Compound feed additive containing potassium bicarbonate and valonia tannin and application of compound feed additive

Abstract

The invention provides a compound feed additive containing potassium bicarbonate and valonia tannin, which can effectively improve the problem of loose stool of chickens caused by replacement of soybean meal by corn protein peptide, and does not influence the growth performance of the chickens.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of poultry nutrition and feed technology, and particularly relates to a compound additive capable of improving the problem of loose stool. BACKGROUND

[0002] As a core protein raw material for poultry feed, soybean meal is heavily dependent on imports in China, and its price is significantly affected by international market fluctuations, which restricts the cost control and sustainable development of the breeding industry. Therefore, it is crucial to develop cheap and readily available local protein sources to replace soybean meal.

[0003] Corn protein peptide is a by-product of the corn processing process, which is made of small molecule peptide mixture through fermentation, resin adsorption, enzymatic hydrolysis and filtration processes. It has the advantages of high protein content, reasonable amino acid composition and low anti-nutritional factors. Moreover, China has abundant corn resources and stable prices, making corn protein peptide a potential high-quality protein source to replace soybean meal. However, during the corn starch processing process, sulfurous acid solution is commonly used as a soaking agent, resulting in a high concentration of sulfite residues in the final product, corn protein peptide. When poultry consumes excessive amounts of sulfite, it can cause loose stool, even diarrhea and other adverse reactions. This problem limits the large-scale application of corn protein peptide in chicken feed.

[0004] Currently, the process methods for removing sulfite residues in corn protein peptide (such as deep water washing, membrane separation, etc.) have problems such as high cost, low efficiency or easy loss of nutrients, making it difficult to be widely used in the feed industry. On the other hand, the industry often uses montmorillonite and other adsorbents to temporarily alleviate the symptoms of diarrhea, but this method only treats the symptoms and does not address the root cause: montmorillonite adsorbs harmful substances while also non-selectively adsorbing nutrients such as vitamins and minerals in the feed, which may lead to nutrient deficiencies and affect chicken growth performance over the long term.

[0005] Therefore, there is a need for an economic and effective solution that can fundamentally solve the problem of loose stool caused by corn protein peptide in chickens, without affecting the overall nutritional value of the feed. SUMMARY

[0006] The present application aims to solve the problems existing in the prior art, and provides a compound feed additive containing potassium bicarbonate and oak tannin, which can effectively improve the problem of loose stool caused by corn protein peptide replacing soybean meal in chickens, without affecting the growth performance of chickens.

[0007] One aspect of the present application provides a compound feed additive containing potassium bicarbonate and oak tannin, which consists of potassium bicarbonate and oak tannin.

[0008] Further, in the compound feed additive, the content of potassium bicarbonate is 0.06% based on the total amount of the basic daily ration, so that the electrolyte balance constant is kept equal to that of the control group (basic daily ration group).

[0009] Further, in the compound feed additive, the content of oak tannin is 0.01%-0.1%, preferably 0.02%, based on the total amount of the basic daily ration.

[0010] Further, in the compound feed additive, the weight ratio of potassium bicarbonate to oak tannin is 3:1.

[0011] Another aspect of the present application provides the use of the compound feed additive in improving the chicken loose stool problem caused by excessive intake of sulfite by chickens.

[0012] Specifically, the excessive intake of sulfite by chickens is caused by the intake of corn protein peptide.

[0013] The present inventors have conducted in-depth research on the root cause of the electrolyte imbalance of chickens caused by excessive sulfite residues in corn protein peptide, which further leads to loose stool. Based on physiological mechanism analysis, a compound regulation idea is innovatively proposed: using potassium bicarbonate as an electrolyte regulator to correct the acid-base and ion balance of the body; at the same time, using oak tannin with significant astringent effect to repair damaged intestinal mucosa and reduce exudation.

[0014] The present inventors have unexpectedly found that when a specific proportion of potassium bicarbonate and oak tannin are compounded, they show significant synergistic effect in improving chicken loose stool caused by corn protein peptide, and the effect is significantly better than that of using any of the components alone. This compounding scheme not only alleviates the symptoms of diarrhea from the root, but also avoids the negative effects of traditional adsorbents such as montmorillonite on nutrients.

[0015] In particular, corn protein peptide is a non-conventional feed raw material, and its application effect in replacing soybean meal in broiler feed is still lacking of research. The loose stool problem caused by it and its solution have not been reported at home and abroad. Therefore, the present application first discloses this specific problem and first proposes the use of potassium bicarbonate and oak tannin in a compound, which is an efficient and safe targeted solution, filling the technical gap in this field and providing a scientific technical solution for the efficient and safe application of corn protein peptide in broiler feed. BRIEF DESCRIPTION OF DRAWINGS

[0016] Figure 1 The average fecal water content of each group of the present application and comparative examples in the early and late growth periods of broilers;

[0017] Figure 2The intestinal permeability indicators for each group in the embodiments and comparative examples of the present invention are: serum diamine oxidase activity and D-lactic acid content;

[0018] Figure 3 The relative mRNA expression levels of the intestinal fluid secretion gene CFTR in each group of the embodiments and comparative examples of the present invention are shown.

[0019] In the attached figures, * indicates a significant difference between the two groups (P<0.05), while no * indicates no significant difference between the two groups (P>0.05). Detailed Implementation

[0020] The present invention will be described below with reference to the accompanying drawings and embodiments to help those skilled in the art understand its objectives, technical solutions, and advantages. It should be noted that the embodiments are merely examples and do not cover all possible situations.

[0021] Unless otherwise specified, all reagents and equipment used in this invention can be obtained by purchasing commercially available products.

[0022] Test design and materials:

[0023] Forty-eighty one-day-old male Arbor Acqua broiler chickens of similar weight (purchased from Jiangsu Jinghai Poultry Co., Ltd.) were randomly selected and randomly divided into four groups, with six replicates in each group and 20 chickens in each replicate.

[0024] in:

[0025] Comparative Example 1, i.e., control group (Group A): fed with a basal diet;

[0026] Comparative Example 2, namely, the 4% corn protein peptide group (Group B): 4% corn protein peptide was used to replace the corresponding proportion of soybean meal in the basal diet;

[0027] Comparative Example 3, namely, the 4% corn protein peptide + 0.06% potassium bicarbonate group (Group C): In the basal diet, 4% corn protein peptide was used to replace the corresponding proportion of soybean meal, and 0.06% potassium bicarbonate was added.

[0028] Example 1, namely, the 4% corn protein peptide + 0.06% potassium bicarbonate + 0.02% acorn tannin group (Group D): In the basal diet, 4% corn protein peptide was used to replace the corresponding proportion of soybean meal, and 0.06% potassium bicarbonate and 0.02% acorn tannin were added.

[0029] The nutritional levels of the four groups were kept consistent, and the trial period was 42 days. The corn protein peptides were purchased from Zhucheng Haotian Pharmaceutical Co., Ltd.

[0030] Animal feed and feeding management:

[0031] The experiment was conducted in accordance with NRC (1994), NY / T 33-2004 Chicken Feeding Standards, and the Chinese Feed Composition and Nutritional Value Table (2022). Pelleted diets were formulated for two stages: 1-21 days of age and 22-42 days of age. The composition and nutritional levels of the basal diets are shown in Table 1 below. The experiment was conducted at the Yizheng Experimental Base of Jiangsu Provincial Poultry Science Research Institute, using stacked cage rearing. The experimental chickens had free access to feed and water. During the experiment, other management measures such as lighting, ventilation, and immunization were implemented in accordance with the feeding management manual.

[0032] [Table 1]

[0033]

[0034] Test results:

[0035] 1. Measurement of fecal moisture content

[0036] Feces were collected from broiler chickens on days 18-20 of the early rearing period and days 38-40 of the later rearing period. The weight of the collected fecal samples was recorded, and the moisture content was determined using a drying method at 100℃. The weight of the dried fecal samples was also recorded. The formula for calculating the moisture content of the feces is:

[0037] Fecal moisture content (%) = (Weight of feces before drying - Weight of dry feces) / Weight of feces before drying * 100

[0038] Figure 1 Example 1 and Comparative Examples 1-3 are shown, namely, the fecal moisture content of broiler chickens in the early and late growth stages of groups A-D.

[0039] Depend on Figure 1 It can be seen that, compared with group A (basal diet group), the water content of broiler feces in group B (4% corn protein peptide group) did not change significantly in the early stage, but the water content of broiler feces in the later stage increased significantly, showing a significant difference (P<0.05). This indicates that the use of corn protein peptide in feed for a period of time will cause chicken feces to become loose.

[0040] Compared with group A, group C (4% corn protein peptide + 0.06% potassium bicarbonate group) also showed a significant increase in fecal water content in the later stage of broiler feeding (P<0.05). However, there was no significant difference between group C and group B (P>0.05), indicating that the additive containing only potassium bicarbonate did not show a significant effect in improving loose stools in chickens.

[0041] Furthermore, it can be seen that although group D (4% corn protein peptide + 0.06% potassium bicarbonate + 0.02% acorn tannin) had no significant effect on the fecal moisture content of broilers in the early stage, the fecal moisture content decreased significantly in the later stage of broiler rearing, showing a significant difference compared with group C (P<0.05). This indicates that adding a certain proportion of potassium bicarbonate and acorn tannin as additives can significantly improve the loose stools induced by corn protein peptide, and potassium bicarbonate and acorn tannin have a synergistic effect.

[0042] 2. Measurement of intestinal permeability indicators

[0043] On day 42 of the experiment, serum from the broilers was isolated, aliquoted into cryovials, and stored at -80°C for later use. The diamine oxidase (D-O-O-Lac) and D-lactate (D-Lac) contents of each group were determined using kits purchased from Nanjing Jiancheng Biotechnology Institute and Wuhan Elite Biotechnology Co., Ltd., respectively. The results are shown in [Figure / Table / Insert Results Here]. Figure 2 middle.

[0044] Depend on Figure 2 As can be seen, compared with group A, the diamine oxidase activity and D-lactic acid content in the serum of broilers in group B were significantly increased (P<0.05), indicating increased intestinal permeability. Increased intestinal permeability is not only a result of diarrhea but also one of the causes of its persistence and worsening, forming a self-amplifying vicious cycle. Therefore, it can be demonstrated that adding corn protein peptides to feed leads to increased intestinal permeability in broilers, thereby causing the exacerbation of diarrhea.

[0045] Compared with group B, there were no significant differences in diamine oxidase activity and D-lactic acid content in the serum of broilers in group C (P>0.05). This indicates that when the additive only contains potassium bicarbonate, it does not significantly improve the permeability of the broiler intestine.

[0046] Compared with group B, group D showed significantly lower levels of diamine oxidase activity and D-lactic acid content in broiler serum (P<0.05). This indicates that the simultaneous addition of potassium bicarbonate and acorn tannins as additives can synergistically reduce intestinal permeability indicators, thereby improving diarrhea induced by corn protein peptides. This is consistent with the results in "1. Measurement of fecal moisture content" above.

[0047] 3. Measurement of mRNA expression level of intestinal fluid secretion gene CFTR

[0048] 3.1 RNA extraction and cDNA preparation

[0049] On day 42 of the experiment, total RNA was extracted from the jejunal mucosa of each group in the examples and comparative examples using the RNAisolater Total RNA Extraction Reagent kit from Nanjing Novizan Biotechnology Co., Ltd. RNA concentration was measured using a nucleic acid quantification instrument. cDNA synthesis was performed according to the instructions of the reverse transcription kit from Shanghai Sangon Biotech Co., Ltd.

[0050] 3.2 Primer pair design for detecting chicken intestinal fluid secretion genes and determination of their mRNA expression levels

[0051] Based on the gene sequence provided by NCBI-GenBank, primers were synthesized by Genewiz Biotechnology Co., Ltd. The primer sequences are shown in Table 2 below. The reaction system and thermal cycling conditions were set according to the manufacturer's instructions (Q-PCR reaction program: 95℃, 15 min; 40 cycles, 94℃, 15 s; 55℃, 30 s; 70℃, 30 s). β-actin was used as an internal reference gene. The relative expression level of the target gene was measured using a 2-1... -ΔΔCT The method of calculation is represented.

[0052] [Table 2]

[0053]

[0054] Figure 3 The results show the relative mRNA expression levels of the intestinal fluid secretion gene CFTR in each group of the examples and comparative examples. Normal expression and function of the CFTR gene are crucial for maintaining chloride ion secretion and mucus hydration in intestinal epithelial cells. Its expression level is positively correlated with diarrhea; that is, the higher the expression level of the CFTR gene, the more severe the diarrhea and the higher the water content of the stool.

[0055] Depend on Figure 3 As can be seen, compared with group A, the relative expression level of the intestinal fluid secretion gene CFTR in broilers in group B was significantly increased, showing a significant difference (P<0.05). This indicates that the use of corn protein peptides in feed increases the expression of genes related to intestinal fluid secretion, thereby increasing fecal water content and aggravating diarrhea.

[0056] Compared with group B, there was no significant difference in the relative expression level of CFTR mRNA in broiler chickens in group C (P>0.05), indicating that when the additive only contains potassium bicarbonate, it has no significant effect on the expression of genes related to intestinal fluid secretion.

[0057] Compared with group B, the relative expression level of the intestinal fluid secretion gene CFTR mRNA in broilers in group D was significantly lower, showing a significant difference (P<0.05). This indicates that when potassium bicarbonate and acorn tannin are added simultaneously in certain proportions as additives, they can significantly reduce the expression of genes related to intestinal fluid secretion through synergistic effects, thereby reducing fecal water content and improving diarrhea induced by corn protein peptides. This is consistent with the results in "1. Fecal water content measurement" and "2. Intestinal permeability index measurement" above.

[0058] 4. Growth performance and meat quality

[0059] On the 42nd day of the experiment, the weight of broilers in both the example and comparative groups was measured on an empty stomach. Feed intake for each replicate in each group was recorded. If any mortality occurred during a replicate, feed was promptly removed, and the new feed intake was updated. The average daily gain (ADG), average daily feed intake (ADFI), and feed conversion ratio (F / G) of broilers in each group from 1 to 42 days of age were calculated. The calculation formulas are as follows:

[0060] Average daily weight gain = (final weight - initial weight) / number of days in the trial

[0061] Average daily feed intake = Total feed intake / Number of days in the experiment

[0062] Feed conversion ratio = Average daily feed intake / Average daily weight gain

[0063] At 42 days of age, one chicken with a weight close to the average was selected from each replicate. After slaughter and dissection, the left pectoralis major muscle was uniformly harvested for meat quality testing. The brightness, redness, and yellowness of the pectoralis major muscle were measured using an automated colorimeter (CR-400, Konica Minolta, Japan). Water loss was measured using the filter paper squeezing method. pH was measured by directly inserting a pH meter (PH-STAR, Metersbonwe, Germany) into the pectoralis major muscle. Three test points were used, and the average value was taken. Specific methods followed the guidelines in NY / T 1333—2017 "Determination of Meat Quality of Livestock and Poultry". Shear force was measured according to NY / T 118—2006 "Determination of Meat Tenderness: Shear Force Test".

[0064] The results of the growth performance and meat quality measurements are shown in Tables 3 and 4 below.

[0065] [Table 3]

[0066]

[0067] [Table 4]

[0068]

[0069] According to Tables 3 and 4, it can be seen that there was no significant difference in broiler growth performance and meat quality between group D and group B (P>0.05). Therefore, based on the above experimental results, the compound additive of potassium bicarbonate and acorn tannin can significantly improve broiler diarrhea caused by corn protein peptides, while having no negative impact on broiler growth performance and meat quality.

Claims

1. A compound feed additive, characterized in that, The compound feed additive consists of potassium bicarbonate and acorn tannins.

2. The compound feed additive according to claim 1, characterized in that, Based on the total amount of the basal diet, the compound feed additive contains, by weight fraction, 0.06% potassium bicarbonate and 0.01-0.1% acorn tannin.

3. The compound feed additive according to claim 2, characterized in that, The content of arabin tannin is 0.02%.

4. The compound feed additive according to claim 1, characterized in that, In the compound feed additive, the weight ratio of potassium bicarbonate to aramid tannin is 3:

1.

5. The application of the compound feed additive according to claim 1 in improving the problem of loose stools in chickens caused by excessive intake of sulfites.

6. In the application according to claim 5, the excessive intake of sulfites in the chicken is caused by the intake of corn protein peptides.

Images