Full-length 16S rRNA gene sequencing and machine learning reveal the bacterial composition of inhalable particles from two different breeding stages in a piggery.

Bacterial loading aggravates the harm of particulate matter (PM) to public health and ecological systems, especially in operations of concentrated animal production. This study aimed to explore the characteristics and influencing factors of bacterial components of inhalable particles at a piggery. The morphology and elemental composition of coarse particles (PM10, aerodynamic diameter ≤ 10 µm) and fine particles (PM2.5, aerodynamic diameter ≤ 2.5 µm) were analyzed. Full-length 16 S rRNA sequencing technology was used to identify bacterial components according to breeding stage, particle size, and diurnal rhythm. Machine learning (ML) algorithms were used to further explore the relationship between bacteria and the environment. The results showed that the morphology of particles in the piggery differed, and the morphologies of the suspected bacterial components were elliptical deposited particles. Full-length 16 S rRNA indicated that most of the airborne bacteria in the fattening and gestation houses were bacilli. The analysis of beta diversity and difference between samples showed that the relative abundance of some bacteria in PM2.5 was significantly higher than that in PM10 at the same pig house (P < 0.01). There were significant differences in the bacterial composition of inhalable particles between the fattening and gestation houses (P < 0.01). The aggregated boosted tree (ABT) model showed that PM2.5 had a great influence on airborne bacteria among air pollutants. Fast expectation-maximization microbial source tracking (FEAST) showed that feces was a major potential source of airborne bacteria in pig houses (contribution 52.64-80.58 %). These results will provide a scientific basis for exploring the potential risks of airborne bacteria in a piggery to human and animal health.

Time-Series Clustering of lncRNA-mRNA Expression during the Adipogenic Transdifferentiation of Porcine Skeletal Muscle Satellite Cells.

Skeletal muscle satellite cells (SMSCs), which are multifunctional muscle-derived stem cells, can differentiate into adipocytes. Long-chain non-coding RNA (lncRNA) has diverse biological functions, including the regulation of gene expression, chromosome silencing, and nuclear transport. However, the regulatory roles and mechanism of lncRNA during adipogenic transdifferentiation in muscle cells have not been thoroughly investigated. Here, porcine SMSCs were isolated, cultured, and induced for adipogenic differentiation. The expressions of lncRNA and mRNA at different time points during transdifferentiation were analysed using RNA-seq analysis. In total, 1005 lncRNAs and 7671 mRNAs showed significant changes in expression at differential differentiation stages. Time-series expression analysis showed that the differentially expressed (DE) lncRNAs and mRNAs were clustered into 5 and 11 different profiles with different changes, respectively. GO, KEGG, and REACTOME enrichment analyses revealed that DE mRNAs with increased expressions during the trans-differentiation were mainly enriched in the pathways for lipid metabolism and fat cell differentiation. The genes with decreased expressions were mainly enriched in the regulation of cell cycle and genetic information processing. In addition, 1883 DE mRNAs were regulated by 193 DE lncRNAs, and these genes were related to the controlling in cell cycle mainly. Notably, three genes in the fatty acid binding protein (FABP) family significantly and continuously increased during trans-differentiation, and 15, 13, and 11 lncRNAs may target FABP3, FABP4, and FABP5 genes by cis- or trans-regulation, respectively. In conclusion, these studies identify a set of new potential regulator for adipogenesis and cell fate and help us in better understanding the molecular mechanisms of trans-differentiation.

Preparation of a Z-Type g-C3N4/(A-R)TiO2 Composite Catalyst and Its Mechanism for Degradation of Gaseous and Liquid Ammonia.

In this study, an (A-R)TiO2 catalyst (ART) was prepared via the sol-gel method, and g-C3N4 (CN) was used as an amendment to prepare the g-C3N4/(A-R)TiO2 composite catalyst (ARTCN). X-ray diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM), Raman spectroscopy, N2 adsorption-desorption curves (BET), UV-Vis diffuse absorption spectroscopy (UV-Vis DRS), and fluorescence spectroscopy (PL) were used to evaluate the structure, morphology, specific surface area, optical properties, and photocarrier separation ability of the catalysts. The results showed that when the modifier CN content was 0.5 g, the dispersion of the ARTCN composite catalyst was better, with stronger light absorption performance, and the forbidden band width was smaller. Moreover, the photogenerated electrons in the conduction band of ART transferred to the valence band of CN and combined with the holes in the valence band of CN, forming Z-type heterostructures that significantly improved the efficiency of the photogenerated electron-hole migration and separation, thus increasing the reaction rate. Gaseous and liquid ammonia were used as the target pollutants to investigate the activity of the prepared catalysts, and the results showed that the air wetness and initial concentration of ammonia had a great influence on the degradation of gaseous ammonia. When the initial concentration of ammonia was 50 mg/m3 and the flow rate of the moist air was 0.9 mL/min, the degradation rate of gaseous ammonia by ARTCN-0.5 reached 88.86%, and it had good repeatability. When the catalytic dose was 50 mg and the initial concentration of NH4+ was 100 mg/L, the degradation rate of liquid ammonia by ARTCN-0.5 was 71.60% after 3 h of reaction, and small amounts of NO3- and NO2- were generated. The superoxide anion radical (·O2-) and hydroxyl radical (·OH) were the main active components in the photocatalytic reaction process.

Removal of microorganisms and antibiotic resistance genes from swine wastewater: a comparison between polyaluminum chloride (PAC), polyaluminum sulfate (LST), and aluminum hydroxide iron (LT).

The presence of antibiotic-resistant bacteria (ARB) and antibiotic-resistance genes (ARGs) in swine wastewater may present a threat to the environment and public health. Conventional swine wastewater treatment processes generally fail to effectively reduce the content of ARGs. Therefore, it is necessary to develop a highly efficient and low-cost treatment method to solve this environmental problem. In doing so, we evaluated the application of three common coagulants in the treatment of swine wastewater. Using metagenomics, we evaluated the removal efficiency of ARG loads, as well as the effect of coagulation on the structure and diversity of swine wastewater, and on the bacterial community. The results showed that the three coagulants could effectively reduce the physicochemical pollution indexes of swine wastewater (e.g., TP, NTU, COD). After treatment, the loads of a variety of antibiotics in the swine wastewater were significantly reduced, with the exception of NFX and SMD, which were all close to 100%. At the same time, in evaluating the total number of microbial colonies and the total number of fecal Escherichia coli bacteria under the three conditions, Polyaluminum Chloride (PAC) ranked first among the three coagulants with 89.18%, 93.07%, 89.92%, 98.76%, 99.60%, and 98.68%. Metagenomic analysis revealed that the abundance of cfcC, tetX, mphE, msrE, tet36, and other ARGs in the water sample after the LST treatment was significantly lower than that of the original swine wastewater sample. These findings demonstrate the feasibility of using coagulants to treat swine wastewater, which is of great significance for improving water quality and reducing the potential impacts of ARGs.

Expressions and Regulatory Effects of P38/ERK/JNK Mapks in the Adipogenic Trans-Differentiation of C2C12 Myoblasts.

BACKGROUND/AIMS: Myoblasts and muscle satellite cells have the potential to transdifferentiate into adipocytes or adipocyte-like cells. Previous studies suggest that mitogen-activated protein kinase (MAPK) is critical to adipogenic trans-differentiation of muscle cells. ERK1/2, P38 and JNK are three major MAPK family members; their activation and regulatory functions during adipogenic trans-differentiation of myoblasts are investigated. METHODS: C2C12 myoblasts were cultured and induced for adipogenic trans-differentiation. Activation patterns of MAPKs were assayed using protein microarray and Western blot. Three specific MAPK blockers, U0126, SB20358 and SP600125, were used to block ERK1/2, P38 and JNK during trans-differentiation. Cellular adipogenesis was measured using staining and morphological observations of cells and expression changes in adipogenic genes. RESULTS: Inhibitors reduced phosphorylation of corresponding MAPK and produced unique cellular effects. Suppressing P38 promoted adipogenic trans-differentiation and intensified adipolytic metabolism in differentiated cells. However, inhibition of ERK1/2 had the opposite effects on adipogenesis and no effect on adipolysis. Blocking JNK weakly blocked trans-differentiation but stimulated adipolysis and induced apoptosis. CONCLUSION: Three MAPKs participate in the regulation of myoblast adipogenic trans-differentiation by controlling adipogenic and adipolysis metabolism.

MicroRNA-199a Targets the Fatty Acid Transport Protein 1 Gene and Inhibits the Adipogenic Trans-Differentiation of C2C12 Myoblasts.

BACKGROUND/AIMS: Muscle cells are able to trans-differentiate into adipocytes with adipogenesis induction. MicroRNAs (miRNAs), a class of small non-coding RNAs, widely participate in the regulation of growth and development of cells. However, the expression and regulatory role of miRNAs in the trans-differentiation of muscle cell are largely unknown. METHODS: C2C12 myoblasts were inducted to adipogenesis trans-differentiation and microarrays were used to assay the changes of expression profile of miRNAs. MiR-199a, a miRNA showed significant change in the trans-differentiation, was selected for the subsequent function study via over- expression and knock down. RESULTS: Dozens of miRNAs showed different changes followed the adipogenesis trans-differentiation of C2C12 cells. In which, miR-199a was decreased in the adipogenic cells and miR-199a over-expression inhibited the trans-differentiation and decreased lipid accumulation in the cells. Moreover, Fatty acid transport protein 1 (Fatp1), a major regulator of trans-membrane transportation and the oxidative metabolism of free fatty acids, was showed to be a target of miR-199a by computational and luciferase reporter assays. Additionally, Fatp1 knock-down by small interfering RNA had similar inhibitory effects on the trans-differentiation in C2C12 cells. CONCLUSION: Our study reveals an important role for miR-199a in the regulation of adipogenic trans-differentiation in muscle cells via suppression of Fatp1 gene.

Effects of Cu (II) on the Growth of Chlorella vulgaris and Its Removal Efficiency of Pollutants in Synthetic Piggery Digestate.

C. vulgaris has a positive effect on the removal of nutrients from pig farm biogas slurry. However, swine wastewater often contains heavy metal ions, such as Cu (II), which may have impacts on the nutrient removal performance of C. vulgaris. Additionally, the heavy metal ions in wastewater can be adsorbed by microalgae. In this study, the stress effect of Cu (II) on the growth of Chlorella vulgaris, the Cu (II) removal by microalgae, and the effect of different concentrations of Cu (II) on the nutrient removal efficiency of C. vulgaris in biogas slurries were explored. The results showed that the microalgae biomass of microalgae on the sixth day of the experiment was the highest in the treatment with a Cu (II) concentration of 0.5 mg/L, which was 30.1% higher than that of the 2.5 mg/L group. C. vulgaris had higher removal efficiencies of Cu (II) at a Cu (II) concentration of 0.1~1.5 mg/L. The-OH, C=O, -COOH, and C-O groups on the surface of the algal cells play a significant role in the removal of Cu (II). The removal rates of COD, NH3-N, TN, and TP by C. vulgaris at a Cu (II) concentration of 0.5 mg/L were the highest, which were 89.0%, 53.7%, 69.6%, and 47.3%, respectively.

Role of akirin in skeletal myogenesis.

Akirin is a recently discovered nuclear factor that plays an important role in innate immune responses. Beyond its role in innate immune responses, Akirin has recently been shown to play an important role in skeletal myogenesis. In this article, we will briefly review the structure and tissue distribution of Akirin and discuss recent advances in our understanding of its role and signal pathway in skeletal myogenesis.

Characteristics and Traceability Analysis of Microbial Assemblage in Fine Particulate Matter from a Pig House.

Fine particulate matter (PM2.5) can carry numerous substances and penetrate deep into the respiratory tract due to its small particle size; associated harmful microorganisms are suspected to increase health risks for humans and animals. To find out the microbial compositions of PM2.5 in piggeries, their interaction and traceability, we collected PM2.5 samples from a piggery while continuously monitoring the environmental indicators. We also identified pathogenic bacteria and allergens in the samples using high-throughput sequencing technology. We analyzed the microbial differences of PM2.5 samples at different heights and during different times of day and investigated the microbial dynamics among the PM2.5 samples. To better understand the interaction between microorganisms and environmental factors among different microbial communities, we applied the network analysis method to identify the correlation among various variables. Finally, SourceTracker, a commonly used microbial traceability tool, was used to predict the source of airborne microorganisms in the pig house. We identified 14 potential pathogenic bacteria and 5 allergens from PM2.5 in the pig houses, of which Acinetobacter was the dominant bacterium in all samples (relative abundance > 1%), which warrants attention. We found that bacteria and fungi directly affected the the microbial community. The bacterial community mainly played a positive role in the microbial community. Environmental variables mainly indirectly and positively affected microbial abundance. In the SourceTracker analysis using fecal matter and feed as sources and PM2.5 sample as sink, we found that fecal matter made the greatest contribution to both bacterial and fungal components of PM2.5. Our findings provide important insights into the potential risks of pathogens in PM2.5 to human and animal health and their main sources.

Pollutant removal from swine wastewater and kinetics in constructed rapid infiltration system (CRI system).

The purpose of this paper is centred on the kinetics of removal of main pollutants in wastewater and to compared different hydraulic loading conditions of the constructed rapid infiltration system (CRI system) in terms of removal efficiencies, effluent concentrations, mass removal rate (MRR), and the first-order removal rate coefficient (k) of COD, TOC, NH4+-N, TN, and TP. The results showed that the higher the hydraulic loading, the higher the effluent concentration. The results that synthesized hydraulic loading, effluent concentrations, removal efficiencies, and other conditions showed that the best hydraulic loading was 40 cm/d. When the hydraulic load was 40 cm/d, the effluent average concentrations of COD, TOC, NH4+-N, TN, TP, Cu2+ and the removal efficiencies were 27.31 ± 16.40 mg/L, 86.11%, 10.55 ± 5.25 mg/L, 84.64%, 0.59 ± 0.87 mg/L, 99.60%, 143.31 ± 14.77 mg/L, 7.04%, 5.64 ± 1.38 mg/L, 79.20%, and 0.13 ± 0.47 mg/L, 97.51%, respectively. According to a kinetic study of the primary pollutants, the MRR increased with an increase in the hydraulic loading, except for ammonia nitrogen. CRI-3, CRI-4 were high significant correlated with ammonia nitrogen (with R2 = 93.65% and R2 = 95.03%, respectively), while CRI-2, CRI-3, and CRI-4 were high significant correlated with total nitrogen (with R2 = 94.56%, R2 = 96.70% and R2 = 96.56% respectively).

Characteristics of PM2.5 and Its Correlation with Feed, Manure and NH3 in a Pig-Fattening House.

Fine particulate matter (PM), including PM2.5 in pig houses, has received increasing attention due to the potential health risks associated with PM. At present, most studies have analyzed PM2.5 in Chinese pig houses utilizing natural ventilation. These results, however, are strongly affected by the internal structure and regional environment, thus limiting their applicability to non-mechanically ventilated pig houses. This experiment was carried out in an environmentally controlled pig house. The animal feeding operation and manure management in the house were typical for Southwest China. To reduce the influence of various environmental factors on PM2.5, the temperature and humidity in the house were maintained in a relatively stable state by using an environmental control system. The concentration of PM2.5 in the pig house was monitored, while the biological contents and chemical composition of PM2.5 were analyzed, and feed, manure, and dust particles were scanned using an electron microscope. Moreover, bacterial and fungal contents and some water-soluble ions in PM2.5 were identified. The results showed that the concentration of PM2.5 in the pig house was strongly affected by pig activity, and a phenomenon of forming secondary particles in the pig house was found, although the transformation intensity was low. The concentration of PM2.5 had negative correlations of 0.27 and 0.18 with ammonia and hydrogen sulfide, respectively. Interestingly, a stronger correlation was observed between ammonia and hydrogen sulfide and ammonia and carbon dioxide concentrations (the concentration of ammonia had stronger positive correlations with hydrogen sulfide and carbon dioxide concentrations at +0.44 and +0.59, respectively). The main potential sources of PM2.5 production were feed and manure. We speculate that manure could contribute to the broken, rough, and angular particles that formed the pig house PM2.5 that easily adhered to other components.

The Preparation of {001}TiO2/TiOF2 via a One-Step Hydrothermal Method and Its Degradation Mechanism of Ammonia Nitrogen.

{001}TiO2/TiOF2 photocatalytic composites with a high activity {001} crystal plane were prepared by one-step hydrothermal methods using butyl titanate as a titanium source and hydrofluoric acid as a fluorine source. X-ray diffraction (XRD), scanning electron microscopy (SEM), high-resolution transmission electron microscopy (HRTEM), raman spectroscopy, N2 adsorption-desorption curve (BET), UV-Vis diffuse absorption spectroscopy (UV-Vis DRS), X-ray photoelectron spectroscopy (XPS), and fluorescence spectroscopy (PL) were used to evaluate the structure, morphology, specific surface area, optical properties, and photocarrier separation ability of {001}TiO2/TiOF2. Ammonia nitrogen was taken as the target pollutant, and the degradation performance of the catalyst was investigated. The results show that hydrofluoric acid improves the content of {001} crystal plane of TiO2 with high activity; it also improves the specific surface area and dispersion of the composite material and adjusts the ratio of {001}TiO2 to TiOF2 in the composite material to enhance the absorption capacity of the composite material and reduce the band gap width of the composite material. The degradation rate of ammonia nitrogen by 100 mg F15 is 93.19% when the initial concentration of ammonia nitrogen is 100 mg/L and pH is 10. Throughout the reaction process, the {001}TiO2/TiOF2 composite produces superoxide anion radical (·O2-) and hydroxyl radical (·OH) to oxidize NH3·H2O and generate N2 accompanied by a small amount of NO3- and NO2-.

Degradation of ammonia gas by Cu2O/{001}TiO2 and its mechanistic analysis.

A heterogeneous composite catalyst Cu2O/{001}TiO2 was successfully prepared by the impregnation-reduction method. With ammonia as the target pollutant, the degradation performance and degradation mechanism analysis of the prepared composite catalyst were investigated, providing technology for the application of photocatalysis technology in ammonia treatment reference. The catalysts were characterized by X-ray diffraction (XRD), transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM), specific surface area (BET), fluorescence spectrum (PL) and UV-visible absorption (UV-Vis). The results showed: compared with single {001}TiO2, the addition of Cu2O to form a composite catalyst can reduce the recombination of electron-hole pairs, resulting in increased absorption intensity in the visible light range, decreased band gap width, and finally improved the degradation performance. When the composite ratio is 1 : 10, the specific surface area is the largest, which is 72.51 m2 g-1, and the degradation rate of ammonia is also the highest maintained at 85%. After repeated use for 5 times, the degradation rate of ammonia decreases gradually due to the loss of catalyst and photo-corrosion. In the whole reaction process, surface adsorbed water and associated hydroxyl radical participate in the ammonia degradation reaction, and finally form free hydroxyl radical and NO3 -. It provides some theoretical support for ammonia gas treatment, which is of great significance to protect the environment.

Comparative study on the removal efficiency of hydrogen sulfide (H2S) using three different packings.

To improve the removal efficiency on hydrogen sulfide (H2S), a biofilter was developed and was made of polyvinyl chloride (PVC) pipes. The effects of three different packings (i.e., packing A, packing B, and packing C), containing different proportions of activated carbon, sawdust, wormcast, perlite, and pig manure compost, based on different biofilter parameters on H2S removal efficiency, were investigated. With the extension of running time, the H2S removal rate of packing A reached up to 90.12%, that of packing B reached a peak at 92.96%, and that of packing C was highest at 87.21%. The contribution rate of each packing at the bottom of the device was significantly greater (p < 0.01) than that of other parts, and those of the top of the devices were all greater than those of the middle of the devices. The H2S removal rate increased with greater filler layer height. The removal rate of group B increased first with humidity, and then declined, with the optimal humidity level for the removal of H2S 50-65% in this study. With the prolongation of the run, the pH of packing A was reduced from 7.1 to 5.91, while the pH of packing B and C remained within the range of 6.53-7.10. An increase was found in the number of bacteria and fungi over time. The count of bacteria in packing B and C and following a decreasing order was bottom > middle > top, whereas that for fungi was the opposite. In conclusion, it is thought that packing B (comprising wormcast + sawdust + activated carbon) is more efficient in the removal of H2S than the other packings and may thus be utilized in biofilters. These results hope to provide useful information for future related research on the removal efficiency of H2S using packings. IMPLICATIONS: Wormcasts use as biological filter packing to remove H2S is limited and needs yet to be explored. A comparative study on the removal efficiency of H2S using three packings showed the packings that included wormcast were more efficient than others, and showed the combined features of physical absorption and biological removal with long sustainability and good efficiency, although these were largely influenced by environmental factors and nutrient content for the microorganisms. In summary, wormcast could be utilized in biological filters in the future in related research beacuse of its good efficiency and low cost.

Characterization of bioactive recombinant antimicrobial peptide parasin I fused with human lysozyme expressed in the yeast Pichia pastoris system.

Parasin I (PI) is a 19 amino acid peptide with potent antimicrobial activities against a broad spectrum of microorganisms and is a good candidate for development as a novel antimicrobial agent. The objective of this study was to express and characterize a codon optimized parasin I peptide fused with human lysozyme (hLY). A 513 bp cDNA fragment encoding the mature hLY protein and parasin I peptide was designed and synthesized according to the codon bias of Pichia pastoris. A 4×Gly flexible amino acid linker with an enterokinase cleavage (DDDDK) was designed to link the PI to the C-terminal of hLY. The codon optimized recombinant hLY-PI was cloned into the pPICZαA vector and expressed in P. pastoris. The over-expressed extracellular rehLY-PI was purified using Ni sepharose affinity column and exhibited a molecular mass of approximately 18 kDa. After digested with enterokinase the rehLY-PI protein release its corresponding rehLY and rePI, with molecular mass of 16 kDa and 2 kDa, respectively, on Tricine-SDS-PAGE. The released rehLY exhibited similar lytical activity against Micrococcus lysodeikticus to its commercial hLY. The digested rehLY-PI product exhibited antimicrobial activities against Bacillus subtilis, Staphylococcus aureus and Escherichia coli, and synergism has been found between the released rePI and rehLY. In conclusion, we successfully optimized a rehLY-PI fusion protein encoding gene and over-expressed the rehLY-PI in P. pastoris. The recombination protein digested with enterokinase released functional hLY and antimicrobial parasin I, which demonstrates a potential for future use as an animal feed additive to partly replace antibiotic.

Selenoprotein Genes Exhibit Differential Expression Patterns Between Hepatoma HepG2 and Normal Hepatocytes LO2 Cell Lines.

The purpose of this study was to compare messenger RNA (mRNA) expression of selenoprotein genes between hepatoma HepG2 and normal hepatocytes LO2 cell lines. Liver HepG2 and LO2 cells were cultured in 12-well plates under the same condition until cells grew to complete confluence, and then cells were harvested for total RNA and protein extraction. The qPCRs were performed to compare gene expression of 14 selenoprotein genes and 5 cancer signaling-related genes. Enzyme activities were also assayed. The results showed that human hepatoma HepG2 cells grew faster than normal hepatocytes LO2 cells. Among the genes investigated, 10 selenoprotein genes (Gpx1, Gpx3, Gpx4, Selx, Sepp, Sepw1, Sepn1, Selt, Seli, Selh) and 3 cancer signaling-related genes (Bcl-2A, caspase-3, and P38) were upregulated (P < 0.05), while Selo and Bcl-2B were downregulated (P < 0.05) in hepatoma HepG2 cells compared to LO2 cells. Significant correlations were found between selenoprotein genes and the cancer signaling-related genes Caspase3, P53, Bc1-2A, and Bc1-2B. Our results revealed that selenoprotein genes were aberrantly expressed in hepatoma HepG2 cells compared to normal liver LO2 cells, which indicated that those selenoprotein genes may play important roles in the occurrence and development of liver carcinogenesis.

Partial optimization of the 5-terminal codon increased a recombination porcine pancreatic lipase (opPPL) expression in Pichia pastoris.

Pancreatic lipase plays a key role in intestinal digestion of feed fat, and is often deficient in young animals such as weaning piglets. The objective of this study was to express and characterize a partial codon optimized porcine pancreatic lipase (opPPL). A 537 bp cDNA fragment encoding N-terminus amino acid residue of the mature porcine pancreatic lipase was synthesized according to the codon bias of Pichia pastoris and ligated to the full-length porcine pancreatic lipase cDNA fragment. The codon optimized PPL was cloned into the pPICZαA (Invitrogen, Beijing, China) vector. After the resultant opPPL/pPICZαΑ plasmid was transformed into P. pastoris, the over-expressed extracellular opPPL containing a His-tag to the C terminus was purified using Ni Sepharose affinity column (GE Healthcare, Piscataway, NJ, USA), and was characterized against the native enzyme (commercial PPL from porcine pancreas, Sigma). The opPPL exhibited a molecular mass of approximately 52 kDa, and showed optimal temperature (40°C), optimal pH (8.0), Km (0.041 mM), and Vmax (2.008 µmol x mg protein(-1) x min(-1)) similar to those of the commercial enzyme with p-NPP as the substrate. The recombinant enzyme was stable at 60°C, but lost 80% (P<0.05) of its activity after exposure to heat ≥60°C for 20 min. The codon optimization increased opPPL yield for ca 4 folds (146 mg x L(-1) vs 36 mg x L(-1)) and total enzyme activity increased about 5 folds (1900 IU x L(-1) vs 367 IU x L(-1)) compared with those native naPPL/pPICZαΑ tranformant. Comparison of gene copies and mRNA profiles between the two strains indicated the increased rePPL yields may partly be ascribed to the increased protein translational efficiency after codon optimization. In conclusion, we successfully optimized 5-terminal of porcine pancreatic lipase encoding gene and over-expressed the gene in P. pastoris as an extracellular, functional enzyme. The recombination enzyme demonstrates a potential for future use as an animal feed additive for animal improvement.

Effects of active immunization against myostatin on carcass quality and expression of the myostatin gene in pigs.

The study was conducted to investigate the effects of active immunization against myostatin on the titer of myostatin antibody, carcass evaluation, activity of creatine kinase and the expression of the myostatin gene in pigs. Eighteen pigs were allotted into three groups (six pigs per group), and pigs in treatment 1, 2 and 3 were immunized with physiological saline, 1 mg or 4 mg myostatin per pig, respectively. Six pigs were killed by electrical stunning followed by exsanguination at BW of 100 kg. The results indicated that the titer of myostatin antibody was increased in treated groups compared to the control group on day 42 (P < 0.01) and d 84 (P < 0.01). The carcass lean percentage was significantly increased in the treatment groups compared to the control group (P < 0.01), and intramuscular fat was significantly decreased in the 4 mg group compared to the control group (P < 0.05). The muscle creatine kinase activity of pigs treated with 1 mg and 4 mg myostatin was lower than the control group. The immunization of myostatin significantly decreased the myostatin gene expression levels in muscle. It was concluded that optimal active immunization against myostatin could increase the content of myostatin antibody, suppress the activity of creatine kinase and the expression of myostatin gene, and therefore improve the carcass lean percentage for pigs.