CN112544816A

CN112544816A - Feed for improving intestinal microbial diversity of chickens and application thereof

Info

Publication number: CN112544816A
Application number: CN202011361445.4A
Authority: CN
Inventors: 黄志朝; 易显凤; 邓素嫒; 史静; 曹慧慧; 杨启晟
Original assignee: Guangxi Zhuang Autonomous Region Institute of Animal Husbandry
Current assignee: Guangxi Zhuang Autonomous Region Institute of Animal Husbandry
Priority date: 2020-11-27
Filing date: 2020-11-27
Publication date: 2021-03-26
Anticipated expiration: 2040-11-27
Also published as: CN112544816B

Abstract

The invention relates to the technical field of microbial diversity feed processing, in particular to a feed for improving the microbial diversity of chicken intestinal tracts and application thereof, and tests on different green feeds and different pastures show that the fermented purple elephant grass has the effect of promoting the formation of microbial diversity flora of the chicken intestinal tracts relative to other pastures and even common varieties of elephant grass, namely Guimin primer, and when the addition amount of the fermented purple elephant grass is 10-25 percent, the microbial diversity of the chicken intestinal tracts is obviously improved; feeding experiments show that the chicken with higher microbial diversity has obvious intestinal tract regulating effect and greatly reduced morbidity, so that the chicken yield and quality are further improved, and the breeding advantages are most obvious due to the addition of 20% of fermented grassiness.

Description

Feed for improving intestinal microbial diversity of chickens and application thereof

[ technical field ] A method for producing a semiconductor device

The invention relates to the technical field of microbial diversity feed processing, in particular to a feed for improving intestinal microbial diversity of chickens and application thereof.

[ background of the invention ]

The Jinling local-top chicken is one of the distinctive chicken varieties bred in Guangxi at present, is praised by people due to delicious taste and fine meat quality, and is still in a relatively extensive stage about breeding of the chicken varieties at present, so that farmers do not research the feed formula of the chicken of the varieties; in the breeding process of the breeder chicken, intestinal diseases are an important factor influencing the growth and development of the breeder chicken, and the incidence rate of the intestinal diseases is slightly higher compared with that of other chicken breeds; at present, when the inventor researches the chicken variety, the inventor finds that the conventional probiotics are added: the intestinal tract disease of the chicken is not obviously relieved when the lactobacillus or EM bacterial liquid is added into the feed, which is probably because different varieties of chicken have different colonization capacities on different strains, and certain probiotics can colonize in certain varieties of chicken, but a good colonization effect cannot be achieved in the varieties of chicken, so that a dominant flora cannot be effectively formed to modify the intestinal tract of the chicken; for this reason, the applicant considered to start with the isolation of microorganisms from the chicken intestine in order to be able to isolate from this chicken intestine microorganisms suitable for colonizing this chicken intestine. Therefore, in order to realize once sampling and realize microbial screening as much as possible, the applicant considers that certain feed additives are added, so that the abundance and the biodiversity of the intestinal flora of the chickens can be improved, the later strain screening work is facilitated, the increase of the abundance of the intestinal microbes of the chickens can also effectively reduce the occurrence of intestinal diseases of the chickens in the research process, and the improvement of the feed formula is far simpler than the strain screening work.

[ summary of the invention ]

The feed improves the abundance of microorganisms in the intestinal tracts of the Jinling local-collar chickens, radically improves the digestion and absorption capacity of the Jinling local-collar chickens on the feed, and improves the nutrient intake, thereby achieving the purposes of improving the chicken yield and improving the meat quality.

Feed for improving the microbial diversity of chicken intestinal tracts, which comprises 10 to 25 weight percent of fermented grassiness.

Further, the feed also comprises 75-90 wt% of conventional chicken feed, and the conventional chicken feed comprises the following components in percentage by weight: 30% of corn flour, 20% of rice bran, 0.1% of salt, 1% of trace elements and the balance of rice flour.

Further, the trace elements consist of the following components: 10% of vitamin A, 35% of vitamin D, 30% of methionine, 5% of ferrous sulfate and 20% of vitamin E.

Further, the fermentation method for fermenting the grassiness comprises the following steps: cleaning and crushing elephant grass, and then composting at room temperature, wherein the moisture content of the grass pile is ensured to be 50% -60% during composting; and (5) after the compost is retted for 7d, drying the elephant grass until the moisture is 5 percent, and finishing the treatment.

Further, the variety of the grassiness is as follows: purple elephant grass.

Further, the breeding method of the purple elephant grass comprises the following steps: the method comprises the steps of performing sexual propagation on Brazilian purple elephant grass, selecting plants with high grass yield and good quality for cuttage vegetative propagation, selecting seed stems with high single-plant yield, high crude protein content, more tillers, large leaf quantity, thick and strong seed stems and no harm of diseases and insects from a propagation group, eliminating bad plants, and performing stable selection and planting for 10 times to obtain plants tending to stability, namely the purple elephant grass.

The application also comprises the application of the feed for improving the intestinal microbial diversity of the chickens in reducing the intestinal morbidity of the chickens.

The application also comprises the application of the feed for improving the intestinal microbial diversity of the chickens in improving the growth speed.

The application also comprises a feeding method for improving the growth speed by applying the feed for improving the intestinal microbial diversity of the chickens, wherein the method is to add fermented grassiness with the weight percentage of 20 percent into the conventional chicken feed.

The invention has the following beneficial effects:

through repeated tests on different green feeds and different pastures, the application finally discovers that the purple elephant grass variety has larger influence on the microbial diversity of the chicken intestinal tract than the conventional elephant grass variety, the purple elephant grass is used after fermentation and can better promote the formation of the microbial diversity flora of the chicken intestinal tract, and probably because the contents of certain trace elements in the purple elephant grass are different from those of common elephant grass, for example, the content of anthocyanin is far higher than that of common elephant grass, and after silage fermentation, the trace elements in the forage grass are further released and chelated, and the nutritional ingredients of the forage grass are greatly different from those of the green feeds, so that the stimulation generating effect of the fermented feeds on microorganisms in the chicken intestinal tract is greater than that of the green feeds; the applicant takes fermented purple elephant grass as a model forage grass for further research, and finally finds that the addition of the fermented elephant grass influences the diversity of intestinal microorganisms of the chicken, the diversity is obviously improved when the addition is 10% -25%, and the effect of stimulating the growth and the reproduction of microorganisms or inhibiting the growth and the reproduction of the microorganisms cannot be achieved when the addition is too much or too little; through feeding experiments, the applicant finds that chickens with higher intestinal microorganism diversity have obvious intestinal regulation effect and greatly reduced morbidity, so that the yield and the quality of chicken are further improved, and the breeding advantages of the fermented elephant grass with the addition of 20% are most obvious.

[ description of the drawings ]

FIG. 1 is a petal plot of the distribution of chicken intestinal microbial species;

FIG. 2 is a heat map of the distribution of the first 30 chicken gut microbial species.

[ detailed description ] embodiments

The invention is further illustrated below with reference to the figures and the examples.

Example 1:

test animals and sites

The experiment is carried out in a chicken farm of Guangxi livestock institute, and 60-day-old chicks are selected and bred to Jinling local-neck chickens.

288 feathers of broilers with similar weights are randomly divided into 6 groups, each group is provided with 4 repetitions, each repetition is 12-feather broilers, and each cage is fed with 3-feather broilers. The pre-test period is 7d, and the test official period is 60 d. Cage culture, free food intake and drinking, natural illumination, environment humidity of about 65 percent, good ventilation and clean and sanitary maintenance.

The feed for raising chickens is prepared from the following components in percentage by weight:

experiment one: 10% of Guiminzhenxiaohong and 90% of conventional chicken feed

The method for treating the elephantopus cinerea and the minjian elephantopus grass in the experiment comprises the following steps: cleaning and crushing the elephant grass, and adding the elephant grass into the chicken feed according to the mass ratio.

Experiment two: 10% of Guiminzhenxiaohong and 90% of conventional chicken feed

The method for treating the elephantopus cinerea and the minjian elephantopus grass in the experiment comprises the following steps: cleaning and crushing elephant grass, and then composting at room temperature, wherein the moisture content of the grass pile is ensured to be 50% -60% during composting; after the stack is retted for 7d, drying the elephant grass until the moisture is 5 percent, and finishing the treatment; and then adding the processed Guimingxiao elephantopus into the chicken feed according to the mass ratio.

Experiment three: 10% of purple elephant grass and 90% of conventional chicken feed

The purple elephant grass treatment method of the experiment comprises the following steps: cleaning and crushing the elephant grass, and adding the elephant grass into the chicken feed according to the mass ratio.

Experiment four: 10% of purple elephant grass and 90% of conventional chicken feed

The purple elephant grass treatment method of the experiment comprises the following steps: cleaning and crushing elephant grass, and then composting at room temperature, wherein the moisture content of the grass pile is ensured to be 50% -60% during composting; after the stack is retted for 7d, drying the elephant grass until the moisture is 5 percent, and finishing the treatment; and then adding the treated purple elephant grass into the chicken feed according to the mass ratio.

Experiment five: 10% alfalfa and 90% conventional chicken feed

The alfalfa processing method of the experiment comprises the following steps: cleaning and crushing the alfalfa, and adding the alfalfa into the chicken feed according to the mass ratio.

Experiment six: 10% alfalfa and 90% conventional chicken feed.

The alfalfa processing method of the experiment comprises the following steps: cleaning and crushing alfalfa, and then composting at room temperature, wherein the moisture content of the grass pile is ensured to be 50% -60% during composting; after the stack is retted for 7d, drying the elephant grass until the moisture is 5 percent, and finishing the treatment; and then adding the treated alfalfa into the chicken feed according to the mass ratio.

Control group: conventional chicken feed.

The purple elephant grass variety of the embodiment is a variety which is selected by the applicant independently and is examined by the autonomous region (or country) of the Guangxi Zhuang nationality at present.

The variety is introduced from Brazil, and the breeding method comprises the following steps:

the purple elephant grass is subjected to sexual propagation, plants with high grass yield and good quality are selected for cuttage vegetative propagation, seed stems with high single-plant yield, high crude protein content, more tillers, large leaf quantity, thick and strong seed stems and no harm of diseases and insects are selected from a propagation population, bad plants are eliminated, the plants with stable performance trend are obtained through continuous 10 times of stable selection, and the purple elephant grass plants can be planted in Guangxi and have the characteristics of large leaf quantity, high crude protein content and high fresh grass yield.

Through detection, 2007-2009 comparison tests are carried out on 3 elephant grass varieties such as purple elephant grass, Guiminyin elephant grass and king grass in Lingyun county and Tian-equant county. The results of 3-year observation and measurement show that the average annual grass production of purple elephant year in Nanning county, Lingyun county and Tian county is 134530.7 kg/hm²～160 763.4kg/hm²In between, the average annual grass production of the three test points is 148829.16 kg/hm²The difference is not significant; the crude protein content of the purple elephant grass is higher by 7.28 percent and is respectively improved compared with that of the Guiminzhen elephant grass and the wangcao11.49 percent and 35.82 percent, has obvious difference with the grassiness of the world of Guijian, and has obvious difference with the king grass; the purple elephant grass has lower crude fiber, the crude ash content is respectively 31.46 percent and 35.17 percent of that of the Guijian Zhenxiang elephant grass and the wangcao, and the difference is extremely obvious; the minimum stem-leaf ratio of the purple elephant grass is 0.91: 1, the amount of the purple elephant grass is 34.49 percent and 18.57 percent lower than that of the Chinese angelica root and the Chinese sage respectively, and the difference obviously indicates that the leaf amount of the purple elephant grass is more abundant than that of the other two types; in addition, the purple elephant grass has unique purple red color, contains rich anthocyanin, has higher oxidation resistance and stronger functionality in feed application. In conclusion, the purple elephant grass has the unique advantages of high crude protein and crude ash content, leafiness and purplish red color, and is a variety worthy of vigorous popularization in Guangxi.

The conventional chicken feed comprises the following components in percentage by weight: 30% of corn flour, 20% of rice bran, 0.1% of salt, 1% of trace elements and the balance of rice flour; the trace elements comprise the following components in parts by weight: 10% of vitamin A, 35% of vitamin D, 30% of methionine, 5% of ferrous sulfate and 20% of vitamin E.

After 60 days of feeding, randomly taking chickens from experiment one to experiment six and a control group, slaughtering and dissecting, sampling chicken intestinal tracts, then carrying out 16s rDNA sequencing, analyzing the microbial community situation in the chicken intestinal tracts, comparing all optimized sequences with OTU representative sequences at the level of genus level, setting the sequences with the similarity of more than 97 percent of the OTU representative sequences as the same OTU, and statistically generating an OTU abundance table, wherein the results are shown in Table 1:

TABLE 1 OTU number of each sample

Group of	Experiment one	Experiment two	Experiment three	Experiment four	Experiment five	Experiment six	Control group
								Number of OTU	257	226	208	277	201	242	210

As can be seen from Table 1, the highest OTU number in experiment four indicates that the fermented purple elephant grass can well improve the OTU abundance of the chicken intestinal tract and improve the diversity of microorganisms; the OTU quantity difference of the experiment I, the experiment III, the experiment V and the control group is not large, even the OTU quantity of the experiment III and the experiment V is lower than that of the control group, so that the green feed has no great influence on the intestinal microorganisms of the chickens, and the diversity of the intestinal microorganisms of the chickens can be reduced by adding some green feed; compared with the control group and the unfermented feed, the OTU number of the second experiment, the fourth experiment and the sixth experiment is obviously increased, so that the microbial content of the green feed is increased after the green feed is subjected to stack retting fermentation, and trace elements are further released, so that the generation of microbial communities in intestinal tracts of chickens can be effectively stimulated, the microbial abundance in the intestinal tracts of chickens is increased, and the fermented purple elephant grass is greater than the fermented Guimin elephant grass is greater than the fermented alfalfa from the improvement result.

Example 2:

on the basis of example 1, the applicant has concluded that fermented purple elephant grass has the greatest effect on intestinal microorganisms in chickens in fermented forage grass, and further conducted the following experiments in order to further study the appropriate amount thereof: the 200 feathers of 60-day-old chicks with similar weight are randomly divided into 10 groups, each group is provided with 4 replicates, each replicate is provided with 5-feather chicks, and each cage is provided with 5-feather chicks. The pre-test period is 7d, and the test official period is 60 d. The chicken is raised in cages, free feeding and drinking are realized, natural illumination is realized, the environment humidity is about 65%, ventilation is good, cleanness and sanitation are kept, the chickens are raised according to the formula shown in the table 2, the slaughtered chickens are dissected at the 60 th day, the intestinal tracts of the chickens are sampled, then 16s rDNA sequencing is carried out, the microbial community condition in the intestinal tracts is analyzed, and the specific result is shown in the table 3.

Table 2 feed ingredient table of this example

In table 2, the preparation method of fermented purple elephant grass is as follows: cleaning and crushing elephant grass, and then composting at room temperature, wherein the moisture content of the grass pile is ensured to be 50% -60% during composting; after the stack is retted for 7d, drying the elephant grass until the moisture is 5 percent, and finishing the treatment; then adding the treated purple elephant grass into the chicken feed according to the mass ratio; the conventional chicken feed of this example was the same as that of example 1.

The sequencing analysis shows that the chicken intestinal microorganism OTU of the embodiment is shown in the table 3:

TABLE 3 Effect of the amount of added fermented elephant grass on the OTU of chicken intestinal tract

Experimental group	A	B	C	D	E	F	G	H	I	J
											OTU	210	224	277	283	295	291	262	275	246	208

As can be seen from table 3, the addition amount of the fermented purple elephant grass and the community of the microorganisms in the chicken intestinal tract are not in a linear relationship, that is, the addition amount of the fermented purple elephant grass is more, the community number of the microorganisms in the chicken intestinal tract is more, 10% -25% of the addition amount of the fermented purple elephant grass can effectively promote the formation of the microbial community in the chicken intestinal tract, from 30% of the addition amount, the OTU number is in a descending trend, even the total fermented elephant grass feeding group (J) has a lower number of the microorganisms in the chicken intestinal tract than the common feed group (a), so that the abundance of the microorganisms can be increased and the abundance of the microorganisms is related to the addition amount of the silage; this is probably because dominant bacteria of certain types of bacteria may be formed in the green feed and can inhibit the growth of other bacteria, thus leading to the reduction of the diversity of the microorganisms, and in the application, the addition amount of the fermented purple elephant grass is 10-25 percent, which can effectively improve the diversity of the intestinal microorganisms of the chickens.

Performing cluster analysis according to the OTU quantity, drawing an OTU petal graph, as shown in FIG. 1, wherein the common OTU quantity of two groups of samples or 10 samples is not identified, and only the OTU quantity of 10 groups of samples is identified, wherein circles with different colors represent different sample groups, the number on the petal represents the OTU quantity specific to the sample, and the middle white circle represents the OTU quantity common to all samples; as can be seen from the figure, the number of OTUs shared among 10 groups of samples reached 193; the special OTU of the group B experiment group is 1, and accounts for 0.45 percent of the total number of the OTUs of the group B experiment group; the specific OTU of the group C experiment group is 8, which accounts for 2.89% of the total number of the OTUs of the group C experiment group; the specific OTU of the group D experiment group is 10, which accounts for 3.53% of the total number of the OTUs of the group; the special OTU of the group E experiment group is 12, which accounts for 4.07% of the total number of the OTUs of the group E experiment group; the specific OTU of the F group of experiment groups is 7, which accounts for 2.41 percent of the total number of the OTUs of the group of experiment groups; the specific OTU of the group G experiment group is 3, which accounts for 1.15% of the total number of the OTUs of the group; the specific OTU of the group H experiment group is 1, and accounts for 0.36% of the total number of the OTUs of the group H experiment group; the characteristic OTU of the groups I-J and A was 0. Therefore, the more OTUs in the experimental group, the corresponding increase of the specific OTU, and the most specific microorganisms in the experimental group E in terms of the specific OTU indicate that the diversity of the experimental group is the highest.

The analysis of the first 30 species of the microorganisms showed that the results are shown in FIG. 2, which shows that the species of the microorganisms most abundant in the chicken intestine are Bacteroides genus (Bacteroides genus), 2 nd unrassified bacteria, 3 rd Megamonas genus (Megamonas genus), 4 th Ruminoccaceae family; the chicken intestinal microorganisms are proved to contain rich germplasm resources, and the method is a research direction for further developing probiotic research work.

Example 3:

feeding experiment:

from the analysis of the example 1 and the example 2, when the addition amount of the purple fermented elephant grass is 10% -25%, and when the addition amount is 20%, the specific OTU content is highest, so that the addition amount of the purple fermented elephant grass is 20% and is used as an experimental group, the addition amounts of the fermented Guiminjian elephant grass and the fermented alfalfa with the same mass ratio are simultaneously selected to prepare corresponding feeds, the feeding experiment is carried out on the Jinling local junipers, and the feed formula for feeding is shown in the table 4:

TABLE 4 feed formula for feeding experiment

The conventional chicken feed of this example is the same as that of example 1, the preparation method of fermented pasture is the same as that of example 1, and the chickens of the Jinling local neck are fed according to the above table, and the feeding grouping method is as follows: the 140 feathers of 60-day-old chickens with similar body weight are randomly divided into 7 groups, wherein each group comprises 20 feathers and half of a male parent and a female parent.

The feeding method comprises the following steps: after chicken groups are grouped, each group is confined in a breeding field, then the feed in the table 4 is put into the field, the feed and the water are sufficient, the chickens can feed freely, and other breeding conditions are consistent; the raising period is 60d, the initial weight of the chickens is counted before the experiment, the final weight of the chickens is counted and calculated after the experiment, and the average weight is calculated by taking an average value finally because the chickens die in the raising process, namely the average weight gain is the total end body weight at the end of the period/the total number of the chickens at the end of the period-the total end body weight at the beginning of the period/the total number of the chickens at the beginning of the period; the incidence of intestinal diseases of chickens is observed in the whole experimental period, the incidence is recorded as 1 time after 1 time until the chickens are cured, if the chickens are cured and relapse, the incidence is recorded as 2, and the final incidence calculation formula is as follows whether the chickens have the diseases or not: incidence (%) ═ number of attacks × 100/total number of initial chickens; and calculating the mortality rate, wherein the mortality rate calculation formula is as follows: mortality (%). number of dead chicks x 100/initial chicken total.

TABLE 5 Effect of feed on Chicken quality

Group of	Average weight gain (g/d)	Incidence (%)	Mortality (%)
				Group 1	17.54	10	0
Group 2	18.06	5	0
				Group 3	19.03	0	0
Group 4	18.52	5	0
				Group 5	17.12	25	5
Group 6	17.04	15	10
				Group 7	15.24	50	15

As can be seen from table 5, from the weight gain, the average weight gain of the groups 1 to 6 is higher than that of the group 7, the morbidity of the groups 1 to 6 is lower than that of the group 7, and the mortality of the groups 1 to 6 is lower than that of the group 7, which indicates that the biological diversity of the chicken intestinal tract is improved by adding the fermented green feed, so that the chicken intestinal tract can be adjusted, the digestive ability of the chicken is improved, the weight gain is larger than that of the control group (the group 7), and the weight gain, the morbidity and the mortality of the group 3 are the largest, so that the influence on the chicken feeding is the best when the adding amount of the fermented purple elephant grass reaches 20%, the weight is increased by 24.87% and the morbidity and mortality are reduced by 100% compared with the control group.

Therefore, the optimum addition amount of the fermented purple elephant grass is 20% in the process of raising the chickens.

In conclusion, the fermented purple elephant grass is applied to the feed, the diversity of intestinal microorganisms of the chickens can be effectively improved, meanwhile, intestinal tracts of the chickens can be adjusted, the effects of increasing weight and reducing morbidity and mortality can be achieved, and the optimal addition amount is 20% in the experiment of feeding the chickens.

The above description is intended to describe in detail the preferred embodiments of the present invention, but the embodiments are not intended to limit the scope of the claims of the present invention, and all equivalent changes and modifications made within the technical spirit of the present invention should fall within the scope of the claims of the present invention.

Claims

1. Feed for improving intestinal microbial diversity of chickens, characterized in that the feed comprises 10-25% by weight of fermented grassiness.

2. The feed for improving intestinal microbial diversity of chickens according to claim 1, further comprising 75-90% by weight of conventional chicken feed, wherein the conventional chicken feed comprises the following components in percentage by weight: 30% of corn flour, 20% of rice bran, 0.1% of salt, 1% of trace elements and the balance of rice flour.

3. The feed for improving intestinal microbial diversity of chickens of claim 1, wherein the trace elements consist of: 10% of vitamin A, 35% of vitamin D, 30% of methionine, 5% of ferrous sulfate and 20% of vitamin E.

4. The feed for improving the intestinal microbial diversity of chickens of claim 1, wherein the fermentation method for fermenting the grassiness is as follows: cleaning and crushing elephant grass, and then composting at room temperature, wherein the moisture content of the grass pile is ensured to be 50% -60% during composting; and (5) after the compost is retted for 7d, drying the elephant grass until the moisture is 5 percent, and finishing the treatment.

5. The feed for improving intestinal microbial diversity of chickens of claims 1-4, wherein the grassiness variety is: purple elephant grass.

6. Use of a feed according to any one of claims 1 to 5 for increasing the microbial diversity of the intestinal tract of chickens for reducing the incidence of intestinal disease in chickens.

7. Use of a feed according to any one of claims 1 to 5 for increasing the intestinal microbial diversity of chickens for increasing the growth rate.

8. A method for increasing the growth rate of chickens using the feed for increasing the intestinal microbial diversity of chickens of any one of claims 1 to 5, wherein the method comprises adding 20 wt% of fermented elephant grass to the conventional chicken feed.