CN113475624A - Poecilobdella manillensis compound feed - Google Patents
Poecilobdella manillensis compound feed Download PDFInfo
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23K10/00—Animal feeding-stuffs
- A23K10/20—Animal feeding-stuffs from material of animal origin
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23K20/00—Accessory food factors for animal feeding-stuffs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/20—Inorganic substances, e.g. oligoelements
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
Abstract
The invention relates to a Poecilobdella manillensis compound feed, which belongs to the technical field of aquaculture and comprises pig serum, egg protein, threonine, methionine, lysine, tryptophan, glutathione, glycine, arginine, salt and water. The invention provides artificial compound feed for poecilobdella manillensis, which fully considers the characteristics of food habit, nutrition requirement, amino acid balance and digestion of the poecilobdella manillensis, selects the conventional feed raw materials which are easy to digest and absorb for reasonable compatibility, and has high protein source amino acid balance, high solubility and good feeding effect of the poecilobdella manillensis. After the compound feed is fed, the survival rate of the poecilobdella manillensis is high, the hematemesis phenomenon does not occur in the feeding process, and the nutritional requirements of the poecilobdella manillensis under the artificial culture condition are met.
Description
Technical Field
The invention belongs to the technical field of aquaculture, and particularly relates to a poecilobdella manillensis compound feed.
Background
Poecilobdella manillensis is a special medicinal animal resource, and natural hirudin secreted by salivary glands of Poecilobdella manillensis is the strongest natural antithrombin substance, and has higher curative effect on treating cardiovascular and cerebrovascular diseases, especially thrombotic diseases. Hirudin is the most important active component in Poecilobdella manillensis, and is also the strongest natural thrombin inhibitor discovered so far, which is mainly an active polypeptide consisting of 65-66 amino acids. The aquatic animals utilize feed protein, mainly utilize amino acid of the feed to synthesize self amino acid, and the level of the feed protein and the composition of the amino acid have close influence on the growth of the aquatic animals.
At present, fresh blood of animals is mainly fed in artificial breeding of poecilobdella manillensis, and the fresh blood is used as a natural feed source of the poecilobdella manillensis, so that the poecilobdella manillensis is easy to pollute and deteriorate, has large pollution to the water environment, and has biological risk. The main feed protein sources in the artificial feed mostly adopt plasma protein powder and blood cell protein powder, and the artificial feed prepared by the two feed protein sources solves the feed problem of the poecilobdella manillensis to a certain extent, but the feeding effect of the artificial feed is lower than that of blood. When the addition of the plasma protein powder and the blood cell protein powder is increased, the ingestion effect is improved, but the dissolution effect is gradually reduced, the Poecilobdella manillensis can cause the phenomenon of hematemesis, and then the phenomenon of body curling and wrinkling is caused, and the phenomenon of increased mortality rate is caused. The existing artificial mixed feed for poecilobdella manillensis does not consider the nutrition requirement and amino acid balance of the poecilobdella manillensis, and the optimal feeding cannot be realized.
Disclosure of Invention
The invention aims to provide artificial compound feed for poecilobdella manillensis aiming at the defects of the existing artificial compound feed for the poecilobdella manillensis, the food habit, the nutritional requirement and the digestion characteristic of the poecilobdella manillensis are fully considered, the nutritional levels of protein, small peptide and amino acid in the artificial compound feed for the poecilobdella manillensis are reasonably formulated, and the conventional feed raw materials which are easy to digest and absorb by the poecilobdella manillensis are selected for reasonable compatibility, so that the nutritional requirement under the artificial breeding condition of the poecilobdella manillensis is met. The utilization rate of the compound feed is high, and the raw materials used by the compound feed are stable.
In order to realize the purpose, the invention is realized by the following technical scheme:
the protein level range of the poecilobdella manillensis compound feed is 9.78-11.17%, and the small peptide level range is 1.48-2.5%.
The Poecilobdella manillensis compound feed comprises pig serum, egg protein, threonine, methionine, lysine, tryptophan, glutathione, glycine, arginine, salt and water. The mixture ratio of each raw material is as follows: 40.00 percent of pig serum, 16.09-30.59 percent of egg protein, 0.49-0.55 percent of threonine, 0.095-0.10 percent of methionine, 0.20-0.33 percent of lysine, 0.06-0.07 percent of tryptophan, 1.48-2.5 percent of glutathione, 0.50 percent of glycine, 0.50 percent of arginine, 0.40 percent of salt and 25.68-39.99 percent of water; the sum of the weight percentages of all the raw materials is 100 percent.
The aquatic animals utilize the feed protein, mainly utilize the amino acid of the feed to synthesize the self amino acid, so the feed protein and the amino acid composition thereof have close influence on the growth performance of the aquatic animals. Protein nutrition is essentially amino acid nutrition, the nutritional value of protein depends on whether the composition of the amino acids is balanced, and the more consistent the amino acid composition of the feed protein source and the amino acid requirement pattern of animals, the better the amino acid balance of the feed protein source.
The invention provides artificial compound feed for poecilobdella manillensis, which fully considers the characteristics of food habit, nutrition requirement, amino acid balance and digestion of the poecilobdella manillensis, reasonably establishes the levels of protein, small peptide and amino acid in the artificial compound feed for the poecilobdella manillensis, selects the conventional feed raw materials which are easy to digest and absorb for compatibility, and has stable used raw materials and easy preparation. The protein source amino acid of the compound feed has high degree of balance and solubility, and the feeding effect of the poecilobdella manillensis is good. After the compound feed is fed, the growth performance and the antithrombin activity of the poecilobdella manillensis are improved, the survival rate is high, and no hematemesis phenomenon occurs in the feeding process, which indicates that the compound feed meets the nutritional requirements of the poecilobdella manillensis under the artificial culture condition.
Drawings
FIG. 1 is a comparative analysis of the solubility of proteins from different feed protein sources;
FIG. 2 is a graph showing the gray correlation between a feed protein source and Poecilobdella manillensis;
FIG. 3 is a comparison of the food intake of different feed protein sources;
FIG. 4 shows the body curling and wrinkling phenomenon of Poecilobdella manillensis;
FIG. 5 is a graph of the effect of different feeds on the total food intake of Poecilobdella manillensis;
FIG. 6 is a graph of the effect of different feeds on the total weight gain of Poecilobdella manillensis;
FIG. 7 is a graph of the effect of different feeds on the average daily weight gain of Poecilobdella manillensis;
FIG. 8 is a graph of the effect of different feeds on the end body weight of Poecilobdella manillensis;
FIG. 9 is a graph of the effect of different feeds on the weight gain of Poecilobdella manillensis;
FIG. 10 is a graph of the effect of different feeds on the feed factor of Poecilobdella manillensis;
FIG. 11 is a graph of the effect of different feeds on the specific growth rate of Poecilobdella manillensis;
FIG. 12 is a graph of the effect of different feeds on the survival rate of Poecilobdella manillensis;
FIG. 13 is a graph of the effect of different feeds on the efficiency of Poecilobdella manillensis protein;
FIG. 14 is a graph of the interaction of protein and small peptide levels on total food intake;
FIG. 15 is a graph of the interaction of protein and small peptide levels on total weight gain;
FIG. 16 is a graph of the interaction of protein and small peptide levels on daily average weight gain;
FIG. 17 is a graph of the interaction of protein and small peptide levels versus end-of-term body weight;
FIG. 18 is a graph of the interaction of protein and small peptide levels on rate of gain;
FIG. 19 is the interaction of protein and small peptide levels for a particular growth rate;
FIG. 20 is a graph of growth performance indicators versus feed protein levels;
FIG. 21 is a graph of growth performance indicators versus feed small peptide levels;
FIG. 22 is a comparative analysis of antithrombin activity in Poecilobdella manillensis in different feeds;
FIG. 23 is a graph of protein and small peptide level interaction with anticoagulant active ingredients;
FIG. 24 is a graph of the relationship of anticoagulant active ingredient to feed protein level;
figure 25 is a graph of anticoagulant activity versus small peptide levels.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, preferred embodiments of the present invention will be described in detail below to facilitate understanding of the skilled person.
Comparative analysis of feed protein source of amino acid balance of Poecilobdella manillensis
1. Comparative analysis of water-soluble crude protein content of different feed protein sources
As can be seen from fig. 1, there is a significant difference in the solubility of the protein of different feed protein sources (P < 0.05); the protein solubility difference among the isolated soybean protein, the soybean meal and the fish meal is not significant (P is more than 0.05); the protein solubility difference among the soybean protein concentrate, the soybean meal and the fish meal is not significant (P is more than 0.05); the protein solubility difference among milk powder, beef and pork is not significant (P is more than 0.05); the difference of protein solubility between pork and chicken is not significant (P is more than 0.05); the protein solubility difference among the pig blood, the blood cell albumen powder and the pig serum is not significant (P is more than 0.05); the solubility difference of the protein among the pig blood, the blood cell albumen powder and the whole egg is not significant (P >0.05), the protein solubility is the highest with egg albumen, and the protein solubility is not significant with the pig serum (P > 0.05). The protein solubility of 5 raw materials of pig blood, blood cell albumen powder, pig serum, egg albumen and whole egg is above 10%, the solubility of plasma albumen powder and egg yolk protein is above 9%, the raw materials are 7 raw materials with high protein solubility, and the protein solubility of the rest 8 raw materials is low.
2. Analysis of amino acid grey correlation degree of different feed protein sources
According to the grey correlation analysis method, the leech body amino acid composition of Poecilobdella manillensis is used as a reference sample, and the amino acid composition of 15 reference feed protein sources is used as greyA color system for calculating the correlation coefficient gamma of each amino acid in the 15 protein feed and the amino acid in the corresponding Poecilobdella manillensis0i(k) See table 1.
TABLE 1 Grey correlation coefficient of the balance of amino acids between feed protein source and Poecilobdella manillensis
As can be seen from Table 1, the correlation coefficient between each amino acid in the protein material and the amino acid of the hirudo is greatly different, and the closer the correlation coefficient is to 1, the higher the correlation coefficient is to the amino acid corresponding to the Poecilobdella manillensis.
The amino acid correlation coefficients of each feed protein source in table 1 were averaged to obtain the grey correlation degree of each feed protein source, see table 2.
TABLE 2 Grey correlation between amino acids of feed protein source and amino acids of Poecilobdella manillensis
As can be seen from table 2 and fig. 2: the amino acid gray correlation degree of the 15 feed protein sources and the Poecilobdella manillensis is as follows from high to low: beef, soybean concentrated protein, plasma protein powder, pork, chicken, fish meal, chicken protein, pig serum, soybean protein isolate, whole egg, pig blood, blood cell protein powder, soybean meal, milk powder and egg yolk. Wherein the correlation degree of the seven raw materials of beef, soybean protein concentrate, plasma protein powder, pork, chicken, fish meal and pig serum and the amino acid of the poecilobdella manillensis is more than 0.8.
3. Comparative analysis of feeding effect of poecilobdella manillensis on feed protein source
Selecting 7 feed protein sources of plasma protein powder, blood cell protein powder, pig blood, pig serum, egg protein, egg yolk and whole egg with high protein solubility and the amino acid relevance degree of the feed protein source, wherein the feed protein source refers to the ratio of water content to dry matter of blood of 2:8, independently preparing the raw materials to ensure that the protein content of the raw materials is close to the protein content of blood, homogenizing all the raw materials by a high-speed homogenizer, filling the raw materials into an artificial sausage casing for feeding, and scoring and evaluating the ingestion effect according to a set complete system, wherein the evaluation method is shown in a table 3. The feeding effect of the 7 feed protein sources was compared as shown in table 4.
TABLE 3 evaluation of feeding effect of Poecilobdella manillensis on feed protein sources
TABLE 4 comparison of the feeding effect of Poecilobdella manillensis on a single feed protein source
In order to compare the food intake of Poecilobdella manillensis on various single feed protein sources, 6 raw materials including plasma protein powder, blood globulin powder, pig blood, pig serum, egg protein and whole egg which have food intake phenomena are respectively compared, and the food intake is shown in figure 3.
As can be seen from fig. 3, the food intake of the poecilobdella manillensis to 7 raw materials is significantly different (P <0.05), and the food intake of the poecilobdella manillensis of each test group is sequentially from large to small: pig serum, plasma protein powder, egg protein, pig blood, blood cell protein powder and whole egg. Relatively high in pig serum, egg protein and plasma protein powder. The food intake of whole eggs is the lowest. The feed intake of the poecilobdella manillensis to pig serum is obviously higher than that of the rest 6 feed protein sources, and is respectively 3.69g (P <0.05), 4.81g (P <0.05), 9.83g (P <0.05), 10.34g (P <0.05) and 14.84g (P <0.05) higher than that of plasma protein powder, egg protein, pig blood, blood cell protein powder and whole egg.
4. Analysis of ingestion effect and addition amount of different feed protein sources
4.1 comparison of intake effects of different amounts of plasma protein powder
The dissolution forms of plasma protein powders with different addition amounts were observed, respectively, and are shown in table 5.
TABLE 5 comparison of plasma protein powder intake
As can be seen from Table 5, the dissolving effect gradually decreases with the increase of the plasma protein powder, and when the addition amount of the plasma protein powder is more than 15%, the ingestion effect is the best, but the feed has obvious caking phenomenon. By feeding plasma protein powder, the phenomenon that the Poecilobdella manillensis suffers from hematemesis after eating the plasma protein powder, and then the phenomenon that the body curls and wrinkles (see figure 4) occurs, and the phenomenon that the death rate is increased occurs.
4.2 comparison of ingestion effects of different pig serum white addition amounts
On the basis of adding 5% of plasma protein powder, 10%, 20%, 30%, 40% and 50% of pig serum are respectively added, and the ingestion effects of the poecilobdella manillensis at different adding concentrations are compared, and the results are shown in Table 6.
TABLE 6 comparison of the feeding effect of different pig serum addition concentrations
As can be seen from Table 6, with the increase of the addition amount of the pig serum, the ingestion effect of the poecilobdella manillensis is gradually enhanced, the ingestion amount is gradually increased, and when the addition amount of the pig serum is more than 40%, the ingestion effect is better.
4.3 comparison of feeding effects of different egg protein addition amounts
On the basis of 40% of pig serum, 10%, 20%, 30%, 40%, 50% and 60% of chicken protein are respectively added, and the ingestion effects of the poecilobdella manillensis at different adding concentrations are compared. See Table 7
TABLE 7 comparison of egg protein feeding Effect
As can be seen from Table 7, the egg protein and the pig serum have better dissolving effect within the range of 10-60% and better ingestion effect, and test Poecilobdella manillensis can quickly gather around the feed and show better ingestion behavior.
In conclusion, the pig serum and the egg protein are suitable for being used as a feed protein source of poecilobdella manillensis, the optimal addition amount of the pig serum is more than or equal to 40%, and the addition amount of the egg protein is between 10% and 60%, so that the pig serum and the egg protein have good ingestion effect.
Examples
On the basis that pig serum and egg protein are used as feed protein sources, glutathione is used as a small peptide source, and the pattern of four amino acids of Lys, Met, Thr and Trp is Lys: Met: Thr: Trp: 100:11.92:81.48:14.00, 25 kinds of poecilobdella manillensis compound feeds (examples 1-25) are prepared. The effect of the levels of feed protein and the addition of small peptide (glutathione) (2X 5) on Poecilobdella manillensis was analyzed. The test adopts a two-factor test design, the protein levels are set to be 5 protein levels in total of 8%, 9%, 10%, 11% and 12%, the small peptide (glutathione) is set to be 5 addition levels in total of 0.5%, 1.0%, 1.5%, 2.0% and 2.5% to carry out a cross grouping test, the cross grouping test is divided into 25 treatments, each treatment is carried out for 3 times, and the cross grouping test is carried out for 75 times, each treatment is carried out for 20 times, and the total number is 1500. (see tables 8 and 9.)
Table 8 test design table
Table 9 feed formulations and compositions (fresh sample%)
When the feed is prepared, the feed is stirred by a glass rod after being weighed, and is homogenized by a high-speed homogenizer until no solid residue exists in the feed, and then the prepared feed is filled into an artificial sausage casing with the length of about 20cm for feeding. When feeding, enough feed is guaranteed, the feed is taken freely, and when the optimal feeding time is 5 hours, the feeding frequency is 10 days/time.
Examples analysis
1. Influence of different feeds on growth performance of Poecilobdella manillensis
From the results analysis between groups (fig. 5-13), different feeds significantly affected the total food intake of poecilobdella manillensis (P <0.05), the total gain (P <0.05), the daily average gain (P <0.05), the end-of-term gain (P <0.05), the feed factor (P <0.05), the specific growth rate (P <0.05), the survival rate (P <0.05), and the protein efficiency (P < 0.05).
The main effect analysis of the effect of protein and small peptide levels on the growth performance index of Poecilobdella manillensis is shown in Table 10.
TABLE 10 analysis of the Main Effect of protein and Small peptide levels on the growth Performance index of Poecilobdella manillensis
Note: data in the same column are annotated with different lower case letters to indicate significant difference (P <0.05), and lower case letters with the same or no letter to indicate insignificant difference (P >0.05) under the same level factor.
As shown by the results analysis between factors (Table 9), the growth performance index of Poecilobdella manillensis is significantly influenced by different protein levels in the feed (P < 0.05). With the increase of the protein level of the feed, the total food intake, the whole-period weight gain, the average daily weight gain, the end-of-period weight gain, the specific growth rate, the survival rate, the weight gain rate and the protein efficiency of the poecilobdella manillensis show the trend of rising and falling, and the feed coefficient shows the trend of falling and rising. The feed small peptide level has obvious influence on the total food intake and survival rate of the poecilobdella manillensis (P is less than 0.05), and has no obvious influence on the whole-period weight gain, average daily weight gain, end-period weight gain, specific growth rate and weight gain rate of the poecilobdella manillensis and the protein efficiency (P is more than 0.05).
The feed small peptide level has obvious influence on the total food intake and survival rate of the poecilobdella manillensis (P is less than 0.05), and has no obvious influence on the whole-period weight gain, average daily weight gain, end-period weight gain, specific growth rate and weight gain rate of the poecilobdella manillensis and the protein efficiency (P is more than 0.05).
From the Eta2 values, the main effects of the protein level, the small peptide level and the protein x small peptide on the total food intake, the whole-term weight gain, the daily average weight gain, the end-of-term weight gain, the specific growth rate, the feed coefficient and the weight gain rate of poecilobdella manillensis are that the feed protein level > the protein x small peptide interaction > the small peptide level, and the influence degree of the survival rate is that the feed small peptide level > the protein x small peptide interaction.
The optimal combination of protein and small peptide levels can be found by plotting the interaction plots for total feed intake, total gain, average daily gain, end of term weight, specific growth rate, and gain rate for the feed protein level and the feed small peptide level, respectively, in order to screen the optimal combination of protein and small peptide levels, see fig. 14-19.
2. Rule analysis of influence of protein level on growth performance of Poecilobdella manillensis
The growth performance index of Poecilobdella manillensis is obviously influenced by different protein levels in the feed (P is less than 0.05). As the feed protein level and each growth performance index present obvious regression relations (P <0.05), the regression analysis can be respectively carried out on the feed protein level and each growth performance index, the optimal regression equation can be found out, 9 growth performance indexes of total food intake, total gain, daily average gain, end-of-term weight, gain rate, feed coefficient, specific growth rate, survival rate and protein efficiency and the feed protein level are subjected to regression analysis, the feed protein level is respectively taken as an independent variable x, the growth performance index of a corresponding protein level point is taken as a dependent variable y, a regression relation mathematical model is established, and the regression equation is fitted and shown in figure 20.
The effects of the feed protein and small peptide level interaction on the total food intake, the whole-term weight gain, the daily average weight gain, the end-of-term weight gain, the weight gain rate and the specific growth rate of the poecilobdella manillensis are consistent, namely the 10% protein level and the 2.0% small peptide level are combined with the total food intake, the whole-term weight gain, the daily average weight gain, the end-of-term weight gain, the weight gain rate and the specific growth rate of the poecilobdella manillensis.
As can be seen from FIG. 20, the feed protein level and the total food intake, the whole-term weight gain, the daily average weight gain, the end-of-term weight gain, the weight gain rate, the feed coefficient, the specific growth rate, the survival rate and the protein efficiency of poecilobdella manillensis all show significant cubic curve relations, and the regression equation of the feed protein level and the total food intake of the poecilobdella manillensis is that y is 4.160+4.416x-2.449(x-10)2-0.166(x-10)3，(R20.781). The highest total food intake of 50.20 g/strip of Poecilobdella manillensis is achieved at a feed protein level of 10.83%;
the regression equation of the feed protein level and the total-period weight gain of poecilobdella manillensis is as follows: y-1.538 +0.619x-0.644(x-10)2-0.007(x-10)3，(R20.781). The highest total gain of 7.88 g/stripe was obtained for Poecilobdella manillensis at a feed protein level of 10.48%.
The regression equation for the feed protein level and the daily average weight gain of poecilobdella manillensis is: y-17.094 +6.877x-7.161(x-10)2-0.075(x-10)3，(R20.781), the highest total weight gain of 87.51 mg/pig was obtained for poecilobdella manillensis at a feed protein level of 10.48%;
the regression equation of the feed protein level and the terminal body weight of poecilobdella manillensis is as follows: y-2.249 +0.646x-0.637(x-10)2-0.001(x-10)3，(R20.819), the highest total gain was obtained for poecilobdella manillensis 8.873g per strip at a feed protein level of 10.51%.
The regression equation of the feed protein level and the weight gain rate of poecilobdella manillensis is as follows: y-433.849 +36.116x-70.874(x-10)2-7.345(x-10)3，(R20.878), the highest weight gain of 799.49% was obtained with poecilobdella manillensis at a feed protein level of 10.25%.
The regression equation of the feed protein level and the feed coefficient of poecilobdella manillensis is as follows: y-5.575 +0.064x-0.361(x-10)2-0.071(x-10)3，(R20.999), the lowest feed factor of 6.21 was obtained for poecilobdella manillensis at a feed protein level of 9.91%.
Feed protein waterThe regression equation for the specific growth rate of Poecilobdella manillensis is: y-5.575 +0.064x-0.361(x-10)2-0.071(x-10)3，(R20.999), the lowest specific growth rate of 1.69 was obtained for poecilobdella manillensis at a feed protein level of 9.78%.
The regression equation of the feed protein level and the survival rate of poecilobdella manillensis is as follows: y-65.841 +2.466x-0.555(x-10)2-0.282(x-10)3，(R20.827), the highest survival rate of 92.17% was obtained for poecilobdella manillensis at a feed protein level of 11.17%.
The regression equation for the levels of feed protein and the protein efficiency of poecilobdella manillensis is: y-218.633-1.643 x-8.526(x-10)2+1.647(x-10)3，(R20.992), poecilobdella manillensis achieved the highest protein efficiency of 202.28% at a feed protein level of 9.91%.
From the regression analysis, it can be seen that the regression relationship between each growth performance index and the feed protein level is strongly correlated, and with 9 items of total food intake, total gain, daily average gain, end-of-term weight, gain rate, feed coefficient, specific growth rate, survival rate and protein efficiency as ordinate, it is found that along with the increase of the protein level in the daily ration, the growth performance indexes of the daily ration protein level and poecilobdella manillensis present a three-item curve model, and when the feed protein level is 9.78% -11.17%, the poecilobdella manillensis can obtain the optimal growth performance index.
3. Rule of influence of small peptide (glutathione) level on growth performance of Poecilobdella manillensis
The total food intake and survival rate growth performance index (P <0.05) of poecilobdella manillensis are obviously influenced by different small peptide levels in the feed. As the feed small peptide level and the total food intake and the survival rate present obvious regression relations (P is less than 0.05), the regression analysis can be respectively carried out on the feed small peptide level and the total food intake and the survival rate and the optimal regression equation is found out, and the regression analysis is carried out on the 2 growth performance indexes of the total food intake and the survival rate and the feed small peptide level, the feed small peptide level is respectively taken as an independent variable x, the growth performance index of the corresponding small peptide level point is taken as a dependent variable y, a regression relation mathematical model is established, and the regression equation is fitted, which is shown in the figure 3-22.
From fig. 21 it can be seen that the relationship between the feed small peptide level and the total food intake of poecilobdella manillensis, y being 52.734-4.828x-12.797(x-1.5)2+5.609(x-1.5)3+10.182(x-1.5)4, (R2 being 0.999), shows a significant four-way curve model, with the highest total food intake obtained at the inflection point of the parabola for the protein content of the poecilobdella manillensis feed by parabolic regression and 52.73 g/bar for the highest total food intake at a feed small peptide level of 1.48%; the relationship between the level of the feed small peptide and the survival rate of the poecilobdella manillensis shows a remarkable positive correlation, y is 86.615+3.181x, R2 is 0.976, and the survival rate of the poecilobdella manillensis is remarkably increased along with the increase of the feed small peptide (P < 0.05). The addition range of the feed small peptide is between 1.48 and 2.5 percent, and the Poecilobdella manillensis can obtain higher food intake and survival rate.
4. Influence of different feeds on antithrombin activity in Poecilobdella manillensis
Hirudin binds thrombin in a ratio of 1: 1 as determined by thrombin titration, whereas thrombin has the standard international unit NIH, so that the activity of hirudin can be expressed in units of antithrombin activity ATU, i.e. 1 ATU equals the amount of hirudin which neutralizes 1 NIH thrombin.
As can be seen from FIG. 22, different feeds significantly affected the anticoagulant enzyme activity in Poecilobdella manillensis (P <0.05), the anticoagulant activity component did not significantly differ among groups 2, 3 and 25 (P >0.05), the anticoagulant activity component did not significantly differ among groups 3 and 4 (P >0.05), the anticoagulant activity component did not significantly differ among groups 4 and 5 (P >0.05), the anticoagulant activity component did not significantly differ among groups 5, 6, 22 and 24 (P >0.05), the anticoagulant activity component did not significantly differ among groups 7, 8 and 20 (P >0.05), the anticoagulant activity component did not significantly differ among groups 7, 8 and 23 (P >0.05), the anticoagulant activity component did not significantly differ among groups 19, 21 and 23 (P >0.05), the anticoagulant activity component did not significantly differ among groups 9, 18, 19 and 21 (P >0.05), the anticoagulant activity component did not significantly differ among groups 10, 11 and 17 (P >0.05), the anticoagulant activity did not significantly differ among groups 10, 11 and 18 (P >0.05), the anticoagulant activity did not significantly differ among groups 12, 14 and 15 (P >0.05), and the highest anticoagulant activity content of the Poecilobdella manillensis is 13, significantly higher than the remaining 24 groups (P < 0.05). The group 1 antithrombin activity was the lowest, significantly lower than the anticoagulant activity of the remaining 24 groups of feed (P < 0.05).
The analysis of the main effect of the influencing factors of the antithrombin active ingredient in the Poecilobdella manillensis is shown in Table 11.
TABLE 11 Main Effect of protein and Small peptide levels on Poecilobdella manillensis antithrombin Activity assay of absolutely dry sample%
From the analysis of the results in table 11, it can be seen that different protein addition levels in the feed significantly affected the antithrombin activity in poecilobdella manillensis (P < 0.05). From the feed protein addition level, the antithrombin activity in the poecilobdella manillensis tends to increase and decrease along with the increase of the feed protein level. The anticoagulant active ingredient was highest in the 10% protein level group (P < 0.05).
The level of the feed small peptide has a remarkable influence on the antithrombin activity in the poecilobdella manillensis (P is more than 0.05), and the antithrombin activity in the poecilobdella manillensis tends to increase and decrease along with the increase of the level of the feed small peptide. The antithrombin activity in the poecilobdella manillensis is remarkably reduced (P < 0.05). The differences in anticoagulant activity between the 0.5%, 1.0% and 2.5% small peptide levels were not significant (P >0.05), the differences in antithrombin activity between the 1.0%, 2.0% small peptide levels were not significant (P >0.05), and the 3 small peptide addition groups had no significant effect on antithrombin activity between the 1.5%, 2.0% small peptide levels (P > 0.05). Antithrombin activity was highest in the group of 1.5% small peptide levels, 11.33ATU/g (P <0.05), 6.66ATU/g (P <0.05), 10.02ATU/g (P <0.05) higher than in the group of 0.5%, 1.0%, 2.5% small peptide levels.
From Eta2It is known that the degree of influence of antithrombin activity in Poecilobdella manillensis is the protein level>Interaction of protein levels with small peptide levels>Small peptide levels.
The interaction of feed protein levels and small peptide levels significantly affected the antithrombin activity in poecilobdella manillensis (P < 0.05). The interaction of the anti-thrombin activity at the feed protein level and the small peptide level was plotted, see figure 23 below.
As can be seen from FIG. 23, the antithrombin activity in Poecilobdella manillensis is highest at a 10% protein level in combination with a 1.5% small peptide addition level, and lowest at 8% protein level in combination with a 0.5% small peptide addition level.
In conclusion, different feeds significantly affected the anticoagulant active ingredient in poecilobdella manillensis (P < 0.05). The feed protein and small peptide levels have influence on the anticoagulant active ingredients in the poecilobdella manillensis to the extent that the protein level is greater than the protein x small peptide level, and when the 10% protein level is combined with the 1.5% small peptide adding level, the content of the anticoagulant active ingredients in the poecilobdella manillensis is the highest. When the feed protein level is 10.21% -10.38%, higher content of anticoagulant active ingredients can be obtained.
5. Law of influence of anticoagulation active ingredients of Poecilobdella manillensis and protein level of feed
The different protein levels in the feed significantly affect the antithrombin activity of poecilobdella manillensis (P < 0.05). And (4) performing regression analysis on the feed protein level and the antithrombin activity respectively, and finding out an optimal regression equation. Respectively taking the feed protein level as an independent variable x and the antithrombin activity of the corresponding protein level point as a dependent variable y, establishing a regression relation mathematical model, and performing regression analysis through antithrombin activity and the feed protein level, wherein a regression equation is fit and shown in figure 24.
6. Prediction model research between anticoagulant active ingredient and feed small peptide level in poecilobdella manillensis
The level of different small peptides in the feed significantly affects the antithrombin activity of poecilobdella manillensis (P < 0.05). Respectively carrying out regression analysis on the feed protein level and the antithrombin activity, finding out an optimal regression equation, respectively taking the feed small peptide level as an independent variable x, taking the antithrombin activity of a corresponding small peptide level point as a dependent variable y, carrying out regression analysis through the antithrombin activity and the feed small peptide level, establishing a regression relation mathematical model, and fitting the regression equation into a figure 25.
As can be seen from fig. 25, the levels of the small peptides in the feed are dispersed compared with antithrombin activity and dispersion points between relative gray values, and after regression analysis, no significant regression relationship exists between the levels (P >0.05), and a regression equation between the levels cannot be established.
In conclusion, different feeds obviously influence the growth performance and the antithrombin activity of the poecilobdella manillensis, the optimal protein level range of the artificial compound feed for the poecilobdella manillensis is 9.78-11.17%, and the small peptide level range is 1.48-2.5%.
In the nutrition level range, the raw materials of the poecilobdella manillensis compound feed comprise pig serum, egg protein, threonine, methionine, lysine, tryptophan, glutathione, glycine, arginine, salt and water. The mixture ratio of various raw materials is as follows: 40.00% of pig serum; 16.09-30.59% of egg protein; 0.49-0.55% of threonine; 0.095-0.10% of methionine; 0.20-0.33% of lysine; 0.06-0.07 of tryptophan; 1.48-2.5% of glutathione; 0.50% of glycine; 0.50 percent of arginine; 0.40 percent of salt; 25.68-39.99% of water; the sum of the weight percentages of all the raw materials is 100 percent.
The invention provides artificial compound feed for poecilobdella manillensis, which fully considers the characteristics of food habit, nutrition requirement, amino acid balance and digestion of the poecilobdella manillensis, reasonably establishes the levels of protein, small peptide and amino acid in the artificial compound feed for the poecilobdella manillensis, selects the conventional feed raw materials which are easy to digest and absorb for compatibility, and has stable used raw materials and easy preparation. The protein source amino acid of the compound feed has high degree of balance and solubility, and the feeding effect of the poecilobdella manillensis is good. After the compound feed is fed, the growth performance and the antithrombin activity of the poecilobdella manillensis are improved, the survival rate is high, and no hematemesis phenomenon occurs in the feeding process, which indicates that the compound feed meets the nutritional requirements of the poecilobdella manillensis under the artificial culture condition.
Finally, it is noted that the above-mentioned preferred embodiments illustrate rather than limit the invention, and that, although the invention has been described in detail with reference to the above-mentioned preferred embodiments, it will be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the scope of the invention as defined by the appended claims.
Claims (3)
1. A Poecilobdella manillensis compound feed is characterized in that: the protein level range of the poecilobdella manillensis compound feed is 9.78-11.17%, and the small peptide level range is 1.48-2.5%.
2. The Poecilobdella manillensis compound feed as claimed in claim 1, which is characterized in that: the Poecilobdella manillensis compound feed comprises pig serum, egg protein, threonine, methionine, lysine, tryptophan, glutathione, glycine, arginine, salt and water.
3. The Poecilobdella manillensis compound feed as claimed in claim 1 or 2, wherein the Poecilobdella manillensis compound feed comprises the following components in parts by weight: the Poecilobdella manillensis compound feed comprises the following components in percentage by weight: 40.00 percent of pig serum, 16.09-30.59 percent of egg protein, 0.49-0.55 percent of threonine, 0.095-0.10 percent of methionine, 0.20-0.33 percent of lysine, 0.06-0.07 percent of tryptophan, 1.48-2.5 percent of glutathione, 0.50 percent of glycine, 0.50 percent of arginine, 0.40 percent of salt and 25.68-39.99 percent of water; the sum of the weight percentages of all the raw materials is 100 percent.
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