Optimization on Production Key Factors of Microbial Fermented Feed and Analysis on Composition Variation During Fermentation

[ Objective] In order to study the production key factors of microbial fermented feed and the composition variation during fermentation. [ Methods ] Water, temperature, energy and protein were selected to study the effects on microbial fermented feed. And the fermentation conditions were optimized. Changes on nutrient composition during fermentation were determined and analyzed. [ Result ] Test results showed that feed formula with high sugar content, low protein, 34% - 36% water content, high fermentation temperature was more conducive to the production and actual production needs of fermentation strains. During the fermentation, strains showed interactions, test tended to be completed at the 144^th h after the fermentation, and number of lactic acid bacteria reached the peak of 1.86×10^9 strain/g, contents of lactic acid could even reach 0.89%. As time prolonged, contents of total energy, fiber and isothiocyanate gradually reduced, while contents of water and protein increased slightly. Contents of vitamin were stable, which in an order of VB〉 VE 〉 Vx 〉 VA 〉 VD. Composition of amino acids showed an better change, contents of glutamic acid and proline decreased slowly, while contents of glycine and phenylalanine increased slowly. [ Conclusion] High quality microbial fermentation feed could improve the palatability and safety of feed and keep the intestinal balance of livestock and poultry, which had broad application prospect.

Effect of Fermented Feed on Intestinal Microbial Community Diversity of Chicks

[Objective]The paper was to study the effects of fermented feed on structure and diversity of intestinal flora in the duodenum,jejunum,ileum and cecum of chicks,and to clarify the mechanism of fermented feed on growth and health of chicks.[Method]The 3601-day-old Hyline brown egg chicks were randomly divided into three treatment groups,six replicates each group,and were fed with basal diet,basal diet+antibiotics and basal diet+fermented feed,respectively.The structure and distribution of intestinal microbial community of egg chicks in each group were determined by Illumina MiSeq sequencing technology.[Result]The Chaol index and Shannon index indicated that the abundance and diversity of microorganisms in unfermented feed group were significantly higher than those in fermented feed group,and the abundance and diversity of flora in the duodenum,jejunum and ileum of each group were significantly higher than those in the cecum.[Conclusion]Feeding fermented feed significantly increases the abundance of Lactobacillus and reduces the number of pathogenic bacteria such as DSV and Bacieroides in the intestinal tract,which also indirectly proves that Lactobacillus inhibits the proliferation of DSV and prevents intestinal diseases.Feeding fermented feed increases the abundance of Firmicutes,significantly decreases the proportion of Bacteroidetes and Firmicutes,and promotes the growth of chicks.

Development of Microbial Fermented Feed and Its Effect on the Production Performance of Holstein Cows

[Objectives]This study was conducted to investigate the development method of microbial fermented feed and its effect on the production performance of Holstein cows.[Methods]Thirty Holstein cows were selected and randomly divided into 2 groups,15 cows in each group.The diet composition of the experimental group was:concentrate supplement added with 30% microbial fermented feed+silage+alfalfa hay,and the diet composition of the control group was:concentrate supplement added with 30% flax cake+silage+alfalfa hay.The experimental period was 60 d.[Results]Compared with the control group,the experimental group increased the average daily milk yield per cow by 2.25%(P>0.05)and the total increase in average milk yield per cow in the experimental period by 43.51%(P<0.01),and reduced the average feed-to-milk ratio by 9.20%(P<0.05).The average gross profit per cow of the experimental group was 40.34 yuan higher than that of the control group,showing an increase of 3.53% in the economic benefit.[Conclusions]Feeding a diet supplemented with the microbial fermented feed could improve the efficiency of Holstein cow farming and is recommended to be promoted.

Advances in research on solid-state fermented feed and its utilization:The pioneer of private customization for intestinal microorganisms

With sustainable development of biotechnology,increasing attention has been placed on utilization of solid-state fermented feed(SFF).Solid-state fermented feed has been a candidate strategy to alleviate the contradiction between supply and demand of feed resources,ensure food hygiene safety,promoting energy conservation,and emission reduction.In production of SFF,a variety of organic acids,enzymes,vitamins,peptides,and other unknown growth factors are produced,which could affect performance of animals.Solid-state fermented feed produced by different fermentation techniques has great instability on different physiological stages of different animals,which hinders the application and standardized production of SFF.Herein,we summarize the current advances in the role of the characteristics of SFF prepared by different manufacturing technique and its research progress in animal experiments on growth performance,gastrointestinal ecology,and immune system,so as to provide references for further acquiring a relatively perfect set of SFF production and evaluation systems.

Fermented liquid feed for pigs： an ancient technique for the future

Fermented liquid feed is feed that has been mixed with water at a ratio ranging from 1：1.5 to 1：4. By mixing with water lactic acid bacteria and yeasts naturally occurring in the feed proliferate and produce lactic acid, acetic acid and ethano which reduces the pH of the mixture. This reduction in pH inhibits pathogenic organisms from developing in the feed. In addition, when this low pH mixture is fed, it reduces the pH in the stomach of pigs and prevents the proliferation of pathogens such as coliforms and Salmonella in the gastrointestinal tract. For piglets, the use of fermented liquid feed offers the possibility of simultaneously providing feed and water, which may facilitate an easier transition from sow＇s milk to solid feed. Secondly, offering properly produced fermented liquid feed may strengthen the role of the stomach as the first line of defense against possible pathogenic infections by lowering the pH in the gastrointestinal tract thereby helping to exclude enteropathogens. Finally, feeding fermented liquid feed to pigs has been shown to improve the performance of suckling pigs, weaner pigs and growing-finishing pigs. In this review, current knowledge about the use of fermented liquid feed in pig diets will be discussed. This will include a discussion of the desirable properties of fermented liquid feed and factors affecting fermentation. In addition, advantages and disadvantages of fermented liquid feed will be discussed including its effects on gastrointestinal health, intestinal pH and the types of bacteria found in the gastrointestinal tract as well as the effects of fermented liquid feeds on pig performance.

Consuming fermented distillers＇ dried grains with solubles (DDGS) feed reveals a shift in the faecal microbiota of growing and fattening pigs using 454 pyrosequencing

The objective of this study was to investigate pig fed by Bacillus coagulans-fermented distillers＇ dried grains with solubles （DDGS） on the faecal microbial composition and diversity using 454 pyrosequencing. Healthy crossbred （Durocx Yorkshirex Landrace） growing and fattening pigs （n=48）, with an average initial body weight of 65 kg, were divided into two groups （24 replicates per group; four pens per group; six pigs per pen）, and given either DDGS feed as the control, or B. coagulans-fermented DDGS feed as the treatment. Faecal samples were collected on day 0, 7, 14, 21, and 28. DNA was extracted, and the V3-V6 region of the 16S rRNA gene was amplified. The fermented DDGS feed affected the relative abundance of bacteria populations at the phylum, genus, and species levels. At the genus level, the consumption of fermented DDGS feed led to higher relative abundances of faecal Prevotella, Lactobacillus, Clostridium, Bifidobacterium, Roseburia, and Bacillus, and lower relative abundances of faecal Escherichia, Ruminococcus, Dialister, unclassified Lachnospiraceae, unclassified Ruminococcaceae, and unclassified Enterobacteriaceae than in the control. At the species level, the consumption of fermented DDGS feed led to higher relative abundances of faecal Prevotella sp., Lactobacillus johnsonii, Lactobacillus fermentum, Lactobacillus mucosae, Lactobacillus reuteri, Clostridium butyricum, Bifidobacterium sp., and Roseburia sp., and lower relative abundances of faecal Prevotella copri, Escherichia coil, Ruminococcus gnavus, Ruminococcus flavefaciens, and Dialister sp. than in the control. Principal coordinates analysis indicated a distinct separation in the faecal microbial communities of pigs that were fed the fermented and unfermented DDGS feed. Fermented DDGS feed significantly increased the average daily gain （ADG） of pigs, and significantly decreased the average daily feed intake （ADFI） of feed and feed/gain （F/G）. Thus, our results demonstrate a beneficial shift in the faecal microbiota of pigs consuming fermented DDGS feed, with potential applications in livestock production.

Supplementation of a lacto-fermented rapeseed-seaweed blend promotes gut microbial-and gut immune-modulation in weaner piglets

Background:The direct use of medical zinc oxide in feed will be abandoned after 2022 in Europe,leaving an urgent need for substitutes to prevent post-weaning disorders.Results:This study investigated the effect of using rapeseed-seaweed blend(rapeseed meal added two brown macroalgae species Ascophylum nodosum and Saccharina latissima)fermented by lactobacilli(FRS)as feed ingredients in piglet weaning.From d 28 of life to d 85,the piglets were fed one of three different feeding regimens(n=230 each)with inclusion of 0%,2.5% and 5% FRS.In this period,no significant difference of piglet performance was found among the three groups.From a subset of piglets(n=10 from each treatment),blood samples for hematology,biochemistry and immunoglobulin analysis,colon digesta for microbiome analysis,and jejunum and colon tissues for histopathological analyses were collected.The piglets fed with 2.5% FRS manifested alleviated intraepithelial and stromal lymphocytes infiltration in the gut,enhanced colon mucosa barrier relative to the 0% FRS group.The colon microbiota composition was determined using V3 and V1-V8 region 16S rRNA gene amplicon sequencing by Illumina NextSeq and Oxford Nanopore MinION,respectively.The two amplicon sequencing strategies showed high consistency between the detected bacteria.Both sequencing strategies indicated that inclusion of FRS reshaped the colon microbiome of weaned piglets with increased Shannon diversity.Prevotella stercorea was verified by both methods to be more abundant in the piglets supplied with FRS feed,and its abundance was positively correlated with colonic mucosa thickness but negatively correlated with blood concentrations of leucocytes and IgG.Conclusions:FRS supplementation relieved the gut lymphocyte infiltration of the weaned piglets,improved the colon mucosa barrier with altered microbiota composition.Increasing the dietary inclusion of FRS from 2.5% to 5% did not lead to further improvements.

Research Advance on Application of Microbial Fermented Fodder in Broilers Production: A Short Review

Recently, interest in animal production and poultry production in particular has increased, because poultry meat and eggs and their products are more widely consumed than other products. Human health is also considered one of the priorities. Nowadays, the fermentation industry has become very popular due to its great benefits in the pharmaceutical industry, the improvement of foodstuffs and the animal feed industry and its improvement. Moreover, microbial fermentation can enhance the content and digestibility of nutrients in nontraditional diets while decreasing the quantity of crude fiber and anti-nutritional elements such as tannins. The functional components (organic acids, bioactive peptides, lactic acid bacteria) found in microbial fermented feed can increase meat production in cattle, pigs, and sheep, and broiler productivity, carcass quality, intestinal health, immunological function, and so on. This paper examines the application study of microbial fermented feed in broiler production and serves as a resource for microbial fermented feed promotion and use in broiler production.

Effects of Microbial Fermentation Feeds on Growth Performance of Weaned Piglets

[ Objective] TO study the effects of microbial fermentation feeds on growth performance of piglets, and provide the basis for their promotion and application, [ Method ] All of 60 Duroc x Landrace x Yorkshire crossbred weaned piglets, weighing 8-10 kg, were randomly housed in six groups, 10 piglets in each group. The piglets in control groups were fed with traditional daily food which contained aureomycin at 70 mg/kg . BW and concentrate feeds without antibiotics. While the piglets in experimental groups were fed with concentrate feeds without antibiotics and microbial fermentation feeds. [ Result] The results indicated that the average daily gain of experimental groups raised by 5.56%, and the feed conversion ratio reduced by 3.53% compared to control group. Moreover, the number of probiotics in the feces of experimental groups increased and pH value decreased significantly compared with the control groups （P 〈 0.05）. [ Conclusion] Microbial fermentation feeds can be used to replace antibiotics feeds, and promote healthy growth of the piglets.

Fermented mixed feed alters growth performance,carcass traits,meat quality and muscle fatty acid and amino acid profiles in finishing pigs

This study was conducted to investigate the effects of fermented mixed feed(FMF)on growth performance,carcass traits,meat quality,muscle amino acid and fatty acid composition and mRNA expression levels of genes related to lipid metabolism in finishing pigs.In the present study,144 finishing pigs(Duroc×Berkshire×Jiaxing Black)were randomly allocated to 3 dietary treatments with 4 replicate pens per group and 12 pigs per pen.The dietary treatments included a basal diet(CON),a basal diet+5%FMF and a basal diet+10%FMF.The experiment lasted 38 d after 4 d of acclimation.The results showed that 5%and 10%FMF significantly increased the average daily gain(ADG)of the females but not the males(P<0.05),but FMF supplementation showed no impact on carcass traits.Moreover,10%FMF supplementation increased the meat color45 min and meat color24 h values,while it decreased the shear force relative to CON(P<0.05).In addition,10%FMF significantly increased the contents of flavor amino acids(FAA),total essential AA(EAA),total non-EAA(NEAA)and total AA relative to CON(P<0.05).Furthermore,the diet supplemented with 10%FMF significantly increased the concentration of n-3 polyunsaturated fatty acids(PUFA),n-6 PUFA and total PUFA,and the PUFA to saturated fatty acids ratio(P<0.05),suggesting that FMF supplementation increased meat quality.Moreover,compared with the CON,10%FMF supplementation increased the mRNA expression of lipogenic genes,including CEBPα,PPARγ,SREBP1 and FABP4,and upregulated the expression of unsaturated fatty acid synthesis(ACAA1 and FADS2).Together,our results suggest that 10%FMF dietary supplementation improved the female pigs’growth performance,improved the meat quality and altered the profiles of muscle fatty acids and amino acids in finishing pigs.This study provides a reference for the production of high-quality pork.

Effects of fermented protein feed on the growth performance of pond-raised crab

This study aimed to elucidate the effect of a pellet feed containing fermented proteins on pond-raised Chinese mitten crabs,Eriocheir sinensis in a 7-month crab feeding experiment in Gucheng Lake,Jiangsu province,China.The results showed that fermented protein could significantly increase mean weight gain,crab size and total yield compared with control group.Moreover,the mean weight of male crabs and the number of large-sized male crabs improved over those of the control group.However,feed efficiency ratio did not differ significantly between the two groups.The amino acid content in the muscles of the female crabs of both the groups was higher than that in the muscles of the male crabs.The present study suggests that fermented compound protein may be one kind of potential protein sources for E.sinensis.

Evaluation of nutritive value and in vitro rumen fermentation gas accumulation of de-oiled algal residues

Background：Algae are widely recognized for their high oil content and for exponentially accumulating biomass with particular potential to provide single cell protein for human consumption or animal feed.It is believed that along with biodiesel from algae,the high protein de-oiled algal residue may become an alternative feed supplement option in the future.This study was conducted to investigate de-oiled algal residue obtained from the common Chlorella species,Thalassiosira weissflogii,Selenarstrum capricornutum,Scenedesmus sp.,and Scenedesmus dimorphus for assessment as potential feed supplements for ruminants by comparing with soybean（Glycine max） meal and alfalfa（Medicago sativa） hay.Results：With the exception of T.weissflogii,algal residue had higher concentrations of Cu,Zn,and Mn and lower concentration of Ca,Mg,and K than soybean meal and alfalfa hay.The algal residue CP（crude protein）concentrations ranged from 140 to 445 g/kg DM and varied among the de-oiled residues.In vitro rumen fermentation gas accumulation curves indicated that algal biomass degradation potential was less than that of soybean meal or alfalfa hay by up to 41.7%.The gas production curve,interpreted with a dual pool logistic model,confirmed that the fraction sizes for fast fermenting and slow fermenting of de-oiled algal residues were smaller than those in soybean meal and alfalfa hay,and the fermenting rate of the fractions was also low.Conclusions：Inferior in vitro rumen gas accumulation from the five de-oiled algal residues suggests that these algal byproducts are less degradable in the rumen.

Pyrosequencing investigation into the influence of Cu^(2+),Zn^(2+),Fe^(2+)and I^- mixtures on fungal diversity and toxigenic fungal growth in a fermented liquid feed

A L_9(3~4) orthogonal experiment was conducted to evaluate the influence of 9 mixtures which consisted of Cu^(2+), Zn^(2+), Fe^(2+) and I^- ions at different ion concentrations on fungal diversity and toxigenic fungal growth in a Bacillus subtilis-fermented liquid feed(FLF) using pyrosequencing. The maximal Chao estimator and Shannon index were achieved in the FLF with a mixture of Cu^(2+)(200 mg/kg), Zn^(2+)(160 mg/kg),Fe^(2+)(150 mg/kg) and I^-(2.4 mg/kg). The minimal relative abundance of Aspergillus was achieved when a mixture of Cu^(2+)(200 mg/kg),Zn^(2+), Fe^(2+) and I^-was added to the FLF. Compared with Zn^(2+), Fe^(2+) and I^-,Cu^(2+) was the most important ion in inhibitingAspergillus growth. Adding Zn^(2+)(160 mg/kg), Cu^(2+), Fe^(2+) and I^- to the FLF minimized the relative abundance ofFusarium. It was Zn^(2+) instead of Cu^(2+) played a critical role in suppressing the growth of Fusarium. The proper use of the mixture of Cu^(2+), Zn^(2+), Fe^(2+) and I^-in FLF contributes to inhibit the growth of mycotoxin-producing fungi during storage. The new findings of this study help farmers properly use the mixture of Cu^(2+),Zn^(2+), Fe^(2+) and I^- to inhibit the growth of mycotoxinproducing fungi in the production of high quality FLF and alleviate mycotoxins damages to animals and humans.

Aquafeed fermentation improves dietary nutritional quality and benefits feeding behavior,meat flavor,and intestinal microbiota of Chinese mitten crab(Eriocheir sinensis)

Normally,proper fermentation can be an efficient and widely used method to improve feed quality in animal rearing;however,the studies on crustaceans,especially Eriocheir sinensis,remain limited.This study aimed to investigate whether feed fermentation could meliorate dietary nutritional value and benefit E.sinensis rearing.First,non-fermented feed(NFD)and fermented feed(FD)were produced and assessed,respectively.Then,the“Y”maze feed choice behavior test(180 times;30 times,6 rounds)was conducted to assess the attractiveness of these 2 feeds for crabs.Finally,a total of 80 crabs(44.10±0.80 g)were randomly assigned into 2 groups with 4 replicates,and fed the experimental diets for 8 weeks to evaluate the effects of each feed on growth,antioxidant capacity,meat flavor,and intestinal microbiota.In this study,FD showed higher levels of crude protein(P<0.01),soluble protein(P<0.01),amino acids(P<0.05),lactic acid(P<0.001),and lower levels of crude fiber(P<0.05)and antinutritional factors(agglutinin,trypsin inhibitor,glycinin,andβ-conglycinin)(P<0.001)than NFD.Additionally,FD was more attractive to crabs than NFD(P<0.01)and it stimulated the appetite of crabs more than NFD(P<0.05).The growth performance,feed efficiency,and digestive enzyme activity of FDfed crabs were significantly higher than those of NFD-fed crabs(P<0.05).The electronic sensory measurements and free amino acid profiles revealed that the FD diet had positive impacts on the meat flavor of crabs,particularly in“sweet”and“umami”tastes.Moreover,the antioxidant capacity of FD-fed crabs was significantly higher than that of NFD-fed crabs(P<0.05).Fermented feed also affected the diversity and composition of intestinal microflora.The functional prediction of microbial communities showed that crabs fed FD had a better microecological environment in the intestine.In conclusion,the fermentation of aquafeed could be an effective approach to enhance feed quality and therefore benefit E.sinensis rearing.