The present invention relates to the Animal Nutrition and Physiology, particularly manufacturing of feed pellets and extruded floating feed pellets for poultry birds and fishes from protein flour and oil obtained from different insects.
Background of invention
Different plant proteins and animal proteins are used for manufacturing of poultry feed and fish feed. Insects are rich sources of proteins, lipids and essential nutrients and may be utilized for manufacturing of livestock, poultry and fish feed.
As it can fulfill protein and fat requirement up to 20%-50% but wholesome nutrition cannot be achieved to carry out physiological activities, as a result there is a need of a definite formula along with other feed component.
US20180317520A1 relates to methods for producing an insect product can include providing a dried cricket powder, providing a dried insect, or grinding wet-grinding, or shearing at least one insect into a paste or powder; adding water or an enzyme to the paste or powder to form a slurry; and filtering the slurry to produce an insect product.
The idea is to produce organic food for poultry birds and fishes using insects which are rich in protein and can be seen as a promising alternative to traditional protein source this will reduce environmental pollution and feed costs ( Bovera et al,2015 ., Hussein et al, 2017).
Insects are considered as a possible replacement as they have protein (30% -70%), fats (35%), minerals and vitamins (Borreli et al.2017). They have nutrient composition similar to soybean or fish meal (Schonborg et al.2017).
These insects flourish on organic waste such as vegetable residue, municipal organic waste and straws etc. This bioconversion reduces organic waste by 50 -60% and turning them into high protein biomass (Zheng et al.2013). This also causes reduction in greenhouse gases. Apart from this it is cost effective, natural and ecofriendly feed for animals. Insects share an evolutionary conserved innate immune system by producing anti-microbial peptides as a natural sustainable alternative to antibiotics in feed supplements, which offers significant benefit to the health of farmed animals and aquaculture. Also, the exoskeleton of insects made up of chitin can stimulate innate immune cells. Lauric acid present in insects have antiviral and antibacterial properties. So, using these insects as an animal feed will lead to involvement of local farmers to rear insects on their green waste which will improve their financial stability.
Summary of the invention
Accordingly, the present invention provides feed pellets and extruded floating feed pellets using insect protein and the process of manufacturing thereof.
The feed pellets and extruded floating feed pellets comprises: 30-70% insect hydrolysate; 5-20% insect frass; 2-5 % chitin biochar; 7-10% insect’s lipid; 20-30% carbohydrate source.
The process for preparing feed pellets and extruded floating feed pellets comprises the following steps:
i. rearing of insect larvae on different organic substrates;
ii. harvesting 5 instar black soldier fly larvae and 10-20 instar mealworm larvae and separating insect larvae from insect frass by means of sieve or screen vibrator; insect larvae is kept for fasting for 24-36 hour to clear their gastrointestinal tract of any residual food;
iii. freezing the reared and harvested insect larvae at -4 degree Celsius for 15-20 minutes;
iv. blanching insect larvae to minimize lipid oxidation and reduce microbial contamination of insects;
v. drying insect larvae at 50-55 degree Celsius to reduce moisture content;
vi. defatting insect larvae at 60-70 degree Celsius temperature;
vii. fermenting defatted insect larvae to obtain insect hydrolysate rich in amino acid and chitosan;
viii. preparing chitin biochar from chitin source by known methods and mixing it with the insect hydrolysate;
ix. mixing insect’s frass with the mixture of insect hydrolysate & chitin biochar;
x. adding carbohydrate source in the mixture;
xi. preparing feed pellets or extruded feed pellets of the mixture obtained;
xii. spraying insect lipid over the feed pellets or extruded feed pellets obtained;
xiii. drying lipid coated pellets at 50-55 degree Celsius to remove moisture for storage of the pellets obtained.
Objectives of the invention
Main objective of the present invention is to provide a composition of poultry and fish feed with high nutritional value and with antimicrobial properties.
Other objective of the present invention is to provide poultry and fish feed comprising protein and oil obtained from different insects.
Other objective of the present invention is to provide method of manufacturing feed pellets and extruded floating feed pellets for poultry birds and fishes.
Other objective of the present invention is to provide method of rearing insects using organic waste as a substrate.
Brief description of the drawings
Figure 1: Organic waste is utilized by different types of insects. In this bioconversion, protein and fat rich larvae are obtained along with excreta of insects which is known as insect frass. In further process protein and lipid are extracted by means of defatting process. Protein meal is further fermented to obtain insect hydrolysate.
Chitin biochar is manufactured by pyrolysis of insect exoskeleton and dead insects.
Insect hydrolysate is mixed with insect frass, carbohydrate and biochar to form extruded pellets or non extruded pellets. Insect lipid is sprayed over feed pellets.
Figure 2: Rearing of insects by utilizing organic waste. These insects are harvested at a specific maturation stage. After harvesting these insects are freezed to slow down the metabolism which is further blanched to reduce lipid oxidation and rancidity.
The blanched larvaes are dried to reduce moisture content. Dried larvae are defatted to separate lipid and protein component. Insect protein is fermented to obtain insect hydrolysate .
During rearing of insects some byproducts are generated, these are insect excreta known as insect frass and insect exoskeleton or dead insects.
Different insect frass is mixed according to their nutritional value. Dead insects or insect exoskeleton is utilized to manufacture chitin biochar by means of pyrolysis.
In final step insect hydrolysate is mixed with insect frass, biochar and carbohydrate source to manufacture extruded or non extruded feed pellets.
Detailed description of the invention
The present invention provides a composition of poultry and fish feed with high nutritional value and with antimicrobial properties. In some other aspects poultry and fish feed comprises of protein and oil obtained from different insects. In some other aspects present invention also provides method of manufacturing feed pellets and extruded floating feed pellets for poultry birds and fishes. Yet in other aspects present invention provides environment friendly method of rearing insects using organic waste as a substrate.
In an embodiment of the present invention organic waste is treated by insects. In another embodiment the protein and fat rich larvae are reared to obtain insect protein and oil. The insect production is recognized as a rich source of protein, fats, essential amino acids and various micro and macro nutrients. Rearing these insects from organic waste is also an environmentally friendly practice. The feed pellets prepared from insect protein and fat has many health benefits, insect protein is easily digestible. Insect oil is rich in omega 3 fatty acids, unsaturated fat and lauric acid which makes it an antimicrobial feed. Lauric acid is effective against many viruses as well.
In some other embodiments present invention also provides method of manufacturing feed pellets and extruded floating feed pellets for poultry birds and fishes. Insect protein and lipids based extruded and non-extruded pellets could be used as a feed material for poultry birds, fishes and other livestock. These pellets are not only economical and sustainable but also possess health benefits as lauric acid can increase immunity of animal.
In an embodiment the present invention provides feed pellets and extruded feed pellets from insect protein and insect lipid which can replace antibiotics in livestock farming.
In an embodiment of the present invention, the basic components in a formulated feeds are insect hydrolysate; insect frass; chitin biochar; insect’s lipid; carbohydrate source.
In another embodiment the present invention provides a process for preparing feed pellets and extruded floating feed pellets.The process of manufacturing of extruded feed pellets for fish using insect protein, includes following steps:-
1- Rearing of insects on different substrates.
2- Harvesting of Insects
3- Freezing of insects
4- Blanching of insect
5- Drying of insects (Dehydration)
6- Defatting of insects
7- Manufacturing of insect hydrolysate from insect meal
8- Manufacturing of Biochar from insect exoskeleton
9- Mixing of insect’s frass/ excreta along with other ingredients.
10-Extrusion of pellets
11-Spraying of insect oil
12-Drying of pellets
In an embodiment of the present invention insect-based protein sources are used. Insects can be grown over different organic waste so this kind of protein source is economical & sustainable. The reason behind utilizing insect protein is presence of antimicrobial peptides which are antibacterial, antiviral and antifungal. Crude protein content of insects depends on the insect species.
In an embodiment the present invention utilizes combination of protein meal obtained from black soldier fly and mealworm to enhance crude protein level in feed pellets and extruded feed pellets similar to the fish meal. The combination of different species of insect meal is also important to balance amino acid profile required for a specific livestock feed. When the fish meal was replaced by whole insect meal (meal worm meal and black soldier fly meal) the growth of the livestock animal was affected and the animals became slightly resistant to diseases compared with control group. Inclusion of black soldier fly and meal worm combination can reduce the need for mineral supplementation in feed formulation.
In another embodiment of present invention fermentation of insect protein is carried out utilizing bacterial enzymes like protease , chitinase and chitosanase released during process of fermentation degrades chitin in to chitosan and other beneficial products which can enhance immunity of the animal . Microbes used in this fermentation are Aspergillus niger, L.buchneri, L.fermentum , Bacillus subtilis. Hydrolysis of protein also causes the release of amino acid, breakage of peptide bonds and release of antimicrobial peptides in insect meal.
In another embodiment of present invention chitin biochar is added to the feed, it helps in better absorption of amino acids and cationic peptides. The chitin biochar is produced by pyrolysis of exoskeleton of insects as they are rich in chitin. The biochar manufactured from chitin have more absorbing capacity. Chitin biochar will aid in absorption of amino acids like lysine which is a cationic charge amino acid and most important for growth. Chitin biochar will also absorb cationic charge antimicrobial peptides like cecropin.
In another embodiment present invention utilizes insect oil/lipids instead of fish oil or vegetable oil. Insect oil is rich in omega fatty acids and Lauric acid which is a potent antiviral agent. The metabolites released from lauric acid can be used as an immediate form of energy by extrahepatic tissues.
Lauric acid is converted in to monolaurin which can reduce stress and inflammation. It is also effective against various viruses, bacteria and fungi. Unsaturated fats are more prone for rancidity and Black soldier fly lipid are rich in saturated fat (70%).
To reduce rancidity Blanching of insects is a better option. Lipid present in black soldier fly is slightly rancid their TBARS value is less than 3.6 MDA/kg (Jennifer Larouche et al 2019). Blanching of insect can reduce this value to 1 MDA /kg.
In another embodiment present invention includes carbohydrate source like rice bran, wheat bran ,maize or de oiled cake can be used. These are important for energy source.
In another embodiment present invention includes insect frass. Insect frass is a left over residue of organic substrate and excreta of insects. It is rich in chitin ,protein and lipids. Insect frass of both insects (BSF and mealworm) is rich in protein. Mealworm excreta is rich in linoleic acid ,which is very much important in fish feed. It is also rich in omega 6 fatty acid which is also necessary for fish nutrition.(Nergui ravzanaadii et al 2012).
Insect frass also acts as a probiotic as Zhao et al (2017) concluded in his study that black soldier fly larvae can transfer actinobacteria ,proteobacteria from gut to biowaste i.e. frass.(storelli et al 2018, vogel et al 2018, cheng Liang Jang, Wei Zheng Jin et al 2019)
Actinobacteria acts as a probiotic in aquaculture. So the frass of black soldier fly will enhance the probiotic effect. Inclusion of frass / excreta of black soldier fly larvae and meal worm can enhance immunity and growth of livestock animals. It will also reduce the production cost of feed formulation.
In another embodiment present invention insect frass and other feed ingredients are mixed with insect hydrolysate/fermented from insect meal.
In another embodiment present invention includes insect lipids. Mold like Aspergillus Flavus contaminate the feed with alfatoxins. Lauric acid and mysteric acid present in black soldier fly lipid is effective against this fungus. So it also enhances shelf life of feed.
In another embodiment present invention organic waste is utilized by insects to convert carbon content into protein and fat. This reduces not only the cost of protein source required for animal feed but also reduces carbon footprint. Different type of insects can be grown in different organic waste substrate.
In another embodiment present invention utilises waste from grain industries i.e. wheat bran for rearing mealworms, fruit & vegetables waste and restaurant waste for rearing black soldier fly larvae (BSFL).
In another embodiment of present invention 5 instar BSF larvae’s are harvested and separated from insect frass by means of sieve or screen vibrator. Insect are kept for fasting for 24-36 hour to clear their gastrointestinal tract of any residual food.
Harvested larvae can’t be kept for long time as they can proceed to their next stage of their life cycle i.e. prepupae. So immediate processing of insects is required. Insects are kept for deep freezing at -4 degree Celsius for 15-20 minutes to slow down the metabolism of insects but prolong freezing can deteriorate the lipid content and protein of insects.
In another embodiment of present invention both insects BSFL and meal worm larvae are freezed by same procedure.
In another embodiment of present invention the insects are blanched, blanching of insects minimizes lipid oxidation and reduces microbial contamination of insects.
In another embodiment of present invention insects are dried at 50-55 degree Celsius to reduce moisture content. Blanched BSF larvae and mealworm larvae are highly perishable due to their high water content and hence insects are dried to reduce moisture content and to increase crude protein. This step is rate limiting as drying insects at high temperature can cause denaturation of protein.
In another embodiment of present invention defatting of insects is done by either chemical process using soxhlet method or by mechanical extraction.
In another embodiment of present invention screw press method is used for defatting at 60-70 degree temperature. The crude lipid extracted from this method is further utilized in production of livestock feed.
In an embodiment of present invention extruded feed pellets are prepared. Extruded pellets are required for proper gelatinization of feed so that it can float in the water. As floating feed is required for various fishes like pangasius and tilapia.
Moisture plays an important role during extrusion when insect protein content is high the moisture requirement is also increased. Generally 12-15 % moisture is required for extrusion. For extrusion process defatting is essential as the lipid content in the mixture can decrease the pressure inside the extrusion screw resulting in degradation of pellet quality.
In another embodiment of present invention, after extrusion of pellets, optionally lipid coating is done according to the animal type by simply spraying of melted lipid/oil over pellets. Coating of insect oil increases palatability, feed intake and floatability of feed pellets
In another embodiment of present invention, lipid coated pellets are dried at 50-55 degree Celsius to remove moisture for storage.
In another embodiment of present invention, non extruded pellets are prepared.
Novelty in the process:- The BSFL and mealworm larvae protein are promising alternative of protein, but their amount of nutritional component along with anti nutritional factor i.e chitin, changes with the maturity of insect . After a certain maturity of age according to their life cycle they undergo metamorphosis and convert into different life stages like pupae and fly/bettle. They need to be processed soon to maintain nutritional quality and to store Insect meal for long time. To maintain nutritional quality and to store insects for long time the life cycle of insects was delayed. To prevent pupation of mealworm they are shifted to lower temperature zone (at 4-6 degree temperatures) to slow down the metabolism of these insects. Further they are shifted to favorable condition few days before their harvesting.
Similarly to store BSF larvae for long time and to slow down the life cycle of black soldier fly larvae of 2 instar to 3 instar stage, fermented fruit juice (FFJ) with fermented vegetables and fruits is used and the container is shifted to dark and cooler zone. In need of more insect meal ,the BSF containers are shifted to warmer zone and substrate rich in protein like wheat bran or restaurant waste is fed until they reach to the size of 5th instar larvae. Larvae is harvested at this stage. BSFL were successfully stored for 3 months by regulating their lifecycle through this process.
The following examples are given by way of illustration of the working of the invention in actual practice and should not be construed to limit the scope of the present invention in any way.
EXAMPLES
EXAMPLE 1 Combination of insect meal:- The crude protein of mealworm is 55 % and crude protein of black soldier fly larvae is 45 % , the combination of both meal at a ratio of 50:50 will result in 50% crude protein in the final meal.
Similarly if the Lysine content of meal worm is 5.4 g and lysine content of black soldier fly meal is 6.6 g the overall lysine content after combination would be 6 g.
Another protein source used in our feed is insect frass or excreta, This is also rich in protein. Crude protein content of insect frass depends on feed substrate utilized for insect rearing and it ranges from 12-20% crude protein content.
Addition of insect frass will reduce feed cost and it has few health benefits too.
EXAMPLE 2 Clinical results:-
Sample size:-1000 Pangasius fish
Groups: In first trial
A) Control group (Market feed containing 40% fish meal protein)
B) Insect meal (black soldier fly and mealworm) (IM1)
C) Insect meal hydrolysate +Chitin biochar (IM2)Description Control IM1 IM2
Maintained Water parameter
DO 5 ppm
Ammonia 0.5-2.5ppm Normal growth

No mortality Poor growth in comparison to control group

No mortality Similar growth like control group

No mortality

Description Control IM1 IM2
Poor water parameter
DO 2-3 ppm
Ammonia 6-8 ppm Reduced feed intake

40% mortality due to bacterial and fungal diseases. Proper feeding

5% mortality Proper feeding

0.5% mortality
No clinical sign of stress or disease present

Second trial groups:-
A) Insect hydrolysate + Chitin Biochar (IM2)
B) Insect hydrolysate + chitin Biochar + insect frass (IM3)
C) Insect hydrolysate + chitin biochar +insect frass + insect oil coating (IM4)
Sample size 400 pangasius fish
Water parameter: - Very poor (dissolved oxygen 2-3 ppm, ammonia 4 -6 ppm)
Benefits IM2 IM3 IM4 Control
Feeding habit No change No change Increased feed intake
More palatable Decreased feed intake
Floating capacity of extruded feed No change No change Floating time increased Better floating time than IM2 & IM3 feed
Growth Similar growth to control group Better than control group Similar to IM3 Normal growth
Energy of fishes Better than control group Better than control group Energy is better than all groups Lethargic , low energy
Mortality 1% mortality No mortality No mortality 30% mortality
Due to bacterial and fungal diseases.

Third Experiment:- Shifting of infected fishes in IM4 group
Result: - No mortality from IM4 group only mortality was from the shifted fishes
Important:- Aeromonas hydrophila is most common pathogen is fish diseases and inclusion of insect oil (black solodier fly) in formulated feed is increasing lauric acid in feed. Lauric acid is a potent anti bacterial agent against aeromonas hydrophila. Additionally probiotic effects of chitin and Antimicrobial peptide effects are increasing immunity of animals.
Results seen in clinical trial:

Poultry :- Better growth , less mortality , better resistance to diseases.

Fishes:-
a. Less than 1% mortality in comparison to control group where 50% mortality was seen when the parameters of water were kept extremely unfavorable(Dissolved oxygen 2-3 ppm, Ammonia 6-8 ppm).
Mortality in control group was due to different fungal ,bacterial and viral diseases which was result of poor water condition. But Insect meal group fishes were resistant to these pathogens and stress despite of poor water quality.

BSF Group fish showed no mortality even after shifting infected fishes from control group to one BSF group. This shows higher immunity of BSF group fishes.

The experiment was done in winter session at a maintained temperature of 20-24 degree Celsius.

b. Feed intake of control group was less than BSF group.
It shows that BSF feed is more palatable and increases feed intake.

c. The fish from control group were lethargic with low energy While fishes from BSF group are more active .

d. The growth of fingerling of BSF group is similar to control group that too when BSF group feed contain 30% protein in comparison to 40% protein feed (fish meal) in control group
Details of Biological Material
Black Soldier Fly (Hermetia illucens)
Family:- Stratiomyidae.
Genus:- Hermetia
Species: H.illucens
Black soldier fly is native of tropical region but in recent decades it is indigenous in all continents. In India black soldier fly larvae can be collected from compost sites of organic waste or manure piles.
Identification:-
Size of adult black soldier fly varies from 15 mm to 20 mm.
Color of adult black soldier fly :- Black with metallic reflection
It has one pair of wings and two translucent window spots located in abdominal region. The legs are black with whitish tarsi
Black soldier fly larvae are more commonly detectable than adult fly from compost sites of organic waste.
Size of larvae varies as the larvae matures and can reach up to 27 mm in length and 6mm in width.
The color of black soldier fly larvae varies from dull white to black (pre pupae) depending on maturation stage
Unlike round shape of house fly larvae the black soldier fly larvae are bigger in size and slightly flat in shape with segmented markings over body surface describing stages of life cycle.
Mealworm Larvae
Mealworms are larvae stage of mealworm beetle (Tenebrio molitor) a species of darkling beetle.
Mealworm larvae are yellowish in color with brownish bands over the body and around 2.5 cm long , have a segmented body with six legs and two antennae.
The adult mealworm is dark brown to black in color and is 12 to 25 mm long.
Mealworms are edible insects and are practiced in entomology in various regions
In the present invention, the black soldier fly larvae and pre pupae were collected from dumped sites for rotten fruits and vegetables at vegetable markets.
Then the pre pupae and mature larvae were placed to a dark and dry place to convert them in to pupae stage.
Pupae were shifted to breeding area which was made up of mosquito net. After pupae stage flies emerges out for breeding. The required environment for breeding are 60-70% humidity, sunlight and rotten food waste. After 6-7 days eggs were collected from corrugated cardboard placed at food waste. Then the eggs were allowed to hatch for further breeding cycle.
Mealworms are edible insects and are used as a live feed for ornamental fishes and birds.
These are available at aquarium shops.
We bought 20,000 larvae from aquarium shop for breeding mealworms.

Claims
I/We claim:
1. Feed pellets and extruded floating feed pellets using insect protein and the process of manufacturing thereof; wherein the feed pellets and extruded floating feed pellets comprises:
a. 30-70% insect hydrolysate;
b. 5-20% insect frass;
c. 2-5 % chitin biochar;
d. 7-10% insect’s lipid;
e. 20-30% carbohydrate source.

2. The feed pellets and extruded floating feed pellets as claimed in claim 1 wherein the insect selected are black soldier fly and mealworm.

3. The feed pellets and extruded floating feed pellets as claimed in claim 1 wherein the carbohydrate source is selected from group comprising maize bran, wheat bran, rice bran or de-oiled cake and the mixture thereof.

4. A process for preparing feed pellets and extruded floating feed pellets as claimed in claim 1characterized in that said method comprises the following steps:
xiv. rearing of insect larvae on different organic substrates;
xv. harvesting 5 instar black soldier fly larvae and 10-20 instar mealworm larvae and separating insect larvae from insect frass by means of sieve or screen vibrator; insect larvae is kept for fasting for 24-36 hour to clear their gastrointestinal tract of any residual food;
xvi. freezing the reared and harvested insect larvae at -4 degree Celsius for 15-20 minutes;
xvii. blanching insect larvae to minimize lipid oxidation and reduce microbial contamination of insects;
xviii. drying insect larvae at 50-55 degree Celsius to reduce moisture content;
xix. defatting insect larvae at 60-70 degree Celsius temperature;
xx. fermenting defatted insect larvae to obtain insect hydrolysate rich in amino acid and chitosan;
xxi. preparing chitin biochar from chitin source by known methods and mixing it with the insect hydrolysate;
xxii. mixing insect’s frass with the mixture of insect hydrolysate & chitin biochar;
xxiii. adding carbohydrate source in the mixture;
xxiv. preparing feed pellets or extruded feed pellets of the mixture obtained;
xxv. spraying insect lipid over the feed pellets or extruded feed pellets obtained;
xxvi. drying lipid coated pellets at 50-55 degree Celsius to remove moisture for storage of the pellets obtained.
6. The process for preparing feed pellets and extruded floating feed pellets as claimed in claim 5 wherein the organic substrate is selected from group comprising waste from grain industries, fruit & vegetables waste and restaurant waste and mixture thereof.
7. The process for preparing feed pellets and extruded floating feed pellets as claimed in claim 5 wherein the microbes used in the fermentation are Aspergillus niger, L.buchneri, L.fermentum , Bacillus subtilis.

8. The process for preparing feed pellets and extruded floating feed pellets as claimed in claim 5 wherein the defatting of insects is done by soxhlet method or mechanical method at 60-70 degree Celsius.

9. The process for preparing feed pellets and extruded floating feed pellets as claimed in claim 5 wherein the chitin source is insect exoskeleton.