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Although the water extract of Eucommia ulmoides leaf (WEE) promotes egg laying in hens, its palatability is poor. To improve the palatability of E. ulmoides leaf, probiotic fermentation was used, and fermented extract E. ulmoides leaf (FEE) was prepared using Lactiplantibacillus plantarum. The safety of FEE was investigated using a long-term toxicity test, and no oxidative damage, inflammatory reactions, or histological lesions were observed in the experimental rats receiving dietary supplementation of FEE at 200 mg/kg, suggesting that FEE is suitable for long-term feeding. Subsequently, dietary supplementation of FEE (group C) in comparison with dietary supplementation of WEE (group B), as well as a control (group A), was applied in the hen industry. Laying performance, egg quality, egg nutrition, egg flavor, and the gut microbiome were analyzed comparatively. Interestingly, the laying rate was observed to be four percentage points higher with dietary supplementation of FEE at 200 mg/kg compared with the control and two percentage points higher compared with the dietary addition of WEE at the same dosage. Simultaneously, a slight upregulation in daily feed consumption was determined in the FEE-supplemented group compared with the blank control and the WEE-supplemented group, indicating that the inclusion of FEE stimulated the hens’ appetite. Moreover, variations in egg amino acids, fatty acids, and volatile components were obtained with either dietary addition, FEE or WEE, implying that dietary supplementation of the fermented and water-extracted E. ulmoides leaf extracts contributed to egg flavor change. Furthermore, variations in the gut microbiota were mediated by FEE, increasing the relative abundance of the genus Lactobacillus. These alterations in gut microbiota were tightly related to improved laying performance and egg flavor changes. Our results indicate that FEE is a better alternative feed additive in the hen industry than WEE.

To improve the palatability of Artemisia argyi, fermented A. argyi (AAF) were prepared by Lactobacillus plantarum and Saccharomyces cerevisiae, which were used in the hen industry subsequently. Six hundred hens were randomly divided into three groups: control (A), dietary supplementation AAF at a low level (B), and dietary supplementation AAF at a high level (C). After feeding for four months, egg production, egg quality, egg nutrition, egg flavor, plasma biochemical parameters, intestinal histology, and microbiome of the gut contents were analyzed among the three tested groups. Interestingly, 5–6 percentage points elevation in the laying rates were observed in the AAF-supplemented groups in comparison to the control, accompanied with a 5 g increase in daily feed consumption. Since no alteration in egg/body weights was detected, laying performance enhancement was the main effect of dietary supplementation AAF. Meanwhile, the compositions of the egg amino acids and fatty acids changed as the feed inclusion AAF changed, e.g., His and linoleic acid decreased almost 0.1 and 0.5 g/100 g, respectively, while oleic acid increased almost 0.4 g/100 g. In addition, although no significant difference was detected (p > 0.05), the β-diversity of the gut microbiota decreased as the diet addition of AAF decreased, and probiotics (Faecalibacterium, Prevotellaceae, Intestinimonas, and Lachnospiraceae) were the dominant keystone species under AAF treatments. These probiotics were well associated with the egg nutrition component variations based on the correlation analysis, as the Sankey plot showed. Furthermore, the results of headspace-gas chromatography-ion mobility spectrometry manifested that the egg volatile components varied (e.g., the contents of acetone, 4-methyl-3-penten-2-one, 1-hydroxy-2-propanone, ethyl acetate, ethyl octanoate, ethanol, and 2-butanol in the B and C groups were higher than in the A group) and separated clearly as daily supplementation AAF, indicating AAF hugely contributed to the egg flavor variation. Due to no significant differences noticed between the B and C groups, dietary supplementation AAF at a relative low level was enough to serve as a feed attractant in the hen industry for real feeding.

Foodborne pathogenic bacteria have become a worldwide threat to human health, and rapid and sensitive bacterial detection methods are urgently needed. In this study, a facile microfluidic chip was developed and combined with recombinase-aided amplification (RAA) for rapid and sensitive detection of Salmonella typhimurium using a non-contact eddy heater for dynamic lysis of bacterial cells and a 3D-printed fan-shaped active mixer for continuous-flow mixing. First, the bacterial sample was injected into the chip to flow through the spiral channel coiling around an iron rod under an alternating electromagnetic field, resulting in the dynamic lysis of bacterial cells by this non-contact eddy heater to release their nucleic acids. After cooling to ~75 °C, these nucleic acids were continuous-flow mixed with magnetic silica beads using the fan-shaped mixer and captured in the separation chamber using a magnet. Finally, the captured nucleic acids were eluted by the eluent from the beads to flow into the detection chamber, followed by RAA detection of nucleic acids to determine the bacterial amount. Under the optimal conditions, this microfluidic chip was able to quantitatively detect Salmonella typhimurium from 1.1 × 102 to 1.1 × 105 CFU/mL in 40 min with a detection limit of 89 CFU/mL and might be prospective to offer a simple, low-cost, fast and specific bacterial detection technique for ensuring food safety.

Salmonella is a main cause of foodborne illnesses and rapid screening of Salmonella is the key to prevent Salmonella outbreaks, however available detection methods either require a long time, or need complex pretreatment, or have low sensitivity. In this study, a microfluidic biosensor was developed for Salmonella detection using viscoelastic inertial microfluidics for separating magnetic bacteria from unbound magnetic nanoparticles (MNPs) and enzyme catalytic colorimetry for amplifying biological signals. The polyclonal antibodies and horseradish peroxidase (HRP) modified MNPs were first used to specifically capture Salmonella to form magnetic HRP-bacteria. Both magnetic HRP-bacteria and unbound MNPs were magnetically separated from background and resuspended in viscoelastic polyvinylpyrrolidone solution as sample flow. When sample flow was injected with polyvinylpyrrolidone sheath flow into a T-shaped microchannel, larger-sized magnetic HRP-bacteria could penetrate the sample flow, however smaller-sized MNPs remained in the sample flow due to weaker inertial lift force and elastic lift force, resulting in continuous-flow separation of magnetic HRP-bacteria. Finally, magnetic HRP-bacteria were collected and concentrated to catalyze tetramethyl benzidine, and absorbance was measured to determine the bacteria. This biosensor was able to detect Salmonella as low as 30 CFU/mL in 1 h and featured the advantages of shorter time due to a one-step immunoreaction, easier extension due to only one antibody and one label, and lower cost due to less expensive materials.

In this paper, a highly sensitive plasmonic enzyme-linked immunosorbent assay (pELISA) was developed for the naked-eye detection of fumonisin B1 (FB1). Glucose oxidase (GOx) was used as an alternative to horseradish peroxidase as the carrier of the competing antigen. GOx catalyzed the oxidation of glucose to produce hydrogen peroxide, which acted as a reducing agent to reduce Au3+ to Au on the surface of gold seeds (5 nm), This reaction led to a color change in the solution from colorless to purple, which was observable to the naked eye. Various parameters that could influence the detection performance of pELISA were investigated. The developed method exhibited a considerably high sensitivity for FB1 qualitative naked-eye detection, with a visible cut-off limit of 1.25 ng/mL. Moreover, the proposed pELISA showed a good linear range of 0.31−10 ng/mL with a half maximal inhibitory concentration (IC50) of 1.86 ng/mL, which was approximately 13-fold lower than that of a horseradish peroxidase- (HRP)-based conventional ELISA. Meanwhile, the proposed method was highly specific and accurate. In summary, the new pELISA exhibited acceptable accuracy and precision for sensitive naked-eye detection of FB1 in maize samples and can be applied for the detection of other chemical contaminants.

Immunomagnetic separation has been widely used for the separation and concentration of foodborne pathogens from complex food samples, however it can only handle a small volume of samples. In this paper, we presented a novel fluidic device for the specific and efficient separation and concentration of salmonella typhimurium using self-assembled magnetic nanoparticle chains. The laminated sawtooth-shaped iron foils were first mounted in the 3D-printed matrix and magnetized by a strong magnet to generate dot-array high gradient magnetic fields in the fluidic channel, which was simulated using COMSOL (5.3a, Burlington, MA, USA). Then, magnetic nanoparticles with a diameter of 150 nm, which were modified with the anti-salmonella polyclonal antibodies, were injected into the channel, and the magnetic nanoparticle chains were vertically formed at the dots and verified using a fluorescence inverted microscope. Finally, the bacterial sample was continuous-flow injected, and the target bacteria could be captured by the antibodies on the chains, followed by gold standard culture plating to determine the amount of the target bacteria. Under the optimal conditions, the target bacteria could be separated with a separation efficiency of 80% in 45 min. This fluidic device could be further improved using thinner sawtooth-shaped iron foils and stronger magnets to obtain a better dot-array magnetic field with larger magnetic intensity and denser dot distribution, and has the potential to be integrated with the existing biological assays for rapid and sensitive detection of foodborne bacteria.