New Horizons of Covalent Complex of Plant-Derived Recombinant Human Lactoferrin (OsrhLF) Combined with Different Polyphenols: Formation, Physicochemical Properties, and Gastrointestinal Fate.

Four typical dietary polyphenols ((-)-epigallocatechin gallate (EGCG), quinic acid (QA), caffeic acid (CA), and ferulic acid (FA)) were covalently prepared with rice recombinant human lactoferrin (OsrhLF) and bovine lactoferrin (bLF), and their structure and physicochemical properties were investigated, different lycopene emulsions were made by ultrasonic emulsification to analyze gastrointestinal fate. The results indicated that the covalent modification polyphenols changed the secondary/tertiary structure of LF, significantly improving the surface hydrophilicity, thermal stability, and antioxidant activity of LF. Compared with the bLF group, the OsrhLF group was more hydrophilic and the thermal denaturation temperature of the OsrhLF-CA reached 104.4 °C. LF-polyphenol emulsions significantly enhanced the photochemical stability and bioavailability of lycopene and achieved effective encapsulation and protection of lycopene compared to free lycopene, and the OsrhLF-EGCG reached 58.94% lycopene bioavailability. In short, OsrhLF does not differ much from bLF in terms of physicochemical properties and has a strong potential in the field of dietary supplements.

Sea Cucumber Body Vesicular Syndrome Is Driven by the Pond Water Microbiome via an Altered Gut Microbiota.

Apostichopus japonicus (sea cucumber) is one of the most valuable aquaculture species in China; however, different diseases can limit its economic development. Recently, a novel disease, body vesicular syndrome (BVS), was observed in A. japonicus aquaculture. Diseased animals displayed no obvious phenotypic characteristics; however, after boiling at the postharvest stage, blisters, lysis, and body ruptures appeared. In this study, a multiomics strategy incorporating analysis of the gut microbiota, the pond microbiome, and A. japonicus genotype was established to investigate BVS. Detailed analyses of differentially expressed proteins (DEPs) and metabolites suggested that changes in cell adhesion structures, caused by disordered fatty acid ß-oxidation mediated by vitamin B5 deficiency, could be a putative BVS mechanism. Furthermore, intestinal dysbacteriosis due to microbiome variations in pond water was considered a potential reason for vitamin B5 deficiency. Our BVS index, based on biomarkers identified from the A. japonicus gut microbiota, was a useful tool for BVS diagnosis. Finally, vitamin B5 supplementation was successfully used to treat BVS, suggesting an association with BVS etiology. IMPORTANCE Body vesicular syndrome (BVS) is a novel disease in sea cucumber aquaculture. As no phenotypic features are visible, BVS is difficult to confirm during aquaculture and postharvest activities, until animals are boiled. Therefore, BVS could lead to severe economic losses compared with other diseases in sea cucumber aquaculture. In this study, for the first time, we systematically investigated BVS pathogenesis and proposed an effective treatment for the condition. Moreover, based on the gut microbiota, we established a noninvasive diagnostic method for BVS in sea cucumber.

Estimation of potential agricultural non-point source pollution for Baiyangdian Basin, China, under different environment protection policies.

To prevent and control non-point source pollution, many policies have been carried out by government in China. However, the effectiveness of these policies has rarely been evaluated. In this study, the potential and spatial distribution of agricultural non-point source pollution in the Baiyangdian Basin are reported. This investigation considers multiple parameters under various policies with county as a basic unit. The results for the potential pollution from chemical oxygen demand (COD), ammonia nitrogen (NH3-N), total nitrogen (TN) and total phosphorus (TP) are 60.89×104, 3.93×104, 87.05×104 and 15.10×104 Mg, with corresponding intensities of 190, 12, 272 and 47 kg ha-1 for the Baiyangdian Basin in 2016. The highest pollution from COD is attributed to livestock and poultry breeding, whereas TN and TP are dominantly produced by rural domestic sources, and NH3-N is mostly derived from planting. Spatially, distribution of the counties producing larger non-point source pollution presented a northeast to southwest direction, consistent with the Taihang mountain alignment in the basin. The counties with high pollution intensities are mostly in the south and east of the basin. Agricultural non-point source pollution control and prevention policies contributed in pollution reduction. Compared with 2016, the total potential pollution of COD, NH3-N, TN and TP in 2020 decrease by 45.1%, 14.7%, 37.9% and 37.4%, respectively, whereas for an assumed future time (F2), the decreases are 59%, 51.4%, 56.2% and 55.7%, respectively. Prevention measures should focus on reducing pollution from livestock and poultry breeding as well as planting.

Reliable LC-MS Multiattribute Method for Biotherapeutics by Run-Time Response Calibration.

A major challenge of a mass-spectrometry-based quantitative multiattribute method (MAM) for biotherapeutics is its high variability between instruments. For reproducible attribute measurements, not only is a similar instrument model required, but the instruments must also be tuned to the same condition. This poses great long-term challenges, considering the rapid development of new instrumentations. In addition, differences in digestion efficiency, peptide recovery, and artificial modifications during sample preparation also contribute to variability between laboratories. To overcome these challenges, new mathematical methods are developed to calculate the attribute abundance in the sample, using the reference standard (RS) material as calibrant. Most quality attributes in the RS remain constant throughout the life of the standard, and therefore, the RS can serve as a calibrant to correct for the difference between instruments or sample preparation procedures. Because RS data are usually collected in a MAM assay, no additional work is required from the analyst. Data from a large number of attributes demonstrated that these methodologies greatly reduced instrument-to-instrument and sample preparation variabilities. With these methodologies, a consistent instrument model and sample preparation procedure is no longer a requirement. As a result, changes in digestion procedure and advances in instrumentations will not significantly affect the assay result.

Bioremediation of Petrochemical Wastewater Containing BTEX Compounds by a New Immobilized Bacterium Comamonas sp. JB in Magnetic Gellan Gum.

In this study, we investigated the bioremediation of petrochemical wastewater containing BTEX compounds by immobilized Comamonas sp. JB cells. Three kinds of magnetic nanoparticles were evaluated as immobilization supports for strain JB. After comparison with Fe3O4 and a-Fe2O3 nanoparticles, r-Fe2O3 nanoparticle was selected as the optimal immobilization support. The highest biodegradation activity of r-Fe2O3-magnetically immobilized cells was obtained when the concentration of r-Fe2O3 nanoparticle was 120 mg L(-1). Additionally, the recycling experiments demonstrated that the degradation activity of r-Fe2O3-magnetically immobilized cells was still high and led to less toxicity than untreated wastewater during the eight recycles. qPCR suggested the concentration of strain JB in r-Fe2O3-magnetically immobilized cells was evidently increased after eight cycles of degradation experiments. These results supported developing efficient biocatalysts using r-Fe2O3-magnetically immobilized cells and provided a promising technique for improving biocatalysts used in the bioremediation of not only petrochemical wastewater but also other hazardous wastewater.

Phenoloxidase from the sea cucumber Apostichopus japonicus: cDNA cloning, expression and substrate specificity analysis.

Phenoloxidase (PO) is a crucial component of the immune system of echinoderms. In the present study, the full-length cDNA of PO (AjPO) was cloned from coelomocytes of the sea cucumber Apostichopus japonicus using 3'- and 5'-rapid amplification of cDNA ends (RACE) PCR method, which is 2508 bp, with an open reading frame (ORF) of 2040 bp encoding 679 amino acids. AjPO contains a transmembrane domain, and three Cu-oxidase domains with copper binding centers formed by 10 histidines, one cysteine and one methionine respectively. Phylogenetic analysis revealed that AjPO was clustered with laccase-type POs of invertebrates. Using the isolated membrane proteins as crude AjPO, the enzyme could catalyze the substrates catechol, L-3,4-dihydroxyphenylalanine (L-DOPA), dopamine and hydroquinone, but failed to oxidize tyrosine. The results described above collectively proved that AjPO was a membrane-binding laccase-type PO. The quantitative real-time PCR (qRT-PCR) analysis revealed that AjPO mRNA was expressed in muscle, body wall, coelomocytes, tube feet, respiratory tree and intestine with the highest expression level in coelomocytes. AjPO could be significantly induced by lipopolysaccharide (LPS), peptidoglycan (PGN), Zymosan A and polyinosinic-polycytidylic acid (PolyI:C), suggesting AjPO is closely involved in the defense against the infection of bacteria, fungi and double-stranded RNA viruses.