Get 20M+ Full-Text Papers For Less Than $1.50/day. Start a 14-Day Trial for You or Your Team.

Learn More →

Dietary sophorolipid accelerates growth by modulation of gut microbiota population and intestinal environments in broiler chickens

Dietary sophorolipid accelerates growth by modulation of gut microbiota population and intestinal... Background: Gut is a crucial organ for the host’s defense system due to its filtering action of the intestinal membrane from hazardous foreign substances. One strategy to strengthen the gut epithelial barrier function is to upregulate beneficial microflora populations and their metabolites. Sophorolipid (SPL), which is a glycolipid bio- surfactant, could increase beneficial microflora and decrease pathogenic bacteria in the gastrointestinal tract. Therefore, herein, we conducted an experiment with broiler chickens to investigate the fortifying effects of SPL on the host’s gut defense system by modulating the microbiota population. Methods: A total of 540 1-day-old chicks (Ross 308) were used, and they were immediately allotted into three treatment groups (6 replications with 30 chicks/pen) according to their initial body weight. The dietary treatments consisted of CON (basal diet), BAM (10 mg/kg bambermycin), and SPL (10 mg/kg SPL). During the experiment, birds freely accessed feed and water, and body weight and feed intake were measured at the end of each phase. On d 35, birds (one bird/pen) were sacrificed to collect jejunum and cecum samples. Results: Dietary SPL and BAM supplementation significantly accelerated birds’ growth and also significantly improved feed efficiency compared to CON. Intestinal microbial community was significantly separated by dietary SPL supplementation from that of CON, and dietary SPL supplementation significantly increased Lactobacillus spp. and Akkermansia muciniphila. Moreover, birds fed with dietary SPL also showed the highest concentration of cecal butyrate among all treatment groups. Gut morphological analysis showed that dietary SPL significantly increased villus height, ratio of villus height to crypt depth, goblet cell numbers, and the gene expression levels of claudin-1 and mucin 2. Additionally, dietary SPL significantly decreased the mRNA expression level of pro-inflammatory cytokine, interleukin-6, and increased that of anti-inflammatory cytokine, interleukin-10, compared to other treatments. * Correspondence: kwhang@korea.ac.kr Min-Jin Kwak and Min-Young Park contributed equally to this work. Department of Biotechnology, Korea University, 145 Anam-ro, Seoul 02841, Republic of Korea Full list of author information is available at the end of the article © The Author(s). 2021 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. Kwak et al. Journal of Animal Science and Biotechnology (2021) 12:81 Page 2 of 9 Conclusions: Dietary SPL increases the beneficial bacterial population and butyrate concentration, which leads to a strengthened gut barrier function. In addition, the intestinal inflammation was also downregulated by dietary SPL supplementation. Keywords: Broilers,Gutmicrobiota,Gutmorphology,Local inflammation, Mucus barrier, Sophorolipids Introduction Materials and methods In the livestock industry, antibiotics have been widely We conducted all of the studies on the birds in accord- used as growth promoters with sub-therapeutic dosage ance with the guidelines and regulations of the Animal due to their outstanding efficacy in feed conversion and Ethics Committee approved by Korea University (Seoul, animal growth [1]. Nonetheless, the use of antibiotic Republic of Korea), and it was carried out at a research growth promoters has been banned because the live- farm in Cheonan, Republic of Korea (Approval number: stock fed with antibiotic growth promoter could serve as KUIACUC-2020-0097). a reservoir of antibiotic-resistant bacteria [2]. Hence, it could threaten human life by transmitting antibiotic- Birds and diets resistant bacteria to humans by direct animal contact or A total of 540 1-day-old male chicks (Ross 308) were indirect environmental contact [3]. used, and they were allotted into three experimental Consequently, the development of novel and eco- treatment groups according to their body weight (BW; friendly materials (e.g., probiotics, prebiotics, organic initial BW: 40.1 g). Each treatment had six replicates acids, essential oils, and enzymes) to replace antibiotic with 30 chicks per pen. The dietary treatments consisted growth promoters is needed [4]. Additionally, various of CON (basal diet), BAM (10 mg/kg of bambermycin- bio-surfactants have been investigated because of their supplemented diet), and SPL (10 mg/kg SPL supple- antibacterial property [5]. Among bio-surfactants, mented diet). The feed composition is shown in Table 1, sophorolipid (SPL) has received much attention in vari- and the feed and SPL were supported by EASY BIO Inc. ous industrial fields, such as medical, hygiene, and (Seoul, Republic of Korea). Birds freely accessed feed pharmaco-dermatological areas, due to their relatively and water during the experiment, and their BW and feed less toxicity and more biodegradability [6]. intake were measured at the end of each phase (Phase 1: SPL is a glycolipid-type amphiphilic compound similar d0–10; phase 2: d 11–20; phase 3: d 21–35) after 8 h of to bambermycin, and it is produced by non-pathogenic feed deprivation to calculate average daily gain (ADG), yeast species, including Candida bombicola [7]. It con- average daily feed intake (ADFI), and feed efficiency sists of a nonpolar fatty acid tail of 16 or 18 carbon (FE). Birds were raised in a controlled experimental atoms and a polar dimetric carbohydrate head, which is room with a rice hull with an average relative humidity linked by a glycosidic bond and binding of non-polar of 60%. Temperature was maintained for 3 d at 30 °C fatty acid and polar carbohydrate determines structure and daily reduced by 0.5 °C to 24 °C, and lightening was of SPL, acidic and lactonic forms [8]. Both forms of SPL provided by artificial light for 24 h/d. are able to be produced by Candida bombicola, and they exert diverse biological properties according to their Sample processing forms [9]. In acidic form, SPL shows high solubility with At the end of the experiment, 18 birds (one bird per foam-forming ability by its higher esterification capacity, pen, randomly selected) were sacrificed, and the weight however, lactonic form of SPL exhibits better surface and length of the small intestine of the birds were tension reducing activity and biological activity [10]. measured. Jejunum and cecum samples were collected, Collectively, SPL exerts diverse biological properties, immediately frozen, and stored at − 80 °C until further including antibacterial activity, immunomodulation cap- analysis. Parts of jejunum samples were fixed in 4% acity, and stimulation of dermal fibroblasts and collagen formalin solution for histological analysis. production, [11, 12]. These properties imply that SPL has a potential to be applied in the animal feed industry Next generation sequencing for improving animal health and growth however, there Quick-DNA™ Fecal/Soil Microbe Microprep Kit (Zymo are few reports on the application of this compound in Research, CA, USA) was used to extract total genomic this field. Hence, we conducted an experiment to evalu- DNA from the cecal samples of chicks according to the ate the efficacy of SPL on gut microbial population and manufacturer’s protocol. For the microbial community their metabolites could lead to improvement of growth structure analysis, V3-V4 regions of 16S rDNA were performance in in broiler chickens. amplified using the following universal primer set Kwak et al. Journal of Animal Science and Biotechnology (2021) 12:81 Page 3 of 9 Table 1 Ingredients and nutritional values of basal diets second PCR was performed to attach the Illumina univer- sal p5/p7 overhang sequence and sample-specific bar- Ingredients, % Starter Grower Finisher codes. Second PCR conditions were as follows; 1-min Corn 60.92 56.29 62.07 initial denaturation at 98 °C, followed by 10 amplification Soybean meal 26.53 18.25 10.63 cycles (30 s at 98 °C, 30 s at 60 °C, 1 min at 72 °C), final ex- Fermented soybean meal 5.00 0.00 0.00 tension at 72 °C 3 min. The sequencing-ready libraries DDGS 0.00 5.00 5.00 were purified using magnetic beads (TopQ XSEP Mag- Unpolished rice 0.00 4.00 3.00 Bead, CELLMICS, USA). Size distribution of the sequen- Rice bran polish 0.00 1.00 1.50 cing ready libraries were determined using the QSep fragment analyzer (Qsep Inc., Taiwan). DNA quantifica- Rapeseed mineral 0.00 4.00 3.00 tion is performed using the Quit dsDNA HS assay kit Sesameseed meal 0.00 0.00 0.50 (Thermo Scientific, USA). Finally, all the libraries were Poultry meal 2.50 5.50 8.00 pooled in equimolar quantities and diluted as necessary. Animal fat 0.00 1.72 2.00 Sequencing was performed on the Illumina MiSeq plat- L-Lysine sulfate (55%) 0.62 0.72 0.75 form (Illumina, USA) using MiSeq Reagent Kit V3 (2 × L-Methionine (90%) 0.47 0.35 0.31 300 PE) (Illumina, USA). Mock DNA libraries were pre- pared with the ZymoBiomics microbial community DNA Threonine (98%) 0.22 0.19 0.19 standard (Zymo Research, USA) using the amplicon li- L-Tryptophan (99%) 0.01 0.03 0.03 brary preparation procedure described above and se- Choline chloride (50%) 0.10 0.11 0.14 quenced together. This control library allowed us to MCP 1.54 1.08 0.81 evaluate potential biases and errors associated with the Limestone 1.45 1.23 1.55 amplification and sequencing steps. Taxonomy profiling Salt 0.25 0.25 0.25 was performed using the open software program Quanti- tative Insights Into Microbial Ecology (QIIME) version NaHCO 0.05 0.05 0.05 a 1.9.0 with NCBI 16S DB and BLASTN v2.3.0 [13]. We also Vitamin premix 0.20 0.14 0.11 performed a closed-reference operational taxonomic unit Mineral premix 0.15 0.12 0.12 picking process to assign taxonomy using NCBI 16S DB Total 100.00 100.00 100.00 and BLASTN (v2.3.0). Calculated value ME, kcal/kg 2851.00 2945.00 3040.00 Gas chromatography–mass spectrometry The concentration of short-chain fatty acids (SCFA) in CP, % 21.85 20.40 19.00 the cecum contents was determined by gas chromatog- Ca, % 1.00 0.90 1.04 raphy–mass spectrometry (GC–MS). Briefly, 10 mg of P, % 0.77 0.70 0.64 cecal contents were homogenized with extraction solution Lysine, % 1.49 1.32 1.19 consisting of 100 μL of internal standard (100 μmol/L Methionine, % 0.75 0.62 0.56 crotonic acid), 100 μL hydrochloric acid, and 200 μLof Threonine, % 1.02 0.94 0.89 ether. After vigorous vortexing for 10 min, the homoge- nates were centrifuged at 1,000×g for 10 min, and 80 μLof Tryptophan, % 0.25 0.23 0.20 a supernatants were transferred into new glass vials. Ali- Provided per kilogram of complete diet: vitamin A, 6300 IU; vitamin D, 2,800 IU; vitamin E, 35 mg; vitamin K , 1.75 mg; vitamin B , 2 mg; vitamin B , 6 mg; quots were mixed with 16 μLof N-tert-butyldimethylsilyl- 3 1 2 vitamin B , 3 mg; vitamin B ,13 μg; biotin, 0.1 mg; calcium pantothenic acid, 6 12 N-methyltrifluoroacetamide (MTBSTFA) and sealed 15 mg; folic acid, 1.5 mg; niacin, 50 mg tightly. The glass vials were heated at 80 °C for 20 min in a Provided per kilogram of complete diet: Mn, 100 mg; Cu, 17 mg; Zn, 92 mg; Fe, 50 mg; I, 1.5 mg; Co, 0.15 mg; Se, 0.3 mg water bath, and then left at room temperature for 48 h for derivatization. The derivatized samples were run through a 6890 N Network GC System with an HP-5MS column (forward 5′-CCTACGGGNGGCWGCAG-3′,reverse 5′- and 5973 N network mass selective detector. Pure helium GACTACHVGGGTATCTAATCC-3′), synthesized by was used as carrier gas and delivered at a 1.2 mL/min flow Integrated DNA Technologies (IDT, Singapore). PCR con- rate. The head pressure was set at 97 kPa with a 20:1 split. ditions for amplifying the V3-V4 regions of 16S rDNA The inlet temperature was 250 °C, and the transfer line were as follows; 5-min initial denaturation at 98 °C, temperature was 260 °C. The temperature program was as followed by 20 amplification cycles (30 s at 98 °C, 30 s at follows: 60 °C for 3 min, 60–120 °C (5 °C per min), and 120– 56 °C, 1 min 30 s at 72 °C), 5 min final extension at 72 °C. 300°C (20°C per min). The run time was 30min, and SCFA The resulting DNA amplicons were purified using mag- concentrations were quantified by comparing their peak netic beads (TopQ XSEP MagBead, CELLMICS, USA). A areas with standards. Kwak et al. Journal of Animal Science and Biotechnology (2021) 12:81 Page 4 of 9 Gut histological assay Results Jejunum samples, fixed in formalin, were embedded into Growth performance paraffin blocks to prepare 5-μm cross sections using a As indicated in Table 3, birds in the BAM and SPL Rotary Microtome CUT 5062 (SLEE MAINZ, Mainz, groups showed significantly higher BW and ADG Germany). These sections were stained with hematoxylin (P < 0.05) compared to those in the CON group. There and eosin and Alican blue staining methods. Total 10 were no significant differences in average feed intake be- villi and 10 crypts were randomly selected per experi- tween treatments, however, feed efficiency was signifi- mental unit. A single observer measured villus height cantly higher (P < 0.05) in the BAM group compared to (VH) and crypt depth (CD) and counted the goblet cells CON group. numbers. Intestinal microbial population qRT-PCR analysis Dietary BAM and SPL supplementation significantly in- Total RNA from jejunum samples was extracted using creased ACE and Chao 1 indexes compared to CON Trizol® (Invitrogen, Grand Island, NY, USA) according group (P < 0.05), however, Shannon and Simpson in- to the manufacturer’s procedure, and the concentration dexes were not different among treatments (Fig. 1a–d). and purity of RNA were determined using Nanodrop Cecal microbial communities of BAM and SPL groups spectrophotometer (Thermo Scientific, Wilmington, DE, were partially and completely separated from that of USA). Subsequently, cDNA samples were synthesized CON group (Fig. 1e). with the High-Capacity cDNA Reverse Transcription kit At phylum level, dietary SPL supplementation in- (Applied Biosystems, Carlsbad, CA, USA) according to creased Firmicutes population and decreased Bacteroi- the manufacturer’s instructions. Target gene expression detes population (Fig. 2a). Additionally, dietary SPL levels were determined using RealHelix™ Premier qPCR supplementation significantly increased (P < 0.05) the kit (NanoHelix, Daejun, Korea) with a StepOnePlus genus level of Lactobacillus and decreased (P < 0.05) that Real-Time PCR System (Applied Biosystems, Carlsbad, of Streptococcs (Fig. 2b). At the species level, dietary SPL CA, USA). Primers for the target genes are listed in supplementation significantly increased (P < 0.05) the Table 2, and the expression level of glyceraldehyde-3- populations of Lactobacillus helveticus and Lactobacillus phosphate dehydrogenase (GAPDH) is used as a house- -ΔΔCT salivarius in the SPL group compared to their popula- keeping gene. The 2 method was used to quantify tions in the CON group (Fig. 2c), and the SPL group relative mRNA expression levels. also showed a significantly higher (P < 0.05) level of Akkermansia muciniphila compared to the other Statistical analysis treatment groups (Fig. 2d). The population of the All data were analyzed using analysis of variance Streptococcus gallolyticus group was significantly de- (ANOVA) with Statistical System 9.4 (SAS Institute, creased (P < 0.05) by dietary BAM and SPL supplemen- Cary, NC, USA). Significant differences between treat- tation (Fig. 2e). ments were determined using Duncan’s multiple-range tests and were defined at the P < 0.05 level. Table 2 Oligonucleotide primers used in jejunal qRT-PCR analysis Table 3 Effects of sophorolipid on growth performance of Gene name Sequence (forward, reverse) Reference 1,2,3 broilers GAPDH 5′-GAGGGTAGTGAAGGCTGCTG-3’ [14] Treatment CON BAM SPL SEM P-value 5′-CCACAACACGGTTGCTGTAT-3’ Initial BW, g 40.16 40.12 40.19 0.045 0.853 CLDN1 5′-TGGAGGATGACCAGGTGAAGA-3’ [15] b a a Final BW, g 1911.8 2011.6 2010.3 18.587 0.035 5′-CGAGCCACTCTGTTGCCATA-3’ b a a ADG, g/d 53.48 56.33 56.29 0.531 0.035 MUC2 5′-TTCATGATGCCTGCTCTTGTG-3’ [15] ADFI, g/d 88.62 87.89 90.45 0.719 0.344 5′-CCTGAGCCTTGGTACATTCTTGT-3’ b a ab FE 0.60 0.64 0.62 0.014 0.010 IL-6 5′-GACGAGGAGAAATGCCTGACG-3’ [16] Treatments: CON, control group fed with basal diet; BAM, group fed with 10 mg/kg of bambermycin supplemented diet; SPL, group fed with 10 mg/kg of 5′-CCGAGTCTGGGATGACCACTTC-3’ sophorolipid supplemented diet IL-10 5′-TCTACACAGATGAGGTCCTGCC-3’ [16] Abbreviations: ADFI average daily feed intake, ADG average daily gain, BW body weight, FE feed efficiency, SEM standard error of means 5′-AGGTGAAGAAGCGGTGACAG-3’ A pen is an experimental unit; 6 replications (pens) per treatment; 30 chicks Abbreviations: CLDN1 claudin-1, GAPDH glyceraldehyde-3-phosphate per pen a, b dehydrogenase, IL-6 interleukin-6, IL-10 interleukin-10, MUC2 mucin 2, OCLD Mean values within a row have different superscript letters were occludin, ZO-1 zonula occludens-1 significantly different (P< 0.05) Kwak et al. Journal of Animal Science and Biotechnology (2021) 12:81 Page 5 of 9 Fig. 1 Cecal microbial community of broiler chickens fed experimental diets. a-b Dietary effects of bambermycin and sophorolipid on species richness indexes (ACE and Chao1); c-d Dietary effects of bambermycin and sophorolipid on diversity indexes (Shannon and Simpson); e Principal component Analysis ordination plots of microbial communities in the CON, BAM, and SPL groups based on the Jensen-Shannon distance metric. *P < 0.05 compared with CON group. Treatment groups: CON, control group fed with basal diet; BAM, group fed with 10 mg/kg of bambermycin supplemented diet; SPL, group fed with 10 mg/kg of sophorolipid-supplemented diet Cecal short-chain fatty acid concentration Intestinal characteristics and histological analysis As listed in Table 4, dietary SPL supplementation signifi- As shown in Table 5, birds fed with the SPL- cantly increased (P < 0.05) the total concentration of supplemented diet (SPL group) showed significantly re- SCFA, and those of acetate and butyrate in the SPL duced (P < 0.05) intestinal weight compared to those of group compared to that in the BAM group. In addition, the other treatment groups; however, gut weight per the ratio of propionate was significantly lowered length was not changed by SPL supplementation. (P < 0.05) in SPL group compared to BAM group. Hence, dietary SPL supplementation significantly Fig. 2 Gut microbiota population of broilers fed with experimental diets. a Intestinal microflora at phylum; b Intestinal microflora at genus level; b–d Specific bacterial populations at the species level in the cecum of birds (Lactobacillus family, Akkermansia muciniphila, and Streptococcus a, b gallolyticus group). *P < 0.05 compared with CON group. Mean values within a row have different superscript letters were significantly different (P < 0.05). Treatment groups: CON, control group fed with basal diet; BAM, group fed with 10 mg/kg of bambermycin supplemented diet; SPL, group fed with 10 mg/kg of sophorolipid-supplemented diet Kwak et al. Journal of Animal Science and Biotechnology (2021) 12:81 Page 6 of 9 IL-6 IL-10 CLDN1 MUC2 a, Fig. 3 mRNA expression levels of genes related to inflammation, tight junction, and mucin in the jejunum of birds fed with experimental diets. Mean values within a row have different superscript letters were significantly different (P < 0.05). Treatment groups: CON, control group fed with basal diet; BAM, group fed with 10 mg/kg of bambermycin supplemented diet; SPL, group fed with 10 mg/kg of sophorolipid-supplemented diet. Abbreviations: IL-6, interleukin-6; IL-10, interleukin-10; CLDN1, claudin-1; MUC2, mucin 2 increased (P < 0.05) VH compared to that of birds in also significantly increased (P < 0.05) in the SPL treat- the other treatment groups without affecting CD ment group compared to that in the other groups. (Table 5). The ratio of VH to CD in the birds fed with the SPL-supplemented diet was the highest (P < Gene expression levels related to inflammation, tight 0.05) compared to that of birds in CON group. More- junction, and mucin secretion over, the goblet cell numbers per 1 μm of villus were As presented in Fig. 3, dietary SPL supplementation sig- nificantly downregulated (P < 0.05) the expression level of interleukin-6 (IL-6) and upregulated (P < 0.05) that of interleukin-10 (IL-10) compared to the other treatments. Table 4 Effects of sophorolipid on cecal SCFA concentration of The expression level of claudin-1 (CLDN1) significantly 1,2,3 broilers increased (P < 0.05) in the SPL group compared to the Treatment CON BAM SPL SEM P-value other groups. Moreover, the BAM and SPL groups Absolute concentration of SCFA, μmol/g showed significantly increased (P < 0.05) expression level a b a of mucin 2 (MUC2) compared to the CON group. Total SCFA 226.43 179.47 249.61 10.802 0.012 a b a Acetate 185.91 143.14 206.06 9.984 0.017 Discussion Propionate 17.62 15.73 17.18 0.676 0.526 First, the results of this study indicated that SPL could iso-butyrate 3.69 3.84 4.01 0.208 0.850 accelerate the growth of broilers as much as bambermy- ab b a Butyrate 19.20 16.76 22.36 0.978 0.049 cin, which is in agreement with the reports of Boontiam Percentage of SCFA, % et al. [17] which demonstrated that surfactant supple- mentation with a low energy and crude protein diet Acetate 82.11 78.98 82.48 0.852 0.187 ab a b could improve average daily gain and feed efficiency dur- Propionate 7.76 8.91 6.88 0.322 0.021 ing the overall experimental period without affecting iso-butyrate 1.66 2.17 1.60 0.130 0.140 average daily feed intake. Furthermore, both glycolipid- Butyrate 8.47 9.95 9.04 0.615 0.647 type antibiotics and biosurfactant increased the Treatments: CON, control group fed with basal diet; BAM, group fed with 10 beneficial microbiota L. heliveticus and L. salivarius and mg/kg of bambermycin supplemented diet; SPL, group fed with 10 mg/kg of decreased the pathogenic bacteria S. gallolyticus. In ac- sophorolipid supplemented diet Abbreviations: SCFA short-chain fatty acid, SEM standard error of means cordance with our results, Abu Hafsa and Ibrahim [18] A chick is an experimental unit; 6 replications per treatment a, b also demonstrated that similar modulation of gut micro- Mean values within a row have different superscript letters were significantly different (P < 0.05) biota (increased Lactobacillus and decreased Kwak et al. Journal of Animal Science and Biotechnology (2021) 12:81 Page 7 of 9 1,2,3 Table 5 Effects of sophorolipid on gut characteristic and histological analysis of broilers Treatment CON BAM SPL SEM P-value a ab b Intestinal, g/100 g body weight 3.18 3.08 2.96 0.037 0.038 Intestinal weight/length, g/m 30.60 31.12 30.54 0.319 0.735 b ab a Villus height, μm 371.27 398.17 422.50 7.483 0.010 Crypt depth, μm 110.58 104.40 105.50 1.366 0.142 b a a Villus height/crypt depth 3.37 3.82 4.01 0.099 0.013 b b a Goblet cells/villus height, /μm 0.22 0.21 0.34 0.016 < 0.001 Treatments: CON, control group fed with basal diet; BAM, group fed with 10 mg/kg of bambermycin supplemented diet; SPL, group fed with 10 mg/kg of sophorolipid supplemented diet Abbreviation: SEM standard error of means A chick is an experimental unit; 6 replications per treatment a, b Mean values within a row have different superscript letters were significantly different (P < 0.05) Sterptococcus) by dietary probiotic supplementation sig- The gastrointestinal tract is the main region that plays nificantly improved the growth of birds. Various reports a role in a protective system because it is the first organ related Lacobacillus species have suggested that L. heli- to meet external substances with a large contact area veticus is known as the beneficial microbes in bone and [28]. Hence, the gut defense strategy employs various mental health and L. salivarius has a positive relation- physiological factors (e.g., mucus barrier, tight junctions, ship with growth of birds [19–21]. Collectively, micro- and immune cytokines) to maintain the intestinal flora communities modulated by bambermycin and SPL homeostatic balance [29]. Our results suggest that were similar, however SPL have seemed to accelerate the dietary SPL supplementation could enhance mucin- shift of the community compared to that of bambermy- presenting capacity and villus turnover balance by in- cin. Also, these results suggested that this microbial shift creasing beneficial bacterial populations. At phylum has a potential to improve the growth and health of level, there were higher portion of Firmicutes, and lower birds by modulation of their metabolites. that of Bacteroidetes in SPL treatment group compared On the other hand, SCFA are the main metabolites of to the other groups. Bacteroides, known as the LPS- microbial fermentation, and they have been widely stud- generating bacteria group when they are lysed, may play ied to elucidate the specific mechanism of antibiotic a critical role to destruct intestinal integrity and morph- growth promoter linking the host and its intestinal ology by lowering intestinal permeability [30], resulted microbiota [22]. In 2019, Guinan et al. demonstrated in the lower Firmicutes/Bacteroidetes ratio is considered that water supply with antibiotic could downregulated as the favorable index of weight loss [31]. At species the SCFA concentration in cecal contents by increased level, Akkermansia muciniphila, has received attention colonization of Candida albicans [23]. In agreement because of its specific biological properties including im- with this study, the results of our study demonstrated mune modulation, wound healing, and SCFA production that dietary bambermycin supplementation significantly [32]. Moreover, various studies have demonstrated that reduced the concentration of total SCFA compared to A. muciniphila has the potential to act as a probiotic be- birds fed control diet and SPL supplemented diet. cause it could exert a glucose-lowering effect by regulat- Additionally, both acetate and butyrate were increased ing gut barrier integrity through increased expression in the SPL group than BAM group, which might be levels of tight junction proteins [33, 34]. Similar to the due to the increased populations of butyrate- results of these studies, we also found an increased producing bacteria, L. helveticus and mucin-degrading population of A. muciniphila and strengthened gut epi- bacteria, A. muciniphila [24, 25]. An in vitro study thelial integrity and mucus secretion capacity. with L. helveticus showed that this probiotics strain Additionally, our results demonstrated that SPL could significantly increased butyrate concentration, and it relieve the local gut immune response and strengthen maybedue to theupregulated conversion ratioof the intestinal epithelial barrier by modulating the gut lactate to butyrate by lactating-utilizing bacteria [26]. microbiota population. S. gallolyticus, a gram-positive And A. muciniphila supplemented with mucin could pathogenic bacterium, which is commonly found in vari- produce acetate and ethanol from mucin fermentation ous animals and humans, and it is a potential transmis- [27]. Collectively, these results suggest that dietary sion bacterium with antibiotic resistance [35, 36]. In SPL treatment maintain the cecal SCFA concentration addition, Li et al. [37] demonstrated that inoculation of by intestinal colonization of SCFA-producing bacteria S. gallolyticus in duckling induced macrophage necrop- unlike bambermycin supplementation. tosis in spleens by bacterial infection and increased the Kwak et al. Journal of Animal Science and Biotechnology (2021) 12:81 Page 8 of 9 expression levels of IL-6, and that decreased population Consent for publication Not applicable. of S. gallolyticus achieved in response to dietary oregano powder has a negative relationship with the increase of Competing interests anti-inflammatory cytokine IL-10 [38]. In accordance The authors declare that they have no conflict of interest. with the results of these previous studies, decreased Author details population of pathogenic bacteria (S. gallolyticus)by Department of Biotechnology, Korea University, 145 Anam-ro, Seoul 02841, sophorolipid supplementation has the potential to allevi- 2 Republic of Korea. Pathway Intermediates, Seoul 02841, Republic of Korea. ate immune responses by improving pro- (IL-6) and Received: 12 January 2021 Accepted: 13 May 2021 anti-inflammatory (IL-10) cytokine production. On the other hands, our results also found that the higher popu- lation of Lactobacillus genus in BAM and SPL groups References compared to CON group. Lactobacillus is the genera of 1. Danzeisen JL, Kim HB, Isaacson RE, Tu ZJ, Johnson TJ. Modulations of the chicken cecal microbiome and metagenome in response to anticoccidial gram positive and facultative anaerobes and it can in- and growth promoter treatment. PLoS One. 2011;6(11):e27949. https://doi. hibit the pathogen growth by establishment of low pH org/10.1371/journal.pone.0027949. environment through lactic acid producing capacity [39]. 2. Bates J, Jordens JZ, Griffiths DT. Farm animals as a putative reservoir for vancomycin-resistant enterococcal infection in man. J Antibicrob Therefore, our results demonstrated that microbial shift Chemother. 1994;34(4):507–14. https://doi.org/10.1093/jac/34.4.507. by SPL might be able to modulate immune response in 3. Munk P, Knudsen BE, Lukjacenko O, Duarte ASR, Luiken RE, Van Gompel L, broilers’ intestine. et al. Abundance and diversity of the fecal resistome in slaughter pigs and broilers in nine European countries. Nature. 2017;3:898–908. 4. Wang JP, Lee JH, Yoo JS, Cho JH, Kim HJ, Kim IH. Effects of phenyllactic acid Conclusions on growth performance, intestinal microbiota, relative organ weight, blood characteristics, and meat quality of broiler chicks. Poult Sci. 2010;89(7):1549– Dietary sophorolipid supplementation in broiler feed 55. https://doi.org/10.3382/ps.2009-00235. could modulate the intestinal microbiota population and 5. Roy A, Haldar S, Mondal S, Ghosh TP. Effects of supplemental exogenous short-chain fatty acid levels. Hence, the promoted gut emulsifier on performance, nutrient metabolism, and serum lipid profile in broiler chickens. Vet Med Int. 2010;11:262604. environment could improve gut defense integrity, in- 6. Chen C, Jung B, Kim WK. Effects of lysophospholipid on growth cluding intensified mucus layer and tight junction, and performance, carcass yield, intestinal development, and bone quality in alleviate local inflammation, resulting in the acceleration broilers. Poult Sci. 2019;98(9):3902–23. https://doi.org/10.3382/ps/pez111. 7. Cho KJ, Kim YB, Kim EK. Production and application of sophorolipid, a of chick growth. Further studies are required to elucidate microbial surfactant. Korean Soc Biotechnol Bioeng J. 1999;14:747–53. the precise relationship between sophorolipids and their 8. Develter DW, Lauryssen LM. Properties and industrial applications of effects on intestinal health. sophorolipid. Eur J Lipid Sci Technol. 2010;112(6):628–38. https://doi.org/10.1 002/ejlt.200900153. Abbreviations 9. Borsaniyiova M, Patil A, Mukherji R, Prabhune A, Bopegamage S. Biological ADFI: Average daily feed intake; ADG: Average daily gain; ANOVA: Analysis of activity of sophorolipids and their possible use as antiviral agents. Folia variations; BW: Body weight; CD: Crypt depth; CLDN1: Claudin-1; FE: Feed Microbiol. 2016;61(1):85–9. https://doi.org/10.1007/s12223-015-0413-z. efficiency; GAPDH: Glyceraldehyde 3-phosphate dehydrogenase; GC–MS: Gas 10. Kwak MJ, Park MY, Kim J, Lee H, Whang KY. Curative effects of sophorolipid chromatography–mass spectrometry; IL-6: Interleukin-6; IL-10: Interleukin-10; on physical wounds: in vitro and in vivo studies. Vet Med Sci. 2021;00:1–9. MTBSTFA: N-tert-butyldimethylsilyl-N-methyltrifluoroacetamide ; MUC2: Mucin https://doi.org/10.1002/vms3.481. 2; QIIME: Quantitative Insights Into Microbial Ecology; SPL: Sophorolipid; 11. Hardin R, Pierre J, Schulze R, Mueller CM, Fu SL, Wallner SR, et al. SCFA: Short-chain fatty acid; VH: Villus height Sophorolipids improve sepsis survival: effects of dosing and derivatives. J Surg Res. 2008;142:314–9. 12. Concaix FB. Use of sophorolipids comprising diacetyl lactones as agent for Acknowledgements stimulating skin fibroblast metabolism. U S Patent No 6,596,265. 2003. Not applicable. 13. Yoon SH, Ha SM, Kwon S, Lim J, Kim Y, Seo H, et al. Introducing EzBioCloud: a taxonomically united database of 16S rRNA and whole genome Authors’ contributions assemblies. Int J Syst Evol Microbiol. 2017;67(5):1613–7. https://doi.org/10.1 MJK, MYP, JGK and KYW conceived and designed the experiments; MJK and 099/ijsem.0.001755. MYP mainly performed the experiments; MJK and YSC analyzed the data; JC, 14. Li X, McFarland DC, Velleman SG. Effect of Smad3-mediated transforming DP, HBL and IGC contributed reagents/materials/analysis tools; MJK wrote growth factor-β1 signaling on satellite cell proliferation and differentiation the manuscript. All authors read and approved the final manuscript. in chickens. Poult Sci. 2008;87(9):1823–33. https://doi.org/10.3382/ps.2008- Funding 15. Song B, Li H, Wu Y, Zhen W, Wang Z, Xia Z, et al. Effect of This work was financially supported by EASYBIO Inc. and Korea University. microencapsulated sodium butyrate dietary supplementation on growth performance and intestinal barrier function of broiler chickens infected with Availability of data and materials necrotic enteritis. Anim Feed Sci Technol. 2017;232:6–15. https://doi.org/10.1 Not applicable. 016/j.anifeedsci.2017.07.009. 16. Heidari M, Wang D, Delekta P, Sun S. Marek’s disease virus Declarations immunosuppression alters host cellular reponses and immune gene expression in the skin of infected chickens. Vet Immunol Immunopathol. Ethics approval 2016;180:21–8. https://doi.org/10.1016/j.vetimm.2016.08.013. All of works related to animal was conducted in accordance with the 17. Boontiam W, Hyun YK, Jung B, Kim YY. Effects of lysophospholipid guidelines and regulations for the care and the use of experimental animals supplementation to reduced energy, crude protein, and amino acid diets was approved by the Korea University Institutional Animal Care & Use on growth performance, nutrient digestibility, and blood profiles in broiler Committee (Permission No. KUIACUC-2020-0097). chickens. Poult Sci. 2019;98(12):6693–701. https://doi.org/10.3382/ps/pex005. Kwak et al. Journal of Animal Science and Biotechnology (2021) 12:81 Page 9 of 9 18. Abu Hafsa SH, Ibrahim SA. Effect of dietary polyphenol-rich grape seed on 38. Bauer BW, Gangadoo S, Bajagai YS, Van TTH, Moore RJ, Stanley D. Oregano growth performance, antioxidant capacity and ileal microflora in broiler powder reduces Streptococcus and increases SCFA concentration in a chicks. J Anim Physiol Anim Nutr. 2018;102(1):268–75. https://doi.org/1 mixed bacterial culture assay of chicken. PloS One. 2019;13(3):e0216853. 0.1111/jpn.12688. https://doi.org/10.1371/journal.pone.0216853. 19. Ohland CL, Kish L, Bell H, Thiesen A, Hotte N, Pankiv E, et al. Effects of 39. Rodriguez-Cebezas ME, Camuesco D, Arribas B, Garrido-Mesa N, Comalada Lactobacillus helveticus on murine behavior are dependent on diet and M, Bailon E, et al. The combination of fructooligosaccharides and resistant genotype and correlate with alterations in the gut microbiome. starch shows prebiotic additive effects in rats. Clin Nutr. 2010;29(6):832–9. Psychoneuroendocrinology. 2013;38(9):1738–47. https://doi.org/10.1016/j. https://doi.org/10.1016/j.clnu.2010.05.005. psyneuen.2013.02.008. 20. Liang S, Wang T, Hu X, Luo J, Li W, Wu X, et al. Administration of Lactobacillus helveticus NS8 improves behavioral, cognitive, and biochemical aberrations caused by chronic restraint stress. Neuroscience. 2015;310:561–77. https://doi.org/10.1016/j.neuroscience.2015.09.033. 21. Zhu NH, Zhang RJ, Wu H, Zhang B. Effects of Lactobacillus cultures on growth performance, xanthophyll deposition, and color of the meat and skin of broilers. J Appl Poult Res. 2009;18(3):570–8. https://doi.org/10.3382/ja pr.2009-00012. 22. Le Poul E, Loison C, Struyf S, Springael JY, Lannoy V, Decobecq ME, et al. Functional characterization of human receptors for short chain fatty acids and their role in polymorphonuclear cell activation. J Biol Chem. 2003; 278(28):25481–9. https://doi.org/10.1074/jbc.M301403200. 23. Guinan J, Wang S, Hazbun TR, Yadav H, Thangamani S. Antibiotic-induced decreases in the levels of microbial-derived short-chain fatty acids correlate with increased gastrointestinal colonization of Candida albicans. Sci Rep. 2019;9:1–11. 24. Parada Venegas D, De la Fuente MK, Landskron G, González MJ, Quera R, Dijkstra G, et al. Short chain fatty acid (SCFA)-mediated gut epithelial and immune regulation and its relevance in inflammatory bowel diseases. Front Immunol. 2019;10:277–92. 25. Taverniti V, Guglielmetti S. Health-promoting properties of Lactobacillus helveticus. Front Microbiol. 2012;3:392–404. 26. Vitali B, Ndagijimana M, Maccaferri S, Blagi E, Guerzoni ME, Brigidi P. An in vitro evaluation of the effect of probiotics and prebioitcs on the metabolic profile of human microbiota. Anaerobe. 2012;18(4):386–91. https://doi.org/10.1016/j.anaerobe.2012.04.014. 27. Derrien M, Vaughan EE, Plugge CM, de Vos WM. Akkermansia muciniphila gen. nov., sp. Sp. Nov., a human intestinal mucin-degrading bacterium. Int J Syst Evol Microbiol. 2004;54:1469–76. 28. Artis D. Epithelial-cell recognition of commensal bacteria and maintenance of immune homeostasis in the gut. Nat Rev Immunol. 2008;8(6):411–20. https://doi.org/10.1038/nri2316. 29. Schroeder BO. Fight them or feed them: how the intestinal mucus layer manages the gut microbiota. Gastroenterol Rep. 2019;7(1):3–12. https://doi. org/10.1093/gastro/goy052. 30. Chen K, Zhao H, Shu L, Xing H, Wang C, Lu C, et al. Effect of resveratrol on intestinal tight junction proteins and the gut microbiome in high-fat diet- fed insulin resistant mice. Int J Food Sci Nutr. 2020;71(8):965–78. https://doi. org/10.1080/09637486.2020.1754351. 31. Qu L, Liu Q, Zhang Q, Tuo X, Fan D, Deng J, et al. Kiwifruit seed oil prevents obesity by regulating inflammation, thermogenesis, and gut microbiota in high-fat diet-induced obese C57BL/6 mice. Food Chem Toxicol. 2019;125: 85–94. https://doi.org/10.1016/j.fct.2018.12.046. 32. Earley H, Lennon G, Balfe Á, Coffey JC, Winter DC, O’Connell PR. The abundance of Akkermansia muciniphila and its relationship with sulphated colonic mucins in health and ulcerative colitis. Sci Rep. 2019;9:1–9. 33. Ashrafian F, Behrouzi A. Comparative study of effect of Akkermansia muciniphila and its extracellular vesicles on toll-like receptors and tight junction. Gastroenterol Hepatol Bed Bench. 2019;12(2):163–8. 34. Su H, Mo J, Ni J, Ke H, Bao T, Xie J, et al. Andrographolide Exerts antihyperglycemic effect through strengthening intestinal barrier function and increasing microbial composition of Akkermansia muciniphila. Oxid Med Cell. 2020;6538930:1–20. https://doi.org/10.1155/2020/6538930. 35. Dumke J, Hinse D, Vollmer T, Schulz J, Knabbe C, Dreier J. Potential transmission pathways of Streptococcus gallolyticus subsp gallolyticus. PLoS One. 2015;10:e0126507. 36. Nomoto R, Le HTT, Sekizaki T, Osawa R. Antimicrobial susceptibility of Streptococcus gallolyticus isolated from humans and animals. Jpn J Infect Dis. 2013;66(4):334–6. https://doi.org/10.7883/yoken.66.334. 37. Li M, Liu B, Gu C, Zhang W, Yang J, Cheng G, et al. Necroptosis of splenic macrophages induced by Streptococcus gallolyticus subsp. pasteurianus. Avian Dis. 2017;61(1):115–22. https://doi.org/10.1637/11449-061216-Reg. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of Animal Science and Biotechnology Springer Journals

Dietary sophorolipid accelerates growth by modulation of gut microbiota population and intestinal environments in broiler chickens

Download PDF
9 pages

Loading next page...
 
/lp/springer-journals/dietary-sophorolipid-accelerates-growth-by-modulation-of-gut-r7yO03UQB0

References (43)

Publisher
Springer Journals
Copyright
Copyright © The Author(s) 2021
eISSN
2049-1891
DOI
10.1186/s40104-021-00606-x
Publisher site
See Article on Publisher Site

Abstract

Background: Gut is a crucial organ for the host’s defense system due to its filtering action of the intestinal membrane from hazardous foreign substances. One strategy to strengthen the gut epithelial barrier function is to upregulate beneficial microflora populations and their metabolites. Sophorolipid (SPL), which is a glycolipid bio- surfactant, could increase beneficial microflora and decrease pathogenic bacteria in the gastrointestinal tract. Therefore, herein, we conducted an experiment with broiler chickens to investigate the fortifying effects of SPL on the host’s gut defense system by modulating the microbiota population. Methods: A total of 540 1-day-old chicks (Ross 308) were used, and they were immediately allotted into three treatment groups (6 replications with 30 chicks/pen) according to their initial body weight. The dietary treatments consisted of CON (basal diet), BAM (10 mg/kg bambermycin), and SPL (10 mg/kg SPL). During the experiment, birds freely accessed feed and water, and body weight and feed intake were measured at the end of each phase. On d 35, birds (one bird/pen) were sacrificed to collect jejunum and cecum samples. Results: Dietary SPL and BAM supplementation significantly accelerated birds’ growth and also significantly improved feed efficiency compared to CON. Intestinal microbial community was significantly separated by dietary SPL supplementation from that of CON, and dietary SPL supplementation significantly increased Lactobacillus spp. and Akkermansia muciniphila. Moreover, birds fed with dietary SPL also showed the highest concentration of cecal butyrate among all treatment groups. Gut morphological analysis showed that dietary SPL significantly increased villus height, ratio of villus height to crypt depth, goblet cell numbers, and the gene expression levels of claudin-1 and mucin 2. Additionally, dietary SPL significantly decreased the mRNA expression level of pro-inflammatory cytokine, interleukin-6, and increased that of anti-inflammatory cytokine, interleukin-10, compared to other treatments. * Correspondence: kwhang@korea.ac.kr Min-Jin Kwak and Min-Young Park contributed equally to this work. Department of Biotechnology, Korea University, 145 Anam-ro, Seoul 02841, Republic of Korea Full list of author information is available at the end of the article © The Author(s). 2021 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. Kwak et al. Journal of Animal Science and Biotechnology (2021) 12:81 Page 2 of 9 Conclusions: Dietary SPL increases the beneficial bacterial population and butyrate concentration, which leads to a strengthened gut barrier function. In addition, the intestinal inflammation was also downregulated by dietary SPL supplementation. Keywords: Broilers,Gutmicrobiota,Gutmorphology,Local inflammation, Mucus barrier, Sophorolipids Introduction Materials and methods In the livestock industry, antibiotics have been widely We conducted all of the studies on the birds in accord- used as growth promoters with sub-therapeutic dosage ance with the guidelines and regulations of the Animal due to their outstanding efficacy in feed conversion and Ethics Committee approved by Korea University (Seoul, animal growth [1]. Nonetheless, the use of antibiotic Republic of Korea), and it was carried out at a research growth promoters has been banned because the live- farm in Cheonan, Republic of Korea (Approval number: stock fed with antibiotic growth promoter could serve as KUIACUC-2020-0097). a reservoir of antibiotic-resistant bacteria [2]. Hence, it could threaten human life by transmitting antibiotic- Birds and diets resistant bacteria to humans by direct animal contact or A total of 540 1-day-old male chicks (Ross 308) were indirect environmental contact [3]. used, and they were allotted into three experimental Consequently, the development of novel and eco- treatment groups according to their body weight (BW; friendly materials (e.g., probiotics, prebiotics, organic initial BW: 40.1 g). Each treatment had six replicates acids, essential oils, and enzymes) to replace antibiotic with 30 chicks per pen. The dietary treatments consisted growth promoters is needed [4]. Additionally, various of CON (basal diet), BAM (10 mg/kg of bambermycin- bio-surfactants have been investigated because of their supplemented diet), and SPL (10 mg/kg SPL supple- antibacterial property [5]. Among bio-surfactants, mented diet). The feed composition is shown in Table 1, sophorolipid (SPL) has received much attention in vari- and the feed and SPL were supported by EASY BIO Inc. ous industrial fields, such as medical, hygiene, and (Seoul, Republic of Korea). Birds freely accessed feed pharmaco-dermatological areas, due to their relatively and water during the experiment, and their BW and feed less toxicity and more biodegradability [6]. intake were measured at the end of each phase (Phase 1: SPL is a glycolipid-type amphiphilic compound similar d0–10; phase 2: d 11–20; phase 3: d 21–35) after 8 h of to bambermycin, and it is produced by non-pathogenic feed deprivation to calculate average daily gain (ADG), yeast species, including Candida bombicola [7]. It con- average daily feed intake (ADFI), and feed efficiency sists of a nonpolar fatty acid tail of 16 or 18 carbon (FE). Birds were raised in a controlled experimental atoms and a polar dimetric carbohydrate head, which is room with a rice hull with an average relative humidity linked by a glycosidic bond and binding of non-polar of 60%. Temperature was maintained for 3 d at 30 °C fatty acid and polar carbohydrate determines structure and daily reduced by 0.5 °C to 24 °C, and lightening was of SPL, acidic and lactonic forms [8]. Both forms of SPL provided by artificial light for 24 h/d. are able to be produced by Candida bombicola, and they exert diverse biological properties according to their Sample processing forms [9]. In acidic form, SPL shows high solubility with At the end of the experiment, 18 birds (one bird per foam-forming ability by its higher esterification capacity, pen, randomly selected) were sacrificed, and the weight however, lactonic form of SPL exhibits better surface and length of the small intestine of the birds were tension reducing activity and biological activity [10]. measured. Jejunum and cecum samples were collected, Collectively, SPL exerts diverse biological properties, immediately frozen, and stored at − 80 °C until further including antibacterial activity, immunomodulation cap- analysis. Parts of jejunum samples were fixed in 4% acity, and stimulation of dermal fibroblasts and collagen formalin solution for histological analysis. production, [11, 12]. These properties imply that SPL has a potential to be applied in the animal feed industry Next generation sequencing for improving animal health and growth however, there Quick-DNA™ Fecal/Soil Microbe Microprep Kit (Zymo are few reports on the application of this compound in Research, CA, USA) was used to extract total genomic this field. Hence, we conducted an experiment to evalu- DNA from the cecal samples of chicks according to the ate the efficacy of SPL on gut microbial population and manufacturer’s protocol. For the microbial community their metabolites could lead to improvement of growth structure analysis, V3-V4 regions of 16S rDNA were performance in in broiler chickens. amplified using the following universal primer set Kwak et al. Journal of Animal Science and Biotechnology (2021) 12:81 Page 3 of 9 Table 1 Ingredients and nutritional values of basal diets second PCR was performed to attach the Illumina univer- sal p5/p7 overhang sequence and sample-specific bar- Ingredients, % Starter Grower Finisher codes. Second PCR conditions were as follows; 1-min Corn 60.92 56.29 62.07 initial denaturation at 98 °C, followed by 10 amplification Soybean meal 26.53 18.25 10.63 cycles (30 s at 98 °C, 30 s at 60 °C, 1 min at 72 °C), final ex- Fermented soybean meal 5.00 0.00 0.00 tension at 72 °C 3 min. The sequencing-ready libraries DDGS 0.00 5.00 5.00 were purified using magnetic beads (TopQ XSEP Mag- Unpolished rice 0.00 4.00 3.00 Bead, CELLMICS, USA). Size distribution of the sequen- Rice bran polish 0.00 1.00 1.50 cing ready libraries were determined using the QSep fragment analyzer (Qsep Inc., Taiwan). DNA quantifica- Rapeseed mineral 0.00 4.00 3.00 tion is performed using the Quit dsDNA HS assay kit Sesameseed meal 0.00 0.00 0.50 (Thermo Scientific, USA). Finally, all the libraries were Poultry meal 2.50 5.50 8.00 pooled in equimolar quantities and diluted as necessary. Animal fat 0.00 1.72 2.00 Sequencing was performed on the Illumina MiSeq plat- L-Lysine sulfate (55%) 0.62 0.72 0.75 form (Illumina, USA) using MiSeq Reagent Kit V3 (2 × L-Methionine (90%) 0.47 0.35 0.31 300 PE) (Illumina, USA). Mock DNA libraries were pre- pared with the ZymoBiomics microbial community DNA Threonine (98%) 0.22 0.19 0.19 standard (Zymo Research, USA) using the amplicon li- L-Tryptophan (99%) 0.01 0.03 0.03 brary preparation procedure described above and se- Choline chloride (50%) 0.10 0.11 0.14 quenced together. This control library allowed us to MCP 1.54 1.08 0.81 evaluate potential biases and errors associated with the Limestone 1.45 1.23 1.55 amplification and sequencing steps. Taxonomy profiling Salt 0.25 0.25 0.25 was performed using the open software program Quanti- tative Insights Into Microbial Ecology (QIIME) version NaHCO 0.05 0.05 0.05 a 1.9.0 with NCBI 16S DB and BLASTN v2.3.0 [13]. We also Vitamin premix 0.20 0.14 0.11 performed a closed-reference operational taxonomic unit Mineral premix 0.15 0.12 0.12 picking process to assign taxonomy using NCBI 16S DB Total 100.00 100.00 100.00 and BLASTN (v2.3.0). Calculated value ME, kcal/kg 2851.00 2945.00 3040.00 Gas chromatography–mass spectrometry The concentration of short-chain fatty acids (SCFA) in CP, % 21.85 20.40 19.00 the cecum contents was determined by gas chromatog- Ca, % 1.00 0.90 1.04 raphy–mass spectrometry (GC–MS). Briefly, 10 mg of P, % 0.77 0.70 0.64 cecal contents were homogenized with extraction solution Lysine, % 1.49 1.32 1.19 consisting of 100 μL of internal standard (100 μmol/L Methionine, % 0.75 0.62 0.56 crotonic acid), 100 μL hydrochloric acid, and 200 μLof Threonine, % 1.02 0.94 0.89 ether. After vigorous vortexing for 10 min, the homoge- nates were centrifuged at 1,000×g for 10 min, and 80 μLof Tryptophan, % 0.25 0.23 0.20 a supernatants were transferred into new glass vials. Ali- Provided per kilogram of complete diet: vitamin A, 6300 IU; vitamin D, 2,800 IU; vitamin E, 35 mg; vitamin K , 1.75 mg; vitamin B , 2 mg; vitamin B , 6 mg; quots were mixed with 16 μLof N-tert-butyldimethylsilyl- 3 1 2 vitamin B , 3 mg; vitamin B ,13 μg; biotin, 0.1 mg; calcium pantothenic acid, 6 12 N-methyltrifluoroacetamide (MTBSTFA) and sealed 15 mg; folic acid, 1.5 mg; niacin, 50 mg tightly. The glass vials were heated at 80 °C for 20 min in a Provided per kilogram of complete diet: Mn, 100 mg; Cu, 17 mg; Zn, 92 mg; Fe, 50 mg; I, 1.5 mg; Co, 0.15 mg; Se, 0.3 mg water bath, and then left at room temperature for 48 h for derivatization. The derivatized samples were run through a 6890 N Network GC System with an HP-5MS column (forward 5′-CCTACGGGNGGCWGCAG-3′,reverse 5′- and 5973 N network mass selective detector. Pure helium GACTACHVGGGTATCTAATCC-3′), synthesized by was used as carrier gas and delivered at a 1.2 mL/min flow Integrated DNA Technologies (IDT, Singapore). PCR con- rate. The head pressure was set at 97 kPa with a 20:1 split. ditions for amplifying the V3-V4 regions of 16S rDNA The inlet temperature was 250 °C, and the transfer line were as follows; 5-min initial denaturation at 98 °C, temperature was 260 °C. The temperature program was as followed by 20 amplification cycles (30 s at 98 °C, 30 s at follows: 60 °C for 3 min, 60–120 °C (5 °C per min), and 120– 56 °C, 1 min 30 s at 72 °C), 5 min final extension at 72 °C. 300°C (20°C per min). The run time was 30min, and SCFA The resulting DNA amplicons were purified using mag- concentrations were quantified by comparing their peak netic beads (TopQ XSEP MagBead, CELLMICS, USA). A areas with standards. Kwak et al. Journal of Animal Science and Biotechnology (2021) 12:81 Page 4 of 9 Gut histological assay Results Jejunum samples, fixed in formalin, were embedded into Growth performance paraffin blocks to prepare 5-μm cross sections using a As indicated in Table 3, birds in the BAM and SPL Rotary Microtome CUT 5062 (SLEE MAINZ, Mainz, groups showed significantly higher BW and ADG Germany). These sections were stained with hematoxylin (P < 0.05) compared to those in the CON group. There and eosin and Alican blue staining methods. Total 10 were no significant differences in average feed intake be- villi and 10 crypts were randomly selected per experi- tween treatments, however, feed efficiency was signifi- mental unit. A single observer measured villus height cantly higher (P < 0.05) in the BAM group compared to (VH) and crypt depth (CD) and counted the goblet cells CON group. numbers. Intestinal microbial population qRT-PCR analysis Dietary BAM and SPL supplementation significantly in- Total RNA from jejunum samples was extracted using creased ACE and Chao 1 indexes compared to CON Trizol® (Invitrogen, Grand Island, NY, USA) according group (P < 0.05), however, Shannon and Simpson in- to the manufacturer’s procedure, and the concentration dexes were not different among treatments (Fig. 1a–d). and purity of RNA were determined using Nanodrop Cecal microbial communities of BAM and SPL groups spectrophotometer (Thermo Scientific, Wilmington, DE, were partially and completely separated from that of USA). Subsequently, cDNA samples were synthesized CON group (Fig. 1e). with the High-Capacity cDNA Reverse Transcription kit At phylum level, dietary SPL supplementation in- (Applied Biosystems, Carlsbad, CA, USA) according to creased Firmicutes population and decreased Bacteroi- the manufacturer’s instructions. Target gene expression detes population (Fig. 2a). Additionally, dietary SPL levels were determined using RealHelix™ Premier qPCR supplementation significantly increased (P < 0.05) the kit (NanoHelix, Daejun, Korea) with a StepOnePlus genus level of Lactobacillus and decreased (P < 0.05) that Real-Time PCR System (Applied Biosystems, Carlsbad, of Streptococcs (Fig. 2b). At the species level, dietary SPL CA, USA). Primers for the target genes are listed in supplementation significantly increased (P < 0.05) the Table 2, and the expression level of glyceraldehyde-3- populations of Lactobacillus helveticus and Lactobacillus phosphate dehydrogenase (GAPDH) is used as a house- -ΔΔCT salivarius in the SPL group compared to their popula- keeping gene. The 2 method was used to quantify tions in the CON group (Fig. 2c), and the SPL group relative mRNA expression levels. also showed a significantly higher (P < 0.05) level of Akkermansia muciniphila compared to the other Statistical analysis treatment groups (Fig. 2d). The population of the All data were analyzed using analysis of variance Streptococcus gallolyticus group was significantly de- (ANOVA) with Statistical System 9.4 (SAS Institute, creased (P < 0.05) by dietary BAM and SPL supplemen- Cary, NC, USA). Significant differences between treat- tation (Fig. 2e). ments were determined using Duncan’s multiple-range tests and were defined at the P < 0.05 level. Table 2 Oligonucleotide primers used in jejunal qRT-PCR analysis Table 3 Effects of sophorolipid on growth performance of Gene name Sequence (forward, reverse) Reference 1,2,3 broilers GAPDH 5′-GAGGGTAGTGAAGGCTGCTG-3’ [14] Treatment CON BAM SPL SEM P-value 5′-CCACAACACGGTTGCTGTAT-3’ Initial BW, g 40.16 40.12 40.19 0.045 0.853 CLDN1 5′-TGGAGGATGACCAGGTGAAGA-3’ [15] b a a Final BW, g 1911.8 2011.6 2010.3 18.587 0.035 5′-CGAGCCACTCTGTTGCCATA-3’ b a a ADG, g/d 53.48 56.33 56.29 0.531 0.035 MUC2 5′-TTCATGATGCCTGCTCTTGTG-3’ [15] ADFI, g/d 88.62 87.89 90.45 0.719 0.344 5′-CCTGAGCCTTGGTACATTCTTGT-3’ b a ab FE 0.60 0.64 0.62 0.014 0.010 IL-6 5′-GACGAGGAGAAATGCCTGACG-3’ [16] Treatments: CON, control group fed with basal diet; BAM, group fed with 10 mg/kg of bambermycin supplemented diet; SPL, group fed with 10 mg/kg of 5′-CCGAGTCTGGGATGACCACTTC-3’ sophorolipid supplemented diet IL-10 5′-TCTACACAGATGAGGTCCTGCC-3’ [16] Abbreviations: ADFI average daily feed intake, ADG average daily gain, BW body weight, FE feed efficiency, SEM standard error of means 5′-AGGTGAAGAAGCGGTGACAG-3’ A pen is an experimental unit; 6 replications (pens) per treatment; 30 chicks Abbreviations: CLDN1 claudin-1, GAPDH glyceraldehyde-3-phosphate per pen a, b dehydrogenase, IL-6 interleukin-6, IL-10 interleukin-10, MUC2 mucin 2, OCLD Mean values within a row have different superscript letters were occludin, ZO-1 zonula occludens-1 significantly different (P< 0.05) Kwak et al. Journal of Animal Science and Biotechnology (2021) 12:81 Page 5 of 9 Fig. 1 Cecal microbial community of broiler chickens fed experimental diets. a-b Dietary effects of bambermycin and sophorolipid on species richness indexes (ACE and Chao1); c-d Dietary effects of bambermycin and sophorolipid on diversity indexes (Shannon and Simpson); e Principal component Analysis ordination plots of microbial communities in the CON, BAM, and SPL groups based on the Jensen-Shannon distance metric. *P < 0.05 compared with CON group. Treatment groups: CON, control group fed with basal diet; BAM, group fed with 10 mg/kg of bambermycin supplemented diet; SPL, group fed with 10 mg/kg of sophorolipid-supplemented diet Cecal short-chain fatty acid concentration Intestinal characteristics and histological analysis As listed in Table 4, dietary SPL supplementation signifi- As shown in Table 5, birds fed with the SPL- cantly increased (P < 0.05) the total concentration of supplemented diet (SPL group) showed significantly re- SCFA, and those of acetate and butyrate in the SPL duced (P < 0.05) intestinal weight compared to those of group compared to that in the BAM group. In addition, the other treatment groups; however, gut weight per the ratio of propionate was significantly lowered length was not changed by SPL supplementation. (P < 0.05) in SPL group compared to BAM group. Hence, dietary SPL supplementation significantly Fig. 2 Gut microbiota population of broilers fed with experimental diets. a Intestinal microflora at phylum; b Intestinal microflora at genus level; b–d Specific bacterial populations at the species level in the cecum of birds (Lactobacillus family, Akkermansia muciniphila, and Streptococcus a, b gallolyticus group). *P < 0.05 compared with CON group. Mean values within a row have different superscript letters were significantly different (P < 0.05). Treatment groups: CON, control group fed with basal diet; BAM, group fed with 10 mg/kg of bambermycin supplemented diet; SPL, group fed with 10 mg/kg of sophorolipid-supplemented diet Kwak et al. Journal of Animal Science and Biotechnology (2021) 12:81 Page 6 of 9 IL-6 IL-10 CLDN1 MUC2 a, Fig. 3 mRNA expression levels of genes related to inflammation, tight junction, and mucin in the jejunum of birds fed with experimental diets. Mean values within a row have different superscript letters were significantly different (P < 0.05). Treatment groups: CON, control group fed with basal diet; BAM, group fed with 10 mg/kg of bambermycin supplemented diet; SPL, group fed with 10 mg/kg of sophorolipid-supplemented diet. Abbreviations: IL-6, interleukin-6; IL-10, interleukin-10; CLDN1, claudin-1; MUC2, mucin 2 increased (P < 0.05) VH compared to that of birds in also significantly increased (P < 0.05) in the SPL treat- the other treatment groups without affecting CD ment group compared to that in the other groups. (Table 5). The ratio of VH to CD in the birds fed with the SPL-supplemented diet was the highest (P < Gene expression levels related to inflammation, tight 0.05) compared to that of birds in CON group. More- junction, and mucin secretion over, the goblet cell numbers per 1 μm of villus were As presented in Fig. 3, dietary SPL supplementation sig- nificantly downregulated (P < 0.05) the expression level of interleukin-6 (IL-6) and upregulated (P < 0.05) that of interleukin-10 (IL-10) compared to the other treatments. Table 4 Effects of sophorolipid on cecal SCFA concentration of The expression level of claudin-1 (CLDN1) significantly 1,2,3 broilers increased (P < 0.05) in the SPL group compared to the Treatment CON BAM SPL SEM P-value other groups. Moreover, the BAM and SPL groups Absolute concentration of SCFA, μmol/g showed significantly increased (P < 0.05) expression level a b a of mucin 2 (MUC2) compared to the CON group. Total SCFA 226.43 179.47 249.61 10.802 0.012 a b a Acetate 185.91 143.14 206.06 9.984 0.017 Discussion Propionate 17.62 15.73 17.18 0.676 0.526 First, the results of this study indicated that SPL could iso-butyrate 3.69 3.84 4.01 0.208 0.850 accelerate the growth of broilers as much as bambermy- ab b a Butyrate 19.20 16.76 22.36 0.978 0.049 cin, which is in agreement with the reports of Boontiam Percentage of SCFA, % et al. [17] which demonstrated that surfactant supple- mentation with a low energy and crude protein diet Acetate 82.11 78.98 82.48 0.852 0.187 ab a b could improve average daily gain and feed efficiency dur- Propionate 7.76 8.91 6.88 0.322 0.021 ing the overall experimental period without affecting iso-butyrate 1.66 2.17 1.60 0.130 0.140 average daily feed intake. Furthermore, both glycolipid- Butyrate 8.47 9.95 9.04 0.615 0.647 type antibiotics and biosurfactant increased the Treatments: CON, control group fed with basal diet; BAM, group fed with 10 beneficial microbiota L. heliveticus and L. salivarius and mg/kg of bambermycin supplemented diet; SPL, group fed with 10 mg/kg of decreased the pathogenic bacteria S. gallolyticus. In ac- sophorolipid supplemented diet Abbreviations: SCFA short-chain fatty acid, SEM standard error of means cordance with our results, Abu Hafsa and Ibrahim [18] A chick is an experimental unit; 6 replications per treatment a, b also demonstrated that similar modulation of gut micro- Mean values within a row have different superscript letters were significantly different (P < 0.05) biota (increased Lactobacillus and decreased Kwak et al. Journal of Animal Science and Biotechnology (2021) 12:81 Page 7 of 9 1,2,3 Table 5 Effects of sophorolipid on gut characteristic and histological analysis of broilers Treatment CON BAM SPL SEM P-value a ab b Intestinal, g/100 g body weight 3.18 3.08 2.96 0.037 0.038 Intestinal weight/length, g/m 30.60 31.12 30.54 0.319 0.735 b ab a Villus height, μm 371.27 398.17 422.50 7.483 0.010 Crypt depth, μm 110.58 104.40 105.50 1.366 0.142 b a a Villus height/crypt depth 3.37 3.82 4.01 0.099 0.013 b b a Goblet cells/villus height, /μm 0.22 0.21 0.34 0.016 < 0.001 Treatments: CON, control group fed with basal diet; BAM, group fed with 10 mg/kg of bambermycin supplemented diet; SPL, group fed with 10 mg/kg of sophorolipid supplemented diet Abbreviation: SEM standard error of means A chick is an experimental unit; 6 replications per treatment a, b Mean values within a row have different superscript letters were significantly different (P < 0.05) Sterptococcus) by dietary probiotic supplementation sig- The gastrointestinal tract is the main region that plays nificantly improved the growth of birds. Various reports a role in a protective system because it is the first organ related Lacobacillus species have suggested that L. heli- to meet external substances with a large contact area veticus is known as the beneficial microbes in bone and [28]. Hence, the gut defense strategy employs various mental health and L. salivarius has a positive relation- physiological factors (e.g., mucus barrier, tight junctions, ship with growth of birds [19–21]. Collectively, micro- and immune cytokines) to maintain the intestinal flora communities modulated by bambermycin and SPL homeostatic balance [29]. Our results suggest that were similar, however SPL have seemed to accelerate the dietary SPL supplementation could enhance mucin- shift of the community compared to that of bambermy- presenting capacity and villus turnover balance by in- cin. Also, these results suggested that this microbial shift creasing beneficial bacterial populations. At phylum has a potential to improve the growth and health of level, there were higher portion of Firmicutes, and lower birds by modulation of their metabolites. that of Bacteroidetes in SPL treatment group compared On the other hand, SCFA are the main metabolites of to the other groups. Bacteroides, known as the LPS- microbial fermentation, and they have been widely stud- generating bacteria group when they are lysed, may play ied to elucidate the specific mechanism of antibiotic a critical role to destruct intestinal integrity and morph- growth promoter linking the host and its intestinal ology by lowering intestinal permeability [30], resulted microbiota [22]. In 2019, Guinan et al. demonstrated in the lower Firmicutes/Bacteroidetes ratio is considered that water supply with antibiotic could downregulated as the favorable index of weight loss [31]. At species the SCFA concentration in cecal contents by increased level, Akkermansia muciniphila, has received attention colonization of Candida albicans [23]. In agreement because of its specific biological properties including im- with this study, the results of our study demonstrated mune modulation, wound healing, and SCFA production that dietary bambermycin supplementation significantly [32]. Moreover, various studies have demonstrated that reduced the concentration of total SCFA compared to A. muciniphila has the potential to act as a probiotic be- birds fed control diet and SPL supplemented diet. cause it could exert a glucose-lowering effect by regulat- Additionally, both acetate and butyrate were increased ing gut barrier integrity through increased expression in the SPL group than BAM group, which might be levels of tight junction proteins [33, 34]. Similar to the due to the increased populations of butyrate- results of these studies, we also found an increased producing bacteria, L. helveticus and mucin-degrading population of A. muciniphila and strengthened gut epi- bacteria, A. muciniphila [24, 25]. An in vitro study thelial integrity and mucus secretion capacity. with L. helveticus showed that this probiotics strain Additionally, our results demonstrated that SPL could significantly increased butyrate concentration, and it relieve the local gut immune response and strengthen maybedue to theupregulated conversion ratioof the intestinal epithelial barrier by modulating the gut lactate to butyrate by lactating-utilizing bacteria [26]. microbiota population. S. gallolyticus, a gram-positive And A. muciniphila supplemented with mucin could pathogenic bacterium, which is commonly found in vari- produce acetate and ethanol from mucin fermentation ous animals and humans, and it is a potential transmis- [27]. Collectively, these results suggest that dietary sion bacterium with antibiotic resistance [35, 36]. In SPL treatment maintain the cecal SCFA concentration addition, Li et al. [37] demonstrated that inoculation of by intestinal colonization of SCFA-producing bacteria S. gallolyticus in duckling induced macrophage necrop- unlike bambermycin supplementation. tosis in spleens by bacterial infection and increased the Kwak et al. Journal of Animal Science and Biotechnology (2021) 12:81 Page 8 of 9 expression levels of IL-6, and that decreased population Consent for publication Not applicable. of S. gallolyticus achieved in response to dietary oregano powder has a negative relationship with the increase of Competing interests anti-inflammatory cytokine IL-10 [38]. In accordance The authors declare that they have no conflict of interest. with the results of these previous studies, decreased Author details population of pathogenic bacteria (S. gallolyticus)by Department of Biotechnology, Korea University, 145 Anam-ro, Seoul 02841, sophorolipid supplementation has the potential to allevi- 2 Republic of Korea. Pathway Intermediates, Seoul 02841, Republic of Korea. ate immune responses by improving pro- (IL-6) and Received: 12 January 2021 Accepted: 13 May 2021 anti-inflammatory (IL-10) cytokine production. On the other hands, our results also found that the higher popu- lation of Lactobacillus genus in BAM and SPL groups References compared to CON group. Lactobacillus is the genera of 1. Danzeisen JL, Kim HB, Isaacson RE, Tu ZJ, Johnson TJ. Modulations of the chicken cecal microbiome and metagenome in response to anticoccidial gram positive and facultative anaerobes and it can in- and growth promoter treatment. PLoS One. 2011;6(11):e27949. https://doi. hibit the pathogen growth by establishment of low pH org/10.1371/journal.pone.0027949. environment through lactic acid producing capacity [39]. 2. Bates J, Jordens JZ, Griffiths DT. Farm animals as a putative reservoir for vancomycin-resistant enterococcal infection in man. J Antibicrob Therefore, our results demonstrated that microbial shift Chemother. 1994;34(4):507–14. https://doi.org/10.1093/jac/34.4.507. by SPL might be able to modulate immune response in 3. Munk P, Knudsen BE, Lukjacenko O, Duarte ASR, Luiken RE, Van Gompel L, broilers’ intestine. et al. Abundance and diversity of the fecal resistome in slaughter pigs and broilers in nine European countries. Nature. 2017;3:898–908. 4. Wang JP, Lee JH, Yoo JS, Cho JH, Kim HJ, Kim IH. Effects of phenyllactic acid Conclusions on growth performance, intestinal microbiota, relative organ weight, blood characteristics, and meat quality of broiler chicks. Poult Sci. 2010;89(7):1549– Dietary sophorolipid supplementation in broiler feed 55. https://doi.org/10.3382/ps.2009-00235. could modulate the intestinal microbiota population and 5. Roy A, Haldar S, Mondal S, Ghosh TP. Effects of supplemental exogenous short-chain fatty acid levels. Hence, the promoted gut emulsifier on performance, nutrient metabolism, and serum lipid profile in broiler chickens. Vet Med Int. 2010;11:262604. environment could improve gut defense integrity, in- 6. Chen C, Jung B, Kim WK. Effects of lysophospholipid on growth cluding intensified mucus layer and tight junction, and performance, carcass yield, intestinal development, and bone quality in alleviate local inflammation, resulting in the acceleration broilers. Poult Sci. 2019;98(9):3902–23. https://doi.org/10.3382/ps/pez111. 7. Cho KJ, Kim YB, Kim EK. Production and application of sophorolipid, a of chick growth. Further studies are required to elucidate microbial surfactant. Korean Soc Biotechnol Bioeng J. 1999;14:747–53. the precise relationship between sophorolipids and their 8. Develter DW, Lauryssen LM. Properties and industrial applications of effects on intestinal health. sophorolipid. Eur J Lipid Sci Technol. 2010;112(6):628–38. https://doi.org/10.1 002/ejlt.200900153. Abbreviations 9. Borsaniyiova M, Patil A, Mukherji R, Prabhune A, Bopegamage S. Biological ADFI: Average daily feed intake; ADG: Average daily gain; ANOVA: Analysis of activity of sophorolipids and their possible use as antiviral agents. Folia variations; BW: Body weight; CD: Crypt depth; CLDN1: Claudin-1; FE: Feed Microbiol. 2016;61(1):85–9. https://doi.org/10.1007/s12223-015-0413-z. efficiency; GAPDH: Glyceraldehyde 3-phosphate dehydrogenase; GC–MS: Gas 10. Kwak MJ, Park MY, Kim J, Lee H, Whang KY. Curative effects of sophorolipid chromatography–mass spectrometry; IL-6: Interleukin-6; IL-10: Interleukin-10; on physical wounds: in vitro and in vivo studies. Vet Med Sci. 2021;00:1–9. MTBSTFA: N-tert-butyldimethylsilyl-N-methyltrifluoroacetamide ; MUC2: Mucin https://doi.org/10.1002/vms3.481. 2; QIIME: Quantitative Insights Into Microbial Ecology; SPL: Sophorolipid; 11. Hardin R, Pierre J, Schulze R, Mueller CM, Fu SL, Wallner SR, et al. SCFA: Short-chain fatty acid; VH: Villus height Sophorolipids improve sepsis survival: effects of dosing and derivatives. J Surg Res. 2008;142:314–9. 12. Concaix FB. Use of sophorolipids comprising diacetyl lactones as agent for Acknowledgements stimulating skin fibroblast metabolism. U S Patent No 6,596,265. 2003. Not applicable. 13. Yoon SH, Ha SM, Kwon S, Lim J, Kim Y, Seo H, et al. Introducing EzBioCloud: a taxonomically united database of 16S rRNA and whole genome Authors’ contributions assemblies. Int J Syst Evol Microbiol. 2017;67(5):1613–7. https://doi.org/10.1 MJK, MYP, JGK and KYW conceived and designed the experiments; MJK and 099/ijsem.0.001755. MYP mainly performed the experiments; MJK and YSC analyzed the data; JC, 14. Li X, McFarland DC, Velleman SG. Effect of Smad3-mediated transforming DP, HBL and IGC contributed reagents/materials/analysis tools; MJK wrote growth factor-β1 signaling on satellite cell proliferation and differentiation the manuscript. All authors read and approved the final manuscript. in chickens. Poult Sci. 2008;87(9):1823–33. https://doi.org/10.3382/ps.2008- Funding 15. Song B, Li H, Wu Y, Zhen W, Wang Z, Xia Z, et al. Effect of This work was financially supported by EASYBIO Inc. and Korea University. microencapsulated sodium butyrate dietary supplementation on growth performance and intestinal barrier function of broiler chickens infected with Availability of data and materials necrotic enteritis. Anim Feed Sci Technol. 2017;232:6–15. https://doi.org/10.1 Not applicable. 016/j.anifeedsci.2017.07.009. 16. Heidari M, Wang D, Delekta P, Sun S. Marek’s disease virus Declarations immunosuppression alters host cellular reponses and immune gene expression in the skin of infected chickens. Vet Immunol Immunopathol. Ethics approval 2016;180:21–8. https://doi.org/10.1016/j.vetimm.2016.08.013. All of works related to animal was conducted in accordance with the 17. Boontiam W, Hyun YK, Jung B, Kim YY. Effects of lysophospholipid guidelines and regulations for the care and the use of experimental animals supplementation to reduced energy, crude protein, and amino acid diets was approved by the Korea University Institutional Animal Care & Use on growth performance, nutrient digestibility, and blood profiles in broiler Committee (Permission No. KUIACUC-2020-0097). chickens. Poult Sci. 2019;98(12):6693–701. https://doi.org/10.3382/ps/pex005. Kwak et al. Journal of Animal Science and Biotechnology (2021) 12:81 Page 9 of 9 18. Abu Hafsa SH, Ibrahim SA. Effect of dietary polyphenol-rich grape seed on 38. Bauer BW, Gangadoo S, Bajagai YS, Van TTH, Moore RJ, Stanley D. Oregano growth performance, antioxidant capacity and ileal microflora in broiler powder reduces Streptococcus and increases SCFA concentration in a chicks. J Anim Physiol Anim Nutr. 2018;102(1):268–75. https://doi.org/1 mixed bacterial culture assay of chicken. PloS One. 2019;13(3):e0216853. 0.1111/jpn.12688. https://doi.org/10.1371/journal.pone.0216853. 19. Ohland CL, Kish L, Bell H, Thiesen A, Hotte N, Pankiv E, et al. Effects of 39. Rodriguez-Cebezas ME, Camuesco D, Arribas B, Garrido-Mesa N, Comalada Lactobacillus helveticus on murine behavior are dependent on diet and M, Bailon E, et al. The combination of fructooligosaccharides and resistant genotype and correlate with alterations in the gut microbiome. starch shows prebiotic additive effects in rats. Clin Nutr. 2010;29(6):832–9. Psychoneuroendocrinology. 2013;38(9):1738–47. https://doi.org/10.1016/j. https://doi.org/10.1016/j.clnu.2010.05.005. psyneuen.2013.02.008. 20. Liang S, Wang T, Hu X, Luo J, Li W, Wu X, et al. Administration of Lactobacillus helveticus NS8 improves behavioral, cognitive, and biochemical aberrations caused by chronic restraint stress. Neuroscience. 2015;310:561–77. https://doi.org/10.1016/j.neuroscience.2015.09.033. 21. Zhu NH, Zhang RJ, Wu H, Zhang B. Effects of Lactobacillus cultures on growth performance, xanthophyll deposition, and color of the meat and skin of broilers. J Appl Poult Res. 2009;18(3):570–8. https://doi.org/10.3382/ja pr.2009-00012. 22. Le Poul E, Loison C, Struyf S, Springael JY, Lannoy V, Decobecq ME, et al. Functional characterization of human receptors for short chain fatty acids and their role in polymorphonuclear cell activation. J Biol Chem. 2003; 278(28):25481–9. https://doi.org/10.1074/jbc.M301403200. 23. Guinan J, Wang S, Hazbun TR, Yadav H, Thangamani S. Antibiotic-induced decreases in the levels of microbial-derived short-chain fatty acids correlate with increased gastrointestinal colonization of Candida albicans. Sci Rep. 2019;9:1–11. 24. Parada Venegas D, De la Fuente MK, Landskron G, González MJ, Quera R, Dijkstra G, et al. Short chain fatty acid (SCFA)-mediated gut epithelial and immune regulation and its relevance in inflammatory bowel diseases. Front Immunol. 2019;10:277–92. 25. Taverniti V, Guglielmetti S. Health-promoting properties of Lactobacillus helveticus. Front Microbiol. 2012;3:392–404. 26. Vitali B, Ndagijimana M, Maccaferri S, Blagi E, Guerzoni ME, Brigidi P. An in vitro evaluation of the effect of probiotics and prebioitcs on the metabolic profile of human microbiota. Anaerobe. 2012;18(4):386–91. https://doi.org/10.1016/j.anaerobe.2012.04.014. 27. Derrien M, Vaughan EE, Plugge CM, de Vos WM. Akkermansia muciniphila gen. nov., sp. Sp. Nov., a human intestinal mucin-degrading bacterium. Int J Syst Evol Microbiol. 2004;54:1469–76. 28. Artis D. Epithelial-cell recognition of commensal bacteria and maintenance of immune homeostasis in the gut. Nat Rev Immunol. 2008;8(6):411–20. https://doi.org/10.1038/nri2316. 29. Schroeder BO. Fight them or feed them: how the intestinal mucus layer manages the gut microbiota. Gastroenterol Rep. 2019;7(1):3–12. https://doi. org/10.1093/gastro/goy052. 30. Chen K, Zhao H, Shu L, Xing H, Wang C, Lu C, et al. Effect of resveratrol on intestinal tight junction proteins and the gut microbiome in high-fat diet- fed insulin resistant mice. Int J Food Sci Nutr. 2020;71(8):965–78. https://doi. org/10.1080/09637486.2020.1754351. 31. Qu L, Liu Q, Zhang Q, Tuo X, Fan D, Deng J, et al. Kiwifruit seed oil prevents obesity by regulating inflammation, thermogenesis, and gut microbiota in high-fat diet-induced obese C57BL/6 mice. Food Chem Toxicol. 2019;125: 85–94. https://doi.org/10.1016/j.fct.2018.12.046. 32. Earley H, Lennon G, Balfe Á, Coffey JC, Winter DC, O’Connell PR. The abundance of Akkermansia muciniphila and its relationship with sulphated colonic mucins in health and ulcerative colitis. Sci Rep. 2019;9:1–9. 33. Ashrafian F, Behrouzi A. Comparative study of effect of Akkermansia muciniphila and its extracellular vesicles on toll-like receptors and tight junction. Gastroenterol Hepatol Bed Bench. 2019;12(2):163–8. 34. Su H, Mo J, Ni J, Ke H, Bao T, Xie J, et al. Andrographolide Exerts antihyperglycemic effect through strengthening intestinal barrier function and increasing microbial composition of Akkermansia muciniphila. Oxid Med Cell. 2020;6538930:1–20. https://doi.org/10.1155/2020/6538930. 35. Dumke J, Hinse D, Vollmer T, Schulz J, Knabbe C, Dreier J. Potential transmission pathways of Streptococcus gallolyticus subsp gallolyticus. PLoS One. 2015;10:e0126507. 36. Nomoto R, Le HTT, Sekizaki T, Osawa R. Antimicrobial susceptibility of Streptococcus gallolyticus isolated from humans and animals. Jpn J Infect Dis. 2013;66(4):334–6. https://doi.org/10.7883/yoken.66.334. 37. Li M, Liu B, Gu C, Zhang W, Yang J, Cheng G, et al. Necroptosis of splenic macrophages induced by Streptococcus gallolyticus subsp. pasteurianus. Avian Dis. 2017;61(1):115–22. https://doi.org/10.1637/11449-061216-Reg.

Journal

Journal of Animal Science and Biotechnology – Springer Journals

Published: Jul 12, 2021

Keywords: Broilers; Gut microbiota; Gut morphology; Local inflammation; Mucus barrier; Sophorolipids

There are no references for this article.