Salmonella can cause serious foodborne diseases, which is a major concern for global public health. The sensitive and fast analysis of Salmonella is of importance for food safety. Herein, we developed a fluorescence sensing array based on photonic crystals (PCs) and recombinase-aided amplification (RAA) for high-throughput and high-sensitive analysis of Salmonella in foods. In this assay, RAA was employed for powerful amplification of the target DNA of Salmonella, and the generated dsDNA amplicon was embedded with nucleic acid dye SYBR Green I (SGI), emitting fluorescence. Furthermore, the fluorescence signal was physically enhanced by PCs array; meanwhile, the high-throughput and sensitive detection was realized by microplate reader. Because of the isothermal reaction and lyophilized regent of RAA, the proposed array possessed gentle operation and stable amplification performance. Besides, a small volume of loading samples with 2 μL was required on the PCs array for powerful fluorescence enhancement. Due to the synergistic signal amplification effect of RAA and photonic crystals, a low limit of detection (LOD) of 10 CFU/mL was achieved. Besides, the proposed array exhibited excellent performance with 100% accuracy in 20 spiked food samples. Therefore, the proposed method with the advantages of sensitivity and high throughput showed great application potential. 

A fluorescence sensing array based on photonic crystals and RAA for sensitive and high-throughput detection of salmonella in foods

High levels of lipids and cholesterol, particularly LDL, in blood are considered the most common risk factors for cardiovascular diseases that threaten human life. Recently, interest has increased in the use of medicinal plants to treat various diseases due to their lack of side effects. The current study aims to investigate the effect of Ajwa date (AD) and germinated barley (GB) on the lipid profile in rats fed a high-fat diet (HFD). Thirty rats were distributed into five groups (six per group) as follows: the negative control group, the positive control group fed a HFD, and the other three groups that were fed a HFD supplemented with a mixture of AD and GB in equal ratios with different proportions of 20, 30, and 40% in the diet. There was an increase in moisture, protein, phenols, and vitamin C content and a decrease in the content of ash, carbohydrates, fats, and beta-glucans in GB. Blood total cholesterol levels decreased significantly (83.53, 70.12, and 73.55 mg/dL) in the groups fed the AD and GB mixtures in different percentages (20, 30, and 40%). Likewise, the AD and GB mixtures recorded a significant decrease in the level of triglycerides in the treated groups, with no significant effect on the high-density lipoprotein and low-density lipoprotein. There was a significant improvement in the level of alanine aminotransferase, while the level of aspartate aminotransferase was not affected by the treatment. In summary, AD and GB mixtures can modulate the lipid profile alterations caused by HFD through their phytochemical constituents, particularly beta-glucans.

/pdf/effect-of-ajwa-date-and-germinated-barley-on-lipid-profile-2nyledrb.pdf

Effect of Ajwa Date and Germinated Barley on Lipid Profile in Rats Fed High-Fat Diet

Natural toxins (NTs) are poisonous secondary metabolites produced by living organisms developed to ward off predators. Especially low molecular weight NTs (MW<∼1 kDa), such as mycotoxins, phycotoxins, and plant toxins, are considered an important and growing food safety concern. Therefore, accurate risk assessment of food and feed for the presence of NTs is crucial. Currently, the analysis of NTs is predominantly performed with targeted high pressure liquid chromatography tandem mass spectrometry (HPLC-MS/MS) methods. Although these methods are highly sensitive and accurate, they are relatively expensive and time-consuming, while unknown or unexpected NTs will be missed. To overcome this, novel on-site screening methods and non-targeted HPLC high resolution mass spectrometry (HRMS) methods have been developed. On-site screening methods can give non-specialists the possibility for broad "scanning" of potential geographical regions of interest, while also providing sensitive and specific analysis at the point-of-need. Non-targeted chromatography-HRMS methods can detect unexpected as well as unknown NTs and their metabolites in a lab-based approach. The aim of this chapter is to provide an insight in the recent advances, challenges, and perspectives in the field of NTs analysis both from the on-site and the laboratory perspective. 

Recent advances and challenges in the analysis of natural toxins

Background: The edible fungus Auricularia delicate (ADe) is commonly employed in traditional medicine for intestinal disorders; however, its inhibitory effect on colitis-associated colorectal cancer (CAC) and the underlying mechanisms remain unexplored. (2) Methods: The inhibitory effect of ADe on CAC was investigated using a mouse model induced by azoxymethane/dextran sulfate sodium. Results: ADe effectively suppressed the growth and number of intestinal tumors in mice. Intestinal microbiota analyses revealed that ADe treatment increased Akkermansia and Parabacteroides while it decreased Clostridium, Turicibacter, Oscillospira, and Desulfovibrio. ADe regulated the levels of 2′-deoxyridine, creatinine, 1-palmitoyl lysophosphatidylcholine, and choline in serum. Furthermore, the levels of these metabolites were associated with the abundance of Oscillospira and Paraacteroides. ADe up-regulated the free fatty acid receptor 2 and β-Arrestin 2, inhibited the nuclear factor kappa B (NF-κB) pathway, and significantly attenuated the levels of inflammatory cytokines, thereby mitigating the inflammatory in CAC mice. Conclusions: The protective effect of ADe in CAC mice is associated with the regulation of intestinal microbiota, which leads to the inhibition of NF-kB pathway and regulation of inflammation.

Intestinal Microbiota and Metabolomics Reveal the Role of Auricularia delicate in Regulating Colitis-Associated Colorectal Cancer

Today, consumption of diets rich in saturated fat and fructose, associated with a variety of metabolic deregulations, has increased. The aim of this study was to evaluate the effect of dietary supplementation with a residue of defatted chia seed on a diet with low nutritional quality. To do this, C57BL/6 male mice were fed with the Control (C), Low-Nutritional-Quality (LNQ), or supplemented-with-chia-defatted-flour (LNQ+C) diets. After 12 weeks, the glucose and lactate levels were determined in the serum, liver, and kidney, along with reactive oxygen species (ROS) levels, antioxidant enzyme activity, reduced glutathione (GSH), and protein oxidation (AOPP). The LNQ diet increased the glucose and lactate levels (+25% and +50% approx. in the liver, with respect to the control group) and generated oxidative stress by modifying the levels of ROS and the activity of antioxidant enzymes, causing oxidative damage to proteins (+12% in the liver, with respect to the control). Chia supplementation helped to restore the glucose to control levels and modulate the endogenous antioxidant system, resulting in a decrease in protein oxidation products with no differences compared to the control group. In conclusion, supplementation with chia showed beneficial effects on the general health of mice, even when fed a low-nutritional-quality diet.

The Effects of Chia Defatted Flour as a Nutritional Supplement in C57BL/6 Mice Fed a Low-Quality Diet

Auricularia delicate (ADe), an edible fungus belonging to the family Auriculariaceae and order Auriculariales, possesses antimicrobial, hepatoprotective, and antioxidant effects. In this study, after systematic analysis of its composition, ADe was administered to high-fat-diet (HFD)-fed mice to investigate its anti-obesity effect. ADe significantly controlled body weight; alleviated hepatic steatosis and adipocyte hypertrophy; reduced aspartate aminotransferase, total cholesterol, insulin, and resistin; and increased adiponectin levels in HFD-fed mice serum. Based on intestinal microbiota and lipidomics analysis, ADe treatment regulated the composition and abundance of 49 intestinal microorganisms and influenced the abundance of 8 lipid species compared with HFD-fed mice. Based on a correlation analysis of the intestinal microbiota and lipids, Coprococcus showed significant negative associations with ceramide (d18:0 20:0+O), phosphatidylserine (39:4), sphingomyelin (d38:4), and zymosterol (20:2). Moreover, ADe treatment decreased the levels of ROS and MDA and increased the levels of Nrf2, HO-1, and three antioxidant enzymes in HFD-fed mice livers. Collectively, the anti-obesity effect of ADe involves the regulation of oxidative stress and is mediated by the intestinal microbiota. Hence, this study provides a reference for the application of ADe as a candidate food for obesity.

/pdf/anti-obesity-effect-of-auricularia-delicate-involves-3chcttk2.pdf

Anti-Obesity Effect of Auricularia delicate Involves Intestinal-Microbiota-Mediated Oxidative Stress Regulation in High-Fat-Diet-Fed Mice

Fusarium temperatum (F. temperatum) causes maize stalk rot disease, reducing grain yield and producing multiple harmful mycotoxins that threaten food safety and quality. However, current detection methods are limited by the requirement of well-trained personnel, complicated operations and expensive instruments. To address this challenge, gold nanoparticles (AuNPs)-based colorimetric assay using loop-mediated isothermal amplification (LAMP) coupled with CRISPR-Cas12a (Au-CRISPR) was tested for F. temperatum detection. The assay utilizes DNA-modified AuNPs as a colorimetric probe and the results can be readily observed by the naked eye. After optimization, the method can specifically detect as low as 100 copies/μL of target DNA or 10−8 ng/μL of extracted DNA from F. temperatum. The detection platform allows high-throughput testing by integrating microtiter plates and microplate reader. Furthermore, a portable smartphone-based device was developed to realize point of care (POC) detection of F. temperatum in resource-limited settings. This simple, inexpensive and sensitive detection platform has great potential for the applications in field detection. 

Visual detection of Fusarium temperatum by using CRISPR-Cas12a empowered LAMP assay coupled with AuNPs-based colorimetric reaction

Osteoporosis (OP) leads to the acceleration of tooth movement and aggravation of periodontal bone loss during orthodontic treatment. Chroogomphus rutilus (CR) is abundant in nutrients and demonstrates remarkable antioxidant and anti-inflammatory properties. In the present study, the components of CR, including 35.00% total sugar, 0.69% reducing sugar, 14.40% crude protein, 7.30% total ash, 6.10% crude fat, 0.51% total flavonoids, 1.94% total triterpenoids, 0.32% total sterol, 1.30% total saponins, 1.69% total alkaloids, and 1.02% total phenol, were first systematically examined, followed by an investigation into its regulatory effects on bone metabolism in order to mitigate bone loss during orthodontic tooth movement in osteoporotic rats. The results of the imaging tests revealed that CR treatment reduced periodontal bone loss and normalized tooth movement in the OP. In conjunction with analyses of intestinal flora and metabolomics, CR enhances the prevalence of anti-inflammatory genera while reducing the production of inflammatory metabolites. Meanwhile, CR reduced the levels of periodontal inflammatory factors, including TNF-α, IL-1β, and IL-6, by activating Wnt/β-catenin signaling, and promoted periodontal bone formation. These findings imply that CR is a potent supplementary therapy for controlling periodontal bone remodeling in patients with OP undergoing orthodontic treatment.

Chroogomphus rutilus Regulates Bone Metabolism to Prevent Periodontal Bone Loss during Orthodontic Tooth Movement in Osteoporotic Rats

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Anti-Obesity Effect of Auricularia delicate Involves Intestinal-Microbiota-Mediated Oxidative Stress Regulation in High-Fat-Diet-Fed Mice

Visual detection of Fusarium temperatum by using CRISPR-Cas12a empowered LAMP assay coupled with AuNPs-based colorimetric reaction

Chroogomphus rutilus Regulates Bone Metabolism to Prevent Periodontal Bone Loss during Orthodontic Tooth Movement in Osteoporotic Rats