A phage-displayed library of variable domain of heavy chain of the heavy chain antibody (VHH) or nanobody (Nb) was constructed after immunizing an alpaca with aflatoxin B1 (AFB1) conjugated with bovine serum albumin (AFB1-BSA). Two AFB1-specific nanobodies were selected. The obtained nanobodies were compared to an aflatoxin-specific monoclonal antibody B5 with respect to stability under organic solvents and high temperature. The two nanobodies could bind antigen specifically after exposure to temperatures as high as 95 °C. Besides, the nanobodies showed better or similar tolerance to organic solvents. A competitive ELISA with nanobody Nb26 was developed for the analysis of AFB1, exhibiting an IC50 value of 0.754 ng/mL (2.4 μM), linear range from 0.117 to 5.676 ng/mL. Due to the high tolerance to methanol, sample extracts were analyzed by nanobody-based ELISA without dilution. The recovery from spiked peanut, rice, corn and feedstuff ranged from 80 to 115%. In conclusion, the isolated nanobodies are excellent candidates for immunoassay application in aflatoxin determination.
Phytosterols are important micronutrients in human diets. Evidence has shown that phytosterols play an essential role in the reduction of cholesterol in blood and therefore decrease cardiovascular morbidity. In this study, the content and composition of phytosterols in different kinds of vegetable oils were analyzed, and the total phytosterol intake and contribution of foods to intake were estimated based on consumption data. The results showed that the phytosterol contents of rice bran oil, corn oil, and rapeseed oil were higher than those of other vegetable oils and the intake of phytosterol in the Chinese diet was about 392.3 mg/day. The main sources of phytosterols were edible vegetable oils (46.3%), followed by cereals (38.9%), vegetables (9.2%), nuts (2.0%), fruits (1.5%), beans and bean products (1.4%), and tubers (0.8%). Among all vegetable oils, rapeseed oil was the main individual contributor to phytosterol intake (22.9%), especially for the southern residents of China.
Chemical composition of secondary metabolites is of great importance for quality control of agricultural products. Black sesame seeds are significantly more expensive than white sesame seeds, because it is thought that black sesame seeds are more beneficial to human health than white sesame seeds. However, the differences in nutrient composition between black sesame seeds and white sesame seeds are still unknown. The current study examined the levels of different metabolites in black and white sesame seeds via the use of a novel metabolomics strategy. Using widely targeted metabolomics data, we obtained the structure and content of 557 metabolites, out of which 217 metabolites were identified, and discovered 30 metabolic pathways activated by the secondary metabolites in both black and white sesame seeds. Our results demonstrated that the main pathways that were differentially activated included: phenylpropanoid biosynthesis, tyrosine metabolism, and riboflavin metabolism. More importantly, the biomarkers that were significantly different between black seeds and white sesame seeds are highly related to the functions recorded in traditional Chinese medicine. The results of this study may serve as a new theoretical reference for breeding experts to promote the genetic improvement of sesame seeds, and therefore the cultivation of higher quality sesame varieties.
Aflatoxins represent a global public health and economic concern as they are responsible for significant adverse health and economic issues affecting consumers and farmers worldwide. Produced by fungal species from the Aspergillus genus, aflatoxins are a toxic, mutagenic, and carcinogenic group of fungal metabolites that routinely contaminate food and agricultural products. Climate and diet are essential factors in the aflatoxin contamination of food and subsequent human exposure process. Countries with warmer climates and staple foods that are aflatoxin‐susceptible shoulder a substantial portion of the global aflatoxins burden. Enactment of regulations, prevention of pre‐ and postharvest contamination, decontamination, and detoxification have been used to prevent human dietary exposure to aflatoxin. Exploiting their chemical and structural properties, means are devised to detect and quantify aflatoxin presence in foods. Herein, recent developments in several important aspects impacting aflatoxin contamination of the food supply, including: fungal producers of the toxin, occurrence in food, worldwide regulations, detection methods, preventive strategies, and removal and degradation methods were reviewed and presented. In conclusion, aflatoxin continues to be a major food safety problem, especially in developing countries where regulatory limits do not exist or are not adequately enforced. Finally, knowledge gaps and current challenges in each discussed aspect were identified, and new solutions were proposed.
The detection of adulteration of high priced oils is a particular concern in food quality and safety. Therefore, it is necessary to develop authenticity detection method for protecting the health of customers. In this study, fatty acid profiles of five edible oils were established by gas chromatography coupled with mass spectrometry (GC/MS) in selected ion monitoring mode. Using mass spectral characteristics of selected ions and equivalent chain length (ECL), 28 fatty acids were identified and employed to classify five kinds of edible oils by using unsupervised (principal component analysis and hierarchical clustering analysis), supervised (random forests) multivariate statistical methods. The results indicated that fatty acid profiles of these edible oils could classify five kinds of edible vegetable oils into five groups and are therefore employed to authenticity assessment. Moreover, adulterated oils were simulated by Monte Carlo method to establish simultaneous adulteration detection model for five kinds of edible oils by random forests. As a result, this model could identify five kinds of edible oils and sensitively detect adulteration of edible oil with other vegetable oils about the level of 10%.
Anti-idiotypic antibodies recognize the antigenic determinants of an antibody, thus can be used as surrogate antigens. Single domain antibodies from camlid heavy chain antibodies with the benefit features of small size, thermostability and ease in expression, are leading candidates to produce anti-idiotypic antibodies. In this work, we constructed an antibody phage library from the mRNA of an alpaca immunized with an anti-aflatoxin monoclonal antibody (MAb) 1C11. Three anti-idiotypic VHH antibodies were isolated and applied to immunoassay towards aflatoxin as a coating antigen. The best immunoassay developed with one of these VHH antibodies shows an IC50 of 0.16 ng/mL towards aflatoxin B1 and cross-reactivity towards aflatoxin B2, G1 and G2 of 90.4%, 54.4% and 37.7%, respectively. The VHH-based immunoassay was successfully applied to the analysis of peanuts, corn and rice, which are the predominant commodities regularly contaminated by aflatoxins. A good correlation (r2=0.89) was found between the data obtained from the conventional ELISA and the ELISA based on a VHH coating antigen for the analysis of aflatoxins in peanuts and feedstuff. The use of biotechnology in developing the surrogate, the absence of standard aflatoxin and organic solvents in the synthesis procedures, and the reproducibility of the VHH antibody makes it an ideal strategy for replacing conventional synthesized antigens.
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