Synergistic benefits of Funneliformis mosseae and Bacillus paramycoides: Enhancing soil health and soybean tolerance to root rot disease.

To explore the response of soil metabolite composition to soybean disease, the effect of the combined inoculation of arbuscular mycorrhizal fungi (AMF) and plant growth-promoting bacteria on soybean root rot caused by Fusarium oxysporum was studied. A factorial completely randomized design with three factors (AMF, Bacillus. paramycoides, and rot disease stress) was conducted, and eight treatments, including normal groups and stress groups, were performed using pot experiments. GC‒MS and enzymatic assays were used to evaluate the soil factors and soybean growth indicators. The results showed that there were significant differences in the composition of metabolites among the different treatment groups, and 23 metabolites were significantly related to soybean biomass. The combined inoculation of Funneliformis mosseae and Bacillus paramycoides resulted in a significant reduction in harmful soil metabolites associated with root rot disease, such as ethylbenzene and styrene. This reduction in metabolites contributed to improving soil health, as evidenced by enhanced soybean defence enzyme activities and microbial activity, and ß-1,3-glucanase, chitinase and phenylalanine ammonia-lyase activities were improved to alleviate plant rhizosphere stress. Furthermore, soybean plants inoculated with the synergistic treatments exhibited reduced root rot disease severity and improved growth indicators compared to control plants. Plant height, root dry weight (RDW), and shoot and root fresh weight (SRFW) were improved by 4.18-53.79%, and the AM fungal colonization rate was also improved under stress. The synergistic application of Funneliformis mosseae and Bacillus paramycoides can effectively enhance soil health by inhibiting the production of harmful soil metabolites and improving soybean tolerance to root rot disease. This approach holds promise for the sustainable management of soil-borne diseases in soybean cultivation.

Screening of symbiotic Streptomyces spp. and optimization of microalgal growth in a microalgae-actinomycetes co-culture system.

Microalgal biodiesel as a substitute for fossil energy has attracted extensive attention. However, the high cost of microalgae cultivation limits the industrial production of microalgal biodiesel. The co-culture system may offer a means to increase microalgae's biomass production. In this study, Streptomyces strains were selected to construct and optimize co-culture systems with Monoraphidium sp. HDMA-11 and the algal cell biomass, lipid content, phycocyanin content, starch content, and fatty acid composition were determined. The results showed that Streptomyces nojiriensis significantly promoted Monoraphidium sp. HDMA-11 growth and a co-culture system were established. Orthogonal experiments showed that the Monoraphidium sp. HDMA-11 biomass was further increased when the initial culture pH was 7.5, the inoculation time of Streptomyces strain supernatants was 36 h, the volume ratio of microalgal actinomycetes was 1:1, and no additional acetic acid was added. Under these conditions, compared with monocultured Monoraphidium sp. HDMA-11, the cell biomass and lipid productivity of the co-culture system increased by 525.8 and 155.1%, respectively. These results suggest that S. nojiriensis supernatant potentially enhances microalgae biomass and may represent a new method to improve microalgae growth.

The response surface optimization of exopolysaccharide produced by Saccharomyces cerevisiae Y3 and its partial characterization.

Response surface methodology (RSM) was used to optimize the conditions of exopolysaccharides (EPSs) by Saccharomyces cerevisiae Y3. The results indicated that the yield of EPS reached 4.52 ± 0.14 g/L with 10.30% (w/v) sucrose, 0.64% (w/v) yeast extract, liquid volume 141.5 mL, which was 2.40 times the original EPS yield. Y3 EPS contained 83.65 ± 0.16% of total sugars, 15.27 ± 0.26% of uronic acid, 0.78 ± 0.02% of protein and 0.30 ± 0.12% of sulfuric acid groups. Y3 EPS maintained a relatively low viscosity, with intrinsic viscosities of 306.58 mL/g (25 °C) and 200.91 mL/g (35 °C), respectively. The EPS had high water solubility index (WSI), high water holding capacity (WHC) and good emulsifying ability (EA). Meanwhile, the EPS could absorb metal ions such as Cu2+, Fe2+ and Zn2+. In addition, Y3 EPS exhibited good antioxidant properties and coagulated skim milk with a concentration-dependent manner. These results indicated that S. cerevisiae Y3 EPS had applicable prospects in medicine, food, especially the dairy industry.

Effects of pyruvate decarboxylase (pdc1, pdc5) gene knockout on the production of metabolites in two haploid Saccharomyces cerevisiae strains.

Saccharomyces cerevisiae has good reproductive ability in both haploid and diploid forms, a pyruvate decarboxylase plays an important role in S. cerevisiae cell metabolism. In this study, pdc1 and pdc5 double knockout strains of S. cerevisiae H14-02 (MATa type) and S. cerevisiae H5-02 (MATα type) were obtained by the Cre/loxP technique. The effects of the deletion of pdc1 and pdc5 on the metabolites of the two haploid S. cerevisiae strains were consistent. In S. cerevisiae H14-02, the ethanol conversion decreased by 30.19%, the conversion of glycerol increased by 40.005%, the concentration of acetic acid decreased by 43.54%, the concentration of acetoin increased by 12.79 times, and the activity of pyruvate decarboxylase decreased by 40.91% compared to those in the original H14 strain. The original S. cerevisiae haploid strain H14 produced a small amount of acetoin but produced very little 2,3-butanediol. However, S. cerevisiae H14-02 produced 1.420 ± 0.063 g/L 2,3-BD. This study not only provides strain selection for obtaining haploid strains with a high yield of 2,3-BD but also lays a foundation for haploid S. cerevisiae to be used as a new tool for genetic research and breeding programs.

Enhanced ß-mannanase production by Bacillus licheniformis by optimizing carbon source and feeding regimes.

Bacillus licheniformis HDYM-04 was isolated in flax retting water and showed ß-mannanase activity. Carbon sources for ß-mannanase production, as well as the fermentation conditions and feeding strategy, were optimized in shake flasks. When glucose or konjac powder was used as the carbon source, the ß-mannanase activity was 288.13 ± 21.59 U/mL and 696.35 ± 23.47 U/mL at 24 h, respectively, which was approximately 4.4- to 10.68-fold higher than the values obtained with wheat powder. When 0.5% (w/v) glucose and 1% (w/v) konjac powder were added together, maximum enzyme activities of 789.07 ± 25.82 U/mL were obtained, an increase of 13.35% compared to the unoptimized cultures with only 1% (w/v) konjac powder. The enzyme activity decreased in the presence of 1% (w/v) konjac powder, but the highest enzyme activity was 1,533.26 ± 33.74 U/mL, a 1.2-fold increase compared with that in nonoptimized cultures; when 0.5% (w/v) glucose was used, the highest enzyme activity was 966.53 ± 27.84 U/mL, an increase in ß-mannanase activity of 38.79% compared with control cultures. In this study, by optimizing fed-batch fermentation conditions, the yield of ß-mannanase produced by HDYM-04 was increased, laying the foundation for the industrial application and further research of B. licheniformis HDYM-04.

The response surface optimization of ß-mannanase produced by Weissella cibaria F1 and its potential in juice clarification.

A ß-mannanase-producing lactic acid bacteria (LAB) was identified as Weissella cibaria F1 according to physiological and biochemical properties, morphological observations, partial sequence of 16S rRNA gene and API 50 CHL test. In order to improve the yield of ß-mannanase, the response surface methodology (RSM) was originally used to optimize the fermentation conditions. The optimization results showed that when the konjac powder, glucose, and initial pH were 9.46 g/L, 14.47 g/L and 5.67, respectively, the ß-mannanase activity increased to 38.81 ± 0.33 U/mL, which was 1.33 times compared to initial yield (29.28 ± 0.26 U/mL). This result was also supported by larger clearance on the konjac powder-MRS agar plate through Congo Red dyeing. The W. cibaria F1 ß-mannanase could improve the clarity of five fruits juice, i.e., apple, orange, peach, persimmon and blue honeysuckle. Among these, peach juice was the most obvious, clarity increasing by 12.8%. These results collectively indicated that W. cibaria F1 ß-mannanase had an applicable potential in food-level fields.

Optimization of cultivation strategy and medium for bacteriocin activity of Enterococcus faecium HDX-2.

A lactic acid bacteria (LAB) isolated from pickled Chinese cucumber was screened for bacteriocin production. The strain was identified to be Enterococcus faecium HDX-2. Based on the Plackett-Burman (PB) experiment, glucose, Ca2+, and initial pH were found to be the most significant parameters of bacteriocin production. Afterward, effects of the three main parameters on bacteriocin activity were further investigated by central composite design (CCD) and the optimum composition was glucose 22 g/L, Ca2+ 0.6 mM, and initial pH 7.2. Optimum results showed that bacteriocin activity was increased to 1337.60 ± 36.71 AU/mL, 2.23-fold higher than in MRS medium without parameters optimization. The bacteriocin also showed significant antimicrobial activity against Listeria monocytogenes in meat and cheese model system.

Bacteriocin production and inhibition of Bacillus subtilis by Lactobacillus paracasei HD1.7 in an indirect coculture system.

A broad-spectrum antimicrobial peptide named Paracin 1.7 was produced by Lactobacillus paracasei HD1.7, which was isolated from Chinese sauerkraut juice. In this study, the influence of cocultivation on the communication mechanism of L. paracasei HD1.7 and Bacillus subtilis was investigated. The two bacterial strains were grown in monoculture and indirect coculture, and the growth of both bacteria and bacteriocin production as well as the transcriptional level of luxS in L. paracasei HD1.7 and spo0A in B. subtilis were monitored. Bacteriocin production and cell numbers were increased significantly when L. paracasei HD1.7 cells were indirectly cocultured with B. subtilis, and bacteriocin-producing L. paracasei HD1.7 can prevent the growth and sporulation of B. subtilis. After indirect coculture with B. subtilis, the expression of luxS in L. paracasei HD1.7 increased in the exponential growth phase and decreased in the stationary phase compared to monoculture. The expression of spo0A in B. subtilis dropped in the indirect coculture compared to the monoculture. It indicate that the upregulation of luxS is due to a response to a secreted compound produced by B. subtilis. The results show L. paracasei HD1.7 has an amensalism on B. subtilis, while B. subtilis has a commensalism on L. paracasei HD1.7.

Disruption of the lactate dehydrogenase and acetate kinase genes in Klebsiella pneumoniae HD79 to enhance 2,3-butanediol production, and related transcriptomics analysis.

OBJECTIVES: 2,3-Butanediol (2,3-BD) is widely used in several chemical syntheses as well as the manufacture of plastics, solvents, and antifreeze formulations, and can be manufactured by microbial glucose fermentation. Conventional (2,3-BD) fermentation typically has low productivity, yield, and purity, and is expensive for commercial applications. We aimed to delete the lactate dehydrogenase and acetate kinase (ldhA and ack) genes in Klebsiella pneumoniae HD79 by using λRed homologous recombination technology, to eliminate by-products and thereby improve (2,3-BD) production. We also analyzed the resulting gene changes by using transcriptomics. RESULTS: The yield of (2,3-BD) from the mutant Klebsiella strain was 46.21 g/L, the conversion rate was 0.47 g/g, and the productivity was 0.64 g/L·h, which represented increases of 54.9%, 20.5%, and 106.5% respectively, compared to (WT) strains. Lactate and acetate decreased by 48.2% and 62.8%, respectively. Transcriptomics analysis showed that 4628 genes were differentially expressed (404 significantly up-regulated and 162 significantly down-regulated). Moreover, the (2,3-BD) operon genes were differentially expressed. CONCLUSION: Our data showed that the biosynthesis of (2,3-BD) was regulated by inducers (lactate and acetate), a regulator (BudR), and carbon flux. Elimination of acidic by-products by ldhA and ack knockdown significantly improved (2,3-BD) production. Our results provide a deeper understanding of the mechanisms underlying (2,3-BD) production, and form a molecular basis for the improvement this process by genetic modification in the future.

The response surface optimization of exopolysaccharide produced by Weissella confusa XG-3 and its rheological property.

The response surface methodology (RSM) was used to optimize the exopolysaccharide (EPS) production by a lactic acid bacteria (LAB) Weissella confusa XG-3. Two-level factorial design screened three significantly influencing factors sucrose, initial pH and sodium acetate. Central composite design (CCD) predicted under the condition of sucrose 80.1 g L-1, initial pH 5.8 and sodium acetate 3.7 g L-1, the maximal EPS yield obtained a 2.9-fold increase, reaching 97.5 ± 1.1 g L-1. This maximal value was far exceeding EPS production by other W. confusa species strains reported so far. The results suggested that W. confusa XG-3 had a potential for large-scale EPS production. The rheological properties of XG-3 EPS was further investigated. It was a typical non-Newtonian fluid, exhibiting pseudo-plastic behavior. The EPS concentration and temperature exerted positive and negative impact on apparent viscosity, respectively. The XG-3 EPS maintained relatively higher viscosity at moderate pH (6-8). The intrinsic viscosity [η] was 409.7 (25 °C) and 201.7 (35 °C), which was relevant to temperature but irrelevant to EPS concentration. This EPS efficiently coagulated sucrose-supplemented milk in a concentration-dependent manner. These results indicated that XG-3 EPS had an applicable potential in food processing fields especially dairy products.

Monoraphidium sp. HDMA-20 is a new potential source of α-linolenic acid and eicosatetraenoic acid.

BACKGROUND: ω-3 polyunsaturated fatty acids (PUFAs) are synthesized from α-Linolenic acid (ALA, C18:3ω3) and play important roles in anti-inflammatory and antioxidant responses in mammal cells. ALA is an essential fatty acid which cannot be produced within the human body and must be acquired through diet. The purpose of this study was to evaluate the potential of a novel microalgal strain (HDMA-20) as a source of ω-3 PUFAs including ALA and eicosatetraenoic acid (ETA, C20:4ω3). METHOD: Phylogenetic Neighbor-Joining analysis based on 18S ribosomal DNA sequence was used to identify the microalga strain HDMA-20. Autotrophic condition was chosen to cultivate HDMA-20 to reduce the cultivation cost. GC-MS was used to determine the fatty acid composition of HDMA-20 lipid. RESULTS: A microalgal strain (HDMA-20) from Lake Chengfeng (Daqing, Heilongjiang province, China) was found to accumulate high content of ω-3 PUFAs (63.4% of total lipid), with ALA and eicosatetraenoic acid (ETA, C20:4ω3) accounting for 35.4 and 9.6% of total lipid, respectively. Phylogenetic analysis based on 18S ribosomal DNA sequences suggested that the HDMA-20 belonged to genus Monoraphidium (Selenastraceae, Sphaeropleales) and its 18S rDNA sequence information turned out to be new molecular record of Monoraphidium species. The biomass productivity and lipid content of HDMA-20 were also investigated under autotrophic condition. The biomass productivity of HDMA-20 reached 36.3 mg L- 1 day- 1, and the lipid contents was 22.6% of dry weight. CONCLUSION: HDMA-20 not only represent an additional source of ALA, but also a totally new source of ETA. The high content of ω-3 PUFAs, especially ALA, of HDMA-20, makes it suitable as a source of nutrition supplements for human health. In addition, HDMA-20 exhibited good properties in growth and lipid accumulation, implying its potential for cost-effective ω-3 PUFAs production in future.

The response surface optimization of ß-mannanase produced by Lactobacillus casei HDS-01 and its potential in juice clarification.

Lactic acid bacteria (LAB) is an ideal mannanase source due to the bio-safety guarantee. LAB can heterogeneously express ß-mannanase or be directly used as ß-mannanase-producing strains. This research originally optimized the fermentation condition for ß-mannanase produced by Lactobacillus casei HDS-01. The applicable potential of the crude enzyme in juice clarification was investigated. Two-factorial design screened out three factors, i.e., fermentation time (p = 0.0001), glucose (p = 0.0013), and initial pH (p = 0.0167), which significantly affected L. casei HDS-01 ß-mannanase activity. Under the predicted conditions resulting from the central composite design (CCD), i.e., fermentation time 18.23 hr, glucose 12.65 g L-1, initial pH 5.18, the model reached maximal ß-mannanase activity of 81.40 U mL-1. This model was validated by conducting six repeated experiments and subsequent t-test (p = 0.6308). RSM optimization obtained a 1.33-fold increase in ß-mannanase activity. This increase could also be qualitatively detected by larger clearance zone on konjac powder-MRS agar through Congo Red dyeing. The yield and clarity of crude ß-mannanase-treated juices from orange, apple, and pear were significantly higher than controls without enzyme treatment. This study conferred a relatively high ß-mannanase-producing LAB strain with a high bio-safety level and easy and economical use in juice clarification as well as other food-level fields.

Biosynthesis regulation of natamycin production from Streptomyces natalensis HDMNTE-01 enhanced by response surface methodology.

Natamycin has been widely applied in medical treatments and food protection widely due to its effective inhibition to the growth of yeast and mold. As polyene macrolide antibiotic, the biosynthesis pathway of natamycin is relatively clear. To regulate the biosynthesis of natamycin, additions of precursors affecting cell growth and natamycin production were investigated. The results showed that 0.003% (w/v) potassium ferrocyanide and sodium propionate: n-butanol at a ratio of 4:1 was added into the broth at 0 and 24 hr, respectively, and they contributed to yield natamycin, reaching 1.62 g L-1 (174.6% higher than control). Furthermore, response surface methodology was undertaken to enhance natamycin production by Streptomyces natalensis HDMNTE-01 (a wild strain). The optimum conditions determined were: glucose 3.97%; soya peptone 2%; yeast extract 0.5%; original pH 7.0; inoculum volume 6%; growth in a 250-mL flask containing 24.68 mL of medium; shaken (220 rpm) at 28°C for 4 days. Under the optimized conditions, the yield was 2.81 g L-1 natamycin in 5-L fermentor when the fermentation was processed.

Effect of small interfering RNA against Paracin 1.7 bacteriocin produced by Lactobacillus paracasei HD1-7.

Lactobacillus paracasei HD1-7 (CCTCCM 205015), isolated from Chinese sauerkraut fermentation broth, contains the bacteriocin Paracin 1.7 which possesses broad-spectrum antibacterial activity. The gene-silencing effect of small interfering RNA (siRNA) is a potential strategy for further understanding the mechanism of production of Paracin 1.7 by L. paracasei HD1-7. In this study, the effect of siRNA on the expression of the most important proteins in the production of Paracin 1.7, sensor kinase (prcK) and response regulator (prcR), was investigated. SiRNA were designed against prcK and prcR, and qRT-PCR was performed to examine the expression of prcK and prcR mRNA. The efficacy of siRNA was determined by comparing the level of antimicrobial activity of the strains. qRT-PCR showed that siRNA-K4 and siRNA-K5 significantly inhibited the expression of prcK mRNA, and siRNA-R4 and siRNA-R6 significantly inhibited the expression of prcR mRNA. The proteins levels and antibacterial activities of mutant strains were lower than the original and control groups, respectively. The results demonstrate that siRNA inhibited both mRNA expression and the production of Paracin 1.7 in L. paracasei HD1-7. Targeting of prcK and prcR with siRNA appears to be a novel strategy for researching the mechanism of Paracin 1.7 production by L. paracasei HD1-7.

[Construction of recombinant baculovirus vectors started by EF1alpha].

OBJECTIVE: To improve the transduction efficiency of baculovirus and exogenous gene expression level, we chose a mammalian cell-specific promoter-human extension factor 1alpha promoter (EF1-alpha), used virus pseudotyped tools--truncated vessicular stomatitis virus protein G (VSV-GED), added woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) and adenovirus inverted terminal repeats (ITRs). METHOD: We constructed two improved recombinant baculoviruses transfer vectors named pWK and pWK-ITR with the pFB-VSV-GED-WPRE. The recombinant transfer vectors pWK-eGFP, pWK-ITR-eGFP and pWK (-)-eGFP were constructed by inserting the Enhanced Green Fluorescent Protein (eGFP) reporter gene into the downstream of EF1alpha promoter. Constructed recombinant plasmid transfected Sf9 insect cells, and observed the expression of green fluorescent protein by using the inverted fluorescence microscope. RESULTS: The fluorescence expression rate of BV-WK-eGFP, BV-WK-ITR-eGFP containing WPRE and ITRs was significantly higher than the negative control, ITRs can effectively extend the expression time of eGFP, the expression time of eGFP in BV-WK-eGFP and BV-WK-ITR-eGFP increased 72 hours compared to the negative control BV-WK (-) -eGFP. The transduction time of VSV-GED pseudotyped baculovirus BV-WK-eGFP, BV-WK-ITR-eGFP was obviously shorten in OL cells, and reduced 24 hours compared to the negative control BV-WK (-) -eGFP, transduction efficiency were higher 25.7% and 36.5% than the negative control BV-WK (-) -eGFP, respectively. CONCLUSION: The experiments proved that the VSV-GED could effectively improve the transduction efficiency of baculovirus, WPRE could enhance the expression efficiency of the exogenous gene significantly, and ITRs extend the expression time. The research will lay a foundation to explore improved recombinant baculovirus express exogenous genes in vertebrate cells and research the new recombinant live vector vaccine.

[Construction of baculovirus vector with WPRE regulatory element to express Newcastle disease virus F gene in primary chicken embryo cells].

OBJECTIVE: To construct the recombinant baculovirus with mammaliancell-specific promoter and woodchuck hepatitis virus post-transcriptional regulatory element (WPRE), to highly express Newcastle disease virus (NDV) F gene in the primary chicken embryo cells. METHOD: We extracted total RNAs from NDV La Sota strain. Then the F gene was amplified by reverse transcription polymerase chain reaction. We constructed the baculoviral vector (pCMV-WPRE-F) with F gene fused with the WPRE near its 3'end, which expressed under the control of the CMV promoter. The F gene recombinant bacmid was obtained by Bac-to-Bac system and transfected into sf9 insect cells to acquire F gene recombinant baculovirus. After amplification of recombinant baculovirus, the recombinant virus was transfected into chicken primary cells with 50 multiplicity of infection, and the proteins were harvested at 72 h after infection. The F protein expression levels mediated by WPRE regulatory element were analyzed. RESULTS: Western blot results show that the F gene was successfully expressed in chicken primary cells. The product was a 56kDa protein and could be recognized by anti-NDV serum. The WPRE fusion significantly improved the F gene expression as 10 mmol/L butyrate did, but different to butyrate, the WPRE regulatory element was nontoxic to cells. CONCLUSION: The optimized recombinant baculovirus could efficiently deliver NDV F gene into chicken primary cells and express the F antigen protein. In addition, the WPRE regulatory element could increase the expression levels of exogenous gene mediated by baculovirus in chicken primary cells. The research provides us a potential basis for the gene engineered vaccines of NDV and other avian infectious disease based on baculovirus vector.

Ecological succession of abundant and rare subcommunities during aerobic composting in the presence of residual amoxicillin.

Aerobic composting increases the content of soluble nutrients and facilitates the safe treatment of livestock manure. Although different taxa play crucial roles in maintaining ecological functionality, the succession patterns of community composition and assembly of rare and abundant subcommunities during aerobic composting under antibiotic stress and their contributions to ecosystem functionality remain unclear. Therefore, this study used 16 S rRNA gene sequencing technology to reveal the response mechanisms of diverse microbial communities and the assembly processes of abundant and rare taxa to amoxicillin during aerobic composting. The results indicated that rare taxa exhibited distinct advantages in terms of diversity, community composition, and ecological niche width compared with abundant taxa, highlighting their significance in maintaining ecological community dynamics. In addition, deterministic (heterogeneous selection) and stochastic processes (dispersal limitation) play roles in the community succession and functional dynamics of abundant and rare subcommunities. The findings of this study may contribute to a better understanding of the relative importance of deterministic and stochastic assembly processes in composting systems, and the ecological functions of diverse microbial communities, ultimately leading to improved ecological environment.

Effect of combined bacteria on the flax dew degumming process: Substance degradation sequence and changes in functional bacteria taxa.

In this study, 16S rDNA high-throughput sequencing, Fourier transform infrared spectroscopy, and two-dimensional correlation spectroscopy techniques were used to analyze the mechanisms driving the sequence of degradation of gummy substances by the microbial community and hydrolytic enzymes during the flax dew degumming process. The results revealed that the inoculation of combined bacteria induced quorum sensing, modulated hydrolytic enzyme production, and reshaped the community structure. Lignin-degraded genera (Pseudomonas and Sphingobacterium) were enriched, and the relative abundances of pectin- and cellulose-degraded genera (Chryseobacterium) decreased in the early degumming stages. Hemicellulose-degraded genera (Brevundimonas) increased over the degumming time. Moreover, the abundance of lignin hydrolytic enzymes improved in the early stages, while the abundance of pectin hydrolytic enzymes increased at the end of degumming. Various types of functional bacteria taxa changed the sequence of substance degradation. Electron scanning microscopy and differential scanning calorimetry results indicated that the degumming, facilitated by the inoculation of combined bacteria, was nearly completed by 21 d. The fibers exhibited smoother and more intact properties, along with higher thermal stability, as indicated by a melting temperature of 71.54 °C. This study provides a reference for selecting precise degumming bacterial agents to enhance degumming efficiency.

[Construction of baculovirus vector with Cytomegaloviruse promoter to express eGFP in primary chicken embryo cells].

OBJECTIVE: Baculovirus is known as a safe vector candidate due to its non-replication in mammalian cells. The tropism to different cells and transduction efficiency can be improved by introducing cell-specific promoter, VSV-GED and different functional regulatory elements. The optimized pseudotyped recombinant baculovirus can express eGFP gene in primary chicken cells, which provides us a new approach to develop engineered poultry vaccines. METHOD: The pseudotyped recombinant baculoviruses were constructed with cytomegaoviyns (CMV) promotor, VSV-GED, woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) and inverted terminal repeats (ITRs). The recombinant baculoviruses contained eGFP reporter gene were transfected chicken primary cells, and the eGFP protein expression levels mediated by different baculoviruses were analyzed. RESULTS: The expression of eGFP was detected at 12 hours after infection. The transduction efficiency of the pseudotyped recombinant baculoviruses increased from 36% to 48.2% by inserting VSV-GED. The expression effect of eGFP in recombinant baculovirus carrying WPRE element was similar to that by adding 10 mmol/L butyrate. However, the WPRE elements are nontoxic to cells. Within 72 hours, the expression intensity of eGFP in the recombinant baculovirus with ITRs increased gradually. CONCLUSION: The VSV-GED element can improve the transduction efficiency and WPRE can increase the reporter gene eGFP expression levels mediated by baculovirus in chicken primary cells. The recombinant baculovirus with the ITRs elements can extend the expression time of eGFP.

Aerobic composting with sauerkraut fermentation waste water: Increasing the stability and complexity of bacterial community and changing bacterial community assembly processes.

Aerobic composting renders the sauerkraut fermentation waste water harmless while adding soluble nutrients. Unravelling the bacterial community assembly processes, changes in community robustness and community cohesion and the relationship between them under composting treatment of sauerkraut fermentation waste water is an interesting topic. Sauerkraut fermentation waste water was used for composting, which increased bacterial linkages, community robustness, competitive behaviour during warming periods and cooperative behaviour during cooling periods, and the control of community assembly processes shifts from deterministic processes (variable selection) to stochastic processes (decentralised limitation). At the same time, the influence of community robustness and community cohesion on community assembly processes was increased. Community cohesion and robustness were significantly correlated with community function. These results indicate that community robustness and community cohesion are critical for the biological handling of hazardous substances.