Effects of Brown Fish Meal Replacement with Fermented Soybean Meal on Growth Performance, Feed Efficiency and Enzyme Activities of Chinese Soft-shelled Turtle, Pelodiscus sinensis

A 120-day feeding experiment was conducted to investigate the effects of partial replacement of brown fish meal (BFM) by fermented soybean meal (FSBM) in diets of Chinese soft-shelled turtle (Pelodiscus sinensis). The turtles (initial mean body weight, (115.52 ± 1.05) g) were fed with three experimental diets, in which 0%, 4.72% and 9.44% BFM protein was replaced by 0%, 3% and 6% FSBM, respectively. Results showed that the feeding rate (FR), specific growth rate (SGR) and feed efficiency ratio (FER) of turtles fed with the diet containing 3% FSBM were not significantly different from the control group (0% FSBM) (P > 0.05). However, FR, SGR and FER of turtles fed with the diet containing 6% FSBM were significantly lower than those of the control group (P < 0.05). No significant differences were observed in the activities of serum glutamic-oxalacetic transaminase and glutamicpyruvic transaminase among dietary treatments (P > 0.05). However, the uric acid concentration in turtles fed with the diet containing 3% or 6% FSBM was significantly lower than that in the control group (P < 0.05). There were no significant differences in the activities of lysozyme, alkaline phosphatase and total superoxide dismutase among dietary treatments (P > 0.05). The results suggested that FSBM could replace 4.72% BFM protein in turtle diets without exerting adverse effects on turtle growth, feed utilization and measured immune parameters.

Comparative effects of soy protein concentrate,enzyme‑treated soybean meal,and fermented soybean meal replacing animal protein supplements in feeds on growth performance and intestinal health of nursery pigs

Background Soy protein supplements,with high crude protein and less antinutritional factors,are produced from soybean meal by different processes.This study evaluated the comparative effects of various soy protein supplements replacing animal protein supplements in feeds on the intestinal immune status,intestinal oxidative stress,mucosaassociated microbiota,and growth performance of nursery pigs.Methods Sixty nursery pigs(6.6±0.5 kg BW)were allotted to five treatments in a randomized complete block design with initial BW and sex as blocks.Pigs were fed for 39 d in 3 phases(P1,P2,and P3).Treatments were:Control(CON),basal diet with fish meal 4%,2%,and 1%,poultry meal 10%,8%,and 4%,and blood plasma 4%,2%,and 1%for P1,P2,and P3,respectively;basal diet with soy protein concentrate(SPC),enzyme-treated soybean meal(ESB),fermented soybean meal with Lactobacillus(FSBL),and fermented soybean meal with Bacillus(FSBB),replacing 1/3,2/3,and 3/3 of animal protein supplements for P1,P2,and P3,respectively.Data were analyzed using the MIXED procedure in SAS 9.4.Results The SPC did not affect the BW,ADG,and G:F,whereas it tended to reduce(P=0.094)the ADFI and tended to increase(P=0.091)crypt cell proliferation.The ESM did not affect BW,ADG,ADFI,and G:F,whereas tended to decrease(P=0.098)protein carbonyl in jejunal mucosa.The FSBL decreased(P<0.05)BW and ADG,increased(P<0.05)TNF-α,and Klebsiella and tended to increase MDA(P=0.065)and IgG(P=0.089)in jejunal mucosa.The FSBB tended to increase(P=0.073)TNF-α,increased(P<0.05)Clostridium and decreased(P<0.05)Achromobacter and alpha diversity of microbiota in jejunal mucosa.Conclusions Soy protein concentrate,enzyme-treated soybean meal,and fermented soybean meal with Bacillus could reduce the use of animal protein supplements up to 33%until 7 kg body weight,up to 67%from 7 to 11 kg body weight,and entirely from 11 kg body weight without affecting the intestinal health and the growth performance of nursery pigs.Fermented soybean meal with Lactobacillus,however,increased the immune reaction and oxidative stress in the intestine consequently reducing the growth performance.

Free Amino Acid Profiles of Fermented Soybean Products Prepared by Bacillus subtilis and Rhizopus oligosporus

Fermented soybean is one of the traditional foods widely consumed in many Asian countries. Most products（i.e., Natto, Jang, Kinema and Thua nao） are produced mainly by the bacterial activity. This study was performed as a part of the program to improve the soybean fermentation, and a use of the co-culture between the bacterium Bacillus subtilis and the fungus Rhizopus oligosporus was selected. For fermentation, the raw soybeans were washed, sterilized by autoclaving, and inoculated with a mixed microbial starter culture of B. subtilis（102 CFU · g-1） and R. oligosporus（102 conidia · g-1）. The Free Amino Acid（FAA） contents were then investigated by using High Performance Liquid Chromatography（HPLC）. The results showed that the amounts of FAAs and essential amino acids in the fermented soybean samples were found at significantly higher concentration when compared to those of the cooked, non-fermented soybean samples（p〈0.05）. These data indicated that an involvement of microbial fermentation helped to accelerate an increase of FAA. The food compositional data were useful as a reference（and/or guidance） for the nutritional value, when compared to other fermented soybeans and related products.

Application of Antimicrobial Bacillus subtilis Strain as a Starter Culture to Improve Qualities and Safety of Fermented Soybean (SIENG) Produced in Cambodia

Fermented foods play a very important role in Cambodian health and nutrition,as well as other developing countries where food preservation methods may be limited.SIENG,a Khmer fermented soybean product,naturally contains both beneficial and pathogenic microorganisms.Traditional fermentation that relies on natural microbial flora and environmental conditions results in variable product quality and can lead to spoilage.A starter culture such as Bacillus subtilis can ensure the safety and stability of the products.The objective of this study is to control the growth of Gram positive pathogens contaminated into traditional fermented soybean(SEING)by using antimicrobial Bacillus subtilis isolated from the same kind of food.Out of 120 SIENG samples,49 B.cereus strains were isolated,and 12 of B.cereus were positively synthesized compared with the lyophilized control enterotoxin.Two of these strains(BTM8-7 and BTM8-8)produced high levels of enterotoxin.We identified five Bacillus strains with the ability to fight against indicator pathogenic microorganisms.Among the five strains,B.CeM6-2 had the highest activity level against Lactobacillus plantrum ATCC 8014 and the largest diameter.B.CeM6-2 tolerated up to 20 h at 30℃ and 22 h at 37℃.In testing the strains with PK and PK-PMSF enzymes,bacteriocin produced by the strain B.CeM6PK untreated and B.CeM6-2PK-PMSF had a significantly stronger ability to suppress all the pathogenic indicators from 0 h to 47 h compared to the B.CeM6-2PK.Moreover,CeM6-2 outperformed the Miyagino strains,as it actively produced bacteriocin that fought against all four indicator strains of Gram positive and lactic acid groups,especially against Listeria monocytogenes,Streptococcus pyrogene,Leuconostoc mesenterids and L.plantarum,from 0 h-58 h and 0 h-40 h at 35℃.CeM6-2(1%)strain had the highest ability to fight against B.cereus at 24 h and at 34 h to 44 h incubation as well.CeM6-2(10:0 mL)and CeM6-2(9:1 mL)have the strongest ability to fight against B.cereus at room temperature(48 h and 72 h).The longer incubation and time at room temperature produce the highest level of bacteriocin.Thus,bacteriocins produced by B.CeM6-2 can be used as a preservative in food processing industries to avoid food spoilage even in higher temperatures and time.

Isolation, Characterization and Bio-Control Activities of Bacillus Subtilis from the Fermented Soybean in Cambodia

The objective of this study was to isolate Bacillus subtilis produced Bacteriocin from Khmer traditional fermented soybean to control the growth of pathogenic bacteria from 120 SIENG samples from 9 different local open markets in Phnom Penh, 1 local open market and 2 local producers in Kandal province in Cambodia. 119 out of 120 samples were positive with Bacillus subtilis strains. Fifteen of these isolated strains exhibited antimicrobial activity to Lactobacillus plantarum by agar well diffusion assay and one of them （CeM6-7 strain） exhibited strongest antimicrobial activity. The filtered supernatant of this strain also suppressed the growth of several Gram-positive pathogens （such as Staphylococcus aureus） or lactic acid bacteria in broth. The filtrate could also suppress the growth of L. plantarum inoculated into soymilk. Antimicrobial B. subtilis strain CeM6-7 is thought to be applicable for making a starter culture to produce much safe fermented soybean foods by suppressing the growth of naturally contaminated B. cereus or S. aureus during production. In addition, the use of this starter culture may contribute to prevent spoilage or quality loss of soybean products caused by the over growth lactic acid bacteria or other Gram-positive bacteria.

Monascus purpureus M-32 fermented soybean meal improves the growth,immunity parameters,intestinal morphology,disease resistance,intestinal microbiota and metabolome in Pacific white shrimp(Litopenaeus vannamei)

This study was conducted to evaluate the effects of Monascus purpureus M-32 fermented soybean meal(MFSM)on growth,immunity,intestinal morphology,intestinal microbiota,and intestinal metabolome of Pacific white shrimp(Litopenaeus vannamei).Four groups of diets were formulated,including control group(30%fish meal and 30%soybean meal[SBM]included in the basal diet)and three experimental groups which MFSM replaced 20%(MFSM20),40%(MFSM40),and 60%(MFSM60)of SBM in control group,respectively.Results showed that the soluble proteins larger than 49 kDa in MFSM were almost completely degraded.Meanwhile,the crude protein,acid-soluble protein,and amino acid in MFSM were increased.The results of shrimp culture experiment showed that the replacement of SBM with MFSM decreased FCR(P<0.001)and content of malondialdehyde(P=0.007)in the experimental groups,and increased weight gain rate(P=0.006),specific growth rate(P=0.002),survival rate(P=0.005),in-testinal villus height(P<0.001),myenteric thickness(P=0.002),the activities of superoxide dismutase(P=0.002),and lysozyme(P=0.006)in experimental groups,as well as increased content of calcium(Ca2+)and phosphorus(PO43-)in blood and muscle,and enhanced resistance to Vibrio parahaemolyticus infection.The gut microbiota of MFSM groups was significantly different from that of the control group,and the abundance of Actinobacteria and Verrucomicrobia increased significantly in the MFSM60 group,whereas Proteobacteria and Firmicutes decreased.Compared with the control group,there were sig-nificant changes in the levels of several intestinal metabolites in the MFSM60 group,including leuko-triene C5,prostaglandin A1,taurochenodeoxycholic acid,carnosine,and itaconic acid.The fermentation of SBM by the strain M.purpureus M-32 has the potential to enhance the nutritional quality of SBM,promote the growth of L.vannamei,boost immune response,improve intestinal morphology and microbiota composition,as well as influence intestinal metabolites.

Fermented soybean meal improved laying performance and egg quality of laying hens by modulating cecal microbiota,nutrient digestibility,intestinal health,antioxidant and immunological functions

Antinutritional factors in feedstuffs may limit their utilization in livestock production,but fermentation process can be used to improve feed quality;however,studies on fermented soybeans for laying hens remain limited.We investigated the effect of fermented soybean meal(FSBM)at various inclusion levels as a partial replacement for soybean meal(SBM)on egg production,egg quality,amino acid digestibility,gut morphology and microbiota,antioxidant capacity and immune response of young laying hens.A total of 360 Hy-line Brown laying hens aged 18 weeks were selected and divided into 5 groups of 6 replicates each and 12 birds per replicate.The control group received a basal diet while the trial group received the basal diet with FSBM included at 2.5%,5.0%,7.5%and 10.0%,respectively,for 12 weeks.Our findings revealed that the nutritional value of FSBM was higher compared to that of SBM in terms of reduced content of trypsin inhibitors and increased contents of crude protein,amino acids and minerals.FSBM enhanced egg production(P<0.05),feed-to-egg ratio(P<0.05),and albumen quality(albumen height and Haugh unit)(P<0.05).Furthermore,FSBM improved apparent fecal amino acid digestibility(P<0.05),gut morphology(increased villus height,villus width,villus height-to-crypt depth ratio and decreased crypt depth)(P<0.05),antioxidant capacity(reduced malondialdehyde and increased catalase,total superoxide dismutase,glutathione peroxidase and total antioxidant capacity)(P<0.05)and immune function(increased concentrations of IgG,IgA,and IgM;increased levels of transforming growth factor beta and Toll-like receptor 2;and reduced levels of interleukin 1βand tumor necrosis factor alpha)(P<0.05).Further analysis showed that FSBM altered the composition of the gut microbiota favoring beneficial microbes.These findings suggest that probiotic fermentation improved the nutritional value of SBM.The inclusion of FSBM in the diets of laying hens at 2.5%or 5.0%improved amino acid digestibility,gut health,immune function,egg production and egg quality.

Fermented soybean meal improves the growth performance, nutrient digestibility, and microbial flora in piglets

In order to increase nutritive values of soybean meal(SBM), 3 species of microbes were used to ferment SBM. Through a 3 x 3 orthogonal design and parameter measurements of soybean peptide and antinutritional factor contents in the fermented soybean meal(FSBM), it was estimated that the best microbial proportion of Bacillus subtilis, Hansenula anomala and Lactobacillus casei was 2:1:2 for SBM fermentation(P < 0.05). The further piglet feeding experiment showed that 10% FSBM substitute for SBM had no significant effect on growth performance of suckling piglets(d 7-28)(P> 0.05). However, newlyweaned piglets(d 28-38) fed 10% FSBM and different levels of plasma protein obtained higher average daily gain(ADG) and feed conversion ratio(FCR), compared with those without FSBM but with 6% plasma protein(P < 0.05). Piglets(d 38-68) fed diets supplemented with FSBM and soybean protein concentrate(SBPC) at 3.75% and 7.5% respectively increased nutrient digestibility,fecal enzyme activity and lactic acid bacteria counts, and decreased fecal Escherichia coli counts(P < 0.05), compared with the control. These data indicated that FSBM had positive effects on nutrient digestibility and fecal microflora for piglets.

Effects of dietary fresh fermented soybean meal on growth performance, ammonia and particulate matter emissions, and nitrogen excretion in nursery piglets

This study was conducted to investigate the effects of fresh fermented soybean meal（FSM） on the growth performance of nursery piglets, nitrogen excretion in feces, and the concentrations of ammonia（NH3） and particulate matter（PM） in the piggery. A total of 472 nursery piglets（Landrace×Yorkshire,（16.3±0.36） kg body weight） were randomly allocated into two treatments with 236 pigs in each treatment. The pigs were fed the basal diet without fresh FSM（control） or diet containing 10%（100 g/kg） fresh FSM（FSM group）, and the crude protein content of the two groups was consistent. The feeding trial lasted for 28 d. The results showed that the pigs fed fresh FSM had increased（P〈0.05） average daily gain（ADG） compared with the control. There was no significant difference（P〈0.05） in feed to gain ratio（F：G） between the two groups. During the whole experiment, the concentration of NH3 in the piggery decreased（P〈0.05） by 19.0%, and the concentrations of PM（PM（10） and PM（2.5）） in the piggery decreased（P〈0.05） by 19.9% and 11.6%, respectively, in the FSM group, compared with the control. The ammonia nitrogen and nitrite content in feces increased（P〈0.05） by 32.9% and 28.4%, respectively, in the FSM group. The fecal p H declined（P〈0.05） significantly in the FSM group compared with the control. At the end of experiment, total protein（TP） concentration was increased（P〈0.05） significantly and blood urea nitrogen（BUN） concentration was decreased（P〈0.05） for pigs fed the diet with fresh FSM. The results indicated that dietary fresh FSM not only improved the growth performance of nursery piglets, but also reduced the NH3 concentration in the piggery due to nitrogen conversion, and decreased the concentrations of PM（10） and PM（2.5） in the piggery.

The Effect of Soybean Meal Fermented by Aspergillus usami on Phosphor Metabolism in Growing Pigs

[Objective] The experiment aimed to check the pigs＇ digestive utilization of phosphor and other nutrient components in fermented soy- bean meal. [ Method] 15 8-week old two-way cross growing pigs （average weight was 23.6 kg） were selected and divided randomly into 3 groups, 5 pigs per group. The method of total feces collection was adopted to compare the digestibility of phosphor and other nutrient components in growing pigs fed respectively by fermented soybean meal by Aspergillus usami and normal soybean meal. The experimental design were normal soybean group （NS group, P： 0.087%）, fermented soybean meal group （FS group, P： 0.089%）, normal soybean ＋ fermented soybean meal mixing group （ MS group, P： 0.089% ）. [ Result] The results indicated that the digestibility of growing pigs in FS group was significantly higher than that of NS group. Meanwhile, the digestibility of crude protein was significantly improved. [ Conclusion] The above results indicated that Feeding with soybean meal fermented by Aspergillus usami not only could increase the digestibility of phosphor and protein, but also decrease the excretion of phosphor and nitrogen.

Determination of Genistein in the Fermented By-product of Soybean Curd, an Indonesian Food, and Its in vivo Assay on Carrageenan-lnduced Mice

In our country, Indonesia, especially in West Java, people frequently consume fermented by-product of soybean curd for their main course. The by-product of curd, usually is discarded, can be fermented using either Neurospora （Neurospora sitophila） or Rhizopus （Rhizopus oligosporus） yeast to make oncom Genistein is a phytoestrogen, an estrogen-like chemical compound present in plants, that has anti-inflammatory property. Genistein can be determined by UV spectrophotometric method based on the UV absorbance of its complex with aluminum chloride. Addition of acid is a critical point in the procedure. The aim of this research was to know whether genistein is still contained in oncom This research also studied about oncom＇s activity on carrageenan-induced mice. Acetosal was used as drug control. Result showed that genistein was detected contained in oncom extract. The in vivo assay indicated that oncom extract （dose 1,000 mg/kg of body weight） reduced oedema in carragenan-induced paw of mice, which potencies were 066 times lower than that of acetosal＇s. Oncom might be useful as food and as a weak inflammation reducer as well.

Antioxidant effect of Lactobacillus fermentum HFY02-fermented soy milk on D-galactose-induced aging mouse model

This study aimed to investigate the antioxidant effect of soybean milk fermented by a new type of Lactobacillus fermentum(LF-HFY02)by using D-galactose induced aging mice model.Firstly,the optimal fermentation conditions was screened out by detecting the effects of different fermentation temperature and time on the active components and antioxidant activity of soybean milk in viro.And then unfermented soybean milk and the soybean milk fermented by different Lactobacillus was given by gavage to D-galactose-induced aging mouse.The activities of GSH,GSH-Px,SOD,CAT and T-AOC in serum,brain and liver of soybean milk fermented by LF-HFY02 were significantly increased,while the content of MDA and the level of AGEs in hippocampal were significantly decreased compared with D-galactose induced group.Further more,the mRNA expression of GSH and SOD in mouse liver were obviously up-regulated by soybean milk fermented by LF-HFY02.The skin tissue structure of mice in the LF-HFY02 fermented soybean milk group was more complete,the collagen fibers were increased and arranged orderly and liver inflammation has improved compared with the model group.And Western blot analysis showed that LF-HFY02 effectively upregulated EGFR,SOD and GSH protein expression in mouse liver.These findings suggest that LF-HFY02 can effectively prevent D-galactose-induced oxidation and aging in mice,and the effect was even better than that of the Lactobacillus delbruechii subsp.bulgaricus and vitamin C.Thus,LF-HFY02 may be potentially employed as a probiotic strain.In conclusion,soybean milk fermented by LF-HFY02 can increase the content of antioxidant factors and the activity of antioxidant enzymes by regulating gene and protein expression,and finally inhibit the process of tissue cell peroxidation,and improve the oxidative damage of mouse skin and liver.The results could provide a basis for the research and development and industrial production of probiotic-related fermented soybean milk products.

A model to reduce tyrosine crystallization in Chinese fermented soybean curd by adjusting NaCl,ethanol and temperature during fermentation

Objectives:White spots,mainly consisting of tyrosine,refer to small white particles that exist on the surface of mature Chinese fermented soybean curd or in the fermentation broth and have affected the appearance of Chinese fermented soybean curd.Materials and Methods:The study determined the composition of the white spots throughout mass spectrometry and high-performance liquid chromatography and investigated the relationship between tyrosine content,amino acid nitrogen content,depth of the proteolysis index and protease activity to interpret the mechanism of white spot formation during fermentation.Results:In this study,the results confrm that tyrosine accounts for 78.22%of the white spots.Excessive tyrosine content can promote the formation of white spots during fermentation.Free tyrosine content,amino acid nitrogen content,depth of the proteolysis index and protease activity during the fermentation process increased during fermentation.Correlation analysis results show that free tyrosine content was positively correlated with amino acid nitrogen content,depth of the proteolysis index and protease activity,which showed that the production of tyrosine is related to the degree of proteolysis.Solubility studies show that high concentrations of NaCl and ethanol affect the production of tyrosine and reduce its solubility,while the increase in temperature increases the solubility of tyrosine.Taking NaCl,ethanol and temperature as infuencing factors,a mathematical model of tyrosine solubility was established.Conclusion:This study lays the groundwork for understanding the mechanism of white spot formation during Chinese fermented soybean curd fermentation and provides solutions for controlling white spot formation.

UPLC-QTOF-MS Analysis on Isoflavones in Douchi

Douchi is a kind of traditional Chinese fermented soybean food.Ultra-high performance liquid chromatography quadrupole-time of flight mass spectrometry(UPLC-QTOF-MS)was applied to separate and identify 12 kinds of isoflavones in Douchi within 16 min.The chromatographic separation was carried out on an ACQUITY UPLC HSS T3 column with a gradient elution program where water containing 0.1%formic acid and acetonitrile containing 0.1%formic acid were used as mobile phases.Detection by using electrospray ionization of positive ion mode was applied in the mass spectrometry.Isoflavones were identified by determining the accurate mass and referring to references in this study.

Poly-γ-glutamic acid-producing Bacillus velezensis fermentation can improve the feed properties of soybean meal

Soybean meal(SBM),a high-protein animal feed,contains several anti-nutritional factors,which limit its utilization.Poly-γ-glutamic acid(γ-PGA),a biological macromolecule,can significantly improve the health and growth benefits of animal feed.However,there are rare reports about SBM fermentation withγ-PGA-producing bacteria to improve its nutritional quality.In this study,we isolated a glutamic acid-independent polyγ-glutamic acid(γ-PGA)producing Bacillus velezensis SCP024-S-4 bacterial strain and used it for solid fermentation of SBM.The results showed that the maximumγ-PGA yield in fermented soybean meal(FSBM)reached up to 89.64 g/kg without adding glutamic acid.Meanwhile,the content of proteins,free amino acids and small peptides increased by 12.83%,347.96μg/g and 108.11 mg/g,respectively.Soy globulin,β-conglutinin and trypsin inhibitor all decreased below 3 mg/g.Mice were fed with CK(control diet),SBM and FSBM respectively.The average daily food intake in the FSBM,SBM,and CK groups were 7.68,5.25,and 6.36 g,respectively.Compared with the CK group,the mice weight growth rate was significantly higher in the FSBM group.Mice blood and spleen tests showed no negative effect of FSBM.The high-throughput sequencing of mice intestinal microflora indicated that FSBM increased the richness of intestinal microbiota;especially,the types and quantities of Lactobacillus were significantly increased.This study provides a feasible strategy to improve the nutritional quality of SBM as an animal feed.