Two novel mollusk short-form ApeC-containing proteins act as pattern recognition proteins for peptidoglycan.

The Apextrin C-terminal (ApeC) domain is a new protein domain largely specific to aquatic invertebrates. In amphioxus, a short-form ApeC-containing protein (ACP) family is capable of binding peptidoglycan (PGN) and agglutinating bacteria via its ApeC domain. However, the functions of ApeC in other phyla remain unknown. Here we examined 130 ACPs from gastropods and bivalves, the first and second biggest mollusk classes. They were classified into nine groups based on their phylogenetics and architectures, including three groups of short-form ACPs, one group of apextrins and two groups of ACPs of complex architectures. No groups have orthologs in other phyla and only four groups have members in both gastropods and bivalves, suggesting that mollusk ACPs are highly diversified. We selected one bivalve ACP (CgACP1; from the oyster Crossostrea gigas) and one gastropod ACP (BgACP1; from the snail Biomphalaria glabrata) for functional experiments. Both are highly-expressed, secreted short-form ACPs and hence comparable to the amphioxus ACPs previously reported. We found that recombinant CgACP1 and BgACP1 bound with yeasts and several bacteria with different affinities. They also agglutinated these microbes, but showed no inhibiting or killing effects. Further analyses show that both ACPs had high affinities to the Lys-type PGN from S. aureus but weak or no affinities to the DAP-type PGN from Bacillus subtilis. Both recombinant ACPs displayed weak or no affinities to other microbial cell wall components, including lipopolysaccharide (LPS), lipoteichoic acid (LTA), zymosan A, chitin, chitosan and cellulose, as well as to several PGN moieties, including muramyl dipeptide (MDP), N-acetylglucosamine (GlcNAc) and N-acetylmuramic acid (MurNAc). Besides, CgACP1 had the highest expression in the gill and could be greatly up-regulated quickly after bacterial challenge. This is reminiscent of the amphioxus ACP1/2 which serve as essential mucus lectins in the gill. Taken together, the current findings from mollusk and amphioxus ACPs suggest several basic common traits for the ApeC domains, including the high affinity to Lys-type PGN, the bacterial binding and agglutinating capacity, and the role as mucus proteins to protect the mucosal surface.

Colicins of Escherichia coli Lead to Resistance against the Diarrhea-Causing Pathogen Enterotoxigenic E. coli in Pigs.

Gut microbes can affect host adaptation to various environment conditions. Escherichia coli is a common gut species, including pathogenic strains and nonpathogenic strains. This study was conducted to investigate the effects of different E. coli strains in the gut on the health of pigs. In this study, the complete genomes of two E. coli strains isolated from pigs were sequenced. The whole genomes of Y18J and the enterotoxigenic E. coli strain W25K were compared to determine their roles in pig adaptation to disease. Y18J was isolated from feces of healthy piglets and showed strong antimicrobial activity against W25K in vitro. Gene knockout experiments and complementation analysis followed by modeling the microbe-microbe interactions demonstrated that the antagonistic mechanism of Y18J against W25K relied on the bacteriocins colicin B and colicin M. Compared to W25K, Y18J is devoid of exotoxin-coding genes and has more secondary-metabolite-biosynthetic gene clusters. W25K carries more genes involved in genome replication, in accordance with a shorter cell cycle observed during a growth experiment. The analysis of gut metagenomes in different pig breeds showed that colicins B and M were enriched in Laiwu pigs, a Chinese local breed, but were scarce in boars and Duroc pigs. IMPORTANCE This study revealed the heterogeneity of E. coli strains from pigs, including two strains studied by both in silico and wet experiments in detail and 14 strains studied by bioinformatics analysis. E. coli Y18J may improve the adaptability of pigs toward disease resistance through the production of colicins B and M. Our findings could shed light on the pathogenic and harmless roles of E. coli in modern animal husbandry, leading to a better understanding of intestinal-microbe-pathogen interactions in the course of evolution.

Antioxidant potential of Pediococcus pentosaceus strains from the sow milk bacterial collection in weaned piglets.

BACKGROUND: In modern animal husbandry, breeders pay increasing attention to improving sow nutrition during pregnancy and lactation to favor the health of neonates. Sow milk is a main food source for piglets during their first three weeks of life, which is not only a rich repository of essential nutrients and a broad range of bioactive compounds, but also an indispensable source of commensal bacteria. Maternal milk microorganisms are important sources of commensal bacteria for the neonatal gut. Bacteria from maternal milk may confer a health benefit on the host. METHODS: Sow milk bacteria were isolated using culturomics followed by identification using 16S rRNA gene sequencing. To screen isolates for potential probiotic activity, the functional evaluation was conducted to assess their antagonistic activity against pathogens in vitro and evaluate their resistance against oxidative stress in damaged Drosophila induced by paraquat. In a piglet feeding trial, a total of 54 newborn suckling piglets were chosen from nine sows and randomly assigned to three treatments with different concentrations of a candidate strain. Multiple approaches were carried out to verify its antioxidant function including western blotting, enzyme activity analysis, metabolomics and 16S rRNA gene amplicon sequencing. RESULTS: The 1240 isolates were screened out from the sow milk microbiota and grouped into 271 bacterial taxa based on a nonredundant set of 16S rRNA gene sequencing. Among 80 Pediococcus isolates, a new Pediococcus pentosaceus strain (SMM914) showed the best performance in inhibition ability against swine pathogens and in a Drosophila model challenged by paraquat. Pretreatment of piglets with SMM914 induced the Nrf2-Keap1 antioxidant signaling pathway and greatly affected the pathways of amino acid metabolism and lipid metabolism in plasma. In the colon, the relative abundance of Lactobacillus was significantly increased in the high dose SMM914 group compared with the control group. CONCLUSION: P. pentosaceus SMM914 is a promising probiotic conferring antioxidant capacity by activating the Nrf2-Keap1 antioxidant signaling pathway in piglets. Our study provided useful resources for better understanding the relationships between the maternal microbiota and offspring. Video Abstract.

Modeling and Structural Optimization of MSF-RO Desalination System.

Based on the mathematical modeling and operational optimization studies of reverse osmosis (RO) and multistage flash (MSF) desalination, the structural optimization of the hybrid process was specially studied in this work with the consideration of reducing comprehensive expenses under given operational conditions. Firstly, according to the process mechanism and flowchart of the RO and MSF seawater desalination technologies, seven hybrid structures with different feed conditions were designed, and their connection equations were established for numerical calculation. Then, in order to evaluate the economic performance of the hybrid systems with different structures, the hourly average operational cost equations of RO and MSF processes were established and formulated as the comprehensive evaluation indicators. Next, with a given water production requirement, simulation calculations of the hybrid system with seven different structures were performed. The results show that the hybrid system with the fourth structure has the lowest operational cost of 4.6834 CNY/m3, and at the same time it has the lowest blowdown. However, if we take GOR or production water temperature as the target, the optimal structure of the hybrid system is the fifth or the seventh option. The obtained results are helpful in structural optimization of the hybrid system with aspects of operational cost reduction, maximum GOR, or minimizing the wastewater discharge.

Effective separation and recovery of Zn, Cu, and Cr from electroplating sludge based on differential phase transformation induced by chlorinating roasting.

Heavy metals in electroplating sludge (ES) are usually amorphous and easily released in the environment. Especially for the ES containing multiple heavy metals, owing to the complex composition and lack of effective disposal method, it has been storage for a long time. In order to avoid environmental pollution, effective treatment methods are very urgent and necessary. Here, chlorinating roasting method was developed to enlarge the phase difference of heavy metals to fulfill the utilization of ES containing multiple heavy metals (Zn, Cr, and Cu). When CaCl2 was used as additive, Zn and Cu were volatilized to the gas phase, while Cr was oxidized to Cr(V)/(VI) and retained in the solid phase with readily leachable state. The recovery percentage of Zn, Cu, and Cr can reach 99%, 98%, and 96% respectively by chlorinating roasting for 4 h at 1000 °C with the CaCl2 addition proportion of 100%. After further extraction and purification, the purity of Cr and Zn can reach 92% and 99% respectively. Moreover, the mechanism of the differential phase transformation induced by chlorinating roasting was analyzed by the method of thermodynamics and kinetics. The kinetic reaction equation of the ZnCl2 and CuCl2 volatilization process can be described by phase boundary reaction and the function is G(α) = 1-(1-α)1/3. This work provides a simple and effective method for the treatment of ES containing multiple heavy metals.

Effects of Phytic Acid-Degrading Bacteria on Mineral Element Content in Mice.

Trace minerals are extremely important for balanced nutrition, growth, and development in animals and humans. Phytic acid chelation promotes the use of probiotics in nutrition. The phytic acid-degrading strain Lactococcus lactis psm16 was obtained from swine milk by enrichment culture and direct plate methods. In this study, we evaluated the effect of the strain psm16 on mineral element content in a mouse model. Mice were divided into four groups: basal diet, 1% phytic acid, 1% phytic acid + psm16, 1% phytic acid + 500 U/kg commercial phytase. Concentrations of acetic acid, propionic acid, butyric acid, and total short-chain fatty acids were significantly increased in the strain psm16 group compared to the phytic acid group. The concentrations of copper (p = 0.021) and zinc (p = 0.017) in liver, calcium (p = 0.000), manganese (p = 0.000), and zinc (p = 0.000) in plasma and manganese (p = 0.010) and zinc (p = 0.022) in kidney were significantly increased in psm16 group, while copper (p = 0.007) and magnesium (p = 0.001) were significantly reduced. In conclusion, the addition of phytic acid-degrading bacteria psm16 into a diet including phytic acid can affect the content of trace elements in the liver, kidney, and plasma of mice, counteracting the harmful effects of phytic acid.

Identification of prognostic long intergenic non-coding RNAs as competing endogenous RNAs with KRAS mutations in colorectal cancer.

Colorectal cancer (CRC) is recognized as a common type of human cancer, and KRAS mutations are correlated with poor CRC survival outcomes. The evaluation and prediction of CRC results remain challenging. In the present study, RNA sequencing and clinical data from The Cancer Genome Atlas were used to identify KRAS mutation-related prognostic long intergenic non-coding RNAs (lincRNAs) in CRC. Significantly dysregulated lincRNAs and independent prognostic lincRNAs with KRAS mutations in CRC were identified. Two lincRNAs with KRAS mutations, LINC00265 and AL390719.2, were selected as key prognostic lincRNAs for both 10- and 5-year survival rates. In addition, competing endogenous (ce)RNA models were constructed to comprehensively assess the oncogenic performance of the two key lincRNAs. The ceRNA models suggested that LINC00265 and AL390719.2 are critical for the cell cycle and cancer pathways. Finally, reverse transcription-quantitative PCR was used to validate the ceRNA models in 12 pairs of CRC tissue samples. These prognostic lincRNAs may provide novel biomarkers for the prognostic prediction of CRC. The ceRNA model may also demonstrate the underlying mechanism of these lincRNAs in CRC.

One-step extraction of high-purity CuCl2·2H2O from copper-containing electroplating sludge based on the directional phase conversion.

The recovery of heavy metals is a vital way to turn electroplating sludge into resources and reduce its environmental hazards. However, the complex compositions of the polymetallic electroplating sludge severely limit the selective recovery of metal resources such as copper. In this study, we took a kind of copper-containing electroplating sludge (C-ES) as an example present and investigated the process of copper extraction. The copper and other metals were directional converted through an accurate phase transformation process carried out by chlorination combined with thermal regulation. Eventually, the copper was selectively recovered in the form of CuCl2·2H2O, while the rest of the metals were converted into stable metal salts or oxides. The HCl solution was the best regulator for selective copper recovery. Under the optimal conditions, the recovery of copper approached 97% and the purity of the CuCl2·2H2O product was about 95%. The kinetic reaction equation of the CuCl2 volatilization process can be described by Power Low, G(α) = α1/15. The economic estimate based on experimentation indicates the profit of recycling CuCl2·2H2O is about $23.2/kg. This work provides a novel, simple, and efficient approach to the selective recovery of heavy metal from polymetallic solid wastes.

Optimization, Characteristics, and Functions of Alkaline Phosphatase From Escherichia coli.

Weaning of piglets could increase the risk of infecting with Gram-negative pathogens, which can further bring about a wide array of virulence factors including the endotoxin lipopolysaccharide (LPS). It is in common practice that the use of antibiotics has been restricted in animal husbandry. Alkaline phosphatase (AKP) plays an important role in the detoxification and anti-inflammatory effects of LPS. This study investigated the protective effects of AKP on intestinal epithelial cells during inflammation. Site-directed mutagenesis was performed to modulate the AKP activity. The enzyme activity tests showed that the activity of the DelSigD153G-D330N mutants in B. subtilis was nearly 1,600 times higher than that of the wild-type AKP. In this study, an in vitro LPS-induced inflammation model using IPEC-J2 cells was established. The mRNA expression of interleukin-(IL-) 6, IL-8, and tumor necrosis factor-α (TNF-α) were extremely significantly downregulated, and that of ASC amino acid transporter 2 (ASCT-2), zonula occludens protein-1 (ZO-1), and occludin-3 (CLDN-3) were significantly upregulated by the DelSigD153G-D330N mutant compared with LPS treatment. This concludes the anti-inflammatory role of AKP on epithelial membrane, and we are hopeful that this research could achieve a sustainable development for the pig industry.