Fermentation Condition Optimization and Quality Evaluation of Feed Fermented by Compound Probiotics

WAN Li; WU Guofang; WANG Lei; ZHANG Xiaowei; FENG Yuzhe

doi:10.3969/j.issn.1006-267x.2022.05.061

Chinese Journal of Animal Nutrition >

2022 , Vol. 34 >Issue 5: 3358 - 3375

DOI: https://doi.org/10.3969/j.issn.1006-267x.2022.05.061

RESEARCH PAPER

Fermentation Condition Optimization and Quality Evaluation of Feed Fermented by Compound Probiotics

WAN Li ,
WU Guofang ,
WANG Lei ,
ZHANG Xiaowei ,
FENG Yuzhe

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1. College of Animal Husbandry and Veterinary Sciences, Qinghai University, Xining 810016, China;
2. Qinghai Provincial Key Laboratory of Animal Nutrition and Feed Science for Plateau Grazing Animals, Xining 810016, China;
3. Yak Engineering Technology Research Center of Qinghai Province, Xining 810016, China;
4. Qinghai Provincial Key Laboratory of Protection and Innovative Utilization of Plateau Livestock Genetic Resources, Xining 810016, China

Received date: 2021-09-29

Online published: 2022-05-14

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Abstract

This experiment was conducted to explore the effects of different fermentation conditions on the quality of feed fermented by compound probiotics. Chose 3 strains of lactic acid bacteria (Lactobacillus rhamnosus, Pediococcus lactis and Lactobacillus casei) and 2 strains of yeasts (Saccharomyces cerevisiae and Pichia pastoris), using 4 factors (inoculation amount, inoculation ratio, temperature and water to material ratio) and 3 levels orthogonal design to optimize the fermentation conditions of concentrate supplement. There were 9 groups, and the ammonia nitrogen (NH₃-N), soluble sugar (SS), starch (ST) contents, pH, 3 kinds of mycotoxins-aflatoxin B₁ (AFB₁), zearalenone (ZEN), deoxynivalenol (DON) contents and microbial flora changes in each group of fermented feed were determined, and using range analysis method to find the optimal fermentation conditions of comprehensive indicators. The concentrate supplement was fermented for different time (0, 1, 3, 5, 7, 10 and 15 d) under optimal fermentation conditions, by determining the conventional nutrient contents, enzyme activities, mycotoxin contents, and aerobic stability of the fermented feed to determine the optimal fermentation time. The results showed as follows:1) AFB₁ and ZEN content optimized groups were G1 (inoculation amount 5%, inoculation ratio 1:1, temperature 20℃ and water to material ratio 0.4:1) and G7 groups (inoculation amount 15%, inoculation ratio 2:1, temperature 20℃ and water to material ratio 1:1), NH₃-N content optimized groups were G3 (inoculation amount 5%, inoculation ratio 2:1, temperature 30℃ and water to material ratio 0.4:1) and G7 groups, ST content optimized group was G6 group (inoculation amount 10%, inoculation ratio 2:1, temperature 30℃ and water to material ratio 1:1), DON content optimized groups were G5 (inoculation amount 10%, inoculation ratio 1:2, temperature 25℃ and water to material ratio 1:1) and G7 groups, and SS content optimized groups were G4 (inoculation amount 10%, inoculation ratio 2:1, temperature 20℃ and water to material ratio 0.6:1) and G6 groups. 2) The 16S sequencing results showed that the α diversity and β diversity of the microorganisms in fermented feed in the G3 group were significantly higher than those in the G1, G2 (inoculation amount 5%, inoculation ratio 1:2, temperature 25℃ and water to material ratio 0.6:1), G4, G6, G7, G8 (inoculation amount 15%, inoculation ratio 1:1, temperature 30℃ and water to material ratio 0.4:1) and G9 groups (inoculation amount 15%, inoculation ratio 1:2, temperature 25℃ and water to material ratio 0.6:1) (P<0.05), and slightly higher than those in the G5 group (P>0.05); the relative abundances of Lactobacillus plantarum and Lactobacillus brevis in the G3 group were significantly higher than those in the G1, G2, G4, G5, G6, G8 and G9 groups (P<0.05), but had no significant difference between G3 and G7 groups (P>0.05). 3) Compared with day 0 of fermentation (no fermentation), the crude protein (CP) content, cellulase (CL), lipase (LPS) and acid protease (ACP) activities were significantly increased on the day 5 of fermentation (P<0.05), the NH₃-N, neutral detergent fiber (NDF) and acid detergent fiber (ADF) contents were significantly reduced on day 5 of fermentation (P<0.05), and the β-amylase (β-AL) activity, pH, SS, ST, AFB₁, ZEN and DON contents were significantly reduced on day 10 of fermentation (P<0.05). 4) After the fermented feed was exposed to oxygen for 20 days, the temperature of the center of the feed was 2℃ higher than the ambient temperature, and the pH increased abnormally. Conclusion:1) the optimal fermentation conditions for concentrate supplement are inoculation amount 5%, inoculation ratio 2:1, temperature 30℃, water to feed ratio 1:1, which can effectively increase the diversity of microorganisms in the fermented feed and increase the relative abundance of Lactobacillus brevis. 2) The optimal fermentation time for concentrate supplement is 5 days, which can increase the nutrient contents of the fermented feed and reduce the content of mycotoxins. 3) After the fermentation is finished and the lid is opened, the fermented feed should be fed within 20 days.

Key words： compound probiotics; concentrate supplement; fermentation; condition optimization; quality identification

Cite this article

WAN Li , WU Guofang , WANG Lei , ZHANG Xiaowei , FENG Yuzhe . Fermentation Condition Optimization and Quality Evaluation of Feed Fermented by Compound Probiotics[J]. Chinese Journal of Animal Nutrition, 2022 , 34(5) : 3358 -3375 . DOI: 10.3969/j.issn.1006-267x.2022.05.061

References

[1] KIM T I, MAYAKRISHNAN V, LIM D H, et al.Effect of fermented total mixed rations on the growth performance, carcass and meat quality characteristics of Hanwoo steers[J].Animal Science Journal, 2018, 89(3):606-615.

[2] 昌捷.发酵饲料是解决饲料无抗生素的金钥匙[J].国外畜牧学(猪与禽), 2020, 40(5):82-85. CHANG J.Fermentation feed is the golden key to solve feed without antibiotics[J].Animal Science Abroad (Pigs and Poultry), 2020, 40(5):82-85.(in Chinese)
[3] 钟晨, 姜世光, 王修启, 等.微生物发酵饲料在畜禽生产中的研究进展[J].动物营养学报, 2020, 32(8):3516-3525. ZHONG C, JIANG S G, WANG X Q, et al.Research progress of microbial fermentation feed in livestock and poultry production[J].Chinese Journal of Animal Nutrition, 2020, 32(8):3516-3525.(in Chinese)
[4] 万里, 冯宇哲, 王磊, 等.微生物发酵饲料的研究进展及其应用[J].饲料研究, 2021, 44(13):142-145. WAN L, FENG Y Z, WANG L, et al.Research progress and application of microbial fermented feed[J].Feed Research, 2021, 44(13):142-145.(in Chinese)
[5] MUCK R E, NADEAU E M G, MCALLISTER T A, et al.Silage review:recent advances and future uses of silage additives[J].Journal of Dairy Science, 2018, 101(5):3980-4000.

[6] MUCK R.Recent advances in silage microbiology[J].Agricultural and Food Science, 2013, 22(1):3-15.

[7] KRISTENSEN N B, SLOTH K H, HØJBERG O, et al.Effects of microbial inoculants on corn silage fermentation, microbial contents, aerobic stability, and milk production under field conditions[J].Journal of Dairy Science, 2010, 93(8):3764-3774.

[8] 邹春悦.酵母菌在食品中的作用研究进展[J].当代化工研究, 2018(3):96-97. ZOU C Y.Research progress in the role of yeast in food[J].Modern Chemical Research, 2018(3):96-97.(in Chinese)
[9] 张政.活性酵母及其发酵饲料对瘤胃发酵及营养物质消化率的影响[D].硕士学位论文.呼和浩特:内蒙古农业大学, 2017. ZHANG Z.Effects of active yeast and its fermented feeds on the rumen fermentation and nutrient digestibility[D].Master's Thesis.Hohhot:Inner Mongolia Agricultural University, 2017.(in Chinese)
[10] JEYANATHAN J, MARTIN C, MORGAVI D P.The use of direct-fed microbials for mitigation of ruminant methane emissions:a review[J].Animal, 2014, 8(2):250-261.

[11] 余淼, 严锦绣, 彭忠利, 等.微生物发酵饲料对肉牛免疫机能的影响[J].中国畜牧兽医, 2013, 40(4):114-117. YU M, YAN J X, PENG Z L, et al.Effects of microbial fermented feed on immune function parameter of beef cattle[J].China Animal Husbandry & Veterinary Medicine, 2013, 40(4):114-117.(in Chinese)
[12] SURYANTO E, BULKAINI B, ASHARI A, et al.Carcass quality, marbling and cholesterol content of male Bali cattle fed fermented cocoa shell[J].Journal of the Indonesian Tropical Animal Agriculture, 2014, 39(4):249-255.
[13] WANAPAT M, TOTAKUL P, VIENNASAY B, et al.Sunnhemp (Crotalaria juncea, L.) silage can enrich rumen fermentation process, microbial protein synthesis, and nitrogen utilization efficiency in beef cattle crossbreds[J].Tropical Animal Health and Production, 2021, 53(1):187.
[14] 魏越波.益生菌发酵配合饲料在蛋鸡无抗养殖上的应用[D].硕士学位论文.保定:河北农业大学, 2020. WEI Y B.Fermentation of compound feed by probiotics and application in laying hens breeding without antibiotic[D].Master's Thesis.Baoding:Agricultural University of Hebei, 2020.(in Chinese)
[15] 周静, 肖嫩群.酵母菌与乳酸菌混合培养条件研究[J].安徽农业科学, 2011, 39(15):8824-8825, 8830. ZHOU J, XIAO N Q.Study on mixed culture conditions of yeast and lactic acid bacteria[J].Journal of Anhui Agricultural Sciences, 2011, 39(15):8824-8825, 8830.(in Chinese)
[16] 吝常华.肉鸡饲料原料发酵工艺优化及其营养价值评定[D].硕士学位论文.北京:中国农业科学院, 2018. LIN C H.The optimization of broiler feed fermentation processing and its nutrition evaluation[D].Master's Thesis.Beijing:Chinese Academy of Agricultural Sciences, 2018.(in Chinese)
[17] 孙安琪.白酒糟与菊芋渣混合青贮发酵品质及微生物多样性研究[D].硕士学位论文.兰州:兰州理工大学, 2019. SUN A Q.Study on fermentation quality and microbial diversity of Jerusalem artichoke residue and grain stillage mixed silage[D].Master's Thesis.Lanzhou:Lanzhou University of Technology, 2019.(in Chinese)
[18] TABACCO E, PIANO S, REVELLO-CHION A, et al.Effect of Lactobacillus buchneri LN4637 and Lactobacillus buchneri LN40177 on the aerobic stability, fermentation products, and microbial populations of corn silage under farm conditions[J].Journal of Dairy Science, 2011, 94(11):5589-5598.

[19] 李可义.单菌种和混合菌种在发酵饲料中乳酸菌数量的动态变化研究[J].江西水产科技, 2017(1):8-9. LI K Y.Study on the dynamic change of the number of lactic acid bacteria in fermented feed by single strain and mixed strain[J].Jiangxi Fishery Sciences and Technology, 2017(1):8-9.(in Chinese)
[20] 周密密, 刘晓宇, 许忠祥, 等.鲜食马铃薯中青枯病菌的检测[J].中国植保导刊, 2021, 41(3):5-10, 40. ZHOU M M, LIU X Y, XU Z X, et al.Detection of Ralstonia solanacearum in potatoes[J].China Plant Protection, 2021, 41(3):5-10, 40.(in Chinese)
[21] 蔡瑾.海带提取物对密执安棒形杆菌环腐亚种的抑菌作用研究[D].博士学位论文.太原:山西大学, 2014. CAI J.Study on the antimicrobial activity of Laminaria japonica extract against Clavibater michiganense subsp. sepedonicum (Spieckermann & Kotthoff) Davis et al[D].Ph.D.Thesis.Taiyuan:Shanxi University, 2014.(in Chinese)
[22] 李娟, 郑姚, 王利.鱼源泡囊短波单胞菌16S rDNA基因及耐药性分析[J].动物医学进展, 2020, 41(12):56-60. LI J, ZHENG Y, WANG L.Analysis of 16S rDNA gene and drug resistance of Brevundimonas vesicularis in fish[J].Progress in Veterinary Medicine, 2020, 41(12):56-60.(in Chinese)
[23] JONES B A, SATTER L D, MUCK R E.Influence of bacterial inoculant and substrate addition to lucerne ensiled at different dry matter contents[J].Grass and Forage Science, 1992, 47(1):19-27.

[24] 贾存辉, 钱文熙, 吐尔逊阿依·赛买提, 等.粗饲料营养价值指数及评定方法[J].草业科学, 2017, 34(2):415-427. JIA C H, QIAN W X, SAIMAITI T E X A Y, et al.Roughage nutritional value evaluation indices and research methods[J].Pratacultural Science, 2017, 34(2):415-427.(in Chinese)
[25] 王啸林, 冯冠智, 张城瑞, 等.乳酸菌和纤维素酶对不同比例马铃薯渣与玉米秸秆混合青贮发酵品质、有氧稳定性和体外发酵参数的影响[J/OL].中国畜牧杂志:1-17.(2021-09-28)[2021-10-31].https://doi.org/10.19556/j.0258-7033.20210402-03.DOI:10.19556/j.0258-7033.20210402-03. WANG X L, FENG G Z, ZHANG C R, et al.Effects of lactic acid bacteria and cellulase on fermentation quality, aerobic stability and in vitro fermentation parameters of potato residue and corn straw mixed silage with different proportion[J/OL].Chinese Journal of Animal Science:1-17.(2021-09-28)[2021-10-31].https://doi.org/10.19556/j.0258-7033.20210402-03.DOI:10.19556/j.0258-7033.20210402-03.(in Chinese)
[26] 孙旸, 地里百尔·努尔, 林晓琼, 等.酵母和乳酸菌混合发酵对玉米秸秆蛋白质含量的影响[J].吉林农业大学学报, 2016, 38(5):557-561. SUN Y, NUER D L B E, LIN X Q, et al.Effects of mixed fermentation of yeast and Lactobacillus on protein content of maize straw[J].Journal of Jilin Agricultural University, 2016, 38(5):557-561.(in Chinese)
[27] MCDONALD P, HENDERSON A R, HERON S J E.The biochemistry of silage[M].2nd ed.Kingston:Chalcombe Publications, 1991.
[28] 罗撄宁, 罗盈, 包锦泽, 等.添加乳酸菌和香草醛对甜玉米籽实青贮饲料品质的影响[J].饲料工业, 2020, 41(19):50-53. LUO Y N, LUO Y, BAO J Z, et al.Effects of adding lactic acid bacteria and vanillin on silage quality of corn seed[J].Feed Industry, 2020, 41(19):50-53.(in Chinese)
[29] 王小平, 王小琪, 李标, 等.复合菌剂对青贮玉米营养成分及其饲喂滩羊效果的研究[J].中国生态农业学报, 2020, 28(8):1258-1264. WANG X P, WANG X Q, LI B, et al.Testing the performance of compound microbial additives in silage maize nutrients and the effect of feeding Tan sheep[J].Chinese Journal of Eco-Agriculture, 2020, 28(8):1258-1264.(in Chinese)
[30] 程方, 李巨秀, 来航线, 等.多菌种混合发酵马铃薯渣产蛋白饲料[J].食品与发酵工业, 2015, 41(2):95-101. CHENG F, LI J X, LAI H X, et al.Research on production of protein feed from mixed-strains fermentatation of potato residue[J].Food and Fermentation Industries, 2015, 41(2):95-101.(in Chinese)
[31] 王志敬, 杨慧山, 葛影影, 等.发酵时间对青贮凤梨渣pH·亚硝酸盐·氨态氮·可溶性糖和有机酸含量的影响[J].安徽农业科学, 2020, 48(21):100-102. WANG Z J, YANG H S, GE Y Y, et al.Effects of fermentation time on pH, contents of nitrite, ammonia nitrogen, soluble sugar and organic acid in silage pineapple residue[J].Journal of Anhui Agricultural Sciences, 2020, 48(21):100-102.(in Chinese)
[32] 王平.不同乳酸菌发酵对玉米淀粉中直链淀粉含量的影响[J].粮油加工与食品机械, 2006(2):86-89. WANG P.Effect on content of amylase in corn starch through lactic acid bacterias fermentation[J].Machinery for Cereals Oil and Food Processing, 2006(2):86-89.(in Chinese)
[33] 宋华文, 王金才.乳酸菌与酵母菌在水产养殖中的应用[J].江西水产科技, 2018(5):12-15. SONG H W, WANG J C.Application of lactic acid bacteria and yeast in aquaculture[J].Jiangxi Fishery Sciences and Technology, 2018(5):12-15.(in Chinese)
[34] 侯然然.酵母细胞壁中葡甘露聚糖的提取及其霉菌毒素吸附效果[D].硕士学位论文.北京:中国农业科学院, 2007. HOU R R.Preparation of glucomannan from Saccharomyces cerevisiae and its effects on adsorption of mycotoxin[D].Master's Thesis.Beijing:Chinese Academy of Agricultural Sciences, 2007.(in Chinese)
[35] 彭俊, 励飞, 刘维, 等.饲料中黄曲霉毒素B₁和呕吐毒素生物法降解研究进展[J].饲料博览, 2020(1):28-32, 36. PENG J, LI F, LIU W, et al.Advances in biodegradation of aflatoxin B₁ and deoxynivalenol in feed[J].Feed Review, 2020(1):28-32, 36.(in Chinese)
[36] BOVO F, FRANCO L T, ROSIM R E, et al.The ability of Lactobacillus rhamnosus in solution, spray-dried or lyophilized to bind aflatoxin B₁[J].Journal of Food Research, 2014, 3(2):35-42.

[37] HERNANDEZ-MENDOZA A, GUZMAN-DE-PEÑA D, GARCIA H S.Key role of teichoic acids on aflatoxin B1 binding by probiotic bacteria[J].Journal of Applied Microbiology, 2009, 107(2):395-403.

[38] 刘畅, 刘阳, 邢福国, 等.黄曲霉毒素B₁吸附菌株的筛选及吸附机理研究[J].核农学报, 2010, 24(4):766-771. LIU C, LIU Y, XING F G, et al.Screening of aflatoxin B₁ binding strains and binding mechanism of yeast strain Y1[J].Journal of Nuclear Agricultural Sciences, 2010, 24(4):766-771.(in Chinese)
[39] NIDERKORN V, BOUDRA H, MORGAVI D P.Binding of Fusarium mycotoxins by fermentative bacteria in vitro[J].Journal of Applied Microbiology, 2006, 101(4):849-856.

[40] REPEDKIENE J, LEVINSKAITE L, PASKEVICIUS A, et al.Toxin-producing fungi on feed grains and application of yeasts for their detoxification[J].Polish Journal of Veterinary Sciences, 2013, 16(2):391-393.

[41] 赵雪芹, 朱风华, 陈甫, 等.6种益生菌对玉米赤霉烯酮和呕吐毒素降解能力的研究[J].黑龙江畜牧兽医, 2020(14):108-111. ZHAO X Q, ZHU F H, CHEN F, et al.Study on the degradation ability of 6 kinds of probiotics on zearalenone and deoxynivalenol[J].Heilongjiang Animal Science and Veterinary Medicine, 2020(14):108-111.(in Chinese)
[42] WANG N, WU W W, PAN J W, et al.Detoxification strategies for zearalenone using microorganisms:a review[J].Microorganisms, 2019, 7(7):208.
[43] HU W, SCHMIDT R J, MCDONELL E E, et al.The effect of Lactobacillus buchneri 40788 or Lactobacillus plantarum MTD-1 on the fermentation and aerobic stability of corn silages ensiled at two dry matter contents[J].Journal of Dairy Science, 2009, 92(8):3907-3914.

[44] BUMBIERIS V H, Jr, DE PIETRO GUIMARÃES V A, DE SOUZA FORTALEZA A P, et al.Aerobic stability in corn silage (Zea mays L.) ensiled with different microbial additives[J].Acta Scientiarum.Animal Sciences, 2017, 39(4):357-362.

[45] TENNANT R K, SAMBLES C M, DIFFEY G E, et al.Metagenomic analysis of silage[J].Journal of Visualized Experiments, 2017(119):e54936.
[46] KLEINSCHMIT D H, SCHMIDT R J, KUNG L, Jr.The effects of various antifungal additives on the fermentation and aerobic stability of corn silage[J].Journal of Dairy Science, 2005, 88(6):2130-2139.

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