Pet feed containing cockroach ground powder

Abstract

The invention discloses pet feed containing cockroach ground powder, and belongs to the technical field of pet food. The content of the cockroach ground powder in the pet feed is 4%-16%. The pet feed added with the cockroach ground powder can adjust the intestinal flora balance of pets, maintain the integrity of intestinal barriers, reduce the permeability of intestinal mucosa and adjust the CD4 + / CD8 + cell proportion of mesenteric lymph nodes, and has a very positive effect on the health of pets.

Description

Technical Field

[0001] The invention relates to the technical field of pet food, in particular to a pet feed containing cockroach powder. Background Art

[0002] With the development of economy, people's living standard has been greatly improved, keeping pets is becoming more and more common, and pets have become close companions of human beings. Pets bring a lot of happiness to human beings, but pets are most likely to have digestive system and immune system diseases in the process of keeping. For example, cats and dogs are very prone to diarrhea, abdominal pain and vomiting, loose and sticky stools, and foul odor. Cats with such diseases have gastrointestinal disorders, poor immunity, slow growth and development, rough and messy fur, and malnutrition. Pet feed can regulate and improve the intestinal flora and immune balance of pets through "diet therapy" to maintain the health of pets.

[0003] The cockroach (Periplaneta americana L.) is one of the insects of the genus Periplaneta in the family Blattidae. It is called cockroach, oil thief, stove pheasant, etc. in folk culture and is considered one of the four pests. However, as a traditional Chinese medicine, cockroach plays an important role in the treatment of diseases. It was recorded as early as more than 2,000 years ago. The "Shennong's Herbal Classic" records that cockroach "treats blood stasis, hard symptoms, cold and heat, breaks up accumulation, and throat and pharyngeal paralysis". The "Compendium of Materia Medica·Insects·Cockroach" records that its main treatments are "blood stasis, hard symptoms, cold and heat, lowering qi, and benefiting blood vessels". Modern Chinese medicine research has found that cockroach ingredients contain active ingredients such as glycoproteins, proteins, polypeptides, amino acids, nucleotides, and trace elements. Cockroach extracts have the effects of protecting the liver, promoting wound healing, anti-oxidation, anti-tumor, anti-inflammatory, and regulating gastrointestinal flora.

[0004] The gastrointestinal tract is a place where complex microorganisms live in animals and humans. The community in the gastrointestinal environment is the intestinal flora. The intestinal flora will have a significant impact on the host during the occurrence of various diseases. Therefore, in the entire life cycle of animals and humans, the intestinal flora plays a vital role in metabolic homeostasis and maintaining immunity. Microorganisms bring many benefits to the host through a series of physiological functions, such as preventing pathogens, regulating the body's immune ability, increasing the integrity of the intestinal barrier, and regulating substance metabolism. Changes in the composition and structure of intestinal microorganisms are related to the pathogenesis of many diseases. The imbalance between beneficial and harmful microorganisms can lead to the development of a variety of chronic diseases, such as obesity, diabetes, colitis and cancer. At the same time, microbial byproducts also have an impact on the integrity of the intestinal epithelium, and the imbalance of byproducts will directly lead to the accelerated onset of chronic diseases. There are currently no reports on the use of cockroaches in pet feed. Summary of the invention

[0005] The purpose of the present invention is to provide a pet feed containing cockroach powder to solve the problems existing in the above-mentioned prior art.

[0006] To achieve the above object, the present invention provides the following solutions:

[0007] One of the technical solutions of the present invention is a pet feed containing cockroach powder, wherein the content of the cockroach powder in the pet feed is 4% to 16%.

[0008] Based on the above technical solution, the present invention has the following technical effects:

[0009] The pet feed added with cockroach powder provided by the present invention can regulate the balance of pet intestinal flora, maintain the integrity of intestinal barrier, reduce intestinal mucosal permeability, regulate mesenteric lymph node CD4 + / CD8 + Cell ratio plays a very positive role in the health of pets. BRIEF DESCRIPTION OF THE DRAWINGS

[0010] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0011] Figure 1 The changes of ALT (A) and AST (B) levels in rat serum liver function indicators. Note: One-way ANOVA was used to compare sample data between groups; compared with the basic feed group, *P<0.05, **P<0.01, ***P<0.001. Same below.

[0012] Figure 2 This is the HE staining result of rat liver tissue (200×).

[0013] Figure 3 The growth performance of rats in each group is shown in Figure 1. A is the final body weight, B is the feed intake, C is the specific growth rate, D is the liver-to-body ratio, and E is the intestine-to-body ratio.

[0014] Figure 4 The effect of cockroach powder on the content of SCFAs in rat feces. Among them, A is acetic acid, B is propionic acid, C is butyric acid, and D is valeric acid.

[0015] Figure 5 The effect of cockroach powder on spleen and thymus index of rats.

[0016] Figure 6 To study the effect of cockroach powder on IgA (A), IgG (B) and IgM (C) in rat serum.

[0017] Figure 7 CD4 + 、CD8+ Flow cytometry plots.

[0018] Figure 8 The effect of cockroach powder on T lymphocyte subsets in rat peripheral blood. + , B is CD8 + , C is CD4 + / CD8 + . DETAILED DESCRIPTION

[0019] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0020] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0021] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0022] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present application description and examples are exemplary only.

[0023] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0024] The technical solutions described in the present invention, unless otherwise specified, are all conventional solutions in the art, and the reagents or raw materials used, unless otherwise specified, are purchased from commercial channels or have been disclosed.

[0025] The embodiment of the present invention provides.

[0026] In some specific embodiments, a pet feed containing cockroach powder, wherein the content of the cockroach powder in the pet feed is 4% to 16%.

[0027] In some specific embodiments, the cockroach powder is contained in the pet feed in an amount of 8% to 16%.

[0028] In some specific embodiments, the cockroach powder is contained in the pet feed in an amount of 12%.

[0029] In some specific embodiments, the pet feed also includes 10% to 16% fish meal, 5% chicken meal, 29% to 35% flour, 5% oatmeal, 30% soy fiber powder, 2% chicken fat, 0.5% multivitamin premix, 2% mineral element premix and 0.5% glycoprotein.

[0030] In some specific embodiments, the pet feed also includes 10% to 12% fish meal, 5% chicken meal, 29% to 35% flour, 5% oatmeal, 30% soy fiber powder, 2% chicken fat, 0.5% multivitamin premix, 2% mineral element premix and 0.5% glycoprotein.

[0031] In some specific embodiments, the pet feed also includes 10% fish meal, 5% chicken meal, 33% flour, 5% oatmeal, 30% soy fiber powder, 2% chicken fat, 0.5% multivitamin premix, 2% mineral element premix and 0.5% glycoprotein.

[0032] The pet food or pet feed added with cockroach powder provided by the present invention is beneficial to regulating the imbalance of pet intestinal flora, improving pet intestinal health, and improving immune function and growth performance. The cockroach-added feed of the present invention can be used in livestock and pet feed, and has the advantages of high protein, amino acid, and nucleotide content and low heavy metal content (consistent with the heavy metal limit of animal medicinal materials in the Chinese Pharmacopoeia).

[0033] The cockroach powder of the present invention has the following requirements for ingredients: total protein content ≥65%, crude fat content ≤15%, chitin content ≤15%, free amino acid content calculated as alanine ≥7%, nucleotides calculated as creatinine, uracil, xanthine and hypoxanthine ≥2 mg / g, total peptide content ≥12%. The main heavy metal limits in the cockroach powder are: arsenic ≤1 mg / kg, mercury ≤0.2 mg / kg, lead ≤1 mg / kg, and cadmium ≤0.3 mg / kg.

[0034] The present invention finds that the medicinal insect cockroach has a good regulating effect on the intestinal flora of animals, and can provide animals with high-quality insect protein, amino acids, nucleotides and other life active substances, so it can be added to animal feed as a high-quality additive. However, the amount of addition directly affects the regulating effect on the intestinal flora and immunity of animals. If the addition ratio is low, the regulating effect is not obvious; if the addition ratio is too high, the cockroach medicinal material contains about 15% chitosan (chitosan is a polymer of N-acetylglucosamine), which will consume a large amount of gastric acid and enzymes during digestion, increasing the gastrointestinal burden of animals. At the same time, the addition of cockroach powder is too high, which will also increase the feed cost. Therefore, the present invention finds the appropriate range of the addition ratio of cockroach powder in pet food through research, which is 4% to 16%.

[0035] Example 1

[0036] 1Animal feeding and feed formulation

[0037] The experiment used SD rats weighing about 200 g, half male and half female. The rats were kept in separate cages, fed with cold boiled water, cleaned regularly and replaced with rat bedding. The experimental animal use and management principles were followed during the experiment.

[0038] After rats were adapted to the basic feed for one week, 48 rats were randomly divided into 6 groups, namely the basic feed group, 4%, 8%, 12%, 16% and 20% groups, with 8 rats in each group. The basic feed group was fed with basic pet food without cockroach powder, and the other 5 groups were fed with cockroach powder at a mass fraction of 4%, 8%, 12%, 16% and 20% on the basis of the basic feed, respectively. The experimental period was 30 days. The pet food formula is shown in Table 1. During the experiment, the changes in the growth performance, intestinal flora, short-chain fatty acids (SCFAs) and immune indicators of the rats were statistically observed.

[0039] Table 1 Feed formula (dry weight, %)

[0040]

[0041]

[0042] All feed raw materials are crushed, passed through a 40-mesh sieve, and evenly mixed according to the feed formula (Table 1). Then, an appropriate amount of purified water is added and fully mixed. A feed pelletizer is used to make pellet feeds of different particle sizes, which are then dried at low temperature and sealed in bags.

[0043] 2 Effects of cockroach powder on rat liver

[0044] ALT and AST, important indicators for clinical evaluation of liver function, are mainly accumulated in hepatocytes. When liver damage occurs, the hepatocyte membrane is destroyed, causing a large amount of ALT and AST to be transferred from hepatocytes to the blood.

[0045] The results are shown in Table 2 and Figure 1 Compared with the basic feed group, there were no significant differences in ALT and AST in the cockroach powder-added groups, indicating that cockroach powder had no effect on the liver function of rats.

[0046] Table 2 Effect of cockroach powder on ALT and AST levels in rat serum liver function indicators

[0047]

[0048] observe Figure 2 The results of HE staining of rat liver pathological tissue showed that the basic feed group and the groups supplemented with cockroach powder showed that the hepatocytes were uniform and clearly structured, arranged in an orderly manner, the liver structure was obvious, there was no inflammatory cell infiltration, and no pseudolobule formation, indicating that cockroach powder had no effect on rat liver tissue.

[0049] Effect of cockroach powder on growth performance of rats

[0050] The palatability of pet feed containing cockroach powder was demonstrated by observing the feeding desire of each rat, and the feeding speed and amount were comprehensively evaluated, with performance such as rapidity, hesitation, and slowness as indicators for evaluation of excellence.

[0051] The results are shown in Table 3 and Figure 3 As shown, the feed intake of the 8%-16% group was 93.7±3.1%, 97.2±2.2% and 91.9±1.3%, respectively, and the feeding desire was evaluated as excellent. The specific growth rate, liver-body ratio and intestine-body ratio were better than those of the basic feed group, and there was no loose stool. The feed intake of rats in the basic feed group was 71.8±4.1%, and 3 of the 8 rats had loose stool. The feed intake of rats in the 4% and 20% groups was 89.9±1.7% and 70.5±3.2%, respectively, and the specific growth rate, liver-body ratio and intestine-body ratio were better than those of the basic feed group. The rats had smooth fur, good activity, and no loose stool, but the feeding desire of the rats was evaluated as good, which was weaker than that of the 8%-16% group. The rats in the 20% group had a good feed intake in the first two days of feeding, but the feed intake began to decline from the third day. The reason was that the amount of cockroach powder added in the 20% group was too high. Cockroaches contain about 15% insect chitosan. Excessive intake of chitosan consumes a large amount of gastric acid, which affects the digestion of animals. This shows that the amount of cockroach powder added in the pet food is too high. Compared with the existing pet feed, the 4%-16% group has a better palatability of the formula of the present invention. The rats have oily fur, are lively and active, have good feces molding, good growth, and no loose stools.

[0052] Table 3 Effects of different contents of cockroach powder on the growth performance of rats in each group

[0053]

[0054] 4. The regulatory effect of cockroach-added feed on intestinal flora

[0055] The intestinal microecology is composed of intestinal flora and its metabolites as well as the intestinal mucosal immune system. Under normal circumstances, the intestinal flora is in a state of dynamic balance. Studies have shown that intestinal flora imbalance may lead to the destruction of the intestinal barrier, causing the body's absorption and metabolic disorders, and inducing various diseases, including chronic obesity, diabetes, and atherosclerosis. The ability of cockroach powder pet feed to regulate intestinal flora and improve intestinal function was studied by measuring the changes in the structure and abundance of intestinal flora in intestinal contents through the 16S rRNA of rat intestinal microorganisms, and studying the effects of cockroach powder addition on rat intestinal flora and its metabolites.

[0056] According to the species composition analysis, the community structure of rat intestinal flora can be analyzed at various taxonomic levels. In this experiment, the type and quantity of flora contained in rat fecal samples at a certain taxonomic level were analyzed at the phylum, family and genus levels.

[0057] At the phylum taxonomic level, there were 24 phyla of bacteria, among which Firmicutes and Bacteroidetes were the main flora in the colonic feces of rats, accounting for more than 90% of the total intestinal microorganisms. In addition to Firmicutes and Bacteroidetes, there were 6 phyla with an abundance of more than 0.03%, which were ranked from large to small in terms of flora abundance: Actinobacteria, Spirochaetae, Proteobacteria, Saccharibacteria, Tenericutes, and Cyanobacteria. The proportions of Firmicutes and Bacteroidetes in the total intestinal microorganisms in the basal diet group were (83.86%) and (1.28%), respectively. After intervention with cockroach powder added to feed, the proportion of Firmicutes and Bacteroidetes in the total intestinal microorganisms in the 4% group was (88.80%), and the proportion of Firmicutes and Bacteroidetes in the 8% group was (91.97%), and the proportion of Bacteroidetes in the 12% group was (93.48%), and the proportion of Bacteroidetes in the 16% group was (88.69%), and the proportion of Bacteroidetes in the 20% group was (78.95%), and the proportion of Bacteroidetes in the 20% group was (78.95%), and the proportion of Bacteroidetes in the 20% group was (1.56%).

[0058] Compared with the basal diet group, the abundance of Firmicutes in the 4%, 8%, 12%, 16% and 20% groups increased by 4.94%, 8.11%, 9.62%, 4.83% and 4.91%, respectively, and the abundance of Bacteroidetes increased by 3.89%, 2.09%, 4.34%, 0.33% and 0.28%, respectively.

[0059] In order to further understand the effect of cockroach addition to feed on the intestinal flora of rats, the intestinal flora structure of rats was analyzed at the family and genus levels. First, at the family taxonomic level, a total of 163 families of microorganisms were obtained. The main dominant bacteria in the intestinal flora of rats were Lactobacillaceae, Ruminococcaceae, Clostridiaceae_1, Peptostreptococcaceae, Bacteroidales_S24-7_group, Lachnospiraceae, Erysipelotrichaceae, Spirochaetaceae, Bifidobacteriaceae, Enterococcaceae, Staphylococcaceae, etc. After the intervention of adding cockroach powder to feed, the relative abundance of probiotics Ruminococcaceae, Bifidobacteriaceae and Lactobacillaceae increased in the 4%, 8%, 12% and 16% groups, and the relative abundance of pathogenic bacteria Enterococcaceae and Staphylococcaceae decreased. This indicates that the abundance of each dominant bacterial community changed after feeding with cockroach powder added to feed, and cockroach powder added to feed can change the structure and abundance of rat intestinal flora at the family level.

[0060] Effect of cockroach powder on SCFAs content in rat feces

[0061] SCFAs are metabolites of intestinal flora and can bind to receptors distributed in the intestine, adipocytes, and immune cells, thereby promoting intestinal balance, increasing fat decomposition, and regulating the immune system. In this experiment, GC-MS was used to determine the content of SCFAs in rat feces.

[0062] The results are shown in Table 4 and Figure 4As shown in the results, the contents of acetic acid, propionic acid, butyric acid and valeric acid in the feces of rats in the 4%, 8%, 12% and 16% groups were significantly higher than those in the basic feed group, and the differences were statistically significant (P<0.05). The contents of acetic acid, propionic acid, butyric acid and valeric acid in the 4% group were also higher than those in the basic feed group, but there was no statistical significance for propionic acid and valeric acid. The contents of acetic acid, propionic acid, butyric acid and valeric acid in the feces of rats in the 20% group showed an increasing trend compared with those in the basic feed, but the differences were not statistically significant. This indicates that a dose of 4% to 16% cockroach powder can increase the content of SCFAs in rat feces and has a certain protective effect on the intestinal barrier of rats.

[0063] Table 4 Effect of cockroach powder on SCFAs content in rat feces ( n=5)

[0064]

[0065] Effects of cockroach powder on immune organ indexes in rats

[0066] The thymus and spleen are the main immune organs of animals. The immune organ index is one of the indicators for evaluating immune function. The thymus index and spleen index reflect the immune capacity of the animal body to a certain extent. The results are shown in Table 5 and Figure 5 As shown in the figure, the spleen and thymus indexes of rats in the five experimental groups supplemented with cockroach powder were all improved compared with those in the basic feed group, and the spleen and thymus indexes of the 8%-16% group were significantly different from those in the basic feed group (P<0.05), indicating that cockroaches can improve the immune function of immunocompromised rats. The spleen and thymus indexes of the 4% and 20% groups also tended to increase compared with the basic feed group, but there was no significant difference. The reason for this was that the amount of cockroach powder added in the 4% group was relatively small, and although there was improvement, the effect was not significant; while the amount of cockroach powder added in the 20% group was too high, and the intake of chitin in cockroaches was excessive, which required a large amount of gastric acid and enzymes to digest, but increased the gastrointestinal burden of rats, causing the rats to reduce their feed intake from the third day, and the immune-promoting effect on the thymus and spleen was not obvious.

[0067] Table 5 Effect of cockroach powder on spleen and thymus index of rats ( n=8)

[0068]

[0069]

[0070] Effect of cockroach powder on serum immunoglobulin levels in rats

[0071] The immunoglobulin content is an important indicator of the humoral immunity level. An increase in the immunoglobulin content indicates that the body's immune function is enhanced. The immunoglobulins secreted by B cells are mainly IgA, IgG, and IgM. IgA accounts for 10%-15% of the total immunoglobulins in serum; IgG is the immunoglobulin with the highest proportion in the body, accounting for about 80% of the total immunoglobulins in serum. In the body's immune response, IgG can activate the complement in humoral immunity and play a role in clearing toxins; IgM is an antibody secreted in the early stage of the immune response. It is the smallest protein in serum immunoglobulins, accounting for about 5%-10%. In this study, compared with the basic feed group, the IgA, IgG, and IgM levels of each group supplemented with cockroach powder increased, among which the difference between the 4% and 16% groups was statistically significant (P<0.05). The experimental results are shown in Tables 6 and Figure 6 The results suggest that a certain amount of cockroach powder can interfere with the body's immunoglobulin level and make it tend to normal level, so as to maintain the balance of the body's immune function.

[0072] Table 6 Effect of cockroach powder on IgA, IgG and IgM in rat serum ( n=8)

[0073]

[0074] 8 Results of T lymphocyte subset detection in rat peripheral blood

[0075] The results of the effects on rat peripheral blood T lymphocyte subsets are shown in Table 7. Figure 7 , 8 CD4 + : Compared with the basic feed group, the cockroach powder groups intervention can increase CD4 + The proportions of CD8 + : Compared with the basic feed group, the CD8 + The proportion of T lymphocytes also increased, but the difference was not statistically significant; CD4 + / CD8 + : Compared with the basic feed group, the CD4 + / CD8 + The ratios were all increased, and the difference in the 12% group was statistically significant (P<0.01).

[0076] Table 7 Effect of cockroach powder on peripheral blood T lymphocyte subsets in rats ( n=5)

[0077]

[0078]

[0079] In summary, the research results of the present invention found that after the intervention of adding cockroach powder to feed, the 4%, 8%, 12%, and 16% groups increased the relative abundance of probiotics Ruminococcaceae, Bifidobacteriaceae, and Lactobacillaceae, and reduced the relative abundance of pathogenic bacteria Enterococcaceae and Staphylococcaceae. It shows that after feeding with cockroach powder added to feed, the abundance of each dominant bacterial community has changed, and cockroach powder added to feed can change the structure and abundance of rat intestinal flora at the family level. Feed added with cockroach powder can significantly increase the level of short-chain fatty acids, a metabolite of the intestine, regulate the structure of the intestinal flora, increase the number of some beneficial bacteria, and reduce the number of some harmful bacteria.

[0080] Compared with the basic feed group, the spleen and thymus indexes, immunoglobulin content, CD4 + 、CD8 + 、CD4 + / CD8 + All of them were improved, among which the 4%, 8%, 12% and 16% groups had the most significant effects. The results suggest that adding a certain proportion of cockroach powder to feed can improve the immunity of rats. The reason may be that the active ingredients such as protein, glycoprotein, amino acid, polyol, nucleotide in cockroach powder can interfere with T lymphocytes, regulate the immune system of rats, and produce immune response.

[0081] Combined with the above research results, the appropriate amount of cockroach powder added to pet feed is in the range of 4% to 16%.

[0082] In summary, pet feed containing cockroach powder can regulate the balance of pet intestinal flora, maintain the integrity of the intestinal barrier, reduce intestinal mucosal permeability, and regulate the CD4 + / CD8 + Cell ratio plays a very positive role in the health of pets.

[0083] Obviously, the above embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the implementation methods of the present invention. For ordinary technical users in the relevant field, other different forms of changes or modifications can be made on the basis of the above description. It is not necessary and impossible to list all the implementation methods here. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the claims of the present invention.

Claims

1. A pet feed containing cockroach powder, characterized in that: The content of the cockroach powder in pet feed is 4% to 16%.

2. The pet food according to claim 1, characterized in that: The content of the cockroach powder in pet feed is 8% to 16%.

3. The pet food according to claim 2, characterized in that: The content of the cockroach powder in the pet feed is 12%.

4. The pet food according to claim 1, characterized in that: The pet feed also includes 10% to 16% of fish meal, 5% of chicken meal, 29% to 35% of flour, 5% of oatmeal powder, 30% of soybean fiber powder, 2% of chicken fat, 0.5% of complex vitamin premix, 2% of mineral element premix and 0.5% of glycoprotein.

5. The pet food according to claim 4, characterized in that: The pet feed also includes 10% to 12% of fish meal, 5% of chicken meal, 29% to 35% of flour, 5% of oatmeal powder, 30% of soybean fiber powder, 2% of chicken fat, 0.5% of complex vitamin premix, 2% of mineral element premix and 0.5% of glycoprotein.

6. The pet food according to claim 5, characterized in that: The pet feed also includes 10% fish meal, 5% chicken meal, 33% flour, 5% oatmeal powder, 30% soybean fiber powder, 2% chicken fat, 0.5% complex vitamin premix, 2% mineral element premix and 0.5% glycoprotein.

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