Evaluation of intervention effects of dietary coenzyme Q10 supplementation on oxidized fish oil-induced stress response in largemouth bass Micropterus salmoides.

The aim of this experiment was to investigate whether dietary coenzyme Q10 could alleviate stress response of Micropterus salmoides caused by oxidized fish oil. Four isonitrogenous and isoenergetic diets were formulated to contain 100% fresh fish oil (FFO), 50% fresh fish oil + 50% oxidized fish oil (BFO), 100% oxidized fish oil (OFO) and 100% oxidized fish oil + 0.1% coenzyme Q10 (QFO) and were fed to Micropterus salmoides (95 ± 0.60 g) for 70 days. Higher weight gain rate was recorded in fish fed diet supplemented with coenzyme Q10 (CoQ10). FFO and BFO significantly increased contents of fat and energy in whole-body, while protein and energy retention significantly decreased in fish fed OFO. Apparent digestibility of energy and fat showed a significant decrease trend with increased the proportion of dietary oxidized fish oil. Fish fed OFO significantly increased activities of superoxide dismutase and catalase, while CoQ10 supplementation significantly reduced activities of alanine aminotransferase and aspartate aminotransferase in plasma. Contents of n-3 polyunsaturated fatty acids and highly unsaturated fatty acids, especially EPA and DHA in liver and muscle significantly decreased in fish fed OFO. Transcriptome analysis indicated that a total of 1238, 1189 and 1773 differentially expressed genes (DEGs, |log2(fold change) | >= 1 and q-value<=0.001) were found in the three comparison groups (FFO vs. OFO, FFO vs. QFO, OFO vs. QFO), respectively. After KEGG enrichment, the main changed pathways in the two comparison groups (FFO vs. OFO, OFO vs. QFO) related to the immune system. Dietary OFO up-regulated the expression of immune-related genes and inflammatory factors, while dietary CoQ10 supplementation reduced these effects.

Molecular characterization and functional analysis of tumor necrosis factor receptor-associated factor 2/7 and tumor necrosis factor receptor 1-associated death domain protein from Larimichthys crocea.

In the present study, we characterized tumor necrosis factor receptor-associated factor 2/7 (lcTRAF2/7) and TNFR1-associated death domain protein (lcTRADD) in Larimichthys crocea (L. crocea) and examined their expression profiles in tissues of Vibrio-challenged and unchallenged fish. The coding sequences of lcTRAF2, lcTRAF7, and lcTRADD were 1488, 2454, and 744 nucleotides, and they encoded proteins of 495, 344, and 248 amino acids, respectively. The results of phylogenetic analysis revealed that lcTRAF2, lcTRAF7, and lcTRADD were closest to Oplegnathus fasciatus (85%), Xiphophorus maculatus (97%), and Acanthochromis polyacanthus (65%), respectively. Multiple sequence alignment showed that lcTRAF2 and lcTRAF7 were highly conserved with other vertebrate TRAFs in their functional domains; however, lcTRADD was poorly conserved. The results of quantitative real-time polymerase chain reaction analysis indicated that lcTRAF2, lcTRAF7, and lcTRADD were constitutively expressed in the spleen, liver, kidney, heart, brain, gill, bladder, skin, fin, eye, and muscle. After challenging fish with Vibrio parahaemolyticus, the mRNA expression levels of lcTRAF2, lcTRAF7, and lcTRADD were upregulated in liver, spleen, and kidney. Immunofluorescence staining revealed that lcTRAF2 and lcTRADD were cytoplasmic in localization, whereas lcTRAF7 targeted both the cytoplasm and nucleus. In addition, the NF-κB protein level was upregulated after lipopolysaccharide stimulation in lcTRAF2, lcTRAF7, or lcTRADD overexpressing cells. Taken collectively, these results have improved our understanding of the functions of TRAF2, TRAF7, and TRADD in pathogenic infections in teleosts.

Deployment strategy of multiple miniaturized near-infrared spectrometers based on spectral transfer for characterizing soil organic matter and nitrogen.

Developing timely, convenient, and low-cost methods for high-frequency characterization of soil nutrients is necessary for implementing precise soil nutrient management. With the current availability of numerous calibration models of laboratory benchtop near-infrared (NIR) spectrometers for rapid soil nutrient characterization and the appearance of low-cost, convenient miniaturized NIR spectrometers, this study proposes an efficient deployment strategy to address model failure due to inter-device variation based on spectral transfer. The strategy involves using Direct Standardization (DS) to migrate the spectra from multiple miniaturized NIR spectrometers with a laboratory benchtop NIR spectrometer and then directly applying the existing calibration models of the laboratory benchtop instrument to the transferred spectra for soil nutrient analysis. The results indicated that the DS method successfully transferred the spectra of miniaturized devices to be consistent with the spectra of the laboratory benchtop instrument. The soil organic matter (SOM) predictions using the transferred spectra and the calibration models of the laboratory benchtop instrument were even more accurate than those using the respective models developed for each miniaturized devices, with root mean square error (RMSE) of 0.177 %, 0.177 %, and 0.150 %, respectively, while the performances of total nitrogen (TN) predictions were comparable to those using the respective models, with RMSE of 0.013 %, 0.012 %, and 0.010 %, respectively. Bland-Altman plots demonstrated good consistency between the strategy proposed in this study and the strategy of developing respective models for each miniaturized device, with no difference in predictions for the independent validation set compared to the laboratory benchtop instrument. This study proved the feasibility of deployment strategy of multiple miniaturized NIR spectrometers based on spectral transfer, offering a new solution for high-frequency on-site soil nutrient characterization.

Rapid and simultaneous detection of multiple illegal additives in feed and food by SERS with reusable Cu2O-Ag/AF-C3N4 substrate.

Illegal additives can bring the economic benefit, resulting in the continuous irregularities in the use of illegal additives. In this study, a method for rapid, sensitive, and simultaneous detection of multiple illegal additives including enrofloxacin, malachite green, nitrofurazone, and Sudan Ⅰ in feed and food samples by surface-enhanced Raman spectroscopy (SERS) with Cu2O-Ag/AF-C3N4 composite substrate was developed. A Cu2O-Ag/AF-C3N4 composite substrate was prepared by reacting Cu2O modified by AF-C3N4 nanosheets with AgNO3 solution. The substrate has a limit of detection (LOD) of 1.29 × 10-6 mg/L, a good linear relationship of between 10-6 and 10-2 mg/L, and an R2 value of 0.95 for Rhodamine B detection. Furthermore, the substrate showed high uniformity and reproducibility, with relative standard deviations (RSD) of 6.74% and 4.85%, respectively. Adding AF-C3N4 nanosheets not only increased the enhancement effect of the substrate, which was 4.4 times of that before addition, but also endowed it with good self-cleaning characteristics owing to its excellent photocatalytic activity. The substrate can be reused, with over 80% of the original Raman signal strength remaining after four repeat uses. The SERS based on the above substrate was used to detect the illegal additives, the LOD of enrofloxacin, malachite green, nitrofurazone, and Sudan Ⅰ can reach 4.67 × 10-4 mg/L, 2.57 × 10-5 mg/L, 5.7 × 10-7 mg/L and 6.92 × 10-5 mg/L. The results reveal that this substrate has great application potential in feed and food safety.

New insights into ß-glucan-enhanced immunity in largemouth bass Micropterus salmoides by transcriptome and intestinal microbial composition.

ß-glucan is widely used in aquaculture due to its immunostimulatory effects, but the specific effect and potential regulatory mechanism on largemouth bass (Micropterus salmoides) are still unclear. Here, we evaluated the effects of ß-glucan on growth, resistance to Aeromonas schubertii, intestinal health, and transcriptome of largemouth bass to reveal the potential regulators, metabolic pathways, and altered differential microbiota. Four experimental diets were designed with ß-glucan supplementation levels of 0 (control), 100 (LA-100), 200 (MA-200), and 300 (HA-300) mg kg-1, and each diet was fed to largemouth bass (79.30 ± 0.50 g) in triplicate for 70 days, followed by a 3-day challenge experiment. Results showed that different ß-glucan supplementations had no significant effects on growth performance and whole-body composition. Fish fed a diet with 300 mg kg-1 ß-glucan significantly increased the activity of lysozyme than those fed diets with 0 and 100 mg kg-1 ß-glucan. In addition, the survival rate of largemouth bass in ß-glucan supplementation groups was significantly higher than the control group at 12- and 24-h challenge by Aeromonas schubertii. Transcriptome analysis showed that a total of 1,245 genes were differentially expressed [|log2(fold change)| ≥1, q-value ≤0.05], including 109 immune-related differentially expressed genes (DEGs). Further analysis revealed that significantly upregulated and downregulated DEGs associated with immunity were mapped into 12 and 24 pathways, respectively. Results of intestinal microflora indicated that fish fed a diet with 300 mg kg-1 ß-glucan had higher bacterial richness and diversity as evaluated by Sobs, Chao, Ace, and Simpson indices, but no significant differences were found in the comparison groups. Furthermore, 300 mg kg-1 ß-glucan significantly increased the relative abundance of Mycoplasma and decreased Proteobacteria (mainly Escherichia-Shigella and Escherichia coli) and Bacillus anthracis in largemouth bass intestinal microflora. The findings of this study provided new insights that will be valuable in future studies to elucidate the mechanism of immunity enhancement by ß-glucan.

New rapid detection method of total chlorogenic acids in plants using SERS based on reusable Cu2O-Ag substrate.

A new method for rapidly detecting of total chlorogenic acids (CGAs) in plants by surface-enhanced Raman spectroscopy (SERS) based on reusable Cu2O-Ag substrate was developed in this study. The Cu2O-Ag substrate prepared by the in-situ growth method had high uniformity with peak intensity relative standard deviation (RSD) of 5.27%, repeatability with peak intensity RSD of 3.58%, and sensitivity with an analytical enhancement factor of 1.27 × 105 for detecting CGAs. Furthermore, the substrate had excellent reusability, after it was reused for seven cycles, the signal strength of CGAs was still above 80% of initial. Compared with the standard method of high-performance liquid chromatography (HPLC), the SERS method can successfully analyze the contents of total CGAs in plants, such as Stevia rebaudiana leaves, coffee beans, Lonicera japonica leaves, and Eucommia ulmoides flowers, with recovery rate from 93.26% to 112.65%, and the limit of detection was 0.13 µg/mL. The total CGAs content of Stevia rebaudiana leaves samples detected by HPLC and SERS have good consistency with R = 0.9760 and RMSE = 3286 mg/kg. Furthermore, the SERS method only needed less than 1 min, one standard and reusable substrate in this study to analyze, which can further reduce the cost of method analysis. Therefore, the SERS method with the appropriate substrate can provide a rapid, accurate, and economic way to detect the total CGAs in plants.