A Chinese medicinal preparation for improving the activity of barleyworms, Chinese medicinal feed and breeding method

Abstract

The invention relates to a Chinese medicine preparation, Chinese medicine feed and a feeding method for improving the activity of barley worms, and belongs to the technical field of barley worm breeding. The Chinese medicine preparation for improving the activity of barley worms comprises the following raw materials: dandelion root, houttuynia cordata, vitamin B and vitamin B1. After the present invention feeds barley worms with the Chinese medicine preparation added with houttuynia cordata and dandelion root, gene detection finds that the barley worms fed with the Chinese medicine feed for improving the activity of barley worms have an upward trend in chitin binding and metabolism, and the digestive ability is also increased, which indicates that the immunity and disease resistance of the barley worms fed with the Chinese medicine feed added with houttuynia cordata and dandelion root are improved, and they are not easy to get sick and die.

Description

Technical Field

[0001] The invention belongs to the technical field of barleyworm breeding, and particularly relates to a traditional Chinese medicine preparation for improving the activity of barleyworm, a traditional Chinese medicine feed and a breeding method. Background Art

[0002] The mealworm is formed by super mealworms. The larvae of the mealworm are rich in protein and fat. The protein content can reach more than 51%, and the fat content is about 29%. It also contains many nutrients, including sugars, amino acids, vitamins and minerals. With the rapid development of my country's economy and the continuous improvement of people's living standards, the demand for rare livestock and poultry and special aquatic animals will inevitably increase. Nowadays, due to the lack of protein feed resources, the development of the breeding industry has been restricted to a large extent. Therefore, the development and utilization of mealworms will inevitably be the main direction for solving the source of protein feed in the future. Meatworms can not only directly provide high-grade protein feed for animals, but also provide protein food for humans. Mealworms also contain rich organic substances, inorganic substances such as various salts, and free amino acids required by the human body. Nutritional health products made from mealworms have the effects of improving human immunity, anti-fatigue, delaying aging, lowering blood lipids, and anti-cancer.

[0003] Mealworms rarely get sick within 45-50 days after hatching. This is mainly because they have strong disease resistance at this stage. After 50 days, their disease resistance begins to decline and they are easily infected by bacteria. The mortality rate reaches 60%, or even all of them die.

[0004] Studies have found that the digestive tract of barley worm larvae is straight and easily infected by bacteria. Houttuynia cordata contains houttuynia cordata, which has a strong inhibitory effect on various bacteria, inhibits bacterial reproduction, prevents bacterial spread, achieves the purpose of sterilization, and prevents bacterial infectious diseases. Houttuynia cordata also has the function of enhancing the activity of white blood cells, which can play a defensive role. It can also participate in immune response, remove harmful and useless substances in the body, and effectively improve immunity. Houttuynia cordata has a significant inhibitory effect on Escherichia coli, Shigella dysenteriae, and Salmonella typhi.

[0005] Dandelion also has a spectrum of antibacterial effects. It has different degrees of inhibition on various ice-field microorganisms such as Gram-negative bacteria, fungi, and spirochetes. Dandelion contains three mushrooms, phytophenols, coumarins, flavonoids, phenolic acids, polysaccharides, carotene, fatty acids, as well as amino acids, proteins and other ingredients. In addition, dandelion also has a variety of vitamins and minerals, and has a wide range of biological activities. It can play a certain role in immune regulation, antibacterial and anti-inflammatory, and promoting gastrointestinal smooth muscle motility. Summary of the invention

[0006] The first object of the present invention is to provide a Chinese medicinal preparation for improving the activity of mealworms to solve the technical problem that the disease resistance of mealworms declines and the mortality rate is high after 50 days of hatching.

[0007] The second object of the present invention is to provide a Chinese medicinal feed for improving the activity of mealworms, wherein the Chinese medicinal feed comprises a Chinese medicinal preparation for improving the activity of mealworms.

[0008] The third object of the present invention is to provide a breeding method for improving the activity of mealworms. When using the breeding method to breed mealworms, it is necessary to feed the mealworms with traditional Chinese medicine feed that improves the activity of mealworms.

[0009] In order to achieve the above purpose, the technical solution adopted by the present invention is: a traditional Chinese medicine preparation for improving the activity of barleyworm, comprising the following raw materials: dandelion root, houttuynia cordata, vitamin B and vitamin B1.

[0010] Furthermore, the weight ratio of dandelion root: houttuynia cordata: vitamin B: vitamin B1 is 98-102:98-102:0.5-1.5:0.5-1.5.

[0011] A Chinese medicinal feed for improving the activity of barleyworms. The Chinese medicinal feed is prepared by processing the above-mentioned Chinese medicinal preparation for improving the activity of barleyworms into 300 ultrafine Chinese medicinal powders and mixing them into feed. The feed comprises wheat bran, corn meal and bone meal, and the mass fraction percentages of the wheat bran, corn meal and bone meal are 83-87%: 8-12%: 3-7%.

[0012] Furthermore, the weight ratio of the feed to the traditional Chinese medicine preparation is 100:19-30.

[0013] A method for raising mealworms for improving their activity comprises feeding mealworms with the Chinese medicinal feed for improving their activity and cabbage leaves, wherein the weight ratio of the feed in the Chinese medicinal feed for improving their activity to the cabbage leaves is 100:10-15.

[0014] Furthermore, the mealworms are placed in a space with a temperature of 23-28° C., an air humidity of 50-70%, and a light intensity of 33-50% for 24 hours, and are fed with a Chinese medicinal feed that improves the activity of the mealworms once a day for one week.

[0015] Beneficial effects of the present invention:

[0016] After the present invention feeds barley worms with the traditional Chinese medicine preparation added with houttuynia cordata and dandelion root, genetic testing finds that, compared with barley worms not fed with the traditional Chinese medicine feed for improving the activity of barley worms, the barley worms fed with the traditional Chinese medicine feed for improving the activity of barley worms have an upward trend in chitin binding and metabolism, and their digestive capacity is also upward regulated, which indicates that the immunity and disease resistance of the barley worms fed with the traditional Chinese medicine feed for adding houttuynia cordata and dandelion root are improved, and they are not easily infected with diseases and die.

[0017] After feeding barley worms with the Chinese medicinal feed for improving the activity of barley worms of the present invention, by comparing the barley worms fed with the Chinese medicinal feed for improving the activity of barley worms with the barley worms not fed with the Chinese medicinal feed for improving the activity of barley worms, it is found that the molting rate of the barley worms fed with the Chinese medicinal feed for improving the activity of barley worms is increased, the mortality rate of the barley worms is reduced, and the pupation of the barley worms is more uniform. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 It is a heatmap diagram of the correlation between samples in the treatment group and the control group of the present invention;

[0019] Figure 2 It is a volcano map of differentially expressed genes between samples in the treatment group and the control group of the present invention;

[0020] Figure 3 This is a diagram showing the difference in gene expression between samples in the treatment group and the control group of the present invention. DETAILED DESCRIPTION

[0021] The present invention will be further described below in conjunction with the embodiments of the present invention and the accompanying drawings.

[0022] Example 1

[0023] The Chinese medicine preparation for improving the activity of barley worms in this embodiment includes dandelion root, houttuynia cordata, vitamin B and vitamin B1, which are all purchased from Dasenlin prescription. The Chinese medicine preparation for improving the activity of barley worms is mixed with feed in a ratio of 100g / kg of houttuynia cordata, 100g / kg of dandelion root, 1g / kg of vitamin B and 1g / kg of vitamin B1, and the Chinese medicine preparation for improving the activity of barley worms is processed into 300-mesh ultrafine Chinese medicine powder.

[0024] The superfine Chinese medicine powder is mixed into the feed to obtain the Chinese medicine feed for improving the activity of the barley worm, the formula of the feed is 85% wheat bran, 10% corn flour, and 5% bone meal, wherein the breeding method for improving the activity of the barley worm in this embodiment is to place the barley worm in a space with a temperature of 25°C, an air humidity of 50%, and a light intensity of 33% for 24 hours, feed the barley worm with the Chinese medicine feed for improving the activity of the barley worm once a day, and feed for one week. When the Chinese medicine feed for improving the activity of the barley worm is used to feed the barley worm, 13g of cabbage leaves are added to every 100g of feed.

[0025] Example 2

[0026] The Chinese medicine preparation for improving the activity of barley worms of the present embodiment comprises dandelion root, houttuynia cordata, vitamin B and vitamin B1, and the dandelion root, houttuynia cordata, vitamin B and vitamin B1 are all purchased from Dasenlin prescription. The Chinese medicine preparation for improving the activity of barley worms is mixed in the ratio of 98g / Kg of houttuynia cordata, 98g / Kg of dandelion root, 0.5g / Kg of vitamin B and 0.5g / Kg of vitamin B1, and the Chinese medicine preparation for improving the activity of barley worms is processed into 300-mesh ultrafine Chinese medicine powder. The ultrafine Chinese medicine powder is mixed into the feed to obtain the Chinese medicine feed for improving the activity of barley worms, and the formula of the feed is 84% ​​of wheat bran, 12% of corn flour and 4% of bone meal.

[0027] The method for raising the activity of mealworms in this embodiment is to place mealworms in a room with a temperature of 23°C, an air humidity of 55%, and a light intensity of 40% for 24 hours, and feed the mealworms with a Chinese medicinal feed for raising the activity of mealworms once a day for one week. When the Chinese medicinal feed for raising the activity of mealworms is used to feed mealworms, 10g of cabbage leaves are added to every 100g of feed.

[0028] Example 3

[0029] The Chinese medicine preparation for improving the activity of barley worms in this embodiment comprises dandelion root, houttuynia cordata, vitamin B and vitamin B1, and the dandelion root, houttuynia cordata, vitamin B and vitamin B1 are all purchased from Dasenlin prescription. The Chinese medicine preparation for improving the activity of barley worms is mixed in a ratio of 102g / Kg of houttuynia cordata, 102g / Kg of dandelion root, 1.2g / Kg of vitamin B and 1.2g / Kg of vitamin B1, and the Chinese medicine preparation for improving the activity of barley worms is processed into 300-mesh ultrafine Chinese medicine powder. The ultrafine Chinese medicine powder is mixed into feed to obtain the Chinese medicine feed for improving the activity of barley worms, and the formula of the feed is 87% of wheat bran, 8% of corn flour and 5% of bone meal.

[0030] The method for raising the activity of mealworms in this embodiment is to place mealworms in a room with a temperature of 24°C, an air humidity of 60%, and a light intensity of 42% for 24 hours, and feed the mealworms with a Chinese medicinal feed for raising the activity of mealworms once a day for one week. When the Chinese medicinal feed for raising the activity of mealworms is used to feed mealworms, 11g of cabbage leaves are added to every 100g of feed.

[0031] Example 4

[0032] The Chinese medicine preparation for improving the activity of barley worms of the present embodiment comprises dandelion root, houttuynia cordata, vitamin B and vitamin B1, and the dandelion root, houttuynia cordata, vitamin B and vitamin B1 are all purchased from Dasenlin prescription. The Chinese medicine preparation for improving the activity of barley worms is mixed in the proportion of 98g / Kg of houttuynia cordata, 98g / Kg of dandelion root, 0.9g / Kg of vitamin B and 0.9g / Kg of vitamin B1, and the Chinese medicine preparation for improving the activity of barley worms is processed into 300-mesh ultrafine Chinese medicine powder. The ultrafine Chinese medicine powder is mixed into the feed to obtain the Chinese medicine feed for improving the activity of barley worms, and the formula of the feed is 85% of wheat bran, 12% of corn flour and 3% of bone meal.

[0033] The method for raising the activity of mealworms in this embodiment is to place mealworms in a room with a temperature of 25°C, an air humidity of 65%, and a light intensity of 45% for 24 hours, and feed the mealworms with a Chinese medicinal feed for raising the activity of mealworms once a day for one week. When the Chinese medicinal feed for raising the activity of mealworms is used to feed mealworms, 12g of cabbage leaves are added to every 100g of feed.

[0034] Example 5

[0035] The Chinese medicine preparation for improving the activity of barley worms in this embodiment comprises dandelion root, houttuynia cordata, vitamin B and vitamin B1, and the dandelion root, houttuynia cordata, vitamin B and vitamin B1 are all purchased from Dasenlin prescription. The Chinese medicine preparation for improving the activity of barley worms is mixed in a ratio of 103g / Kg of houttuynia cordata, 103g / Kg of dandelion root, 1.3g / Kg of vitamin B and 1.3g / Kg of vitamin B1, and the Chinese medicine preparation for improving the activity of barley worms is processed into 300-mesh ultrafine Chinese medicine powder. The ultrafine Chinese medicine powder is mixed into feed to obtain the Chinese medicine feed for improving the activity of barley worms, and the formula of the feed is 83% of wheat bran, 10% of corn flour and 7% of bone meal.

[0036] The method for raising the activity of mealworms in this embodiment is to place mealworms in a room with a temperature of 26°C, an air humidity of 70%, and a light intensity of 50% for 24 hours, and feed the mealworms with a Chinese medicinal feed for raising the activity of mealworms once a day for one week. When the Chinese medicinal feed for raising the activity of mealworms is used to feed mealworms, 15g of cabbage leaves are added to every 100g of feed.

[0037] 1. Basic Information

[0038] 600 2-year-old mealworms were divided into a treatment group and a control group. The treatment group contained 3 samples, namely Treat_1, Treat_2 and Treat_3, each containing 100 mealworms; the control group contained 3 samples, namely Control_1, Control_2 and Control_3, each containing 100 mealworms. The mealworms in the treatment group and the control group were fed in a room with a 24h temperature of 25°C, an air humidity of 50%, and a light intensity of 33%. The mealworms in the treatment group were fed with the prepared Chinese medicinal feed and cabbage leaves that improve the activity of mealworms, and the mealworms in the control group were fed with normal feed and cabbage leaves. After one week, the molting rate, mortality rate and pupation of the mealworms in the treatment group and the control group were observed. Among them, the weight ratio of feed to cabbage leaves in the Chinese medicinal feed that improves the activity of mealworms is 100:15. Every 100 mealworms are fed with 150g of feed, the formula of which is 85% of wheat bran, 10% of corn flour and 5% of bone meal. The Chinese medicinal preparation for improving the activity of mealworms is mixed with the feed in a ratio of 100g / Kg of houttuynia cordata, 100g / Kg of dandelion root, 1g / Kg of vitamin B and 1g / Kg of vitamin B1. The Chinese medicinal preparation for improving the activity of mealworms is processed into 300-mesh ultrafine Chinese medicinal powder.

[0039] After the barleyworms in the treatment group were fed with the Chinese medicine preparations that improve the activity of barleyworms for one week, the barleyworms in the treatment group and the control group were sequenced using the Illumina second-generation high-throughput sequencing platform and the PE150 sequencing strategy. A total of 291,296,346 raw reads were measured, and 289,259,914 clean reads were obtained after quality control. Trinity and bowtie2 software were used to complete sequence assembly and alignment, and then all genes were quantitatively analyzed. According to the set difference combination, differential genes were identified, and a total of 4,478 differential genes were identified, of which 2,018 were upregulated genes and 2,460 were downregulated genes. Functional enrichment analysis was performed on these differential genes to explore their functions.

[0040] 2. Sequencing Data Statistics

[0041] Data statistics were performed on the original data and the high-quality data (clean reads) obtained by filtering to obtain basic data information, which is shown in Table 1.

[0042] Table 1 Data statistics

[0043]

[0044] As can be seen from Table 1, the percentage of bases with base quality values ​​≥ 30 (Q30) of all samples was ≥ 90.87%, and an average of 48,209,986 high-quality reads were obtained for each sample.

[0045] 3. Trinity Assembly

[0046] For transcriptome studies without a reference genome, after obtaining high-quality RNA-seq sequencing data, all sequencing reads need to be assembled de novo to generate contigs and singletons. The results are shown in Table 2.

[0047] Table 2

[0048]

[0049]

[0050] 4. Comparison results statistics

[0051] Clean reads data were aligned to the assembly “Trinity.fasta” using bowtie2 software (version: v2.2.9), and the results are shown in Table 3.

[0052] Table 3 Comparison results statistics

[0053]

[0054] As can be seen from Table 3, the alignment efficiency of reads of each sample with the reference genome sequence is 65.24% to 71.52%, the alignment efficiency with the unique corresponding position of the reference genome is 31.06% to 31.50%, and the alignment efficiency with multiple corresponding positions of the reference genome is 33.91% to 40.03%.

[0055] 5. Quantitative analysis of gene expression

[0056] The abundance of transcripts reflects the expression level of genes. The higher the transcript abundance, the higher the gene expression level. In RNA_seq analysis, the expression level of genes is calculated by the number of sequences (clean reads) located in the genomic region. Based on the comparison results of all samples with the reference genome, the expression value of each gene transcript in the sample is calculated. Some results are shown in Table 4 below.

[0057] Table 4

[0058]

[0059]

[0060] 6. Correlation analysis between samples

[0061] According to the expression of each gene in each sample, the correlation between samples is calculated using the Pearson correlation coefficient and a heatmap is made. The higher the correlation, the darker the color. The heatmap is as follows Figure 1 shown.

[0062] VII. Statistics of differentially expressed genes

[0063] In order to compare the differences in gene expression between different samples, differential expression analysis was performed using edgeR, and genes with a FDR value less than 0.05 and a FoldChange absolute value greater than 2 were considered to be significantly differentially expressed genes. The results are shown in Table 5. Figure 2 as well as Figure 3 As shown, the results showed that a total of 4478 significantly differentially expressed genes were mined, including 2018 up-regulated genes and 2460 down-regulated genes.

[0064] Table 5

[0065]

[0066] 8. GO enrichment and KEGG enrichment of differentially expressed genes

[0067] GO enrichment of differentially expressed genes

[0068] GO annotation was performed on the differentially expressed genes, and enrichment analysis was performed on the up-regulated genes and down-regulated genes corresponding to the GO annotations. According to qvalue<0.05 and the enrichment feature Enrich Direct as Over, significant enrichment results were screened out. Some of the results are shown in Tables 6 and 7 below.

[0069] Table 6 Upregulated genes

[0070]

[0071]

[0072] Table 7 Down-regulated genes

[0073]

[0074] Through GO enrichment analysis of differentially expressed genes, compared with the control group, the up-regulated genes in the treatment group were mainly concentrated in the following functional categories, such as chitin binding and metabolism (chit in binding, chit in metabolic process), epidermal structure composition (structural constituent of cuticle), digestion ability (digestion), etc.; the down-regulated genes were mainly concentrated in RNA binding and shearing (RNA binding, nuclear acid binding, mRNA splicing, via spliceosome, RNA splicing, mRNA processing), actin filament binding (actinfilament binding), etc.

[0075] Chitin, also known as chitosan, is an important component of the hard shell of insects. It can make the shell of mealworms more solid and reliable. Chitin is also found in its internal structure (including the digestive tract, tracheal system, reproductive ducts and the inner epidermis of various dermal glands). This shows that the growth and development of mealworms are closely related to the synthesis of chitin. In addition, chitin can activate and repair cell functions, enhance encouragement and regulation functions, prevent diseases, improve disease resistance and accelerate recovery functions. It also has the detoxification function of excreting toxic and harmful substances from the body and the function of regulating victory and balance. Therefore, the upregulation of chitin binding and metabolism can improve the vitality of mealworms.

[0076] KEGG enrichment of differentially expressed genes

[0077] KEGG annotation was performed on the differentially expressed genes, and the up-regulated genes and down-regulated genes corresponding to the KEGG annotated map numbers were enriched and analyzed respectively. According to qvalue < 0.05 and the enrichment feature Enrich Direct as Over, significant enrichment results were screened out. Some of the results are shown in Tables 8 and 9.

[0078] Table 8 Upregulated genes

[0079]

[0080] Table 9 Downregulated genes

[0081]

[0082]

[0083] Through KEGG enrichment analysis of differentially expressed genes, compared with the control group, the up-regulated genes in the treatment group were concentrated in the following pathways, such as pyruvate metabolism, pentose phosphate pathway, purine metabolism, etc.; the down-regulated genes were concentrated in RNA transport, messenger RNA biogenesis, nicotinamide and nicotinamide metabolism, etc. In addition, some pathways have both up-regulated and down-regulated genes, such as spliceosome, proteasome, exosome, etc. These are complex and synergistic metabolic pathways.

[0084] Pyruvate is a weakly acidic organic acid with both carbonyl and carboxyl functional groups in its molecule. In addition to the properties of carboxylic acid and ketone, it also has the properties of α-keto acid, and is the simplest α-keto acid (belonging to carbonyl acid). Pyruvate can also achieve the mutual conversion between sugar, fat and amino acids in the body through acetyl CoA and tricarboxylic acid cycle. Therefore, pyruvate plays an important pivotal role in the metabolic connection of the three major nutrients. Studies have shown that pyruvate can inhibit the oxidation of oxygen free radicals in mice. At the same time, as a hydrogen peroxide scavenger, it has the effect of preventing free radical damage. It has been proven to protect the body from functional damage in cardiac reperfusion injury and acute renal failure. Pyruvate can play an antioxidant role through two mechanisms, delaying aging, protecting eyes and vision, reducing the effects of aging on the skin, and preventing chronic inflammatory diseases. Therefore, the upregulation of pyruvate metabolism can improve the vitality of mealworms.

Claims

1. A Chinese medicine preparation for improving the activity of barley worms, characterized in that: The invention is composed of the following raw materials: dandelion root, houttuynia cordata, vitamin B and vitamin B1; the weight ratio of dandelion root: houttuynia cordata: vitamin B: vitamin B1 is 98-102:98-102:0.5-1.5:0.5-1.

5.

2. A Chinese medicinal feed for improving the activity of barley worms, characterized in that: The Chinese medicinal feed is prepared by processing the Chinese medicinal preparation for improving the activity of barleyworms as claimed in claim 1 into 300-mesh ultrafine Chinese medicinal powder and mixing it into feed, wherein the feed comprises wheat bran, corn meal and bone meal, and the mass fraction percentages of the wheat bran, corn meal and bone meal are 83-87%: 8-12%: 3-7%.

3. The Chinese medicinal feed for improving the activity of barley worms according to claim 2, characterized in that: The weight ratio of the feed to the traditional Chinese medicine preparation is 100:19-30.

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