Phosphorus recovery from biogas fermentation liquid by Ca-Mg loaded biochar

Shortage in phosphorus （P） resources and P wastewater pollution is considered as a serious problem worldwide. The application of modified biochar for P recovery from wastewater and reuse of recovered P as agricultural fertilizer is a preferred process. This work aims to develop a calcium and magnesium loaded biochar （Ca-Mg/biochar） application for P recovery from biogas fermentation liquid. The physico-chemical characterization, adsorption efficiency, adsorption selectivity, and postsorption availability of Ca-Mg/biochar were investigated. The synthesized Ca-Mg/biochar was rich in organic functional groups and in CaO and MgO nanoparticles. With the increase in synthesis temperature, the yield decreased, C content increased, H content decreased, N content remained the same basically, and BET surface area increased. The P adsorption of Ca-Mg/biochar could be accelerated by nano-CaO and nano-MgO particles and reached equilibrium after 360min. The process was endothermic, spontaneous, and showed an increase in the disorder of the solid-liquid interface. Moreover, it could be fitted by the Freundlich model. The maximum P adsorption amounts were 294.22, 315.33, and 326.63 mg/g. The P adsorption selectivity of Ca-Mg/biochar could not be significantly influenced by the typical pH level of biogas fermentation liquid. The nano-CaO and nano-MgO particles of Ca-Mg/biochar could reduce the negative interaction effects of coexisting ions. The P releasing amounts of postsorption Ca-Mg/biochar were in the order of Ca-Mg/B600 〉 Ca-Mg/B4S0 〉 Ca-Mg/B300. Results revealed that postsorption Ca-Mg/biochar can continually release P and is more suitable for an acid environment.

Sludge fermentation liquid addition attained advanced nitrogen removal in low C/N ratio municipal wastewater through short-cut nitrification-denitrification and partial anammox

Biological nitrogen removal of wastewater with low COD/N ratio could be enhanced by the addition of wasted sludge fermentation liquid(SFL),but the performance is usually limited by the introducing ammonium.In this study,the process of using SFL was successfully improved by involving anammox process.Real municipal wastewater with a low C/N ratio of 2.8–3.4 was treated in a sequencing batch reactor(SBR).The SBR was operated under anaerobic-aerobic-anoxic(AOA)mode and excess SFL was added into the anoxic phase.Stable short-cut nitrification was achieved after 46d and then anammox sludge was inoculated.In the stable period,effluent total inorganic nitrogen(TIN)was less than 4.3 mg/L with removal efficiency of 92.3%.Further analysis suggests that anammox bacteria,mainly affiliated with Candidatus_Kuenenia,successfully reduced the external ammonia from the SFL and contributed approximately 28%–43%to TIN removal.Overall,this study suggests anammox could be combined with SFL addition,resulting in a stable enhanced nitrogen biological removal.

Microbial inoculants and garbage fermentation liquid reduced root-knot nematode disease and As uptake in Panax quinquefolium cultivation by modulating rhizosphere microbiota community

Objective:To find a suitable ecological cultivation measure to solve the problem of root-knot nematode disease of Panax quinquefolium(Panacis Quinquefolii Radix)and the heavy metals accumulating in its roots.Methods:Three-year-old P.quinquefolium was treated with four different combinations of microbial inoculant(MI)and garbage fermentation liquid(GFL)[the joint application of‘TuXiu’MI and Fifty potassium MI(TF),the combination use of‘No.1'MI and Fifty potassium MI(NF),‘Gulefeng’poly-γ-glutamic acid MI(PGA),GFL],and the untreated control(CK).Here,high-throughput sequencing,ICP-MS and UPLC were employed to systematically characterize changes of microbial diversity and structure composition,heavy metals(As,Cd and Pb)content and ginsenoside content among different treatments.Results:The results revealed that different MIs and GFL could increase the root dry weight of P.quinquefolium,PGA enhanced it by 83.24%,followed by GFL(49.93%),meanwhile,PGA and GFL were able to lessen root-knot nematode disease incidence by 57.25%and 64.35%.The treatment of PGA and GFL can also effectively reduce heavy metals in roots.The As content in GFL and PGA was decreased by 52.17%and 43.48%respectively,while the Cd and Pb contents of GFL and PGA was decreased somewhat.Additionally,the content of total ginsenosides was increased by 42.14%and 42.07%,in response to TF and NF,respectively.Our metagenomic analysis showed that the relative abundance of particular soil microbial community members related to the biocontrol of root-knot nematode disease and plant pathogen(i.e.,Chaetomium in NF,Xylari in GFL,and Microascus in PGA),heavy metal bioremediation(Hyphomacrobium in PGA and Xylaria in GFL),and nitrogen fixation(Nordella and Nitrospira in TF)was significantly increased;notably,potential harmful microflora,such as Plectosaphaerella and Rhizobacter,were more abundant in the control group.Conclusion:MI and GFL could improve the quality of P.quinquefolium by modifying its rhizosphere microbial community structure and composition,both of them are beneficial to the development of ecological cultivation of P.quinquefolium.

Purification of four strains of endophytic fungi from Astragalus and their optimized liquid fermentations

This study was designed to isolate endophytic fungi from A. mongholicus （growing in northeast China） to determine whether they can produce bioactive metabolites. Four strains of endophytic fungi （strains 16, 17, 23 and 75） were successfully isolated from A. mongholicus using the surface disinfection method. According to ITS-rDNA sequences analysis, strains 16 and 75 were identified as Fusarium oxysporum, and strains 17 and 23 were identified as Bionectria ochroleuca. We applied the Box-Behnken design （BBD） to optimize the liquid fermentation conditions and obtain the maximum cell dry weight （CDW） yield. Opti-mal parameters were obtained under the following experimental condi-tions：temperature of 28＆#176;C, potato dextrose agar （PDA） liquid medium of 80 mL and rotation speed of 150 rpm. The four isolated endophytic fungi did not produce astragalosides I-IV, flavonoids or polysaccharides. Iso-lation of additional species of endophytic fungi from A. mongholicus and determination of their capacity to produce biologically active substances are subjects in need of further research.

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.

Fermentation Conditions on the Production of Single Cell Protein Feed by Water Chestnut Peel

In order to improve the comprehensive utilization value of water chestnut peel and the income of farmers. Contents of crude protein （ CP）, total sugar and reducing sugar were taken as indicators. Effects of initial pH, ratio of yeast to fungi species, incubation time and liquid volume on production of single cell pro- tein （SCP） feed was studied, and technological conditions on production of SCP feed by water chestnut peel were optimized by orthogonal test. Results showed that the production of SCP feed by water chestnut peel was optimal when pH was 5.0, ratio of yeast to fungi species was 2： 1, fermentation time was 2 d and the liquid volume was 70 mL / 250 mL. Under the optimum conditions, content of fermentation CP was 64.25%, content of total sugar was 19.8%, content of reducing sugar was 5.0%, content of coarse fibre was 0.0% and content of ash was 8.04%.

Matrix-assisted laser desorption ionization mass spectrometry based quantitative analysis of cordycepin from Cordyceps militaris

Cordycepin,which has great immunomodulatory activities such as anticancer,antifungal,antivirus,antileukemia and lipid-lowering ones,is the secondary metabolite of Cordyceps militaris(C.militaris).Liquid submerged fermentation is the common cultivation process to produce cordycepin.To optimize the fermentation process and improve production,monitoring the cordycepin secretion in the fermentation is essential.The measurement based on chromatography-mass spectrometry methods is generally involved in the complex sample pretreatments and time-consuming separation,so more rapid and convenient methods are required.Matrix-assisted laser desorption ionization mass spectrometry(MALDI-MS)is more attractive for faster and direct detection.Therefore,MALDI-MS detection combined with isotope-labeled internal standard was applied to the measurement of cordycepin content in the fermentation broth and mycelium.This method made accurate quantification of cordycepin in the range of 5-400μg/mL with a relative standard deviation of 5.6%.The recovery rates of fermentation samples after the 1,13,and 25 days were 90.15%,94.27%,and 95.06%,respectively.The contents of cordycepin in the mycelium and fermentation broth were 136 mg/g and 148.39 mg/mL on the 20 th culture day,respectively.The cordycepin secretion curve of the liquid fermentation of C.militaris was real-time traced over 25 days.

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.

Identification of differential metabolites in liquid diet fermented with Bacillus subtilis using gas chromatography time of flight mass spectrometry

Growth and health responses of pigs fed fermented liquid diet are not always consistent and causes for this issue are still not very clear. Metabolites produced at different fermentation time points should be one of the most important contributors. However, currently no literatures about differential metabolites of fermented liquid diet are reported. The aim of this experiment was to explore the difference of metabolites in a fermented liquid diet between different fermentation time intervals. A total of eighteen samples that collected from Bacillus subtilis fermented liquid diet on days 7, 21 and 35 respectively were used for the identification of metabolites by gas chromatography time of flight mass spectrometry(GCTOF-MS). Fifteen differential metabolites including melibiose, sortitol, ribose, cellobiose, maltotriose,sorbose, iso maltose, maltose, fructose, D-glycerol-1-phosphate, 4-aminobutyric acid, beta-alanine, tyrosine, pyruvic acid and pantothenic acid were identified between 7-d samples and 21-d samples. The relative level of melibiose, ribose, maltotriose, D-glycerol-1-phosphate, tyrosine and pyruvic acid in samples collected on day 21 was significantly higher than that in samples collected on day 7(P < 0.01),respectively. Eight differential metabolites including ribose, sorbose, galactinol, cellobiose, pyruvic acid,galactonic acid, pantothenic acid and guanosine were found between 21-d samples and 35-d samples.Samples collected on day 35 had a higher relative level of ribose than that in samples collected on day 21(P < 0.01), In conclusion, many differential metabolites which have important effects on the growth and health of pigs are identified and findings contribute to explain the difference in feeding response of fermented liquid diet.

Chemical characteristics changes associated with extracted protein and salt addition and effect of different additives in the curing process on sensory acceptance of liquid fermented fish from tilapia frame

Liquid fermented fish(Nam pla ra)is a traditional local condiment and popular consumption in Thailand.According to previous research,tilapia frame could be used as a raw material for the production,but the amount of protein received is still below the standard.In addition,there is no information of additives in curing process on consumer preferences.In this research,response surface methodology was used to study the influence of extracted protein from soybean meal(5-15%)and salt(15-20%)contents on chemical characteristics of Nam pla ra after 4 months of fermentation.The amount of roasted rice and roasted rice bran addition in curing process on sensory preferences were also studied.The results showed the linear effect of extracted protein and salt contents on NaCl content(P<0.05).When extracted protein increased,NaCl content of sample decreased,while increased salt concentration,NaCl content increased.Extracted protein showed linear relationship with autolytic degradation products and formaldehyde nitrogen of sample(P<0.05).Increase in extracted protein concentration caused an increment to both autolytic degradation products and formaldehyde nitrogen.In addition,salt content showed linear relationship with protein,total nitrogen,ammonia nitrogen and amino nitrogen(P<0.05).When increased salt concentration,protein and all kind of nitrogen tested in this study decreased.The statistical models for protein and NaCl contents(as an index of product quality)and our experimental validation,indicated that the models were appropriate to predict values.For acceptability test,there was no significant different in all attributes scores of cured samples with 10%roasted rice,roasted rice bran and mixture of both.In addition,their showed a significant different in all attributes scores lower than the commercial sample.In conclusion,the highest protein content was found in a treatment prepared by added salt content 15%by weight of digested material.Addition of proteins extracted from soybean meal did not lead to increase in protein content of finished product.However,chemical properties of all treatments could meet Thai community product standard.

Spraying compound probiotics improves growth performance and immunity and modulates gut microbiota and blood metabolites of suckling piglets

One factor that shapes the establishment of early neonatal intestinal microbiota is environmental microbial exposure,and probiotic application has been shown to promote health and growth of piglets.Thus,this study hypothesized that environmental probiotic application in early days of life would be beneficial to newborn piglets.This study aimed to investigate the effect of spraying a compound probiotic fermented liquid(CPFL)into the living environment of piglets on their early growth performance and immunity.This work included 68 piglets,which were randomized into probiotic and control groups.Blood and fecal samples were collected at 0,3,7,14,and 21 days of age.Spraying CPFL significantly reshaped the microbiota composition of the delivery room environment,increased piglets’daily weight gain and weaning weight(P<0.001),and modulated piglets’serum cytokine levels(increases in Ig A,Ig G,and IL-10;decrease in IFN-γ;P<0.05 in each case)in piglets.Additionally,spraying CPFL during early days of life modified piglets’gut microbiota structure and diversity,increased the abundance of some potentially beneficial bacteria(such as Bacteroides uniformis,Butyricimonas virosa,Parabacteroides distasonis,and Phascolarctobacterium succinatutens)and decreased the abundance of Escherichia coli(P<0.05).Interestingly,CPFL application also significantly enhanced the gut microbial bioactive potential and levels of several serum metabolites involved in the metabolism of vitamins(B2,B3,B6,and E),medium/long-chain fatty acids(caproic,tetradecanoic,and peptadecanoic acids),and dicarboxylic acids(azelaic and sebacic acids).Our study demonstrated that spraying CPFL significantly could improve piglets’growth performance and immunity,and the beneficial effects are associated with changes in the gut microbiota and host metabolism.Our study has provided novel data for future development of probiotic-based health-promoting strategies and expanded our knowledge of probiotic application in animal husbandry.

Effect of the addition of organic carbon sources on nitrous oxide emission in anaerobic-aerobic(low dissolved oxygen)sequencing batch reactors

The effect of additional organic carbon sources on the production of nitrous oxide(N_(2)O)in anaerobicaerobic(low dissolved oxygen)real wastewater treatment system was investigated.In this paper,three laboratoryscale sequencing batch reactors(SBRs)(SBR-1,SBR-2 and SBR-3)were operating under an anaerobic-aerobic(low dissolved oxygen,0.15-0.45 mg·L^(-1))configuration.The SBRs were‘long-term cultured’respectively with a single municipal wastewater sample,sodium acetate,and a waste-activated sludge alkaline fermentation liquid as the additional carbon sources of real wastewater.Off-gas analysis showed that N_(2)O was emitted into the atmosphere during the aerobic(low dissolved oxygen)period in the three SBRs,and the order of N_(2)O emission rate was SBR-2>SBR-1>SBR-3.It was observed that the higher poly-β-hydroxyvalerate fraction of polyhydroxyalkanoates,the lower glycogen transformation and less nitrite accumulation was in SBR-3,while the opposite behavior was observed in SBR-2.Further research indicated that the interaction of the factors above potentially affected the N_(2)O emission in the anaerobic-aerobic(low dissolved oxygen)system.