentration of lactic acid throughout the fermentation period. The high lactic acid fermentation mode helps to improve the total yield of larvae, by increasing the feed bioconversion efficiency percentage nearly two times referencing from the high butyric acid and high acetic acid fermentation modes.

[0086] In a particular trial which investigated the effect of different inoculant on the black soldier fly larvae rearing temperature, it was observed that “Experiment 2” which used inhouse lactic acid inoculant recorded an average temperature fluctuation from 32.95 °C on the second day until 37.95 °C on the eighth, with the highest average temperature recorded on the seventh day at 41.25 °C. This group with the in-house lactic acid inoculant had the most hostile environment with less temperature fluctuations compared to the other group of experiments such as the Control with Commercial inoculant 1 and Experiment 1 with commercial inoculant 2 as shown in Figure 8. This temperature range is also suitable for the beneficial lactic acid bacteria growth compared to the other groups. When ingested, the higher beneficial lactic acid bacteria improved larvae performance and improved pupae performance.

[0087] Example 6: Description of benefits

[0088] There are an array of benefits of using a lactic acid inoculant for the larvae feed fermentation process. Firstly, the in house developed lactic acid inoculant was homofermentative thus, it rapidly reduces the feed pH through lactic acid production. Lactic acid fermentation prolongs feed stability by inhibiting feed pathogenic bacteria. Pathogenic bacteria such as Enterobacteriace, Clostridium perfringens, E.coli and Salmonella are undesirable in animal feed and the inclusion of lactic acid inoculant can inhibit the growth of these bacteria thus improving feed safety. The lactic acid fermentation also inhibits the growth of undesirable mold and yeast growth. This helps to improve the aerobic stability of the feed when exposed to air. The feed raw materials are predigested by the lactic acid bacteria and hence it is easier for the larvae to utilize the feed nutrition after the fermentation process. Besides that, through the fermentation process, the population of lactic acid bacteria will increase and contribute to increased bacterial biomass in the larvae feed. Bacterial biomasses are rich in nutrients such as crude protein, carbohydrate, and lipids which can be used by the larvae. The combined effects of various benefits help to improve the larvae growth as shown in Figure 9. Larvae fed fermented soy showed significantly higher average larvae weight than larvae fed unfermented fresh soy.

[0089] Example 7 : Characterization of finished feed product

[0090] The fermented larvae feed was characterized as shown in Table 3 below. The purpose of fermenting the raw material prior to feeding the larvae is to produce a stable feed with a high recovery of dry matter, energy, and highly digestible nutrients compared with the feed before fermentation. Microbial fermentation of the raw materials produces a wide range of beneficial end products such as organic acids and feed digesting enzymes and can prevent the nutrient deterioration. A well fermented larvae feed retains high moisture content, nutrient content and reduces the feed pH. It also has a slight acidic odor as a result of the fermentation process. An “as fed basis” refers to feed as normally fed to larvae.

[0091] Table 3: Characterization of fermented larvae feed as fed basis

[0092] Example 8: Description of black soldier fly feeding biology and rearing requirements

[0093] The lifecycle of black soldier flies lasts approximately 30-35 days: eggs (3 days), larvae stage (13-18 days), Pupae stage (10-14 days), and adult stage (5-8 days). The adult black soldier fly does not need to feed on anything during their short lifespan. Apart from the pupal stage, the larvae stage is the longest phase of the BSF life cycle and during this phase the larvae feed voraciously on a wide range of substrates. They efficiently convert the waste nutrients into larval biomass. The larval biomass nutritional values have been evaluated and compared to the animal protein sources used in animal feed. They rely on the fat stored during its larval stage for energy source after emerging into adult flies.

[0094] The voracious feeding nature and extraordinary ability to transform a wide range of organic waste materials into valuable protein and fat sources allows these larvae to become increasingly desirable for waste management purposes. The black soldier fly larvae have simple mouth parts which resemble a “tunnel boring machine”, where the hypopharynx separates finer organic particles from coarser and inorganic ones. High moisture diets which are semiliquid are most suitable to cater the simple feed ingesting mechanism of the black soldier fly larvae. Pretreatment processes such as grinding, crushing, and fermentation of the raw materials improves the feed structure and increases the feed digestibility by the black soldier fly larvae. The black soldier fly larvae adapt well to both continuous feeding and batch feeding methods. Ideal larvae rearing conditions such as a temperature range between 23 to 32 °C and 60-70% of relative humidity enhances the waste bioconversion process into larvae biomass.

[0095] Example 9: Feeding black soldier fly larvae

[0096] Figure 4 shows a feed dosing machine and black soldier fly tray. After fermentation of input materials in a fermentation silo is completed, the fermented feed is carried by a screw conveyor to be transferred to a feed input hopper (401). The feed dose pump (402) doses the feed with a predetermined feed dose value. Finally, larvae were placed on the surface of the dispensed feed in the growing tray (403) to begin their feeding activity.

[0097] 7 kg of the fermented feed was discharged from the dosing machine and filled into a 2160 cm ² (60 cm x 36 cm) plastic tray. Then, 20 grams of 5 days old larvae were added to the tray. The plastic trays filled with fermented feed were stacked up to 10-15 trays per stack and kept at ambient temperature for up to 9 days. The larvae were harvested after 9 days where the feed would have been fully explored and consumed by the larvae, leaving only the feed residues after 9 days. [0098] In the above detailed description, reference is made to the accompanying drawings, which form a part hereof. In the drawings, similar symbols typically identify similar components, unless context dictates otherwise. The illustrative embodiments described in the detailed description, drawings, and claims are not meant to be limiting. Other embodiments may be used, and other changes may be made, without departing from the spirit or scope of the subject matter presented herein. It will be readily understood that the aspects of the present disclosure, as generally described herein, and illustrated in the Figures, can be arranged, substituted, combined, separated, and designed in a wide variety of different configurations, all of which are explicitly contemplated herein.

[0099] The present disclosure is not to be limited in terms of the particular embodiments described in this application, which are intended as illustrations of various aspects. Many modifications and variations can be made without departing from its spirit and scope, as will be apparent to those skilled in the art. Functionally equivalent methods and apparatuses within the scope of the disclosure, in addition to those enumerated herein, will be apparent to those skilled in the art from the foregoing descriptions. Such modifications and variations are intended to fall within the scope of the appended claims. The present disclosure is to be limited only by the terms of the appended claims, along with the full scope of equivalents to which such claims are entitled. It is to be understood that this disclosure is not limited to particular methods, reagents, compounds, compositions or biological systems, which can, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

[00100] As used in this document, the singular forms “a,” “an,” and “the” include plural references unless the context clearly dictates otherwise. Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art. Nothing in this disclosure is to be construed as an admission that the embodiments described in this disclosure are not entitled to antedate such disclosure by virtue of prior invention. As used in this document, the term “comprising” means “including, but not limited to.”

[00101] While various compositions, methods, and devices are described in terms of "comprising" various components or steps (interpreted as meaning "including, but not limited to"), the compositions, methods, and devices can also "consist essentially of" or "consist of" the various components and steps, and such terminology should be interpreted as defining essentially closed-member groups.

[00102] With respect to the use of substantially any plural and/or singular terms herein, those having skill in the art can translate from the plural to the singular and/or from the singular to the plural as is appropriate to the context and/or application. The various singular/plural permutations may be expressly set forth herein for sake of clarity.

[00103] It will be understood by those within the art that, in general, terms used herein, and especially in the appended claims (for example, bodies of the appended claims) are generally intended as “open” terms (for example, the term “including” should be interpreted as “including but not limited to,” the term “having” should be interpreted as “having at least,” the term “includes” should be interpreted as “includes but is not limited to,” etc.). It will be further understood by those within the art that if a specific number of an introduced claim recitation is intended, such an intent will be explicitly recited in the claim, and in the absence of such recitation no such intent is present. For example, as an aid to understanding, the following appended claims may contain usage of the introductory phrases "at least one" and "one or more to introduce claim recitations. However, the use of such phrases should not be construed to imply that the introduction of a claim recitation by the indefinite articles "a" or "an" limits any particular claim containing such introduced claim recitation to embodiments containing only one such recitation, even when the same claim includes the introductory phrases "one or more" or "at least one" and indefinite articles such as "a" or "an" (for example, “a” and/or “an” should be interpreted to mean “at least one” or “one or more”); the same holds true for the use of definite articles used to introduce claim recitations. In addition, even if a specific number of an introduced claim recitation is explicitly recited, those skilled in the art will recognize that such recitation should be interpreted to mean at least the recited number (for example, the bare recitation of "two recitations," without other modifiers, means at least two recitations, or two or more recitations). Furthermore, in those instances where a convention analogous to “at least one of A, B, and C, etc.” is used, in general such a construction is intended in the sense one having skill in the art would understand the convention (for example, “ a system having at least one of A, B, and C” would include but not be limited to systems that have A alone, B alone, C alone, A and B together, A and C together, B and C together, and/or A, B, and C together, etc.). In those instances where a convention analogous to “at least one of A, B, or C, etc.” is used, in general such a construction is intended in the sense one having skill in the art would understand the convention (for example, “ a system having at least one of A, B, or C” would include but not be limited to systems that have A alone, B alone, C alone, A and B together, A and C together, B and C together, and/or A, B, and C together, etc.). It will be further understood by those within the art that virtually any disjunctive word and/or phrase presenting two or more alternative terms, whether in the description, claims, or drawings, should be understood to contemplate the possibilities of including one of the terms, either of the terms, or both terms. For example, the phrase “A or B” will be understood to include the possibilities of “A” or “B” or “A and B.” [00104] In addition, where features or aspects of the disclosure are described in terms of Markush groups, those skilled in the art will recognize that the disclosure is also thereby described in terms of any individual member or subgroup of members of the Markush group. [00105] As will be understood by one skilled in the art, for any and all purposes, such as in terms of providing a written description, all ranges disclosed herein also encompass any and all possible subranges and combinations of subranges thereof. Any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, tenths, etc. As a non-limiting example, each range discussed herein can be readily broken down into a lower third, middle third and upper third, etc. As will also be understood by one skilled in the art all language such as “up to,” “at least,” and the like include the number recited and refer to ranges which can be subsequently broken down into subranges as discussed above. Finally, as will be understood by one skilled in the art, a range includes each individual member. Thus, for example, a group having 1-3 cells refers to groups having 1, 2, or 3 cells. Similarly, a group having 1-5 cells refers to groups having 1, 2, 3, 4, or 5 cells, and so forth.

[00106] Various of the above-disclosed and other features and functions, or alternatives thereof, may be combined into many other different systems or applications. Various presently unforeseen or unanticipated alternatives, modifications, variations or improvements therein may be subsequently made by those skilled in the art, each of which is also intended to be encompassed by the disclosed embodiments.