AU2021102073A4

AU2021102073A4 - Solid-state fermented feed for improving meat quality of fattening pigs as well as preparation method and application thereof

Info

Publication number: AU2021102073A4
Application number: AU2021102073A
Authority: AU
Inventors: Kaiguo Gao; Youjun HU; Zongyong JIANG; Yueqin Qiu; Li Wang; Xiaolu Wen; Qiwen Wu; Xuefen Yang
Original assignee: Institute of Animal Science of Guangdong Academy of Agricultural Sciences
Current assignee: Institute of Animal Science of Guangdong Academy of Agricultural Sciences
Priority date: 2021-04-20
Filing date: 2021-04-20
Publication date: 2021-06-03
Anticipated expiration: 2029-04-20

Abstract

Austracy The present invention provides a solid-state fermented feed for improving meat quality of fattening pigs as well as a preparation method and an application thereof. The solid-state fermented feed is prepared by scientific compatibility of conventional high-quality feed raw materials and multi-bacterium mixed solid state fermentation of the complete feed with aerobic bacteria and anaerobic bacteria according to growth needs of the fattening pigs. The solid-state fermented feed has the advantages of low antigen content, low content of infectious microbes and toxins, excellent palatability and high digestion and utilization ratio. The present invention further provides a preparation method of the solid-state fermented feed, which is simple and convenient in operation, short in needed fermentation time, simple in a preservation method of a finished product, long in preservation time and capable of greatly reducing manual cost in the production process. The solid-state fermented feed is particularly applicable to feeding of the fattening pigs at a pre-slaughtering stage, needs no extra enzyme formulations, acidifiers or other feed additives, is short in use cycle, excellent in feeding effect and high in input-output ratio, may significantly improve the meat quality, and has obvious social and ecological significances and wide market space. 1

Description

Austracy

The present invention provides a solid-state fermented feed for improving meat quality of fattening pigs as well as a preparation method and an application thereof. The solid-state fermented feed is prepared by scientific compatibility of conventional high-quality feed raw materials and multi-bacterium mixed solid state fermentation of the complete feed with aerobic bacteria and anaerobic bacteria according to growth needs of the fattening pigs. The solid-state fermented feed has the advantages of low antigen content, low content of infectious microbes and toxins, excellent palatability and high digestion and utilization ratio. The present invention further provides a preparation method of the solid-state fermented feed, which is simple and convenient in operation, short in needed fermentation time, simple in a preservation method of a finished product, long in preservation time and capable of greatly reducing manual cost in the production process. The solid-state fermented feed is particularly applicable to feeding of the fattening pigs at a pre-slaughtering stage, needs no extra enzyme formulations, acidifiers or other feed additives, is short in use cycle, excellent in feeding effect and high in input-output ratio, may significantly improve the meat quality, and has obvious social and ecological significances and wide market space.

Description

SOLID-STATE FERMENTED FEED FOR IMPROVING MEAT QUALITY OF FATTENING PIGS AS WELL AS PREPARATION METHOD AND APPLICATION THEREOF

Technical Field

The present invention belongs to the technical field of feed, and particularly relates to a solid-state fermented feed for improving meat quality of fattening pigs as well as a preparation method and an application thereof.

Background

In recent years, China has attached great importance to ecological civilization construction and encourages the research, development and application of efficient, safe and healthy breeding technologies of commercial pigs. Meanwhile, the demands of people on high-quality safe meat products are increased. With the target directions of green, efficiency, safety and high quality, researching and developing a preparation technology of pig feed that is high in digestion and utilization ratio, less in emission of heavy metals and other manure contaminants, no residue of antibiotic and heavy metal and high in meat quality becomes a focus of concern in the pig nutrition research and pig production industry. Part of macromolecular proteins in the feed can be decomposed into high-quality micromolecular proteins or polypeptides by microbial fermentation, thereby degrading anti-nutritional factors, generating short-chain fatty acids and other microbial metabolites, providing high-quality bacterial proteins and producing aromatic substances having inducing effect. A microbial fermentation feed may have a certain effect of improving pork pig growth and meat quality, and thus, is increasingly concerned by pig producers. However, preparation and application technologies of the microbial fermentation feed may have many problems as follows: (1) fermentation is performed by taking unconventional feed raw

Description

materials (such as rapeseed meal, cottonseed meal, folium mori and paper mulberry, containing high anti-nutritional factor content) as the substrates in production, for the major purposes that on one hand, defects of national feed resources, particularly protein feed, are overcome and on the other hand, the anti-nutritional factors are degraded through fermentation and certain viable bacteria and bacterial proteins are provided. However, in the fermentation with the unconventional raw materials, the nutrient composition of compound feed (particularly protein content) may not meet current actual nutritional demands of lean-type pigs in China, thereby limiting productive potential of the fattening pigs. (2) Reported fermented feed products are short in storage life and easy to mildew; and fermented raw materials and unfermented raw materials may easily cause problems of contamination and mixing non-uniformity in the process of mixing, thereby going against wide applications in actual production. (3) When the fermented feed is applied in the whole feeding process, production cost is increased; and an input-output ratio is low.

Summary To overcome defects and disadvantages of the prior art, a primary purpose of the present invention is to provide a solid-state fermented feed for improving meat quality of fattening pigs. Firstly, a formula of the solid-state fermented feed is designed according to nutritional demands of fattening pigs at a pre-slaughtering stage (2-4 weeks) (see NRC pig nutrition demands, Version 2012); conventional high-quality feed materials of corn, bean pulp and soybean hull are preferred for compatibility; and the prepared feed meets growth needs of the pigs at this stage and has the characteristics of low antigen content, low toxin contamination and high digestion and utilization ratio; and secondly, the solid-state fermented feed is not added with antibiotics or heavy metals of copper, zinc and arsenic, and is safe

Description

and green. The solid-state fermented feed is applied to feeding the fattening pigs, and improves growth performance and meat quality safety of the fattening pigs. Another purpose of the present invention is to provide a preparation method of the solid-state fermented feed for improving meat quality of fattening pigs. In the preparation method, the complete feed is subjected to multi-bacterium mixed solid state fermentation with preferred aerobic bacteria and anaerobic bacteria having probiotic characteristics. The preparation method has the advantages as follows: fermentation time is short (the fermentation may be completed within 3 days in summer or 5 days in winter); fermentation quality is excellent (nutritional quality of the feed is increased, and hybrid bacteria contamination is avoided); preservation time is long (the feed may be preserved for more than half a year at room temperature); the feed is convenient to be fed (the complete feed replaces the conventional feed by 100%, and risk of introducing hybrid bacteria in an extra mixing process is decreased), and the like. Another purpose of the present invention is to provide an application of the solid-state fermented feed for improving meat quality of fattening pigs. The solid-state fermented feed is used at the stage of 2-4 weeks before slaughtering, and is short in use cycle, excellent in feeding effect and high in input-output ratio. The purposes of the present invention are realized by technical solutions as follows: The solid-state fermented feed for improving meat quality of fattening pigs is prepared by performing solid state fermentation on complete feed for pigs with compound probiotics by utilizing the complete feed for pigs as a fermentation substrate; and the compound probiotics include Lactobacillus reuteri, Bacillus subtilis and Saccharomyces cerevisiae. The complete feed for pigs is preferably the complete feed for pigs used at a later fattening period, further preferably includes the following components in percentage by weight: 60-70% of corn, 10-14% of dehulled soybean meal, 10-18%

Description

of soybean hull, 0-0.5% of L-lysine hydrochloride, 1-3% of soybean oil, 1-3% of molasses, 0-1.5% of mountain flour, 0-2% of calcium hydrophosphate, 0-1.5% of salt and 1% of complex vitamin and mineral premix, and more preferably includes the following components in percentage by weight: 65% of corn, 12.5% of dehulled soybean meal (including crude protein content of 46%), 14.5% of soybean hull, 0.2% of L-lysine hydrochloride, 2% of soybean oil, 2% of molasses, 0. 8 5 % of mountain flour, 1% of calcium hydrophosphate, 0. 9 5 % of salt and 1% of complex vitamin and mineral premix. The complex vitamin and mineral premix provides 115 mg of iron, 9 mg of copper, 112 mg of zinc, 32 mg of manganese, 0.25 mg of iodine, 0.2 mg of selenium, 7000U of vitamin A, 1000U of vitamin D3, 15 mg of vitamin E, 2.0 mg of vitamin K3, 2.0 mg of vitamin B1, 5.0 mg of vitamin B2, 3.5 mg of vitamin B6, 0.02 mg of vitamin B12, 20 mg of nicotinic acid, 10 mg of calcium pantothenate, 1.0 mg of folic acid, 0.2 mg of biotin, 140 mg of oxicap and the balance of carrier for the feed per kilogram, wherein the carrier is preferably zeolite powder. The time of the solid state fermentation is preferably 3-5 days, e.g., 3 days in summer or 5 days in winter. The Lactobacillus reuteri is preferably Lactobacillus reuteri LR1, is isolated from healthy animal intestines by the applicant, is collected in China General Microbiological Culture Collection Center, has a collection number of CGMCC No.11154, and has been disclosed in a patent "LactobacillusReuteri Strain having Probiotic Characteristic and Application thereof' (patent No.: ZL 201510751130.3). The Bacillus subtilis is preferably Bacillus subtilis GIMI.372. The Saccharomyces cerevisiae is preferably Saccharomyces cerevisiae CCTCC No.Y200007.

Description

Living bacterium concentrations of the Lactobacillus reuteri, Bacillus subtilis and Saccharomyces cerevisiae in the compound bacterium solution are preferably 1x107-5x108 CFU/mL, 1x107-1x108 CFU/mL and 5x107-5x108 CFU/nL respectively, and further preferably 1.8x108 CFU/nL, 2x107 CFU/mL and 1.5x10 CFU/mL. The preparation method of the solid-state fermented feed for improving meat quality of fattening pigs includes the following steps: (1) preparation of the complete feed for pigs fully and uniformly mixing various components according to a formula ratio to obtain the complete feed for pigs as a fermentation base material; (2) preparation of fermentation bacterium solution adding activated strains of the Lactobacillus reuteri, Bacillus subtilis and Saccharomyces cerevisiae into a nutrient solution according to needed parts; and uniformly mixing the strains and the nutrient solution to obtain the fermentation bacterium solution; (3) uniformly injecting the fermentation bacterium solution in the step (2) into the fermentation base material in the step (1); fully mixing the fermentation base material and the fermentation bacterium solution to obtain an inoculated base material; then sealing the inoculated base material in a breathing bag; and placing the breathing bag in an environment of 32-37°C for performing solid state fermentation for 3-5 days to obtain the finished solid-state fermented feed. The preparation of the complete feed for pigs in the step (1) preferably specifically includes: O preparation of the complex vitamin and mineral premix accurately weighing vitamin and mineral raw materials required in the formula; uniformly mixing the vitamin and mineral raw materials with part of the carrier to obtain a vitamin mixture and a mineral mixture; fully mixing the vitamin mixture and the mineral mixture; finally adding the residual carrier; and fully and

Description

uniformly mixing with the vitamin mixture and the mineral mixture to obtain the complex vitamin and mineral premix; preparation of a compound complete feed selecting mildew-free odorless corn, dehulled soybean meal and soybean hull; crushing and screening the materials; weighing the materials according to the parts required in the formula and uniformly mixing the materials; then adding L-lysine hydrochloride, soybean oil, molasses, mountain flour, calcium hydrophosphate, salt and the complex vitamin and mineral premix obtained in the step @; and fully and uniformly mixing the materials to obtain the compound complete feed. The nutrient solution in the step (2) includes the following components in percentage by mass: 2% of sucrose, 0.3% of K2HPO 4 and 0.2%of KH 2PO 4

. A temperature of the nutrient solution in the step (2) is preferably 301°C. A ratio of the fermentation base material to the fermentation bacterium solution in the step (3) is preferably as follows: 350-450 liters of the fermentation bacterium solution are added into the fermentation base material per tonnage, and further preferably, 420 liters of the fermentation bacterium solution are added. The sealing operation in the step (3) preferably includes: filling the inoculated base material into the breathing bag; and immediately sealing the breathing bag by a sealing machine to obtain the fermentation complete feed bag. An application of the solid-state fermented feed for improving meat quality of fattening pigs is provided. The solid-state fermented feed for improving meat quality of fattening pigs is applied to feeding the fattening pigs at a pre-slaughtering stage, and preferably applied to feeding the fattening pigs at the stage of 2-4 weeks before slaughtering. Thus, the purposes of improving growth performance, protein retention and meat quality safety of the fattening pigs can be realized. Compared with the prior art, the present invention has advantages and effects as follows:

Description

(1) In the present invention, the complete feed is fermented; the preparation method is simple and convenient in operation, short in needed fermentation time (the fermentation may be completed within 3 days in summer or 5 days in winter), long in finished product preservation time, simple in preservation method and capable of greatly reducing the manual cost in the production process. (2) The preparation method of the fermentation complete feed in the present invention significantly increases nutritional quality of the feed (including increase of protein and hydrolyzed amino acid levels of the feed, decrease of the pH and degradation of crude fibers, see data in embodiment 1), saves the use of feed additives such as enzyme formulations and acidifiers and increases the feed utilization efficiency (see data in embodiment 4); and the input-output ratio is high. (3) The fermentation complete feed in the present invention is applied to feeding the fattening pigs at the pre-slaughtering stage, and has the advantages of short feeding time (2-4 weeks) and excellent feeding effect (significantly improving the meat quality, see data in embodiment 3). In conclusion, the solid-state fermented feed for improving meat quality of fattening pigs as well as the preparation method and the application thereof provided by the present invention meet the requirements and goals of green, efficiency, safety and high quality in the pig development in China, and have obvious social and ecological significances and wide market space.

Detailed Description

The present invention will be further described in detail below in combination with embodiments. However, the embodiments of the present invention are not limited herein. Those skilled in the art shall understand that, modifications or replacements may be made to details and forms of the technical solutions in the present invention without departing from the spirit and scope of the present invention.

'7

Description

However, these modifications and replacements fall within the protection scope of the present invention. Embodiment 1 Preparation of solid-state fermented feed and influence of the solid-state fermented feed on nutritional quality of complete feed for fattening pigs The base material of the solid-state fermented feed for improving meat quality of fattening pigs in the present embodiment includes the following components in percentage by weight: 65% of corn, 12.5% of dehulled soybean meal (including crude protein content of 4 6 %), 1 4 .5% of soybean hull, 0. 2 % of L-lysine hydrochloride, 2% of soybean oil, 2% of molasses, 0.85% of mountain flour, 1% of calcium hydrophosphate, 0.95% of salt and 1% of complex vitamin and mineral premix. The base material in the present embodiment includes the following major nutritional indexes: 3345.61 kcal/kg of digestible energy, 12.61% of crude protein, 0. 6 8 % of calcium, 0.55% of total phosphorus, 0. 3 0% of available phosphorus, 0. 6 9 % of lysine, 0. 2 1% of methionine, 0.54% of mthionine +cystine, 0. 4 9 % of threonine and 0.15% of tryptophan. Further, the complex vitamin and mineral premix provides 115 mg of iron, 9 mg of copper, 112 mg of zinc, 32 mg of manganese, 0.25 mg of iodine, 0.2 mg of selenium, 7000U of vitamin A, 1000U of vitamin D3, 15 mg of vitamin E, 2.0 mg of vitamin K3, 2.0 mg of vitamin B1, 5.0 mg of vitamin B2, 3.5 mg of vitamin B6, 0.02 mg of vitamin B12, 20 mg of nicotinic acid, 10 mg of calcium pantothenate, 1.0 mg of folic acid, 0.2 mg of biotin, 140 mg of oxicap and the balance of carrier (zeolite powder) for the base material per kilogram. The solid-state fermented feed for improving meat quality of fattening pigs is prepared by inoculating 420 liters of fermentation bacterium solution into the base material per tonnage to perform multi-bacterium mixed solid state fermentation for 3-5 days.

Description

Further, the fermentation bacterium solution includes 1.8x108 CFU/nL of Lactobacillus reuteri, 2x107 CFU/mL of Bacillus subtilis and 1.5x108 CFU/mL of Saccharomyces cerevisiae. The Lactobacillus reuteri is preferably Lactobacillus reuteri LR1, is isolated from healthy animal intestines by the applicant, is collected in China General Microbiological Culture Collection Center, and has a collection number of CGMCC No.11154. A preparation method of the Lactobacillus reuteri may refer to an authorized patent ZL201510751130.3. The Bacillus subtilis is preferably Bacillus subtilis GIM1.372 (purchased from Guangdong Microbiological Culture Collection Center). The Saccharomyces cerevisiae CCTCC No.Y200007 has been disclosed in a paper [doi:10.1016/S2095-3119(16)61581-2] published by the applicant and is collected by the entity of the inventor. A preservation method of the Saccharomyces cerevisiae strain may refer to the above paper. A preparation method of the solid-state fermented feed for improving meat quality of fattening pigs includes the following steps: (1) preparation of a complex vitamin and mineral premix vitamin and mineral raw materials required in the formula were accurately weighed; the vitamin and mineral raw materials were uniformly mixed with part of the carrier (zeolite powder); the vitamin and mineral mixtures were fully mixed; and finally the residual carrier was added to be fully and uniformly mixed with the vitamin and mineral mixtures to obtain the complex vitamin and mineral premix; (2) preparation of a compound complete feed mildew-free odorless corn, dehulled soybean meal and soybean hull were selected; the materials were crushed and screened; the materials were weighed according to the parts required in the formula and then uniformly mixed; then L-lysine hydrochloride, soybean oil, molasses, mountain flour, calcium hydrophosphate, salt and the complex vitamin and mineral premix obtained in the step (1) were added in sequence; and the materials were fully and uniformly mixed

Description

to obtain compound complete feed to serve as a fermentation base material for later use; (3) preparation of a fermentation bacterium solution activated strains of Lactobacillus reuteri, Bacillus subtilis and Saccharomyces cerevisiae were added into a nutrient solution (including 2% of sucrose, 0.3% of K2HPO4 and 0.2% of KH 2PO 4 , and the solvent of water) according to needed parts; and the strains and the nutrient solution were uniformly mixed to obtain the fermentation bacterium solution; (4) fermentation of the base material 420 liters of the fermentation bacterium solution in the step (3) were uniformly injected into the base material in the step (2) per tonnage; the base material and the fermentation bacterium solution were fully mixed to obtain an inoculated base material by a fermentation mixer; then the inoculated base material was filled in a breathing bag; the breathing bag was immediately sealed by a sealing machine to obtain a fermentation complete feed bag; and the breathing bag was placed in an environment of 32-37C for performing solid state fermentation for 3-5 days (3 days in summer or 5 days in winter) to obtain the finished fermentation complete feed). Experimental design and methods: a complete feed before fermentation (named as a control group), a complete feed fermented for 5 days (named as fermented for 5 days), a complete feed fermented for 30 days (named as fermented for 1 month) and a complete feed fermented for 180 days (named as fermented for 6 months) were respectively collected by 8 parts; and crude fibers, crude proteins and free amino acid levels and pH value of the feed were determined. Experimental results were shown as Tables 1-2. The crude fibers and the crude proteins were respectively determined by respectively referring to "filtration--semi-automatic operation methods" in PRC National Standard GB/T 6434-2006 and "kjeldahl determination" in GB/T

1i

Description

6432-1994. The amino acids were determined according to "acid hydrolyzation" in the PRC National Standard GB/T 18246-2000. Sulfur containing amino acids were determined according to "ion exchange chromatography" in the PRC National Standard GB/T 15399-94. A pH value determining method: 3 g of the sample was dissolved into 30 mL of double distilled water and centrifuged at 6000r/min for 5 min; the supernatant was taken; and the pH value of the feed was determined by a pH meter. It can be seen from Table 1 that, compared with the control group, the fermentation significantly decreases the crude fiber content of the complete feed (P<0.05), increases the crude protein content (P<0.0001) and decreases the pH value (P<0.0001); and differences among the three groups, i.e., the fermented for 5 days, the fermented for 1 month and the fermented for 6 months, are not significant. It can be seen from Table 2 that, compared with the control group, the three groups, i.e., the fermented for 5 days, the fermented for 1 month and the fermented for 6 months, significantly increase the content of free lysine, threonine, valine, leucine, isoleucine, glutamic acid, glycine, serine, alanine, phenylalanine, histidine, proline and aspartic acid (totally P<0.05); and the differences among the three groups, i.e., the fermented for 5 days, the fermented for 1 month and the fermented for 6 months, are not significant. Table 1Effects of Solid State Fermentation on pH Value and Chemical Components of Complete Feed (88% of Dry Matter Base) Indexes Control group Fermented for 5 d Fermented for 1 month Fermented for 6 months P value

Crude fiber(%) 6.72+0.52' 6.29+0.41b 6.13+0.34b 6.07+0.65b <0.05

Crude protein(%) 12.94+0.08' 13.77±0. 1 0 b 1 3 .7 6 1 0 .0 9 b 13.76±0. 14b <0.0001

pH value 6.11+0.00a 4 .8 81 0. 0 3b 4.52+0.01b 4.51+0.07b <0.0001

Notes: all the data are represented by the mean value mean standard error.

Description

P<0.05 represents significant difference; P<0.01 represents extremely significant difference; and P>0.05 represents no significant difference, similarly hereinafter. Table 2 Effects of Solid State Fermentation on Free Amino Acid Composition of Complete Feed (88% of Dry Matter Base) Amino acid (mg/100 g) Control group Fermented for 5 d Fermented for 1 month Fermented for 6 months P value

Lysine 0.87±0.012a 0 . 9 2 ±0. 0 0 8 0.99±0.01 3 0.98±0.0 1b 0.0173

Threonine 0.22±0.007a 0 . 3 5 ±0. 0 0 7 0 .3 3 ±0. 0 6 0 .3 4±0. 0 5 0.0002

Methionine 0.086±0.003 0.088±0.003 0.089±0.002 0.087±0.003 0.6609

Cystine 0.16±0.003a 0 . 2 2 ±0. 0 0 6 0.21±0.005' 0 .2 1±0. 0 0 4 0.0033

Valine 0.61±0.011a 0 . 6 7 ±0. 0 0 7 0 .6 7±0. 0 0 8 0.68±0.007' 0.0002

Isoleucine 0.52±0.005a 0 . 5 8 ±0. 0 0 6 0.59±0.005' 0.59±0.004' 0.0002

Leucine 1.18±0.001a 1.30±0.0 13 1.3 1±0.0 0 9 1. 3 2±0. 0 0 2 0.0005

Glutamic acid 2.33±0.033' 2 . 4 8 ±0. 0 2 9 ' 2 .4 6 ±0. 0 2 4 ' 2.45±0.009 0.0148

Glycine 0.51±0.0064' 0 . 6 6 ±0. 0 0 6 5 ' 0 .6 0 ±0. 0 0 9 0 .6 2±0. 0 0 7 0.002

Tyrosine 0.52±0.004 0.53±0.006 0.49±0.007 0.48±0.011 0.4273

Serine 0.62±0.006a 0 . 6 6 ±0. 0 0 7 0.68±0.009' 0.68±0.012' 0.0075

Alanine 0.73±0.007a 0.83±0.01 1P 0.840.005 0.84±0.010' 0.0002

Phenylalanine 0.61±0.008 0 . 6 6 ±0. 0 0 8 0.68±0.004' 0 .6 6±0. 0 10 0.012

Histidine 0.35±0.004a 0 . 3 8 ±0. 0 0 5 0.40±0.001' 0.39±0.012' 0.0079

Arginine 0.67±0.012 0.65±0.012 0.68±0.051 0.64±0.009 0.2448

Proline 0.82±0.009a 0 . 9 0 ±0. 0 0 7 0.92±0.010' 0.91±0.004' 0.0007

Aspartic acid 1.17±0.01a 1. 2 7 ±0. 0 2 1.31±0.008' 1.29±0.012' 0.0028

Embodiment 2 Preparation of solid-state fermented feed Base material of the solid-state fermented feed for improving meat quality of fattening pigs in the present embodiment includes the following components in percentage by weight: 60% of corn, 12% of dehulled soybean meal, 16.6% of soybean hull, 0.5% of L-lysine hydrochloride, 3% of soybean oil, 3% of molasses,

11)

Description

1.28% of mountain flour, 1.5% of calcium hydrophosphate, 1.12% of salt and 1% of complex vitamin and mineral premix. Further, the complex vitamin and mineral premix provides 115 mg of iron, 9 mg of copper, 112 mg of zinc, 32 mg of manganese, 0.25 mg of iodine, 0.2 mg of selenium, 7000U of vitamin A, 1000IU of vitamin D3, 15 mg of vitamin E, 2.0 mg of vitamin K3, 2.0 mg of vitamin B1, 5.0 mg of vitamin B2, 3.5 mg of vitamin B6, 0.02 mg of vitamin B12, 20 mg of nicotinic acid, 10 mg of calcium pantothenate, 1.0 mg of folic acid, 0.2 mg of biotin, 140 mg of oxicap and the balance of carrier (zeolite powder) for the base material per kilogram. The solid-state fermented feed for improving meat quality of fattening pigs is prepared by inoculating 420 liters of fermentation bacterium solution into the base material per tonnage to perform multi-bacterium mixed solid state fermentation for 3-5 days. The fermentation bacterium solution includes 1x107 CFU/mL of Lactobacillus reuteri LR1, 1x108 CFU/mL of Bacillus subtilis GIMI.372 and x108 CFU/mL of Saccharomyces cerevisiae CCTCC No.Y200007. The finished fermentation complete feed is prepared by referring to the preparation method of the solid-state fermented feed for improving meat quality of fattening pigs in embodiment 1. Embodiment 3 Preparation of solid-state fermented feed Base material of the solid-state fermented feed for improving meat quality of fattening pigs in the present embodiment includes the following components in percentage by weight: 70% of corn, 10% of dehulled soybean meal (including crude protein content of 4 6 %), 13.92% of soybean hull, 0.15% of L-lysine hydrochloride, 1. 7 % of soybean oil, 1% of molasses, 0.05% of mountain flour, 0.98% of calcium hydrophosphate, 1.2% of salt and 1% of complex vitamin and mineral premix.

Description

Further, the complex vitamin and mineral premix provides 115 mg of iron, 9 mg of copper, 112 mg of zinc, 32 mg of manganese, 0.25 mg of iodine, 0.2 mg of selenium, 7000U of vitamin A, 1000U of vitamin D3, 15 mg of vitamin E, 2.0 mg of vitamin K3, 2.0 mg of vitamin B1, 5.0 mg of vitamin B2, 3.5 mg of vitamin B6, 0.02 mg of vitamin B12, 20 mg of nicotinic acid, 10 mg of calcium pantothenate, 1.0 mg of folic acid, 0.2 mg of biotin, 140 mg of oxicap and the balance of carrier (zeolite powder) for the base material per kilogram. The solid-state fermented feed for improving meat quality of fattening pigs is prepared by inoculating 420 liters of fermentation bacterium solution into the base material per tonnage to perform multi-bacterium mixed solid state fermentation for 3-5 days. The fermentation bacterium solution includes 5x108 CFU/mL of Lactobacillus reuteri LR1, 5x107 CFU/mL of Bacillus subtilis GIMI.372 and x107 CFU/mL of Saccharomyces cerevisiae CCTCC No.Y200007. The finished fermentation complete feed is prepared by referring to the preparation method of the solid-state fermented feed for improving meat quality of fattening pigs in embodiment 1. Embodiment 4 Digestion experiment of fattening pigs To verify utilization efficiency of nutritive materials in the solid-state fermented feed in the present invention, a digestion experiment of fattening pigs is designed in the present embodiment, and used for determining digestibility of nutrients after the fattening pigs at a pre-slaughtering stage are fed with the solid-state fermented feed in embodiment 1. 1. Experimental design A total of 16 castrated male pigs (Durocx(YorkshirexLandrace))with consistent genetic backgrounds, excellent physical conditions and weight of (90±1.3) kg were randomly assigned to the control or fermentation group each

1A

Description

containing 8 replicates, with 1 pig per replicate. The pigs were fixed in metabolism cages to be fed in single cages. The experiment totally included three periods: (1) adaptation period: the period totally lasted 7 days; all the pigs were fed with the complete feed provided in embodiment 1 (that is, the complete feed before fermentation in embodiment 1); and the pigs were free to feed and drink; (2) preliminary feeding period: the period totally lasted 7 days; all the pigs were fed with the control complete feed and the fermentation complete feed provided in embodiment 1 respectively; the pigs were free to feed and drink; the food intake was counted; and the average daily feed intake was calculated; (3) Experimental period: the period totally lasted 7 days; all the pigs were fed with the control complete feed and the fermentation complete feed provided in embodiment 1 respectively (for the two kinds of feeds, chromium oxide having a weight ratio of 0.25% was added in advance to serve as an exogenous indicator); the manure was collected when color of the manure changed; the manure was collected when discharged; the manure was stopped collecting when colored manure was totally discharged on Day 7 of the experiment; and then the experiment was ended. The feed was provided according to 2% of the weight during the experiment; the food intake was counted; and the pigs were free to drunk. 2. Sample collection during experiment (1) 200 g of the feed sample in each group was respectively reserved when the experiment was started and ended and cryopreserved in a refrigerator at -20°C. (2) The total fresh manure of each pig on that day was placed on a stainless steel disc and fully and uniformly mixed; according to a principle that more manure was collected when more manure was discharged and that less manure was collected when less manure was discharged, 20% of the total fresh manure on that day was weighed; the accurately weighed manure was put in an oven at 65°C and dried for more than 3 days; initial moisture content was weighed and recorded;

Description

then the manure was dampened for 24 hours to reach the constant weight; the dried manure samples of each pig collected within 7 days were uniformly mixed, ground by a high-speed crusher and screened through a 40-mesh sieve; and the samples were filled in a bag for later experiment. (3) Urine samples of the various pigs within 24 hours per day were accurately collected; volumes of the urine samples were weighed and recorded; the urine samples were collected in a collecting barrel containing 50 mL of 6mol/L hydrochloric acid for fixing nitrogen; the urine samples were uniformly mixed and filtered with gauze; and 10% of the total urine samples on that day were preserved in the refrigerator at -20°C for later experiment. 3. Index determination (1) Feed sample: crude proteins, crude fat, crude fibers, neutral detergent fibers, acid detergent fibers and total energy were determined. (2) Manure sample: crude proteins, crude fibers, neutral detergent fibers, acid detergent fibers and total energy were determined. (3) Urine sample: the total energy was determined. (4) Determination methods of the crude proteins, crude fat, crude fibers and neutral detergent fibers in the (1) to (3) respectively referred to PRC National Standards such as GB/T 6432-1994, GB/T 6433-2006, GB/T6434-2006 and 20806-2006. The acid detergent fibers were determined according to a method in PRC Agriculture Standard NY/T 1459-2007. A determination method of the total energy referred to an international standard IS09831:1998; and the total energy was determined by an oxygen bomb calorimeter. (5) Used calculation formulas were as follows: Urinary energy=192+31x50O% of digestible urinary nitrogen or 40% of total nitrogen Apparent digestible energy of the feed (MJ/kg)=(ingested total energy MJ-discharged total fecal energy MJ)/the total amount kg of the ingested feed

1A

Description

Apparent metabolic energy of the feed (MJ/kg)=(ingested total energy MJ-discharged total fecal energy MJ-discharged total urinary energy MJ)/the total amount kg of the ingested feed Apparent digestibility (%) of the total energy--(ingested total energy-discharged total fecal energy)/the ingested total energy 4. Result and analysis Statistics results of digestibility of the fattening pigs on the nutritive materials in the solid-state fermented feed were shown in Table 3. Compared with the control group, the digestibility of the total energy, crude proteins, crude fibers and neutral detergent fibers in the fermentation complete feed on the fattening pigs had no significant change (P>0.05), but the digestibility of the acid detergent fibers was significantly increased (P=0.04). Both the apparent digestible energy and the apparent metabolic energy of the fermentation complete feed were significantly higher than that of the control complete feed (P<0.05). Thus, it can be seen that the digestibility of the solid-state fermented feed provided by the present invention on the fattening pigs is higher than that of the unfermented complete feed, which indicated that the solid state fermentation method provided by the present invention has an effect of increasing the digestibility of the complete feed for the fattening pigs. Table 3 Energy Content and Digestibility of the Solid-state Fermented Feed by Fattening Pigs Indexes Control group Fermentation group P value

Total energy digestibility, % 86.53±1.26 87.77±0.43 0.38

Crude protein digestibility, % 97.00±0.26 97.54±0.00 0.10

Crude fiber digestibility, % 78.14±4.05 80.28±1.84 0.64

Neutral detergent fiber digestibility, % 68.60±3.31 71.62±1.50 0.43

Acid detergent fiber digestibility, % 64.34±3.45 73.53±0.63 0.04

Apparent digestible energy (kcal/kg) 3253±47.52 3379±16.66 <0.05

1 '7

Description

Apparent metabolic energy (kcal/kg) 3163±46.13 3285±17.18 <0.05

Embodiment 5 Application of the solid-state fermented feed for improving meat quality of fattening pigs pre-slaughtering To verify the application effect of the solid-state fermented feed for improving meat quality of fattening pigs in the present invention, the fermentation complete feed prepared in embodiment 1 is applied to feeding the fattening pigs having the weight of 90-120 kg. Experimental materials and methods: A total of 96 castrated male pigs Durocx(YorkshirexLandrace) with consistent genetic backgrounds, excellent physical conditions and weight of (90+2.3) kg were randomly assigned to the complete feed before fermentation (named as a control group) and the fermentation complete feed group (named as a fermentation group); each containing 8 replicates, with 3 pigs per replicate. The pigs in the two groups were respectively fed with the control complete feed and the fermentation complete feed in embodiment 1. The experiment lasted for 21 days; and the pigs only freely drunk water and ate the feed during the experiment. The food intake was recorded per day; body weight gain was recorded per week; and a feed-weight ratio was calculated. When the experiment was ended, one pig having moderate weight in each repeat was selected to be slaughtered and sampled to detect the meat quality. The specific determination method referred to PRC Agriculture Standard NY/T 821-2004. Experiment results were shown as Tables 4-7. It can be seen from Table 4 that, compared with the control group, the fermentation complete feed group had no significant influence on the average daily feed intake and the daily gain of the fattening pigs, but tended to decrease the feed-weight ratio (P=0.06). It can be seen from Table 5 that, compared with the control group, the fermentation complete feed group significantly decreased the thickness of backfat at the last rib of the fattening pigs (P=0.03), and increased a lean meat percentage (P=0.006) and a fat free lean growth rate of the carcass

1Q

Description

(P=0.028, Table 5). The determination results of the meat quality showed that, the fermented feed had no significant influence on the back longissimus color L*, a* and b* (the data is not shown) and intramuscular fat content, but significantly decreased back longissimus drip loss within 48 hours after slaughtering (P=0.01) and pH value (P<0.01), and back longissimus shear force (P<0.01, Table 6). Post-slaughtering back longissimus cooking degustation experimental results (Table 7) showed that, the fermentation complete feed group significantly increased tenderness, flavor and aroma of the back longissimus of the fattening pigs after cooking and broth freshness (totally P<0.05). In addition, antibiotic residues in pork in the control group and the fermentation group were lower than a lower limit of detection (the data is not shown); and no antibiotic residue was found. To sum up, the fermentation complete feed in the present invention fed at the pre-slaughtering stage may significantly improve carcass composition of the fattening pigs by promoting protein retention and decreasing body fat retention, and significantly improve the pork quality. The produced pork meets market demands for high quality and safety. Table 4 Effects of Solid State Fermented Feed on Growth Performance of Fattening Pigs Indexes Control group Fermentation group P value

Average daily feed intake (kg/d, 88% 3.34±0.12 3.06±0.11 0.14

of dry matter base)

Average daily gain (kg/d) 0.84±0.03 0.86±0.03 0.70

Feed : gain ratio 4.05±0.15 3.65±0.08 0.06

Table 5 Effects of Fermentation Complete Feed on Carcass Composition of Fattening Pigs Indexes Control group Fermentation group P value

Backfat First rib 4.36±0.34 4.49±0.31 0.63

Description

thickness (cm) Tenth rib 3.13±0.34 3.01±0.23 0.78

Last rib 2.67±0.15 2.11±0.16 0.03

Eye muscle area (cm2 ) 65.01±3.58 70.61±2.25 0.20

Lean meat percentage(%) 53.79±1.51 61.32±1.72 0.006

Fat free lean growth rate of the carcass (g/d) 343.8±25.97 551.5±43.95 0.028

Table 6 Effects of Fermentation Complete Feed on Meat Quality of Fattening Pigs Indexes Control group Fermentation group P value

Intramuscular fat content (%) 1.22±0.11 1.51±0.26 0.21

Drip loss (%) within 24 h after slaughtering 2.55±0.15 2.43±0.07 0.46

Drip loss (%) within 48 h after slaughtering 3.85±0.14 3.08±0.23 0.01

Shear force (N) 66.78±4.50 49.96±3.10 <0.01

pH value within 45 min after slaughtering 5.94±0.11 6.07±0.06 0.36

pH value within 24 h after slaughtering 5.51±0.03 5.53±0.03 0.92

pH value within 48 h after slaughtering 5.61±0.03 5.51±0.03 <0.01

Table 7 Influence of Fermentation Complete Feed on Sensory Indexes of Back Longissimus (Cooking Degustation Experiment) Indexes Control group Fermentation group P value

Juiciness (1-8 scores represent that the meat is extremely 4.52±0.10 4.48±0.13 0.7

dry-extremely juicy)

Tenderness (1-8 scores represent that the meat is extremely 4.26±0.22 4.73±0.11 0.04

difficultly-extremely easily chewed by teeth)

Flavor (1-8 scores represent that the meat has extremely 4.72±0.07 4.96±0.10 0.06

weak-extremely strong flavor, through gustation)

Aroma (1-8 scores represent that the meat has extremely 4.79±0.07 5.18±0.10 0.01

weak-extremely strong aroma, through smell)

Odor (1-8 scores represent that the meat has extremely 5.13±0.07 5.27±0.17 0.27

weak-extremely strong odor)

Description

Broth freshness (1-8 scores represent extremely 4.85±0.07 5.31±0.10 0.04

weak-extremely strong broth freshness)

Meat color (1-8 scores represent extremely poor-extremely 4.89±0.09 5.15±0.1 0.47

excellent meat color)

The above embodiments are preferred embodiments of the present invention. However, the embodiments of the present invention are not limited by the above embodiments. Any other change, modification, replacement, combination and simplification made without departing from the spiritual essence and principles of the present invention shall be equivalent substitutes and included in the protection scope of the present invention.

Claims

1. A solid-state fermented feed for improving meat quality of fattening pigs, which is prepared by performing solid state fermentation on a complete feed for pigs with compound probiotics by utilizing the complete feed for pigs as a fermentation substrate, wherein the compound probiotics comprise Lactobacillus reuteri, Bacillus subtilis and Saccharomyces cerevisiae.

2. The solid-state fermented feed for improving meat quality of fattening pigs according to claim 1, wherein the complete feed for pigs is the complete feed for pigs used at a later fattening period; time of the solid state fermentation is 3-5 days.

3. The solid-state fermented feed for improving meat quality of fattening pigs according to claim 1, wherein the complete feed for pigs comprises the following components in percentage by weight: 60-70% of corn, 10-14% of dehulled soybean meal, 10-18% of soybean hull, 0-0.5% of L-lysine hydrochloride, 1-3% of soybean oil, 1-3% of molasses, -1.5% of mountain flour, 0-2% of calcium hydrophosphate, 0-1.5% of salt and 1% of complex vitamin and mineral premix; the Lactobacillus reuteri is Lactobacillus reuteri LR1, and has a collection number of CGMCC No. 11154; the Bacillus subtilis is Bacillus subtilis GIMI.372; the Saccharomyces cerevisiae is Saccharomyces cerevisiae CCTCC No. Y200007.

4. The solid-state fermented feed for improving meat quality of fattening pigs according to claim 1, wherein the complete feed for pigs comprises the following components in percentage by weight: 65% of corn, 12.5% of dehulled soybean meal comprising crude protein content of 46%, 14.5% of soybean hull, 0.2% of L-lysine hydrochloride, 2% of

Claims

soybean oil, 2% of molasses, 0.85% of mountain flour, 1% of calcium hydrophosphate, 0.95% of salt and 1% of complex vitamin and mineral premix; living bacterium concentrations of the Lactobacillus reuteri, Bacillus subtilis and Saccharomyces cerevisiae are 1x1O7-5x108 CFU/mL, 1x107-1x108 CFU/nL and 5x107-5x108 CFU/mL respectively.

5. The solid-state fermented feed for improving meat quality of fattening pigs according to claim 3, wherein the complex vitamin and mineral premix provides 115 mg of iron, 9 mg of copper, 112 mg of zinc, 32 mg of manganese, 0.25 mg of iodine, 0.2 mg of selenium, 7000U of vitamin A, 1000IU of vitamin D3, 15 mg of vitamin E, 2.0 mg of vitamin K3, 2.0 mg of vitamin B1, 5.0 mg of vitamin B2, 3.5 mg of vitamin B6, 0.02 mg of vitamin B12, 20 mg of nicotinic acid, 10 mg of calcium pantothenate, 1.0 mg of folic acid, 0.2 mg of biotin, 140 mg of oxicap and the balance of carrier for the feed per kilogram; the living bacterium concentrations of the Lactobacillus reuteri, Bacillus subtilis and Saccharomyces cerevisiae are 1.8x108 CFU/mL, 2x107 CFU/mL and 1.5x108 CFU/nL.

6. A preparation method of the solid-state fermented feed for improving meat quality of fattening pigs of any one of claims 1-5, comprising the following steps: (1) preparation of the complete feed for pigs fully and uniformly mixing various components according to a formula ratio to obtain the complete feed for pigs as a fermentation base material; (2) preparation of fermentation bacterium solution adding activated strains of the Lactobacillus reuteri, Bacillus subtilis and Saccharomyces cerevisiae into a nutrient solution according to needed parts; and uniformly mixing the strains and the nutrient solution to obtain the fermentation bacterium solution;

Claims

(3) uniformly injecting the fermentation bacterium solution in the step (2) into the fermentation base material in the step (1); fully mixing the fermentation base material and the fermentation bacterium solution to obtain an inoculated base material; then sealing the inoculated base material in a breathing bag; and placing the breathing bag in an environment of 32-37°C for performing solid state fermentation for 3-5 days to obtain the finished solid-state fermented feed.

7. The preparation method of the solid-state fermented feed for improving meat quality of fattening pigs according to claim 6, wherein the preparation of the complete feed for pigs in the step (1) specifically comprise: D preparation of the complex vitamin and mineral premix accurately weighing vitamin and mineral raw materials required in the formula; uniformly mixing the vitamin and mineral raw materials with part of the carrier to obtain a vitamin mixture and a mineral mixture; fully mixing the vitamin mixture and the mineral mixture; finally adding the residual carrier; and fully and uniformly mixing with the vitamin mixture and the mineral mixture to obtain the complex vitamin and mineral premix; @ preparation of a compound complete feed selecting mildew-free odorless corn, dehulled soybean meal and soybean hull; crushing and screening the materials; weighing the materials according to the parts required in the formula and uniformly mixing the materials; then adding L-lysine hydrochloride, soybean oil, molasses, mountain flour, calcium hydrophosphate, salt and the complex vitamin and mineral premix obtained in the step @; and fully and uniformly mixing the materials to obtain the compound complete feed; a ratio of the fermentation base material to the fermentation bacterium solution in the step (3) is: 350-450 liters of the fermentation bacterium solution are added into the fermentation base material per tonnage.

8. The preparation method of the solid-state fermented feed for improving meat quality of fattening pigs according to claim 6, wherein

Claims

the nutrient solution in the step (2) comprises the following components in percentage by mass: 2% of sucrose, 0.3% of K2HPO 4 and 0.2% of KH 2PO 4 ; a temperature of the nutrient solution in the step (2) is 301°C; a ratio of the fermentation base material to the fermentation bacterium solution in the step (3) is: 420 liters of the fermentation bacterium solution are added into the fermentation base material per tonnage.

9. An application of the solid-state fermented feed for improving meat quality of fattening pigs of any one of claims 1-5, wherein the solid-state fermented feed for improving meat quality of fattening pigs is applied to feeding the fattening pigs at a pre-slaughtering stage.

10. The application of the solid-state fermented feed for improving meat quality of fattening pigs according to claim 9, wherein the solid-state fermented feed for improving meat quality of fattening pigs is applied to feeding the fattening pigs at the stage of 2-4 weeks before slaughtering.