Preparation, purification, and biochemical of fat-degrading bacterial enzymes from pig carcass compost and its application.

BACKGROUND: A lot of kitchen waste oil is produced every day worldwide, leading to serious environmental pollution. As one of the environmental protection methods, microorganisms are widely used treating of various wastes. Lipase, as one of the cleaning agents can effectively degrade kitchen waste oil. The composting process of pig carcasses produces many lipase producing microorganisms, rendering compost products an excellent source for isolating lipase producing microorganisms. To our knowledge, there are no reports isolating of lipase producing strains from the high temperature phase of pig carcass compost. METHODOLOGY: Lipase producing strains were isolated using a triglyceride medium and identified by 16S rRNA gene sequencing. The optimal fermentation conditions for maximum lipase yield were gradually optimized by single-factor tests. The extracellular lipase was purified by ammonium sulfate precipitation and Sephadex G-75 gel isolation chromatography. Amino acid sequence analysis, structure prediction, and molecular docking of the purified protein were performed. The pure lipase's enzymatic properties and application potential were evaluated by characterizing its biochemical properties. RESULTS: In this study, a lipase producing strain of Bacillus sp. ZF2 was isolated from pig carcass compost products, the optimal fermentation conditions of lipase: sucrose 3 g/L, ammonium sulfate 7 g/L, Mn2+ 1.0 mmol/L, initial pH 6, inoculum 5%, temperature 25 ℃, and fermentation time 48 h. After purification, the specific activity of the purified lipase reached 317.59 U/mg, a 9.78-fold improvement. Lipase had the highest similarity to the GH family 46 chitosanase and molecular docking showed that lipase binds to fat via two hydrogen bonds at Gln146 (A) and Glu203 (A). Under different conditions (temperature, metal ions, organic solvents, and surfactants), lipase can maintain enzymatic activity. Under different types of kitchen oils, lipase has low activity only for 'chicken oil', in treating other substrates, the enzyme activity can exceed 50%. CONCLUSIONS: This study reveals the potential of lipase for waste oil removal, and future research will be devoted to the application of lipase.

The hidden costs of emotional labor on withdrawal behavior: the mediating role of emotional exhaustion, and the moderating effect of mindfulness.

BACKGROUND: Employees' withdrawal behavior concerns organization leaders and policymakers in many countries. However, the specific mechanism which emotional labor affects withdrawal behavior has yet to be thoroughly discussed. There needs to be systematic research on how different emotional labor strategies affect work withdrawal, whether directly or through individual perception, and how to respond. METHODS: A total of 286 hotel and catering service employees participated in our study. A series of hierarchical moderated regression analyses were performed to test the hypothesis. RESULTS: The results indicated that surface acting positively affected withdrawal behavior, while deep acting had a negative effect. Emotional exhaustion mediated in this relationship of surface acting with withdrawal behavior and deep acting with withdrawal behavior. Mindfulness showed moderation effects between emotional exhaustion and withdrawal behavior. CONCLUSIONS: Emotional labor and emotional exhaustion are significant in predicting employees' intentions to withdraw, given that emotional exhaustion partially mediates the effects of emotional labor on withdrawal behavior. Significantly, the relationship between emotional exhaustion and withdrawal behavior is weakened by mindfulness.

Recombineering: Genetic Engineering in Escherichia coli Using Homologous Recombination.

The bacterial chromosome and bacterial plasmids can be engineered in vivo by homologous recombination using either PCR products or synthetic double-stranded DNA (dsDNA) or single-stranded DNA as substrates. Multiple linear dsDNA molecules can be assembled into an intact plasmid. The technology of recombineering is possible because bacteriophage-encoded recombination proteins efficiently recombine sequences with homologies as short as 35 to 50 bases. Recombineering allows DNA sequences to be inserted or deleted without regard to the location of restriction sites and can also be used in combination with CRISPR/Cas targeting systems. © 2023 Wiley Periodicals LLC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA. Basic Protocol: Making electrocompetent cells and transforming with linear DNA Support Protocol 1: Selection/counter-selections for genome engineering Support Protocol 2: Creating and screening for oligo recombinants by PCR Support Protocol 3: Other methods of screening for unselected recombinants Support Protocol 4: Curing recombineering plasmids containing a temperature-sensitive replication function Support Protocol 5: Removal of the prophage by recombineering Alternate Protocol 1: Using CRISPR/Cas9 as a counter-selection following recombineering Alternate Protocol 2: Assembly of linear dsDNA fragments into functional plasmids Alternate Protocol 3: Retrieval of alleles onto a plasmid by gap repair Alternate Protocol 4: Modifying multicopy plasmids with recombineering Support Protocol 6: Screening for unselected plasmid recombinants Alternate Protocol 5: Recombineering with an intact λ prophage Alternate Protocol 6: Targeting an infecting λ phage with the defective prophage strains.

Extraction, Purification, Characterization and Application in Livestock Wastewater of S Sulfur Convertase.

Sulfide is a toxic pollutant in the farming environment. Microbial removal of sulfide always faces various biochemical challenges, and the application of enzymes for agricultural environmental remediation has promising prospects. In this study, a strain of Cellulosimicrobium sp. was isolated: numbered strain L1. Strain L1 can transform S2-, extracellular enzymes play a major role in this process. Next, the extracellular enzyme was purified, and the molecular weight of the purified sulfur convertase was about 70 kDa. The sulfur convertase is an oxidase with thermal and storage stability, and the inhibitor and organic solvent have little effect on its activity. In livestock wastewater, the sulfur convertase can completely remove S2-. In summary, this study developed a sulfur convertase and provides a basis for the application in environmental remediation.

Development of a microbial protease for composting swine carcasses, optimization of its production and elucidation of its catalytic hydrolysis mechanism.

BACKGROUND: Dead swine carcass composting is an excellent method for harmless treatment and resource utilization of swine carcass. However, poor biodegradation ability of traditional composting results in poor harmless treatment effect. Researches report that the biodegradation ability of composting can be improved by inoculation with enzyme-producing microorganisms or by inoculation with enzyme preparations. At present, the researches on improving the efficiency of dead swine carcass composting by inoculating enzyme-producing microorganisms have been reported. However, no work has been reported on the development of enzyme preparations for dead swine carcass composting. METHODOLOGY: The protease-producing strain was isolated by casein medium, and was identified by 16 S rRNA gene sequencing. The optimal fermentation conditions for maximum protease production were gradually optimized by single factor test. The extracellular protease was purified by ammonium sulfate precipitation and Sephadex G-75 gel exclusion chromatography. The potential for composting applications of the purified protease was evaluated by characterization of its biochemical properties. And based on amino acid sequence analysis, molecular docking and inhibition test, the catalytic hydrolysis mechanism of the purified protease was elucidated. RESULTS: In this study, a microbial protease was developed for swine carcass composting. A protease-producing strain DB1 was isolated from swine carcass compositing and identified as Serratia marcescen. Optimum fermentation conditions for maximum protease production were 5 g/L glucose, 5 g/L urea, 1.5 mmol/L Mg2+, initial pH-value 8, inoculation amount 5%, incubation temperature 30 °C and 60 h of fermentation time. The specific activity of purified protease reached 1982.77 U/mg, and molecular weight of the purified protease was 110 kDa. Optimum pH and temperature of the purified protease were 8 and 50 °C, respectively, and it had good stability at high temperature and in alkaline environments. The purified protease was a Ser/Glu/Asp triad serine protease which catalyzed substrate hydrolysis by Glu, Arg, Ser, Asp and Tyr active residues. CONCLUSIONS: In general, the microbial protease developed in this study was suitable for industrial production and has the potential to enhance composting at thermophilic stage. Moreover, the catalytic hydrolysis mechanism of the protease was further analyzed in this study.

Study on generation, migration and accumulation of CO in the mining goaf of shallow-buried close distance coal seam group.

There is complex air leakage in the mining of shallow buried close distance coal seam group, which affects the generation and migration of CO in the goaf, and easily leads to exceeding safety limits of CO in the return corner of the working face, which threatens the safety of underground production. To examine this problem, taking Lijiahao Coal Mine as an example, this study analyses the generation law of CO gas, the distribution law of overburden fractures, the characteristics of air leakage in the goaf, the sources of CO in the return corner, and the migration and accumulation law of CO in the goaf under multi-source air leakage in the mining of shallow buried close distance coal seam group through experiment tests, numerical simulations, observations and theoretical analyses. The results indicated that there is an exponential growth relationship between the CO generation rate and the coal temperature, and the critical temperature for rapid oxidation of coal samples is between 70 and 80 °C. The 31,115 working face has complicated air leakage from the working face and ground surface and the goaf of this coal seam. The surface air converges to the return corner through the mining fissure of overburden and 2-2 coal goaf, and the air leakage of the working face flows out from the return roadway through the goaf. The gas leakage in the overlying goaf and the oxidation of residual coal are the main sources of CO in the return corner. The CO generated during the coal mining process and the CO generated by the trackless rubber-tired vehicle operation will increase the CO concentration in the return corner to varying degrees. Under the effect of multi-source air leakage, CO from the overlying goaf and the residual coal in the goaf of this coal seam are migrated to the air return side of the goaf, resulting in the accumulation of CO in the return corner, and both of them have a linear positive correlation with the CO concentration in the return corner. The results of the study have scientific guidance for the control of air leakage and the prevention of CO excess in the goaf.

How Does Inclusive Leadership Curb Workers' Emotional Exhaustion? The Mediation of Caring Ethical Climate and Psychological Safety.

The COVID-19 pandemic has transformed the politics, economy, and society of the world, which has dealt the most severe blow to the hospitality industry. Meanwhile, the pandemic and government control policies have brought high psychological pressure to hospitality front-line employees, resulting in emotional exhaustion. As a part of burnout syndrome, emotional exhaustion poses a threat to employees' mental health, career sustainability, and well-being. Therefore, the purpose of this paper was to investigate the curb effectiveness of inclusive leadership on emotional exhaustion and to explore the mediation roles of ethical climate and psychological safety between them. Time-lagged data were collected from 65 teams and 358 hospitality front-line employees working in Chinese hotels in two stages with a one-month gap. This research verified that inclusive leadership has a negative impact on emotional exhaustion both indirectly through the mediation roles of ethical climate and psychological safety. And the ethical climate and psychological safety played partial mediation roles between inclusive leadership and emotional exhaustion. In theory, the findings explored the dual mediation mechanism of the inhibitory effect of inclusive leadership on emotional exhaustion. In practice, we provided the training and correct guidance to develop inclusive leadership for hotel enterprises and to resolve the emotional exhaustion of employees, which can enhance sustainability in careers.

Study on the air leakage characteristics of a goaf in a shallow coal seam and spontaneous combustion prevention and control strategies for residual coal.

Air leakage from surface mining-induced fissures can easily cause spontaneous combustion of residual coal in the goaf, which threatens the safe production of the underground working face. In order to study the air leakage law of the goaf under the surface air leakage and the prevention and control technology of spontaneous combustion of residual coal. Based on engineering data from the 6104 working face of the Chuancao Gedan coal mine, this study uses a combination of theoretical analyses, numerical simulations, and field observations to study the dynamic distribution characteristics of the air leakage velocity of surface mining-induced fissures in shallow coal seams, the distribution characteristics of relative pressure, the air leakage velocity, the air leakage flow field, the distribution ranges for the "three zones" of spontaneous combustion in the goaf, and a reasonable range for the pressurized ventilation of the working face. The results show that there is a quadratic relationship between the air leakage speed from the surface mining-induced fissures in shallow coal seams and the distance from the working face. The air leakage speed decreases as the distance from the working face increases, and the air leakage speed in the middle of the working face is slower than the air leakage on either side of the goaf. The pressure difference between the goaf and the surface mining-induced fissures is the root cause of air leakage into the goaf, and a change in the pressure difference has a significant impact on the air leakage flow field and the distributions of the "three zones" of spontaneous combustion in the goaf. When the pressure difference between the ground surface and the working face is maintained within the range of 200~-200 Pa, air leakage is effectively reduced, and the spontaneous combustion of residual coal is inhibited. The research results reveal the air leakage mechanism in the goaf of shallow coal seams and provide a reference for the prevention and control of spontaneous combustion of residual coal in the goaf.

Homologs of the Escherichia coli F Element Protein TraR, Including Phage Lambda Orf73, Directly Reprogram Host Transcription.

Bacterial cells and their associated plasmids and bacteriophages encode numerous small proteins of unknown function. One example, the 73-amino-acid protein TraR, is encoded by the transfer operon of the conjugative F plasmid of Escherichia coli. TraR is a distant homolog of DksA, a protein found in almost all proteobacterial species that is required for ppGpp to regulate transcription during the stringent response. TraR and DksA increase or decrease transcription initiation depending on the kinetic features of the promoter by binding directly to RNA polymerase without binding to DNA. Unlike DksA, whose full activity requires ppGpp as a cofactor, TraR is fully active by itself and unaffected by ppGpp. TraR belongs to a family of divergent proteins encoded by proteobacterial bacteriophages and other mobile elements. Here, we experimentally addressed whether other members of the TraR family function like the F element-encoded TraR. Purified TraR and all 5 homologs that were examined bound to RNA polymerase, functioned at lower concentrations than DksA, and complemented a dksA-null strain for growth on minimal medium. One of the homologs, λ Orf73, encoded by bacteriophage lambda, was examined in greater detail. λ Orf73 slowed host growth and increased phage burst size. Mutational analysis suggested that λ Orf73 and TraR have a similar mechanism for inhibiting rRNA and r-protein promoters. We suggest that TraR and its homologs regulate host transcription to divert cellular resources to phage propagation or conjugation without induction of ppGpp and a stringent response. IMPORTANCE TraR is a distant homolog of the transcription factor DksA and the founding member of a large family of small proteins encoded by proteobacterial phages and conjugative plasmids. Reprogramming transcription during the stringent response requires the interaction of DksA not only with RNA polymerase but also with the stress-induced regulatory nucleotide ppGpp. We show here that five phage TraR homologs by themselves, without ppGpp, regulate transcription of host promoters, mimicking the effects of DksA and ppGpp together. During a stringent response, ppGpp independently binds directly to, and inhibits the activities of, many proteins in addition to RNA polymerase, including translation factors, enzymes needed for ribonucleotide biosynthesis, and other metabolic enzymes. Here, we suggest a physiological role for TraR-like proteins: bacteriophages utilize TraR homologs to reprogram host transcription in the absence of ppGpp induction and thus without inhibiting host enzymes needed for phage development.

Genomic and Transcriptomic Analysis of Bovine Pasteurella multocida Serogroup A Strain Reveals Insights Into Virulence Attenuation.

Pasteurella multocida is one of the primary pathogens of bovine respiratory disease (BRD), and causes huge losses in the cattle industry. The Pm3 strain was a natural isolate, which is a strong form of pathogen and is sensitive to fluoroquinolones antibiotics. A high fluoroquinolone resistant strain, Pm64 (MIC = 64 µg/mL), was formed after continuous induction with subinhibitory concentration (1/2 MIC) of enrofloxacin, with the enhanced growth characteristics and large attenuation of pathogenicity in mice. This study reports the whole genome sequence and the transcription profile by RNA-Seq of strain Pm3/Pm64. The results showed an ineffective difference between the two strains at the genome level. However, 32 genes could be recognized in the gene islands (GIs) of Pm64, in which 24 genes were added and 8 genes were lost. Those genes are involved in DNA binding, trehalose metabolism, material transportation, capsule synthesis, prophage, amino acid metabolism, and other functions. In Pm3 strain, 558 up-regulated and 568 down-regulated genes were found compared to Pm64 strain, from which 20 virulence factor-related differentially expressed genes (DEGs) were screened. Mainly differentially transcribed genes were associated with capsular polysaccharide (CPS), lipopolysaccharide (LPS), lipooligosaccharide (LOS). Iron utilization, and biofilm composition. We speculated that the main mechanism of virulence attenuation after the formation of resistance of Pm64 comes from the change of the expression profile of these genes. This report elucidated the toxicity targets of P. multocida serogroup A which provide fundamental information toward the understanding of the pathogenic mechanism and to decreasing antimicrobial drugs resistance.

Efficient decomposition of lignocellulose and improved composting performances driven by thermally activated persulfate based on metagenomics analysis.

In this study, fresh dairy manure and bagasse pith were used as raw materials to study the effect of potassium persulfate in the aerobic composting process. The influence of sulfate radical anion (SO4-·) generated by thermally activated persulfate on physicochemical parameters, lignocellulose degradation, humic substance (HS) formation, microbial community succession, and carbohydrate-active enzymes (CAZymes) composition were assessed during composting. Experimental results showed that the degradation rates of cellulose, hemicellulose and lignin in the treatment group with potassium persulfate (PS) (61.47%, 74.63%, 73.1%) were higher than that in blank control group (CK) (59.98%, 71.47%, 70.89%), respectively. Additionally, persulfate additive promoted dynamic variation of dissolved organic matter (DOM) and accelerated the formation of HS. Furthermore, metagenomics analysis revealed that persulfate changed the structure of the microbial community, and the relative abundances of Actinobacteria and Proteobacteria increased by 17.64% and 34.09% in PS, whereas 12.09% and 29.96% in CK. Glycoside hydrolases (GHs) and auxiliary activities (AAs) families were crucial to degrade lignocellulose, and their abundances were more in PS. Redundancy analysis (RDA) manifested that Actinobacteria and Proteobacteria were closely associated with lignocellulosic degradation. In brief, persulfate could accelerate the degradation of organic components, promote the formation of HS, optimize the structure of microbial community, and improve the compost quality.

Hydrogen peroxide plus ascorbic acid enhanced organic matter deconstructions and composting performances via changing microbial communities.

This work aims to investigate the influence of hydrogen peroxide (H2O2) and ascorbic acid (ASCA) on the physicochemical characteristics, organic matter (OM) deconstructions, humification degree and succession of bacterial communities for co-composting of bagasse pith and dairy manure. The results indicated that H2O2 and ASCA accelerated the degradation of lignocellulose, improved the transformation of dissolved organic matter (DOM), and enhanced the content of humic substance (HS) and the degree of its aromatization. The bacterial communities were significantly changed in the presence of additives, in which the relative abundances of Firmicutes and Actinobacteria significantly increased. Redundancy analysis (RDA) indicated that the degradation of OM and lignocellulose more influenced the bacterial community compositions. Conclusively, adding H2O2 and ASCA accelerated lignocellulose degradation efficiency, and improved the composting process, which provided an optimized method to dispose of lignocellulose wastes and livestock manure.

Biochar reinforced the populations of cbbL-containing autotrophic microbes and humic substance formation via sequestrating CO2 in composting process.

The quality of compost is drastically reduced due to the loss of carbon, which negatively impacts the environment. Carbon emission reduction and carbon dioxide (CO2) fixation have attracted much attention in composting research. In this study, the relationship between CO2 emission, humic substances (HS) formation and cbbL-containing autotrophic microbes (CCAM) was analyzed by adding biochar during cow manure composting. The results showed that biochar can facilitate the degradation of organic matter (OM) and formation of HS, as well as reinforce the diversity and abundance of CCAM community, thereby promoting CO2 fixation and reducing carbon loss during composting. High-throughput sequencing analysis revealed significant increase in Actinobacteriota and Proteobacteria abundance by 30.97 % and 10.48 %, respectively, thus increasing carbon fixation by 32.07 %. Additionally, Alpha diversity index increased significantly during thermophilic phase, while Shannon index increased by 143.12 % and Sobs index increased by 51.62 %. Redundancy analysis (RDA) indicated that CO2 was positively correlated with C/N, temperature, HS and dissolved organic matter (DOM), while the abundance of Paeniclostridium, Corynebacterium, Bifidobacterium, Clostridium, Turicibacter and Romboutsia were positively correlated with temperature, CO2, C/N and E2/E4 (p <  0.01).

[Advances in Polymer Hydrophilic Lubricating Coatings for Medical Catheters].

Polymer hydrophilic lubricating coatings for medical catheters refer to highly hydrophilic coating films fixed on the surface of catheters with binding force, which can reduce the surface friction with human tissues during the use of interventional catheters, improve the patient comfort of and effectively reduce the incidence of infection. Based on the development process of medical catheter coating, this review summarizes recent advances in the field of polymer hydrophilic lubricating coatings for medical catheters from types of hydrophilic coating polymer, development of coating technology and establishment of coating performance evaluation method. Main problems in this field are analyzed and development trends in the future are prospected.

Impacts of red mud on lignin depolymerization and humic substance formation mediated by laccase-producing bacterial community during composting.

The aim of this study was to investigate the impacts of red mud on lignin degradation, humic substance formation and laccase-producing bacterial community in composting to better improve composting performances. The results indicated that the organic matter contents of final compost products in the treatment group with red mud (T) decreased by 25.74%, which was more than the control group without red mud (CK) (12.09%). The final lignin degradation ratio and humic substance concentration of the T were 18.67% and 22.80% higher than that of the CK, respectively. The final C/N values of compost in the CK and T were 11.32 and 10.66, respectively, which were both less than 15, suggesting that compost reached maturity. Redundancy analysis showed that temperature was the main factors driving the variation of laccase-producing bacterial community. Pearson analysis suggested that Pseudomonas, Phenylobacterium, and Caulobacter were the most significantly correlated with lignin degradation and humification in the T.

Overproduction of a Dominant Mutant of the Conserved Era GTPase Inhibits Cell Division in Escherichia coli.

Cell growth and division are coordinated, ensuring homeostasis under any given growth condition, with division occurring as cell mass doubles. The signals and controlling circuit(s) between growth and division are not well understood; however, it is known in Escherichia coli that the essential GTPase Era, which is growth rate regulated, coordinates the two functions and may be a checkpoint regulator of both. We have isolated a mutant of Era that separates its effect on growth and division. When overproduced, the mutant protein Era647 is dominant to wild-type Era and blocks division, causing cells to filament. Multicopy suppressors that prevent the filamentation phenotype of Era647 either increase the expression of FtsZ or decrease the expression of the Era647 protein. Excess Era647 induces complete delocalization of Z rings, providing an explanation for why Era647 induces filamentation, but this effect is probably not due to direct interaction between Era647 and FtsZ. The hypermorphic ftsZ* allele at the native locus can suppress the effects of Era647 overproduction, indicating that extra FtsZ is not required for the suppression, but another hypermorphic allele that accelerates cell division through periplasmic signaling, ftsL*, cannot. Together, these results suggest that Era647 blocks cell division by destabilizing the Z ring.IMPORTANCE All cells need to coordinate their growth and division, and small GTPases that are conserved throughout life play a key role in this regulation. One of these, Era, provides an essential function in the assembly of the 30S ribosomal subunit in Escherichia coli, but its role in regulating E. coli cell division is much less well understood. Here, we characterize a novel dominant negative mutant of Era (Era647) that uncouples these two activities when overproduced; it inhibits cell division by disrupting assembly of the Z ring, without significantly affecting ribosome production. The unique properties of this mutant should help to elucidate how Era regulates cell division and coordinates this process with ribosome biogenesis.

Molecular mechanisms underlying lignocellulose degradation and antibiotic resistance genes removal revealed via metagenomics analysis during different agricultural wastes composting.

In this study, the differences on the physico-chemical parameters, lignocellulose degradation, dynamic succession of microbial community, gene expression of carbohydrate-active enzymes and antibiotics resistance genes were compared during composting systems of bagasse pith/pig manure (BP) and manioc waste/pig manure (MW). The results revealed that biodegradation rates of organic matter, cellulose, hemicellulose and lignin (29.14%, 17.53%,45.36% and 36.48%) in BP were higher than those (15.59%, 16.74%, 41.23% and 29.77%) in MW. In addition, the relative abundance of Bacillus, Luteimonas, Clostridium, Pseudomonas, Streptomyces and expression of genes encoding carbohydrate- active enzymes in BP were higher than those in MW based on metagenomics sequencing. During composting, antibiotics and antibiotic resistance genes were substantially reduced, but the removal efficiency was divergent in the both samples. Taken together, metagenomics analysis was a potential method for evaluating lignocellulose's biodegradation process and determining the elimination of antibiotic and antibiotic resistance genes from different composting sources of biomass.

Roles of non-ionic surfactant sucrose ester on the conversion of organic matters and bacterial community structure during composting.

The purpose of this study was to investigate the effect of sucrose ester (SE) addition on the physico-chemical parameters, organic matter (OM) biodegradation and related bacterial communities structure in dairy manure and cassava residuals co-composting. The biodegradation rates of OM, dissolved organic matters (DOM) and lignocellulose in SE (16.34%, 44.11% and 26.73%) group were higher than those in CK (14.71%, 39.11% and 19.90%). In addition, the content of humic substances (HS) (36.34%) in SE was obviously higher than that in CK (17.68%). The relative abundance of bacterial community in SE changed, in which the abundance of Firmicutes and Actinobacteria increased, while the amount of Blastomonas decreased. Redundancy analysis indicated Bacillus and Acinetobacter were positively correlated with the temperature and OM, whereas Azomonas and Luteimonas showed a positive relation with pH. In conclusion, the amendment of SE accelerated the degradation and conversion of organic matters, enhanced the formation of HS and improved the quality of compost.

Investigating the variation of dissolved organic matters and the evolution of autotrophic microbial community in composting with organic and inorganic carbon sources.

The main purpose of this study was to analyze the effects of different carbon source (Na2CO3, NC; sugarcane molasses, SM) additives on dissolved organic matter (DOM), cbbL-containing autotrophic microbes (CCAM), and the relationship among physico-chemical parameters, DOM and CCAM to better understand carbon transformation in composting. The results showed that SM or NC additive could promote the degradation and transformation of OM and DOM. After adding SM or NC, the Simpson index decreased by 2.03% and 0.51%, respectively, and Luteimonas and Thermomonspora were detected using high throughput sequencing, indicating that SM and NC increased the diversity of CCAM community. Additionally, both NC and SM contributed to improve the abundance of cbbL gene (45.91% and 2.15%) based on fluorescence quantitative PCR (qPCR) analysis at the cooling phase of composting. Pearson correlation analysis revealed that Proteoobacteria, Firmicutes, Acidobacteria and Nematoda were positively related with C/N, OM and DOM (0.5 < R < 0.9, P < 0.05).

Transcriptomic profile of early zebrafish PGCs by single cell sequencing.

Single cell RNA-seq is a powerful and sensitive way to capture the genome-wide gene expression. Here, single cell RNA-seq was utilized to study the transcriptomic profile of early zebrafish PGCs (primordial germ cells) at three different developmental stages. The three stages were 6, 11 and 24 hpf (hours post fertilization). For each developmental stage, three zebrafish PGCs from one embryo were collected, and 9 samples in total were used in this experiment. Single cell RNA-seq results showed that 5099-7376 genes were detected among the 9 samples, and the number of expressed genes decreased as development progressed. Based on the gene expression pattern, samples from 6 and 11 hpf clustered closely, while samples from 24 hpf were more dispersed. By WGCNA (weighted gene co-expression network analysis), the two biggest modules that had inverse gene expression patterns were found to be related to PGC formation or migration. Functional enrichment analysis for these two modules showed that PGCs mainly conducted migration and cell division in early development (6/11 hpf) and translation activity became active in late development (24 hpf). Differentially expressed gene analyses showed that more genes were downregulated than upregulated between two adjacent stages, and genes related to PGC formation or migration reported by previous studies decreased significantly from 11 to 24 hpf. Our results provide base knowledge about zebrafish PGC development at the single cell level and can be further studied by other researchers interested in biological development.