Skip to main content
Download PDF

Comparative effects of enzymatic soybean, fish meal and milk powder in diets on growth performance, immunological parameters, SCFAs production and gut microbiome of weaned piglets

Journal of Animal Science and Biotechnology volume 12, Article number: 106 (2021) Cite this article

Abstract

Background

The objective of this study was to evaluate the replacement effects of milk powder (MK) and fish meal (FM) by enzymatic soybean (ESB) in diets on growth performance, immunological parameters, SCFAs production and gut microbiome of weaned piglets.

Methods

A total of 128 piglets with initial body weight at 6.95 ± 0.46 kg, were randomly assigned into 4 dietary treatments with 8 replicates per treatment and 4 piglets per replicate for a period of 14 d. Piglets were offered iso-nitrogenous and iso-energetic diets as follows: CON diet with MK and FM as high quality protein sources, ESB plus FM diet with ESB replacing MK, ESB plus MK diet with ESB replacing FM, and ESB diet with ESB replacing both MK and FM.

Results

No significant differences were observed in growth performance among all treatments (P > 0.05). However, piglets fed ESB plus FM or ESB diet had increased diarrhea index (P<0.01), and lower digestibility of dry matter (DM), gross energy (GE) or crude protein (CP), relative to piglets fed CON diet (P <  0.01). Moreover, the inclusion of ESB in diet markedly decreased the plasma concentration of HPT and fecal concentration of butyric acid (BA) (P<0.01). The High-throughput sequencing of 16S rRNA gene V3−V4 region of gut microbiome revealed that the inclusion of ESB in diet increased the alpha diversity, and the linear discriminant analysis effect size (LEfSe) showed that piglets fed with ESB plus FM or ESB diet contained more gut pathogenic bacteria, such as g_Peptococcus, g_Veillonella and g_Helicobacter.

Conclusion

The inclusion of ESB in diet did not markedly affect growth performance of piglets, but the replacement of MK or both MK and FM by ESB increased diarrhea index, which could be associated with lower nutrients digestibility and more gut pathogenic bacteria. However, piglets fed diet using ESB to replace FM did not markedly affect gut health-related parameters, indicating the potential for replacing FM with ESB in weaning diet.

Background

Weaning is a critical period for the growth and development of piglets, during which weaned piglets have to cope with a series of problems such as dysfunction of intestinal barrier and systematic inflammation induced by weaning stress, thereby aggravate diarrhea, morbidity and mortality, and poor growth performance [1]. During the suckling period, piglets ingest high digestible milk of sows as the major food. However, the newly weaned piglets are abruptly forced to adapt to the nutritional and environmental changes, especially digest solid diets containing high contents of plant proteins with the immature digestive and immune systems, which has been demonstrated to aggravate the weaning stress [2, 3]. Thus, it is extremely urgent to incorporate the high quality protein sources into diet to prevent weaning stress in piglets [4].

Fish meal (FM) and milk powder (MK) are extensively used in creep feed as high quality protein sources due to the higher digestibility, greater palatability and appropriate composition of amino acids [5, 6]. However, the exorbitant cost of FM and MK have necessitated the identification of alternative cheaper protein sources for the weaning diets [7, 8]. Although soybean has become the primary protein source in swine diet due to the excellent balance of essential amino acids and lower price [9], soybean is not recommendable to be directly used in weaning diets due to its anti-nutritional factors (ANFs), which can cause hypersensitivity [10,11,12].

It is confirmed that bioprocessing of soybean is an effective way to eliminate ANFs and improve the bioavailability of diet [13, 14]. The enzymatic soybean (ESB), produced by fermentation and enzymatic hydrolysis of soybean, is an excellent protein source with less trypsin inhibitors and antigen proteins [15, 16]. In addition, It has been reported that growth performance, antioxidant capacity, immune function and nutrients digestibility of weaned piglets could be improved as the ESB was incorporated into diet to replace some other dietary protein sources such as soybean meal (SBM), soybean protein concentrate (SPC), fermented soybean meal (FSBM) or FM [17,18,19]. However, to the best of our knowledge, there is short of researches regarding the comparison of employing ESB to completely substitute for FM, MK or both FM and MK in weaning diets. Hence, the objective of the study was to assess the comparative effects of FM, MK or both MK and FM replacing with ESB on growth performance, nutrients digestibility, immunological parameters, gut microbiome and short-chain fatty acids (SCFAs) in weaned piglets.

Materials and methods

The experiment followed the animal protection law (Ethic Approval Code: SCAUAC201308–2) and was performed in accordance with the Guide for the Animal Care and Use approved by Sichuan Agricultural University Institutional Animal Care and Use Committee.

Preparation of ingredients

The MK was obtained from Fonterra, New Zealand. The FM was produced by Pesquera diamante, Peru. The ESB, which was obtained from Fatide, Jiangsu Fuhai Biology Co., Ltd., contained 40.00% crude protein (CP), 18.00% fat, 2.80% fiber, and lower ANFs (0.13% stachyose, 0.39% raffinose, 126.32 TIU/g trypsin inhibitor, β-conglycinin and glycinin are less than 1.4 mg/g and 2.8 mg/g respectively) compared with unprocessed soybean.

Animals, diets, and experimental design

A total of 128 piglets ((Landrace × Yorkshire × Duroc) × Yorkshire; 21d ± 2d) with an initial body weight at 6.95 ± 0.46 kg were randomly assigned into 4 dietary treatments in a randomized complete block design according to body weight: (1) CON diet with MK and FM as high quality protein sources; (2) ESB plus FM diet with ESB replacing MK; (3) ESB plus MK diet with ESB replacing FM; (4) ESB diet with ESB replacing both MK and FM. Each treatment group had 32 piglets with 8 pens and 4 piglets (2 barrows and 2 gilts) per pen for the 14-d experiment. As presented in Table 1, all the diets were formulated to be iso-nitrogenous and iso-energetic and meet or exceed the recommendation of NRC (2012) [20]. Piglets were housed in pens (1.5 m × 1.5 m) with infrared lamps hanged above and the temperature was kept between 26 and 28 °C. Piglets had free access to feed and water during the experimental period.

Table 1 The ingredient composition and analyzed nutrient levels of diets (as fed basis)
Full size table

Growth performance

The feed supply and feed refusals were recorded every day, and piglets were individually weighted every week to calculate average daily gain (ADG), average daily feed intake (ADFI) and the ratio of ADFI and ADG (F:G). Diarrhea scores were visually assessed three times a day as previous described [21]. Briefly, firm and well-formed feces were scored as 0; soft and formed feces were scored as 1; fluid and usually yellowish feces were scored as 2; and watery and projectile feces were scored as 3. Diarrhea index was calculated according to the following equation: diarrhea index = the sum of diarrhea scores / (numbers of piglets per pen × experimental days × assessed times per day).

Sample collection

Blood samples (10 mL, n = 8) were collected from jugular vein into sodium heparinized tubes at 08:00 of d 8 after an overnight fast. Plasma was obtained by centrifuging at 3000 × g for 15 min at 4 °C and stored immediately at − 20 °C for later analysis. At the same day, fresh fecal samples (n = 8) were collected by rectal stimulation, then snap frozen at − 80 °C for the gut microbiome analysis.

To determine the nutrients digestibility of piglets, 0.5% chromic oxide was additional added to the diets as an exogenous indicator on d 8. After 4-d adaptation period, fresh fecal samples were collected during d 12 to d 14. The diet and fecal samples for nutrients digestibility determination were stored at − 20 °C until analysis.

Chemical analysis

The diets and feces samples were dried at 65 °C for 72 h, ground through a 0.42-mm screen and analyzed according to methods of AOAC for dry matter (DM) [22]. CP was determined by copper catalyst Kjeldahl method and GE was determined by an automatic adiabatic oxygen bomb calorimeter (Parr 6400, Parr Instrument Co., Moline, IL, USA). Amino acids, except tryptophan, were measured by an automatic amino acid analyzer (L-8900, Hitachi, Tokyo, Japan) after acidolysis for 24 h. Chromium was determined by a flame atomic absorption spectrophotometer (ContrAA 700, Analytikjena, Jena, Germany). The Apparent total tract digestibility (ATTD) was calculated according to the following equation: ATTDnutrient = 1 − (Crdiet × Nutrientfeces) / (Crfeces × Nutrientdiet) [23].

Measurement of plasma parameters

Plasma samples were thawed on the ice before analysis. The 300 μL of supernatant were obtained to determine the concentrations of plasma immunoglobulin A (IgA) and immunoglobulin G (IgG) via automatic biochemical analyzer (Hitachi 3100, Hitachi High-Technologies Corporation, Tokyo, Japan) with corresponding kits (Sichuan Maker Biotechnology Co. Ltd). The levels of haptoglobin (HPT) and pig major acute-phase protein (Pig-MAP) were measured by spectrophotometric methods (Spectra Max M2; Molecular Devices, California, USA), according to the kit instructions (Nanjing Jiancheng Bioengineering Institute, Nanjing, China). There was less than 5% variation of intra-assay and inter-assay coeffcients for each assay.

Quantification of SCFAs

The concentrations of SCFAs were determined by gas chromatography (Varian CP-3800, manual injection, flame ionization detector, FID, 10 μL micro-injector). Approximately 0.7 g of fecal samples were thawed and diluted with 1.5 mL of ultrapure water, and 1.0 mL supernatant was obtained by centrifuging at 3000 × g for 15 min. Then the supernatant was mixed with 0.2 mL of 25% metaphosphoric acid solution and 23.3 μL of 210 mmol/L crotonic acid and the mixed solution was placed at 4 °C for 30 min before centrifuging at 4000 × g for 10 min, afterwards the 0.3 mL of supernatant was mixed with 0.9 mL of methanol, filtered by 0.22-μm filter (Millipore Co., Bedford, MA) after centrifuging at 3500 × g for 5 min.

Sequencing of gut microbiome

The total genomic DNA was extracted from fecal samples (n = 8) using the QIAamp DNA stool Mini Kit (Qiagen, GmbH Hilden, Germany). The concentration and purity of the extracted genomic DNA were measured using a NanoDrop ND-1000 Spectrophotometer (NanoDrop Technologies Inc., Wilmington, DE, USA). The integrity of the extracted genomic DNA was determined by electrophoresis on 1% (w/v) agarose gels. Extracted fecal DNA samples were sent to Majorbio Bio-pharm Technology Co., Ltd. (Shanghai, China) to perform amplicon pyrosequencing on the Illumina MiSeq platforms. The V3−V4 hypervariable region of the 16S rRNA gene was amplified by PCR with primers 338F (5′-ACTCCTACGGGAGGCAGCAG-3′) and 806R (5′-GGACTACHVGGTWTCTAAT-3′). The Uparse 7.0.1090 was used to clustered Operational taxonomic units (OTUs) at 97% sequence similarity. Representative sequences were selected and assigned by the Ribosomal Database Project (RDP) classifier Version 2.11. The relative abundance of each OTU was examined at different taxonomic levels. Diversity within communities (Alpha diversity) calculations and taxonomic community assessments were performed by Mothur 1.30.2 and Qiime 1.9.1. Principal coordinates analysis (PCoA) plots were produced using unweighted UniFrac metrics. The linear discriminant analysis (LDA) effect size (LEfSe) method was performed to elucidate the difference among treatments.

Statistical analysis

Data were analyzed using PROC MIXED of SAS 9.4 (SAS Inst. Inc., Cary, NC, USA). The data were considered as outlier when the student residue was greater than three. The UNIVARIATE procedures of SAS were used to analyze the variance homogeneity and normality, respectively. Least squares means were calculated using the LSMEANS procedure in SAS, and significant differences among treatments were separated using PDIFF option with the Tukey adjustment for the performance data. For growth performance, diarrhea index, ATTD and SCFAs, pens were regarded as the experimental unit and piglet data were reported as a mean for the pen. The different taxa microbes among lines were identified using LEfSe analysis with LDA score > 3. Data are presented as the least squares means and pooled standard error. Results were considered significant when P < 0.05 and tendency toward significance at 0.05 ≤ P < 0.10.

Results

Growth performance

As presented in Table 2, piglets fed CON or ESB plus MK diet had lower diarrhea index (P <  0.01) during d 1–7 and the whole experimental period, relative to piglets fed ESB plus FM or ESB diet. Besides, piglets fed CON or ESB plus MK diet had lower diarrhea index (P <  0.01) during d 8–14, when compared with piglets fed ESB diet. There was no significant difference among dietary treatments in ADG, ADFI and F:G at all phase.

Table 2 Effects of dietary protein sources on growth performance and diarrhea index in weaned piglets
Full size table

The ATTD of nutrients

As presented in Table 3, the digestibility of DM, CP and GE were significantly lower in the piglets fed ESB plus FM diet (P < 0.01), when compared with piglets fed CON diet. The DM, CP and GE digestibility did not markedly differ between piglets fed ESB plus MK diet and CON diet (P > 0.05), but piglets fed ESB plus MK diet had significantly higher digestibility of DM and GE than that of ESB plus FM group (P <  0.01). Besides, piglets fed ESB diet had markedly decreased digestibility of DM and GE, relative to piglets fed CON diet (P <  0.01).

Table 3 Effects of dietary protein sources on ATTD of nutrients in weaned piglets
Full size table

Immunological parameters

As presented in Fig. 1, the plasma concentrations of Pig-MAP, IgG and IgM were not markedly affected by dietary treatments (P > 0.05). Piglets fed ESB plus FM, ESB plus MK and ESB had markedly decreased plasma concentration of HPT when compared with piglets fed CON diet (P <  0.01).

Fig. 1
figure 1

Effects of dietary protein sources on immunological parameters in weaned piglets

Full size image

Gut microbiome

A total of 41,562, 46,965, 53,874 and 50,331 effective sequences in fecal samples from CON, ESB plus FM, ESB plus MK and ESB groups were identified, respectively. From the Venn analysis of OTUs, 818, 901, 908 and 905 unique OTUs were identified in CON, ESB plus FM, ESB plus MK and ESB groups, respectively (Fig. 2A). For beta diversity analysis, unweighted Unifrac PCoA was performed to demonstrate the separation of bacterial community composition among treatments by using the first two principal component scores of PC1 and PC2 (30.38% and 8.1%) of the explained variance, respectively (Fig. 2B).

Fig. 2
figure 2

Effects of dietary protein sources on gut microbiome structure in weaned piglets. Venn diagram showing the unique and shared OTUs in the different groups (A). Unweighted Unifrac PCoA analysis based on OTUs of gut microbiome (B)

Full size image

As shown in Table 4, dietary treatments did not markedly affect Shannon, Simpson and ACE indexes, but piglets fed ESB or ESB plus FM diets had significantly increased Chao 1 index when compared with piglets fed CON diet (P <  0.05).

Table 4 The alpha diversity in the fecal microbiome of weaned piglets
Full size table

The relative abundances at phylum level among treatments are presented in Fig. 3A, suggesting that the top 6 dominated phyla were Firmicutes, Bacteroidota, Actinobacteriota, Proteobacteria, Spirochaetota and Desulfobacterota. Firmicutes occupied the maximal portion of gut microbiome in all samples, with a relative abundance of 50%. At the genus level, a total of 248 genera were identified among all samples, and the top 26 genera (> 1.5% in at least one group) are shown in Fig. 3B.

Fig. 3
figure 3

Effects of dietary protein sources on phyla and genus of gut microbiome in weaned piglets. Relative abundances of phyla (A) and genus (B). The abundance is expressed in terms of a percentage of the total effective bacterial sequences in the fecal samples

Full size image

LEfSe was used to analyze microbial community from phylum to genus level. There were 4, 8, 3 and 17 kinds of dominant bacteria in fecal samples of piglets fed CON, ESB plus FM, ESB plus MK and ESB diets respectively (Fig. 4). The most abundant phylotypes in fecal samples of piglets fed ESB plus FM diet were o_Clostridia_vadinBB60_group, f_Erysipelotrichaceae, g_Peptococcus, and g_Veillonella. And o_RF39, f_Oscillospiraceae, f_Nocardiaceae and g_Helicobacter were enriched in fecal samples of piglets fed ESB diet.

Fig. 4
figure 4

Effects of dietary protein sources on the phylotypes distribution in weaned piglets. LefSe cladogram representing differential abundant taxa in pig gut microbiome (A). LefSe bar representing differential abundant taxa in pig gut microbiome (B)

Full size image

Concentrations of SCFAs in feces

The results pertaining to the SCFAs in feces are presented in Table 5. Compared with piglets fed ESB plus FM, ESB plus MK and ESB diets, piglets fed CON diet had significantly increased level of butyric acid (BA) in feces (P <  0.01) and tended to have higher level of acetic acid (AA) (P = 0.09). The level of propionic acid (PA) in feces was not significant different across the dietary treatments (P > 0.05).

Table 5 Effects of dietary protein sources on the SCFAs levels in fecal samples in weaned piglets
Full size table

Discussion

Soybean is commonly used in the diets of pigs because of its high content of proteins and greater digestibility [24, 25]. However, ANFs in soybean may have negative impact on the immature gastrointestinal tract of weaned piglets, which may result in severe diarrhea [26]. Bio-processed soybean products, such as SPC, FSBM and ESB, have been demonstrated to remove ANFs effectively and improve nutrients digestibility, which leads to a better growth performance in weaned piglets [27, 28]. In addition to decrease ANFs, the fermentation and enzymatic hydrolysis process of ESB also produces more small peptides, which has various physiological function in piglets or other mammals [29,30,31].

In the present study, the growth performance of weaned piglets did not markedly differ among treatments, however, we did observe the growth performance of piglets fed ESB plus FM or ESB diet had been numerically decreased, as indicated by the average 14% decrease in whole period ADG.

Protein digestion has been proposed to be a major dietary factor affecting growth and diarrhea incidence of weaned piglets [32], as the undigested dietary protein enters into the hindgut leading to altered gut microbiome [33]. In this study, we observed the diarrhea index was markedly increased by feeding ESB plus FM diet or ESB diet, which could be partially ascribed to the poor nutrient digestibility.

For weaning piglets, weaning stress is a vital factor that causing immunological and intestinal impairments [34]. The immunoglobulins and acute phase proteins, such as IgG, IgM, Pig-MAP and HPT, could regulate immunity by inhibiting the release of IL-1 and TNF-α [35, 36]. As an inflammatory signaling factor, the plasma HPT concentration could be particularly elevated during the occurrence of inflammation and injury [37]. Our study showed that piglets fed ESB plus FM, ESB plus MK and ESB diets had dramatically decreased plasma concentration of HPT, which is consistent with previous study that piglets fed enzymolytic soybean meal had improved immune function, as indicated by the higher levels of CD4+ and CD8+ in peripheral blood [38]. The immunoregulatory effect of bio-processed soybean may be related to the functional bio-active peptides. Supportively, the small peptides account for up to 33.58% of total protein in ESB we used in this study. Similarly, recent studies have reported that the size of soybean peptide could be reduced to 100 ~ 1000 Da by microbial fermentation and proteolysis during the production of ESB, containing abundant bio-active peptides, such as QRPR and lunasin [39,40,41].

Gut microbiome has been shown to play an important role in development and function of weaned piglets [42]. In our study, the 16S rRNA sequencing was used to investigate the gut microbiome responses to dietary protein sources in weaned piglets. Our results showed that the piglets fed ESB plus MK diet contained the most OTUs. Besides, piglets fed ESB plus FM diet or ESB diet increased Chao 1 index (P = 0.03), and piglets fed ESB diets increased ACE index by 44% ~ 67%, indicating that the inclusion of ESB in diet increased gut microbial richness and diversity in weaned piglets .

To elucidate the difference in microbiome among treatments, LefSe method was conducted to analyze the enriched bacteria in each group. In the present study, piglets fed ESB plus FM or ESB diet had increased the abundances of some pathogenic bacteria in feces, such as o_Clostridia_vadinBB60_group, o_RF39, f_Erysipelotrichaceae, f_Oscillospiraceae, f_Nocardiaceae, g_Peptococcus, g_Veillonella and g_Helicobacter. It has been reported that o_Clostridia_vadinBB60_group and f_Erysipelotrichaceae were enriched in the lumen of colorectal cancer patients [43, 44]. In addition, g_Peptococcus is a classic pathogenic bacteria colonized in animals with gastrointestinal disease, and g_Veillonella was negatively correlated with the nutritional index [45, 46]. The higher abundances of o_Oscillospiraceae and g_Helicobacter have been found to be related to intestinal inflammation [47, 48]. The o_RF39, which belongs to phylum Tenericutes, class Mollicutes, is associated with intestinal disorders [49, 50]. Furthermore, f_Nocardiaceae belongs to phylum Actinobacteria, is a strong predictor of diarrhea in piglets [51]. Taken together, the significantly higher diarrhea index in piglets fed ESB plus FM or ESB diet may be attributed to the intestinal damage induced by the enriched pathogenic bacteria.

SCFAs, the main metabolites produced by bacterial fermentation of dietary fiber and protein in the large intestine, can regulate the absorption of various nutrients and provide nearly 30% of the energy requirements for maintenance in pigs and then improve piglets performance [52,53,54]. In the current study, we found that piglets fed CON diet had higher abundance of g_Bifidobacterium in feces, which has been demonstrated to suppress the colonization of pathogenic bacteria and contribute to the production of SCFAs [55,56,57]. Supportively, we did observe the increased AA and BA levels in piglets fed CON diet.

Conclusion

In this study, the inclusion of ESB in weaning diet did not markedly affect growth performance of piglets, but the substitution of MK or both MK and FM with ESB in diet leaded to higher diarrhea index, which could be ascribed to the lower nutrients digestibility and more gut pathogenic bacteria, such as g_Veillonella, g_Helicobacter and g_Peptococcus. However, piglets fed diet using ESB to replace FM did not markedly affect gut health-related parameters, indicating the potential for replacing FM with ESB in weaning diet.

Availability of data and materials

The datasets analyzed in the current study are available from the corresponding author on reasonable request.

Abbreviations

MK:

Milk powder

FM:

Fish meal

ESB:

Enzymatic soybean

SCFAs:

Short-chain fatty acids

DM:

Dry matter

CP:

Crude protein

GE:

Gross energy

LEfSe:

Linear discriminant analysis effect size

LDA:

Linear discriminant analysis

ANFs:

Anti-nutritional factors

ADG:

Average daily gain

ADFI:

Average daily feed intake

F:G:

Feed to gain ratio

ATTD:

Apparent total tract digestibility

HPT:

Haptoglobin

Pig-MAP:

Pig major acute-phase protein

OTUs:

Operational taxonomic units

RDP:

Ribosomal Database Project

PCoA:

Principal coordinates analysis

AA:

Acetic acid

PA:

Propionic acid

BA:

Butyric acid

References

  1. Boudry G, Peron V, Le Huerou-Luron I, Lalles JP, Seve B. Weaning induces both transient and long-lasting modifications of absorptive, secretory, and barrier properties of piglet intestine. J Nutr. 2004;134(9):2256–62. https://doi.org/10.1093/jn/134.9.2256.

    Article  CAS  PubMed  Google Scholar 

  2. Heo J, Opapeju F, Pluske J, Kim J, Hampson D, Nyachoti C. Gastrointestinal health and function in weaned pigs: a review of feeding strategies to control post-weaning diarrhoea without using in-feed antimicrobial compounds. J Anim Physiol Anim Nutr. 2013;97(2):207–37. https://doi.org/10.1111/j.1439-0396.2012.01284.x.

    Article  CAS  Google Scholar 

  3. Rist V, Weiss E, Eklund M, Mosenthin R. Impact of dietary protein on microbiota composition and activity in the gastrointestinal tract of piglets in relation to gut health: a review. Animal. 2013;7(7):1067–78. https://doi.org/10.1017/S1751731113000062.

    Article  CAS  PubMed  Google Scholar 

  4. Ren M, Liu C, Zeng X, Yue L, Mao X, Qiao S, et al. Amino acids modulates the intestinal proteome associated with immune and stress response in weaning pig. Mol Biol Rep. 2014;41(6):3611–20. https://doi.org/10.1007/s11033-014-3225-3.

    Article  CAS  PubMed  Google Scholar 

  5. Kim S, Van Heugten E, Ji F, Lee C, Mateo R. Fermented soybean meal as a vegetable protein source for nursery pigs: I. effects on growth performance of nursery pigs. J Anim Sci. 2010;88(1):214–24. https://doi.org/10.2527/jas.2009-1993.

    Article  CAS  PubMed  Google Scholar 

  6. Rault JL, Ferrari J, Pluske J, Dunshea F. Neonatal oxytocin administration and supplemental milk ameliorate the weaning transition and alter hormonal expression in the gastrointestinal tract in pigs. Domest Anim Endocrinol. 2015;51:19–26. https://doi.org/10.1016/j.domaniend.2014.11.001.

    Article  CAS  PubMed  Google Scholar 

  7. Jones A, Woodworth J, DeRouchey J, Dritz S, Tokach M, Goodband R. Effect of enzymatically fermented soybean meal and lactobacillus Plantarum on nursery pig performance. Kansas Agr Expt Sta Res Rep. 2016;2(8):14.

    Google Scholar 

  8. Zhao P, Park J, Heo J, Yoo J, Jeong J, Kim I. Partial fish meal replacement with fermented or enzymatically prepared soybean meal in weaned pig diets. Anim Prod Sci. 2015;55(12):1551. https://doi.org/10.1071/ANv55n12Ab009.

    Article  Google Scholar 

  9. Mukherjee R, Chakraborty R, Dutta A. Role of fermentation in improving nutritional quality of soybean meal−a review. Asian Austral J Anim. 2016;29(11):1523.

    Article  CAS  Google Scholar 

  10. Wang T, Qin GX, Sun ZW, Zhao Y. Advances of research on Glycinin and β-Conglycinin: a review of two major soybean allergenic proteins. Crit Rev Food Sci. 2014;54(7):850–62. https://doi.org/10.1080/10408398.2011.613534.

    Article  CAS  Google Scholar 

  11. Wu JJ, Cao CM, Ren DD, Zhang Y, Kou YN, Ma LY, et al. Effects of soybean antigen proteins on intestinal permeability, 5-hydroxytryptamine levels and secretory IgA distribution in the intestine of weaned piglets. Ital J Anim Sci. 2016;15(1):174–80. https://doi.org/10.1080/1828051X.2016.1148559.

    Article  CAS  Google Scholar 

  12. Hao Y, Zhan Z, Guo P, Piao X, Li D. Soybean β-conglycinin-induced gut hypersensitivity reaction in a piglet model. Arch Anim Nutr. 2009;63(3):188–202. https://doi.org/10.1080/17450390902860026.

    Article  CAS  Google Scholar 

  13. Jeong JS, Park JW, Lee SI, Kim IH. Apparent ileal digestibility of nutrients and amino acids in soybean meal, fish meal, spray-dried plasma protein and fermented soybean meal to weaned pigs. Anim Sci J. 2016;87(5):697–702. https://doi.org/10.1111/asj.12483.

    Article  CAS  PubMed  Google Scholar 

  14. Szécsi G, Csehi B, Mednyánszky Z, Kiskó G, Bányai É, Dernovics M, et al. Production of hypoallergenic antibacterial peptides from defatted soybean meal in membrane bioreactor: a bioprocess engineering study with comprehensive product characterization. Food Technol Biotechnol. 2017;55(3):308–24.

    PubMed  PubMed Central  Google Scholar 

  15. Hong KJ, Lee CH, Kim SW. Aspergillus oryzae GB-107 fermentation improves nutritional quality of food soybeans and feed soybean meals. J Med Food. 2004;7(4):430–5. https://doi.org/10.1089/jmf.2004.7.430.

    Article  CAS  PubMed  Google Scholar 

  16. Li S, Ding G, Song F, Sang C, Wang A, Chen N. Comparison of dehulled, fermented and enzyme-treated soybean meal in diets for largemouth bass, Micropterus salmoides: effects on growth performance, feed utilization, immune response and intestinal morphology. Anim Feed Sci Technol. 2020;267:114548. https://doi.org/10.1016/j.anifeedsci.2020.114548.

    Article  CAS  Google Scholar 

  17. Ma XK, Shang QH, Wang QQ, Hu JX, Piao XS. Comparative effects of enzymolytic soybean meal and antibiotics in diets on growth performance, antioxidant capacity, immunity, and intestinal barrier function in weaned pigs. Anim Feed Sci Technol. 2018;248:47–58.

    Article  Google Scholar 

  18. Ma X, Shang Q, Hu J, Liu H, Brøkner C, Piao X. Effects of replacing soybean meal, soy protein concentrate, fermented soybean meal or fish meal with enzyme-treated soybean meal on growth performance, nutrient digestibility, antioxidant capacity, immunity and intestinal morphology in weaned pigs. Livest Sci. 2019;225:39–46. https://doi.org/10.1016/j.livsci.2019.04.016.

    Article  Google Scholar 

  19. Zhou S, Sun Z, Ma L, Yu J, Ma C, Ru Y. Effect of feeding enzymolytic soybean meal on performance, digestion and immunity of weaned pigs. Asian Australas J Anim Sci. 2010;24(1):103–9. https://doi.org/10.5713/ajas.2011.10205.

    Article  CAS  Google Scholar 

  20. NRC. Nutrient requirements of swine, 11th rev. ed. Washington, DC: National Academy Press; 2012.

    Google Scholar 

  21. Che L, Hu L, Zhou Q, Peng X, Liu Y, Luo Y, et al. Microbial insight into dietary protein source affects intestinal function of pigs with intrauterine growth retardation. Eur J Nutr. 2020;59(1):327–44. https://doi.org/10.1007/s00394-019-01910-z.

    Article  CAS  PubMed  Google Scholar 

  22. AOAC. Official methods of analysis. 18th ed. Association of Official Analytical Chemists: Gaithersburg; 2007.

    Google Scholar 

  23. Yu J, Yu G, Yu B, Zhang Y, He J, Zheng P, et al. Dietary protease improves growth performance and nutrient digestibility in weaned piglets fed diets with different levels of soybean meal. Livest Sci. 2020;241:104179. https://doi.org/10.1016/j.livsci.2020.104179.

    Article  Google Scholar 

  24. Barnett KL, Kornegay ET, Risley CR, Lindemann MD, Schurig GG. Characterization of creep feed consumption and its subsequent effects on immune response, scouring index and performance of weanling pigs. J Anim Sci. 1989;67(10):2698–708. https://doi.org/10.2527/jas1989.67102698x.

    Article  CAS  PubMed  Google Scholar 

  25. Fan MZ, Sauer WC, De Lange CFM. Amino acid digestibility in soybean meal, extruded soybean and full-fat canola for early-weaned pigs. Anim Feed Sci Technol. 1995;52(3–4):189–203.

    Article  CAS  Google Scholar 

  26. Wu Y, Jiang Z, Zheng C, Wang L, Zhu C, Yang X, et al. Effects of protein sources and levels in antibiotic-free diets on diarrhea, intestinal morphology, and expression of tight junctions in weaned piglets. Anim Nutr. 2015;1(3):170–6. https://doi.org/10.1016/j.aninu.2015.08.013.

    Article  PubMed  PubMed Central  Google Scholar 

  27. Zheng L, Li D, Li Z, Kang LN, Jiang YY, Liu XY, et al. Effects of Bacillus fermentation on the protein microstructure and anti-nutritional factors of soybean meal. Lett Appl Microbiol. 2017;65(6):520–6. https://doi.org/10.1111/lam.12806.

    Article  CAS  PubMed  Google Scholar 

  28. Sangwoo P, Wook LJ, Cowieson AJ, Guenter P, Awori WT. Soybean meal allergenic protein degradation and gut health of piglets fed protease-supplemented diets. J Anim Sci. 2020;10:10.

    Google Scholar 

  29. Yang YX, Kim YG, Lohakare JD, Yun JH, Chae BJ, Ji P, et al. Comparative efficacy of different soy protein sources on growth performance, nutrient digestibility and intestinal morphology in weaned pigs. Asian Australas J Anim Sci. 2007;20(5):775–83. https://doi.org/10.5713/ajas.2007.775.

    Article  CAS  Google Scholar 

  30. Koepke J, Kaushik R, Gibbons W, Brown M, Levesque C. Evaluation of a bioprocessed soybean meal on nursery pig performance and immune status. J Anim Sci. 2017;95(11):5030–9. https://doi.org/10.2527/jas2017.1679.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Yoshikawa M. Bioactive peptides derived from natural proteins with respect to diversity of their receptors and physiological effects. Peptides. 2015;72:208–25. https://doi.org/10.1016/j.peptides.2015.07.013.

    Article  CAS  PubMed  Google Scholar 

  32. Li J, Yin L, Wang L, Li J, Huang P, Yang H, et al. Effects of vitamin B6 on growth, diarrhea rate, intestinal morphology, function, and inflammatory factors expression in a high-protein diet fed to weaned piglets. J Anim Sci. 2019;97(12):4865–74. https://doi.org/10.1093/jas/skz338.

    Article  PubMed  PubMed Central  Google Scholar 

  33. Jha R, Berrocoso JFD. Dietary fiber and protein fermentation in the intestine of swine and their interactive effects on gut health and on the environment: a review. Anim Feed Sci Technol. 2016;212:18–26. https://doi.org/10.1016/j.anifeedsci.2015.12.002.

    Article  CAS  Google Scholar 

  34. Moeser AJ, Pohl CS, Rajput M. Weaning stress and gastrointestinal barrier development: implications for lifelong gut health in pigs. Anim Nutr. 2017;3(4):313–21. https://doi.org/10.1016/j.aninu.2017.06.003.

    Article  PubMed  PubMed Central  Google Scholar 

  35. Arthington JD, Eicher SD, Kunkle WE, Martin F. Effect of transportation and commingling on the acute-phase protein response, growth, and feed intake of newly weaned beef calves. J Anim Sci. 2003;81(5):1120–5. https://doi.org/10.2527/2003.8151120x.

    Article  CAS  PubMed  Google Scholar 

  36. Ren M, Zhang S, Zeng X, Liu H, Qiao S. Branched-chain amino acids are beneficial to maintain growth performance and intestinal immune-related function in weaned piglets fed protein restricted diet. Asian Australas J Anim Sci. 2015;28(12):1742–50. https://doi.org/10.5713/ajas.14.0131.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Liu Y, Song M, Che TM, Almeida JAS, Lee JJ, Pettigrew JE, et al. Dietary plant extracts alleviate diarrhea and alter immune responses of weaned pigs experimentally infected with a pathogenic Escherichia coli. J Anim Sci. 2013;91(11):5294–306. https://doi.org/10.2527/jas.2012-6194.

    Article  CAS  PubMed  Google Scholar 

  38. Zhou SF, Sun ZW, Ma LZ, Yu JY, Ma CS, Ru YJ. Effect of feeding Enzymolytic soybean meal on performance, digestion and immunity of weaned pigs. Asian Australas J Anim Sci. 2011;24(1):103–9.

    Article  CAS  Google Scholar 

  39. Pan FG, Wang L, Cai Z, Wang YN, Wang YF, Zhang X, et al. Soybean peptide QRPR activates autophagy and attenuates the inflammatory response in the RAW264.7 cell model. Protein Pept Lett. 2019;26(4):301–12. https://doi.org/10.2174/0929866526666190124150555.

    Article  CAS  PubMed  Google Scholar 

  40. Osho SO, Xiao WW, Adeola O. Response of broiler chickens to dietary soybean bioactive peptide and coccidia challenge. Poult Sci. 2019;98(11):5669–78. https://doi.org/10.3382/ps/pez346.

    Article  CAS  PubMed  Google Scholar 

  41. Fernandez-Tome S, Hernandez-Ledesma B. Current state of art after twenty years of the discovery of bioactive peptide lunasin. Food Res Int. 2019;116:71–8. https://doi.org/10.1016/j.foodres.2018.12.029.

    Article  CAS  PubMed  Google Scholar 

  42. Maltecca C, Bergamaschi M, Tiezzi F. The interaction between microbiome and pig efficiency: a review. J Anim Breed Genet. 2020;137(1):4–13. https://doi.org/10.1111/jbg.12443.

    Article  PubMed  Google Scholar 

  43. Chen W, Liu F, Ling Z, Tong X, Xiang C, Moschetta A. Human intestinal lumen and mucosa-associated microbiota in patients with colorectal Cancer. PLoS One. 2012;7(6):e39743. https://doi.org/10.1371/journal.pone.0039743.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Kaakoush NO. Insights into the role of Erysipelotrichaceae in the human host. Front cell infect Mi. 2015;5:84.

    Google Scholar 

  45. Li Y, Guo B, Wu Z, Wang W, Li C, Cai H, et al. Effects of fermented soybean meal supplementation on the growth performance and Cecal microbiota Community of Broiler Chickens. Animals. 2020;10(6):1098. https://doi.org/10.3390/ani10061098.

    Article  PubMed Central  Google Scholar 

  46. Million M, Diallo A, Raoult D. Gut microbiota and malnutrition. Microb Pathog. 2016;106:127–38.

    Article  Google Scholar 

  47. Pan X, Xue F, Nan X, Tang Z, Wang K, Beckers Y, et al. Illumina sequencing approach to characterize thiamine metabolism related Bacteria and the impacts of thiamine supplementation on ruminal microbiota in dairy cows fed high-grain diets. Front Microbiol. 2017;8:1818. https://doi.org/10.3389/fmicb.2017.01818.

    Article  PubMed  PubMed Central  Google Scholar 

  48. Gonciarz W, Lechowicz Ł, Urbaniak M, Kaca W, Chmiela M. Use of Fourier-transform infrared spectroscopy (FT-IR) for monitoring experimental Helicobacter pylori infection and related inflammatory response in Guinea pig model. Int J Mol Sci. 2021;22(1):281.

    Article  CAS  Google Scholar 

  49. Qiu JJ, Liu Z, Zhao P, Wang XJ, Li YC, Sui H, et al. Gut microbial diversity analysis using Illumina sequencing for functional dyspepsia with liver depression-spleen deficiency syndrome and the interventional Xiaoyaosan in a rat model. World J Gastroenterol. 2017;23(5):810–6. https://doi.org/10.3748/wjg.v23.i5.810.

    Article  PubMed  PubMed Central  Google Scholar 

  50. Umu ÖC, Frank JA, Fangel JU, Oostindjer M, da Silva CS, Bolhuis EJ, et al. Resistant starch diet induces change in the swine microbiome and a predominance of beneficial bacterial populations. Microbiome. 2015;3(1):1–15.

    Article  Google Scholar 

  51. Karasova D, Crhanova M, Babak V, Jerabek M, Brzobohaty L, Matesova Z, et al. Development of piglet gut microbiota at the time of weaning influences development of postweaning diarrhea−a field study. Res Vet Sci. 2021;135:59–65. https://doi.org/10.1016/j.rvsc.2020.12.022.

    Article  CAS  PubMed  Google Scholar 

  52. Hu J, Lin S, Zheng B, Cheung PCK. Short-chain fatty acids in control of energy metabolism. Crit Rev Food Sci Nutr. 2018;58(8):1243–9. https://doi.org/10.1080/10408398.2016.1245650.

    Article  CAS  PubMed  Google Scholar 

  53. Dalile B, Oudenhove LV, Vervliet B, Verbeke K. The role of short-chain fatty acids in microbiota–gut–brain communication. Nat Rev Gastroenterol Hepatol. 2019;16(8):461–78. https://doi.org/10.1038/s41575-019-0157-3.

    Article  PubMed  Google Scholar 

  54. Zhao JB, Liu P, Huang CF, Liu L, Li EK, Zhang G, et al. Effect of wheat bran on apparent total tract digestibility, growth performance, fecal microbiota and their metabolites in growing pigs. Anim Feed Sci Technol. 2018;239:14–26. https://doi.org/10.1016/j.anifeedsci.2018.02.013.

    Article  CAS  Google Scholar 

  55. Chen J, Kang B, Jiang Q, Han M, Zhao Y, Long L, et al. Alpha-ketoglutarate in low-protein diets for growing pigs: effects on cecal microbial communities and parameters of microbial metabolism. Front Microbiol. 2018;9:1057. https://doi.org/10.3389/fmicb.2018.01057.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  56. Salazar N, Dewulf EM, Neyrinck AM, Bindels LB, Cani PD, Mahillon J, et al. Inulin-type fructans modulate intestinal Bifidobacterium species populations and decrease fecal short-chain fatty acids in obese women. Clin Nutr. 2015;34(3):501–7. https://doi.org/10.1016/j.clnu.2014.06.001.

    Article  CAS  PubMed  Google Scholar 

  57. Alessandri G, Ossiprandi MC, Macsharry J, Sinderen DV, Ventura M. Bifidobacterial dialogue with its human host and consequent modulation of the immune system. Front Immunol. 2019;10:2348. https://doi.org/10.3389/fimmu.2019.02348.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We would like to thank the staff at our laboratory for their ongoing assistance.

Funding

This work was supported by overseas Expertise Introduction Project for Discipline Innovation (111 Project) and Sichuan Agricultural University Shuangzhi plan for discipline construction project.

Author information

Author notes

  1. Yingjie Li and Yang Liu contributed equally to this work.

Authors and Affiliations

  1. Key Laboratory for Animal Disease-Resistance Nutrition of China Ministry of Education, Institute of Animal Nutrition, Sichuan Agricultural University, No. 211, Huimin Road, Wenjiang District, Chengdu, 611130, Sichuan, People’s Republic of China

    Yingjie Li, Yang Liu, Jiangnan Wu, Qiuhong Chen, Qiang Zhou, Fali Wu, Ruinan Zhang, Zhengfeng Fang, Yan Lin, Shengyu Xu, Bin Feng, Yong Zhuo, De Wu & Lianqiang Che

Authors
  1. Yingjie Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yang Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jiangnan Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Qiuhong Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Qiang Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Fali Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Ruinan Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Zhengfeng Fang
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Yan Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Shengyu Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Bin Feng
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Yong Zhuo
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. De Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Lianqiang Che
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

In this work, Lianqiang Che and Yingjie Li designed the study. Yingjie Li and Jiangnan Wu carried out the animal and laboratory experiments. Yang Liu, Qiuhong Chen, Qiang Zhou, Fali Wu, Ruinan Zhang, Zhengfeng Fang, Yan Lin, Shengyu Xu, Bin Feng, Yong Zhuo, De Wu, and Lianqiang Che analyzed the data. Yingjie Li and Yang Liu wrote the manuscript and Lianqiang Che helped to revise the manuscript. The authors read and approved the final manuscript.

Corresponding author

Correspondence to Lianqiang Che.

Ethics declarations

Ethics approval and consent to participate

The experiment followed the animal protection law (Ethic Approval Code: SCAUAC201308–2) and was performed in accordance with the Guide for the Animal Care and Use approved by Sichuan Agricultural University Institutional Animal Care and Use Committee.

Consent for publication

Not applicable.

Competing interests

The authors declare that they have no conflicts of interest.

Rights and permissions

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Li, Y., Liu, Y., Wu, J. et al. Comparative effects of enzymatic soybean, fish meal and milk powder in diets on growth performance, immunological parameters, SCFAs production and gut microbiome of weaned piglets. J Animal Sci Biotechnol 12, 106 (2021). https://doi.org/10.1186/s40104-021-00625-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1186/s40104-021-00625-8

Keywords

Journal of Animal Science and Biotechnology

ISSN: 2049-1891