RESUMEN
An imidacloprid colloidal gold immunochromatographic strip was developed in this work, and systematic analytical conditions were deeply investigated. The test strips were used for rapid screening of imidacloprid residues in Chinese herbal medicines. The performance of the colloidal gold test strips was investigated by using five selected Chinese herbal medicines (malt, Coix seed, lotus seed, dried ginger and honeysuckle). As a result, the developed imidacloprid colloidal gold immunochromatographic test strips could be used for rapid screening of imidacloprid residues in 60 kinds of different herbs (including 26 kinds of root/rhizome medicines, 20 kinds of seed/fruit/pericarp medicines, 11 kinds of flower/leaf/whole herb medicines, and 3 kinds of bark/aboveground issues of herb medicines), and the cut-off value was 50 µg/kg. The development of this method can achieve the goal of on-site, rapid and low-cost screening of imidacloprid residues in different herbs, which is of great significance for the quality assurance of herbs.
Asunto(s)
Cromatografía de Afinidad , Medicamentos Herbarios Chinos , Oro Coloide , Límite de Detección , Neonicotinoides , Nitrocompuestos , Neonicotinoides/análisis , Nitrocompuestos/análisis , Medicamentos Herbarios Chinos/química , Medicamentos Herbarios Chinos/análisis , Oro Coloide/química , Cromatografía de Afinidad/métodos , Reproducibilidad de los Resultados , Residuos de Plaguicidas/análisis , Modelos LinealesRESUMEN
In this study, we proposed a novel method utilizing polyethyleneimine (PEI)-modified halloysite nanotubes (HNTs)-based hybrid silica monolithic spin tip to analyze hydrophilic ß-lactam antibiotics and ß-lactamases inhibitors in whole blood samples for the first time. HNTs were incorporated directly into the hybrid silica monolith via a sol-gel method, which improved the hydrophilicity of the matrix. The as-prepared monolith was further modified with PEI by glutaraldehyde coupling reaction. It was found that the PEI-modified HNTs-based hybrid silica monolith enabled a large adsorption capacity of cefoperazone at 35.7 mg g-1. The monolithic spin tip-based purification method greatly reduced the matrix effect of whole blood samples and had a detection limit as low as 0.1 - 0.2 ng mL-1. In addition, the spiked recoveries of sulbactam, cefuroxime, and cefoperazone in blank whole blood were in the range of 89.3-105.4 % for intra-day and 90.6-103.5 % for inter-day, with low relative standard deviations of 1.3-7.2 % and 4.9-10.5 %, respectively. This study introduces a new strategy for preparing nanoparticles incorporated in a hybrid silica monolith with a high adsorption capacity. Moreover, it offers a valuable tool to monitor sulbactam, cefoperazone, and cefuroxime in whole blood from pregnant women with the final aim of guiding their administration.
Asunto(s)
Cefoperazona , Cefuroxima , Interacciones Hidrofóbicas e Hidrofílicas , Límite de Detección , Nanotubos , Dióxido de Silicio , Extracción en Fase Sólida , Sulbactam , Cefoperazona/sangre , Cefoperazona/química , Humanos , Sulbactam/sangre , Sulbactam/química , Extracción en Fase Sólida/métodos , Dióxido de Silicio/química , Nanotubos/química , Cefuroxima/sangre , Cefuroxima/química , Arcilla/química , Adsorción , Antibacterianos/sangre , Antibacterianos/química , Polietileneimina/química , Cromatografía Líquida de Alta Presión/métodos , Reproducibilidad de los ResultadosRESUMEN
Here, we report a novel wheat-infecting marafivirus, tentatively named "Triticum aestivum marafivirus" (TaMRV). The full-length genome sequence of TaMRV comprises 6,437 nucleotides, excluding the poly(A) tail. Pairwise sequence comparisons and phylogenetic analysis revealed that TaMRV may represent a novel species within the genus Marafivirus in the family Tymoviridae. We also observed a mass of isometric particles with a diameter of about 30 nm in ultrathin sections of infected wheat leaf tissue. In addition, the leafhopper Psammotettix alienus was identified as a vector for this virus. This is the first report of the occurrence of a wheat-infecting marafivirus.
Asunto(s)
Hemípteros , Tymoviridae , Animales , Tymoviridae/genética , Triticum , ARN Viral/genética , Filogenia , Genoma Viral , GenómicaRESUMEN
Guanidinate homometallic rare-earth ethyl complexes [LLn(µ2-η1:η2-Et)(Et)]2 (Ln = Y(1-Y), Lu(1-Lu)) and heterobimetallic rare-earth ethyl complexes LLn(Et)(µ2-η1:η2-Et)(µ2-η1-Et)(AlEt2) (Ln = Y(2-Y), Lu(2-Lu)) have been synthesized by the treatment of LLn(CH2C6H4NMe2-o)2 (L = (PhCH2)2NC(NC6H3iPr2-2,6)2) with different equivalents of AlEt3 in toluene at ambient temperature. Interestingly, the unprecedented rare-earth ethyne complex [LY(µ2-η1-Et)2(AlEt)]2(µ4-η1:η1:η2:η2-C2H2) (3-Y) containing a [C2H2]4- unit was afforded from 2-Y. The formation mechanism study on 3-Y was carried out by DFT calculations. Furthermore, the nature of the bonding of 3-Y was also revealed by NBO analysis. The reactions of LLn(CH2 C6H4NMe2-o)2 (Ln = Y, Lu) with AlEt3 (4 equiv.) in toluene at 50 °C produced firstly the non-Cp rare-earth ethylene complex LY(µ3-η1:η1:η2-C2H4)[(µ2-η1-Et)(AlEt2)(µ2-η1-Et)2(AlEt)] (4-Y), and the Y/Al ethyl complex LY[(µ2-η1-Et)2(AlEt2)]2 (5-Y) as an intermediate of 4-Y was isolated from the reaction of LY(CH2C6H4NMe2-o)2 with AlEt3 (4 equiv.) in toluene at -10 °C.
RESUMEN
Neonicotinoids are broad-spectrum and highly effective insecticides that work by affecting neural activity in insects. Neonicotinoids are systemic pesticides that are absorbed by plants, transported, and accumulated in plant tissues, including nectar and pollen. Currently, there is a lack of a comprehensive assessment of the level of neonicotinoid contamination and the associated health risks to non-targeted organisms in commercial honey and pollen produced in China. This study collected 160 batches of honey and 26 batches of pollen from different regions and plant sources in China, analyzed the residue patterns of neonicotinoid pesticides, and comprehensively evaluated the exposure risks to non-targeted organisms including bees (adults and larvae) and humans. Furthermore, this study addresses this imperative by establishing a high-throughput, rapid, and ultra-sensitive indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on broad-spectrum monoclonal antibodies to detect and quantify neonicotinoids, with validation conducted using the LC-MS/MS method. The findings indicated that 59.4 % of honey samples contained at least one of eight neonicotinoids, and the ic-ELISA rapid detection and calculation method could detect all the samples containing neonicotinoids. Additionally, the dietary risk assessment for humans and honeybees indicates that the consumption of a specific quantity of honey may not pose a health risk to human due to neonicotinoid intake. However, the Risk Quotient values for imidacloprid to adult bees and bee larvae, as well as clothianidin to bee larvae, were determined to be 2.22, 5.03, and 1.01, respectively-each exceeding 1. This highlights the elevated risk of acute toxicity posed by imidacloprid and clothianidin residues to honey bees. The study bears significant implications for the safety evaluation of non-targeted organisms in the natural food chain. Moreover, it provides scientific guidance for protecting the diversity and health of the ecosystem.
Asunto(s)
Ecosistema , Guanidinas , Insecticidas , Tiazoles , Humanos , Abejas , Animales , Cromatografía Liquida , Espectrometría de Masas en Tándem , Neonicotinoides/toxicidad , Neonicotinoides/análisis , Nitrocompuestos/análisis , Insecticidas/toxicidad , Insecticidas/análisis , Polen/química , Plantas , Medición de RiesgoRESUMEN
Corydalis Rhizoma (CR, Yanhusuo in Chinese) has been widely used as an analgesic in herbal medicine and functional food. Cases of fungal and mycotoxin contamination in CR have been reported. In this study, the composition and diversity of fungal microbiome in CR samples from four herbal markets and two processing methods were investigated by DNA metabarcoding. Variations of the fungal microbiome in CR during cold and conventional storage were monitored. Results showed that Aspergillus was the dominant genus and saprotroph was the dominant trophic mode. Six potential toxigenic fungi, namely, Aspergillus fumigatus, Aspergillus ostianus, Aspergillus terreus, Penicillium citrinum, Penicillium oxalicum, and Trichothecium roseum, were detected. Differences in fungal composition and diversity among various groups based on collection areas and processing methods were also observed. Moreover, the relative abundance of dominant genera in CR samples stored at different temperatures was significantly different and changed with storage time. This study is the first to reveal the influence of collection areas, processing methods, and storage conditions on the fungal microbiome in CR, which was expected to provide a basis for control strategies of fungal contamination in the industrial chain of CR.
Asunto(s)
Corydalis , Micobioma , Micotoxinas , Plantas Medicinales , Corydalis/química , Micotoxinas/análisisRESUMEN
The co-occurrence of fungi and mycotoxins in various foods has been frequently reported in many countries, posing a serious threat to the health and safety of consumers. In this study, the mycobiota in five types of commercial bee pollen samples from China were first revealed by DNA metabarcoding. Meanwhile, the content of total aflatoxins in each sample was investigated by high-performance liquid chromatography with fluorescence detection. The results demonstrated that Cladosporium (0.16 %-89.29 %) was the most prevalent genus in bee pollen, followed by Metschnikowia (0-81.12 %), unclassified genus in the phylum Ascomycota (0-81.13 %), Kodamaea (0-73.57 %), and Penicillium (0-36.13 %). Meanwhile, none of the assayed aflatoxins were determined in the 18 batches of bee pollen samples. In addition, the fungal diversity, community composition, and trophic mode varied significantly among five groups. This study provides comprehensive information for better understanding the fungal communities and aflatoxin residues in bee pollen from different floral origins in China.
Asunto(s)
Aflatoxinas , Micotoxinas , Penicillium , Animales , Abejas , Aflatoxinas/análisis , Micotoxinas/análisis , Penicillium/genética , Cromatografía Líquida de Alta Presión/métodos , Polen/microbiología , Contaminación de Alimentos/análisis , HongosRESUMEN
Transcription-activator-like effector (TALE)-based tools for base editing of nuclear and organellar DNA rely on double-stranded DNA deaminases, which edit substrate bases on both strands of DNA, reducing editing precision. Here, we present CyDENT base editing, a CRISPR-free, strand-selective, modular base editor. CyDENT comprises a pair of TALEs fused with a FokI nickase, a single-strand-specific cytidine deaminase and an exonuclease to generate a single-stranded DNA substrate for deamination. We demonstrate effective base editing in nuclear, mitochondrial and chloroplast genomes. At certain mitochondrial sites, we show editing efficiencies of 14% and strand specificity of 95%. Furthermore, by exchanging the CyDENT deaminase with one that prefers editing GC motifs, we demonstrate up to 20% mitochondrial base editing at sites that are otherwise inaccessible to editing by other methods. The modular nature of CyDENT enables a suite of bespoke base editors for various applications.
RESUMEN
The purpose of this study was to prepare a highly sensitive and specific zearalenone (ZEN) monoclonal antibody, which was then used to develop an indirect enzyme-linked immunosorbent assay (ic-ELISA) and a colloidal gold immunochromatographic assay (GICA). These techniques were used for the detection of Coicis Semen and related products (Coicis Semen flour, Yimigao, and Yishigao). Immunogens were synthesized by oxime active ester techniques and characterized via ultraviolet spectrophotometry. Immunogens were injected subcutaneously into the abdominal cavities and backs of mice. Using the prepared antibodies, we developed ic-ELISA and GICA rapid detection methods, which were then applied for the rapid detection of ZEN and its analogues from Coicis Semen and related products. For ic-ELISA, the half maximal inhibitory concentration (IC50 ) values for ZEN, α-zearalenol (α-ZEL), ß-zearalenol (ß-ZEL), zearalanone (ZAN), α-zearalanol (α-ZAL), and ß-zearalanol (ß-ZAL) were determined to be 1.13, 1.69, 2.06, 0.66, 1.20, and 0.94 ngâ¢mL-1 , respectively. For GICA, the cutoff values of ZEN, α-ZEL, ß-ZEL, α-ZAL, and ß-ZAL on test strips were 0.5 ngâ¢mL-1 in phosphate buffer saline (0.01 M, pH 7.4), while ZAN was found to be 0.25 ngâ¢mL-1 . Furthermore, the cutoff values of test strips were between 10 and 20 µgâkg-1 in Coicis Semen and related products. The results of these two detection methods were in good agreement with results from liquid chromatography-tandem mass spectrometry. This study provides technical support for the preparation of broad-specificity monoclonal antibodies against ZEN and lays the foundation for the simultaneous detection of multiple mycotoxins from food and herbal medicines.
Asunto(s)
Coix , Zearalenona , Animales , Ratones , Zearalenona/análisis , Espectrometría de Masas en Tándem/métodos , Ensayo de Inmunoadsorción Enzimática/métodos , Anticuerpos MonoclonalesRESUMEN
INTRODUCTION: Zearalenone-14-glucoside (Z14G) is a modified mycotoxin that widely contaminates food across the world. Our preliminary experiment showed that Z14G degrades to zearalenone (ZEN) in the intestine exerting toxicity. Notably, oral administration of Z14G in rats induces intestinal nodular lymphatic hyperplasia. OBJECTIVES: To investigate the mechanism of Z14G intestinal toxicity and how it differs from ZEN toxicity. We conducted a precise toxicology study on the intestine of rats exposed to Z14G and ZEN using multi-omics technology. METHODS: Rats were exposed to ZEN (5 mg/kg), Z14G-L (5 mg/kg), Z14G-H (10 mg/kg), and pseudo germ free (PGF)-Z14G-H (10 mg/kg) for 14 days. Histopathological studies were performed on intestines from each group and compared. Metagenomic, metabolomic, and proteomic analyses were performed on rat feces, serum, and intestines, respectively. RESULTS: Histopathological studies showed that Z14G exposure resulted in dysplasia of gut-associated lymphoid tissue (GALT) compared to ZEN exposure. The elimination of gut microbes in the PGF-Z14G-H group alleviated or eliminated Z14G-induced intestinal toxicity and GALT dysplasia. Metagenomic analysis revealed that Z14G exposure significantly promoted the proliferation of Bifidobacterium and Bacteroides compared to ZEN. Metabolomic analysis showed that Z14G exposure significantly reduced bile acid, while proteomic analysis found that Z14G exposure significantly reduced the expression of C-type lectins compared to ZEN. CONCLUSIONS: Our experimental results and previous research suggest that Z14G is hydrolyzed to ZEN by Bifidobacterium and Bacteroides promoting their co-trophic proliferation. This leads to inactivation of lectins by hyperproliferative Bacteroides when ZEN caused intestinal involvement, resulting in abnormal lymphocyte homing and ultimately GALT dysplasia. It is noteworthy that Z14G is a promising model drug to establish rat models of intestinal nodular lymphatic hyperplasia (INLH), which is of great significance for studying the pathogenesis, drug screening and clinical application of INLH.
Asunto(s)
Productos Biológicos , Zearalenona , Ratas , Animales , Zearalenona/metabolismo , Zearalenona/toxicidad , Hiperplasia , ProteómicaRESUMEN
Neonicotinoid insecticides (NNIs) are extensively used across the agricultural products and foods. In order to meet the rapid detection requirements, a novel broad-specificity monoclonal antibody against NNIs was developed for the first time using a multi-immunogen strategy. The antibody's high affinity and its ability to bind target molecules were verified by ic-ELISA. Furthermore, molecular docking was used to evaluate the pivotal forces affecting binding affinity and to determine binding sites. Subsequently, a highly sensitive gold nanoparticle-based immunochromatographic assay was established for the rapid detection of eight NNIs and the IC50 values were 0.03-1.61 ng/mL. The limits of detection for ginseng and tomato ranged from 0.76 to 30.19 µg/kg and 0.87 to 31.57 µg/kg, respectively. The spiked recovery ranged from 72.04% to 120.74%, and the coefficient of variation were less than 9.0%. This study provides a new direction for the development of multiple NNIs residue immunoassays.
Asunto(s)
Anticuerpos Monoclonales , Insecticidas , Nanopartículas del Metal , Ensayo de Inmunoadsorción Enzimática/métodos , Inmunoensayo , Insecticidas/análisis , Simulación del Acoplamiento Molecular , Neonicotinoides/química , Panax , Oro/químicaRESUMEN
In this work, a green analytical method was established for the simultaneous extraction and detection of 20 analytes-10 neonicotinoid insecticides and their 10 major toxic metabolites in edible herbs. QuEChERS and LC-MS/MS were used to analyze the 20 analytes in five edible herbs. The residues of the 20 neonicotinoid insecticides and their metabolites in 109 herbal samples were detected, of which 90 samples were positive, and the residue of total neonicotinoid insecticides ranged from 0.26 to 139.28 µg/kg. Acetamiprid (77.06 %, ≤85.95 µg/kg), imidacloprid (67.89 %, ≤32.49 µg/kg) and their metabolites (N-desmethyl-acetamiprid (44.04 %, ≤18.42 µg/kg) and desnitro imidacloprid (48.62 %, ≤16.55 µg/kg) were most frequently detected in herbs. Significant positive correlations were found between imidacloprid/acetamiprid and their metabolites in Lycii fructus and Citri reticulatae pericarpium. Therefore, more attention may be given to the neonicotinoid insecticide residues in edible herbs in the future.
RESUMEN
BACKGROUND: Cancer cells including cancer stem cells exhibit a higher rate of ribosome biogenesis than normal cells to support rapid cell proliferation in tumors. However, the molecular mechanisms governing the preferential ribosome biogenesis in glioma stem cells (GSCs) remain unclear. In this work, we show that the novel INHAT repressor (NIR) promotes ribosomal DNA (rDNA) transcription to support GSC proliferation and glioblastoma (GBM) growth, suggesting that NIR is a potential therapeutic target for GBM. METHODS: Immunoblotting, immunohistochemical and immunofluorescent analysis were used to determine NIR expression in GSCs and human GBMs. Using shRNA-mediated knockdown, we assessed the role and functional significance of NIR in GSCs and GSC-derived orthotopic GBM xenografts. We further performed mass spectrometry analysis, chromatin immunoprecipitation, and other biochemical assays to define the molecular mechanisms by which NIR promotes GBM progression. RESULTS: Our results show that high expression of NIR predicts poor survival in GBM patients. NIR is enriched in the nucleoli of GSCs in human GBMs. Disrupting NIR markedly suppresses GSC proliferation and tumor growth by inhibiting rDNA transcription and pre-ribosomal RNA synthesis. In mechanistic studies, we find that NIR activates rDNA transcription to promote GSC proliferation by cooperating with Nucleolin (NCL) and Nucleophosmin 1 (NPM1), 2 important nucleolar transcription factors. CONCLUSIONS: Our study uncovers a critical role of NIR-mediated rDNA transcription in the malignant progression of GBM, indicating that targeting this axis may provide a novel therapeutic strategy for GBM.
Asunto(s)
Neoplasias Encefálicas , Glioblastoma , Glioma , Humanos , Glioblastoma/patología , ADN Ribosómico/metabolismo , Neoplasias Encefálicas/patología , Línea Celular Tumoral , Glioma/patología , Células Madre Neoplásicas/metabolismo , Proliferación CelularRESUMEN
The development of the medicinal tea (MT) system has promoted the health awareness in the whole world, and the nutritional elements are also an important resource of health care delivery except for the medicinal components. Among various medicinal teas, Astragalus membranaceus (AM), Zingiberaceae rhizome (ZR), and Lonicera japonica (LJ) were the most popular ingredients in China. However, except for the nutrition value, MT was inevitably contaminated with heavy metals due to the special planting environment and processing system. This study was aimed to investigate the distribution characteristics of nutrition elements and combined health risk of heavy metals in MT sample, referring to the maximum residue limit (MRL), estimated daily intake (EDI), total target hazard quotients (TTHQs), and lifetime cancer risk (LCR). Furthermore, the bioaccessibility of gastrointestinal phase and bioavailability of human colon adeno carcinoma cell line were selected for elaborating the exact damage degree to human digestive system. The results showed that, the nutritional elements of Na, Se, K, Ca, and Mn were very rich in MT, but a total of 50% of MT were contaminated by Cr, Hg, and Cd in raw material. Although the cumulative lifetime cancer risk can be accepted under the bioaccessibility (26.62-99.27%), the heavy metals of Cr, As, Hg, and Fe in AM and LJ posed a slight threaten of non-carcinogenic risk to consumers. This study will give an exactly assessment of multiple elements in digestive system, thus further to predict the potential health risk under the consumption of MT products.
Asunto(s)
Mercurio , Metales Pesados , Contaminantes del Suelo , Humanos , Tés Medicinales , Medición de Riesgo , Metales Pesados/análisis , Mercurio/análisis , China , Monitoreo del Ambiente , Contaminantes del Suelo/análisisRESUMEN
Contemporarily, there has been a growing consumption rate of areca nut (AN) products worldwide, despite the fact that both fresh and processed AN contain various hazardous ingredients, including toxic alkaloids and carcinogenetic aflatoxins. However, there is a dearth of toxicity and potential cancer risk information regarding toxic alkaloids and aflatoxins via consuming AN products. The present study conducted a comprehensive assessment of the combined hazardous effects of AN alkaloids and aflatoxins towards human digestive system, by methods of HPLC analysis, cell study and in vitro digestive system study. The results revealed a synergetic effect of arecoline and aflatoxins was on human gingival normal fibroblast cell of HGF-1 and a proliferation effect on human tongue squamous carcinoma cell of CAL-27. Specifically, the residual arecoline was as high as 91.08 µg·ml-1 in oral phase and 72.41 µg·ml-1 in gastric phase, which could be an evidence of oral cancer. More importantly, 25.93 % of AN products were contaminated with aflatoxins and the maximum value was three times the MRLs. Under these circumstances, the cytotoxic and MOE values raised a considerable health concern in terms of malignancy risk for children that consume processed AN product, especially compared to scenarios that involve adults and/or fresh AN samples. This study would give rise to a better understanding of the hazards associated with AN alkaloids and aflatoxins towards digestive system, and thus to predict the potential carcinogenic risk of AN products.
Asunto(s)
Aflatoxinas , Alcaloides , Adulto , Niño , Humanos , Areca/efectos adversos , Arecolina/toxicidad , Aflatoxinas/toxicidad , Nueces , Alcaloides/toxicidad , Carcinogénesis , Sistema DigestivoRESUMEN
Relevant studies have demonstrated that urban green spaces composed of various types of plants are able to alleviate the morbidity and mortality of respiratory diseases, by reducing air pollution levels. In order to explore the relationship between the spatial pattern of urban green spaces and air pollutant concentrations, this study takes 37 garden cities with subtropical monsoon climate in China as the research object and selects the urban air quality monitoring data and land use type data in 2019 to analyze the relationship between the spatial pattern and the air pollutant concentration through the landscape metrics model and spatial regression model. Moreover, the threshold effect of the impact of green space on air pollutant concentrations is estimated, as well. The results showed that the spatial pattern of urban green space was significantly correlated with the concentrations of PM2.5 (PM with aerodynamic diameters of 2.5 mmor less), NO2 (Nitrogen Dioxide), and SO2 (Sulfur dioxide) pollutants in the air, while the concentrations of PM10 (PM with aerodynamic diameters of 10 mmor less) pollutants were not significantly affected by the green space pattern. Among them, the patch shape index (LSI), patch density (PD) and patch proportion in landscape area (PLAND) of forest land can affect the concentration of PM2.5, NO2, and SO2, respectively. The PLAND, PD, and LSI of grassland and farmland can also have an additional impact on the concentration of SO2 pollutants. The study also found that there was a significant threshold effect within the impact mechanism of urban green space landscape pattern indicators (LSI, PD, PLAND) on the concentrations of PM2.5, NO2, and SO2 air pollutants. The results of this study not only clarified the impact mechanism of the spatial pattern of urban green space on air pollutant concentrations but also provided quantitative reference and scientific basis for the optimization and updating of urban green space to promote public health.
RESUMEN
Dimethomorph (DMM) is a widely used high-efficiency fungicide which poses unpredictable threats to the ecological environment and public health. It is significant to establish a sensitive and robust analytical method for DMM detection. Here, we fabricated a catalytic hairpin assembly (CHA) - based immunofluorescence (IMF) biosensor by using single-strand DNA and DMM antibody co-modified gold nanoparticles (H0-Ab-Au) as anti-interference probes and DMM antigen coated 96-well plate as the immune recognition element and CHA reaction vessel. Parameters relevant to AuNP probes preparation and CHA reaction environment were optimized. After optimization, the LOD of 0.002 ng/mL was calculated, with a linear correlation in inverse proportion to DMM concentration ranging from 0.01 ng/mL to 50 ng/mL. In addition, the developed biosensor was successfully applied to a variety of complex matrix samples, with satisfactory recoveries over a range of 86.74%-118.60%. Moreover, the detection results of IMF biosensor have a good correlation with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Therefore, our proposed IMF biosensor exhibits ultra-high sensitivity and excellent specificity, as well as great potential for application to other hazards.
Asunto(s)
Técnicas Biosensibles , Nanopartículas del Metal , Técnicas Biosensibles/métodos , Cromatografía Liquida , Técnicas Electroquímicas/métodos , Técnica del Anticuerpo Fluorescente , Oro/química , Nanopartículas del Metal/química , Morfolinas , Espectrometría de Masas en TándemRESUMEN
Morindae Officinalis Radix (MOR) and Alpiniae Oxyphyllae Fructus (AOF) have been widely used as dietary supplements and traditional herbal medicines for centuries. Fungal and mycotoxin contamination in MOR and AOF has been reported recently. In this study, fungi in MOR and AOF are first investigated using DNA metabarcoding, and the differences in fungal microbiome between moldy and non-moldy samples are analyzed. The results show that Ascomycota is the most prevailing fungus at the phylum level in MOR and AOF with relative abundances of 49.53-94.32% and 14.81-81.85%, respectively. Penicillium (1.86-76.14%), Cladosporium (1.82-56.65%), and Trichoderma (0.12-19.71%) are the dominant genera in MOR. Penicillium (0.27-56.06%), Papiliotrema (0.04-51.71%), and Cladosporium (3.08-44.41%) are the dominant genera in AOF. Two potential toxigenic fungi were detected, namely, Trichoderma atroviride and Fusarium equiseti. Moreover, the differences in fungal communities between moldy and non-moldy samples were monitored. In conclusion, DNA metabarcoding can be used to assess the fungal microbiome in edible medicinal herbs, thereby providing a basis for ensuring food safety and drug efficacy.
RESUMEN
AIMS: Lycii Fructus (LF) is considered as a 'superfood' due to its health benefits and delicious tastes, which has gained popularity worldwide. However, LF is also a proper host for fungal growth due to its abundant nutrients. Fungal contamination seriously affects the quality and safety of LF and poses threats to consumer health. METHODS AND RESULTS: In this study, a total of 15 LF samples were collected from five provinces in China, and were divided into five groups based on the collection areas. Fungal contamination in LF was investigated by targeting the internal transcribed spacer 2 region using Illumina Miseq PE300 platform, and the differences of fungal community in groups based on collection areas were compared. Results showed that the fungal contamination was detected in all the 15 LF samples. Ascomycota, Dothideomycetes, Pleosporales and Pleosporaceae were dominant at the phylum, class, order and family levels, respectively. At the genus level, Alternaria, Cladosporium and Fusarium were the three dominant genera. In all, 24 fungal species were identified. Among which, two species, namely Penicillium oxalicum and Trichothecium roseum, were potentially toxigenic. CONCLUSIONS: All 15 LF samples were detected with fungal contamination. The differences of fungal community in LF samples collected from different areas were observed. DNA metabarcoding was demonstrated as an efficient method to monitor the fungal contamination in LF. SIGNIFICANCE AND IMPACT OF THE STUDY: This work comprehensively reveals the fungal diversity and composition in LF and provides early warning for potential mycotoxin contamination.
Asunto(s)
Ascomicetos , Micobioma , Micotoxinas , Ascomicetos/genética , Código de Barras del ADN Taxonómico/métodos , ADN de Hongos/genética , Frutas/microbiología , Hongos/genética , Micotoxinas/análisisRESUMEN
Crataegi Fructus, a medicinal and edible herb in China, has been considered a popular dietary supplement globally. It is used for the treatment of dyspepsia and chronic heart failure according to the Chinese Pharmacopoeia (2020). However, fungal contamination in Crataegi Fructus affects its quality and safety, thus preventing its global promotion. In this study, we comprehensively studied the fungal community in processed products of Crataegi Fructus by high-throughput sequencing. A total of 21 Crataegi Fructus samples were collected from five provinces in China, and the samples were divided into five groups based on collection areas, as well as into three groups based on processing methods. We then targeted the internal transcribed spacer 2 sequence through the Illumina Miseq PE300 platform to investigate fungal composition and diversity. Results showed that all 21 samples were detected with fungal contamination, and Ascomycota was dominant at the phylum level. In the groups based on collection areas, Dothideomycetes, Pleosporaceae, and Alternaria were dominant at the class, family, and genus levels, respectively. In the groups based on processing methods, Dothideomycetes, Aspergillaceae, and Alternaria were the most abundant at the class, family, and genus levels, respectively. Differences in fungal communities between various groups were also observed. Furthermore, a total of 115 species were identified, among which seven were potential toxigenic, namely, Trichothecium roseum, Alternaria tenuissima, Aspergillus carbonarius, Penicillium brevicompactum, Aspergillus fumigatus, Rhizopus microspores, and Pichia fermentans. In conclusion, this study reveals great fungal richness and diversity of Crataegi Fructus, providing references for the prevention and control of fungal contamination of Crataegi Fructus in practical production.