Implementation of a food safety management system: a practical guide for micro-companies / Implementación de un sistema de gestión de seguridad alimentaria: una guía práctica para microempresas

Food system is continuously exposed to several safety hazards throughout manufacturing processes. Consequently, the understanding of these mechanisms, the likelihood of their occurrence, and the consequences involves the implementation of an appropriate risk assessment program. Hazard Analysis and Safety Critical Control Points (HACCP) is a powerful tool for self-control system which is fundamental to adapt the requirements of each sector and size of a company but however, is currently limited it at industrial level. In fact, as a result to its flexible application, the recommendations at European level for companies with less than 10 workers focused on a food safety management system (FSMS) based on the principles of good hygiene and manufacturing practices. Therefore, in order to help companies implementing self-monitoring systems, several reference guides have been created under the supervision of health authorities. For this purpose, the aim of these study was to create an abbreviated guide as an introduction to self-control in food safety for spanish micro-enterprises. It is presented as a summary of each prerequisite focused solely on understanding the meaning and documentation necessary to implement it. To carry out the research, European legislation and the main guides at national level, particularly from autonomous communities were took into account. Starting from this information, was provided a classification of the main plans related to prerequisites for hygiene and traceability fulfilling. Moreover, indications concerning the documentation which should be prepared for each one of the prerequisites was detailed. (AU)

El sistema alimentario está continuamente expuesto a varios riesgos de seguridad a lo largo de los procesos de fabricación. En consecuencia, la comprensión de estos mecanismos, la probabilidad de su ocurrencia y las consecuencias implica la implementación de un programa adecuado de evaluación de riesgos. El Análisis de Peligros y Puntos Críticos de Control de Seguridad (APPCC) es una poderosa herramienta para el sistema de autocontrol que es fundamental para adaptar los requisitos de cada sector y tamaño de una empresa pero que, sin embargo, actualmente está limitado a nivel industrial. De hecho, como consecuencia de su aplicación flexible, las recomendaciones a nivel europeo para empresas de menos de 10 trabajadores se centraron en un sistema de gestión de la seguridad alimentaria (SGSA) basado en los principios de buenas prácticas de higiene y fabricación. Por ello, con el fin de ayudar a las empresas a implementar sistemas de autocontrol, se han elaborado varias guías de referencia bajo la supervisión de las autoridades sanitarias. Para ello, el objetivo de este estudio fue crear una guía abreviada como introducción al autocontrol en seguridad alimentaria para microempresas españolas. Se presenta como un resumen de cada requisito previo enfocado únicamente en comprender el significado y la documentación necesaria para implementarlo. Para realizar la investigación se tuvo en cuenta la legislación europea y las principales guías a nivel nacional, en particular de las comunidades autónomas. A partir de esta información, se proporcionó una clasificación de los principales planes relacionados con los requisitos previos para el cumplimiento de la higiene y la trazabilidad. Además, se detallaron indicaciones sobre la documentación que se debe preparar para cada uno de los requisitos previos. (AU)

Assessment of single-nucleotide variant discovery protocols in RNA-seq data from human cells exposed to mycotoxins.

Food and feed contamination by nonlegislated mycotoxins beauvericin (BEA) and enniatin B (ENB) is a worldwide health concern in the present. The principal objective of this work is to assess some of the existing protocols to discover the single nucleotide variants (SNVs) in transcriptomic data obtained by RNA-seq from Jurkat cells in vitro samples individually exposed to BEA and ENB at three concentration levels (1.5, 3 and 5 µM). Moreover, previous transcriptomic results will be compared with new findings obtained using a different protocol. SNVs rs201003509 in BEA exposed cells and the rs36045790 in ENB were found in the differentially expressed genes in all doses compared to controls by means of the Genome Analysis Toolkit (GATK) Best Practices workflow. SNV-RNA-seq complementary pipeline did not show any SNV. Concerning gene expression, discrepant results were found for 1.5 µM BEA exposed cells compared with previous findings. However, 354 overlapped differentially expressed genes (DEGs) were identified in the three ENB concentrations used, with 147 matches with respect to the 245 DEGs found in the previous results. In conclusion, the two discovery SNVs protocols based on variant calling from RNA-seq used in this work displayed very different results and there were SNVs found manually not identified by any pipeline. Additionally, the new gene expression analysis reported comparable but non identical DEGs to the previous transcriptomic results obtained from these RNA-seq data.

The importance of transcriptomics and proteomics for studying molecular mechanisms of mycotoxin exposure: A review.

This review aims to highlight recent advances where transcriptomics and proteomics have been used as a key tool to understand molecular toxicity of mycotoxins. The most studied mycotoxin by using transcriptomic approach is deoxynivalenol (DON), followed by aflatoxins (AFs) and zearalenone (ZEA). Instead, proteomics mostly focuses on AFs but also in this case, mildly to ZEA and DON. However, in both omics approaches, fewer studies investigated the toxicological effect of emerging mycotoxins, patulin, ochratoxin A, T-2 toxin, alternariol and amino-14,16-dimethyloctadecan-3-ol. The study of changes in the expression of genes involved in immune system are the most common purposes for transcriptomics whereas cellular processes in proteomics field. Concerning the techniques used to perform the experiments, RT-qPCR is the most employed in gene expression analysis whereas liquid chromatography coupled with mass spectrometry is the master technique for proteomics assays. The gathered data have reported that the interest in using these omic approaches has increased in the last five years. However, in vitro models take precedence over the in vivo and ex vivo ones. Therefore, there is a need to enhance the use of in vivo models and alternative methods to better understand mycotoxins mode of action on animal and human health.

Bioaccessibility and bioavailability of bioactive compounds from yellow mustard flour and milk whey fermented with lactic acid bacteria.

Microbial fermentation with lactic acid bacteria (LAB) is a natural food biopreservation method. Yellow mustard and milk whey are optimum substrates for LAB fermentation. The aim of the present study was to evaluate the bioaccessibility and bioavailability of bioactive compounds from yellow mustard flour and milk whey both with and without LAB fermentation. All extracts were subjected to a simulated digestion process. Total polyphenols, DL-3-phenyllactic acid (PLA), lactic acid, and the antioxidant activity were determined in the studied matrices before and after simulated digestion. Yellow mustard flour was significantly richer in total polyphenols, whereas significantly higher concentrations of PLA and lactic acid were observed in milk whey. Similar antioxidant activity was determined in both ingredients being in all cases strongly reduced after in vitro digestion. Higher bioaccessibility was found for polyphenols and PLA in milk whey. Transepithelial transport of total polyphenols was higher in yellow mustard flour compared to milk whey, reaching bioavailability values between 3-7% and 1-2%, respectively. PLA transepithelial transport was only significant in both fermented matrices with bioavailability around 4-6%. Transepithelial transport of lactic acid reached values of 31-34% (bioavailability ∼ 22%) and 15-78% (bioavailability ∼ 3%) in milk whey and yellow mustard flour, respectively. LAB fermentation showed beneficial effects on enriching extracts with PLA, lactic acid, and antioxidant activity, as well as increasing bioaccessibility of these acids in yellow mustard flour and total polyphenol bioavailability in milk whey. Results pointed to yellow mustard flour and milk whey as natural preservative ingredients used in the food industry, especially when fermented with LAB.

In vitro blood brain barrier exposure to mycotoxins and carotenoids pumpkin extract alters mitochondrial gene expression and oxidative stress.

Food and feed are daily exposed to mycotoxin contamination which effects may be counteracted by antioxidants like carotenoids. Some mycotoxins as well as carotenoids penetrate the blood brain barrier (BBB) inducing alterations related to redox balance in the mitochondria. Therefore, the in vitro BBB model ECV304 was subcultured for 7 days and exposed to beauvericine, enniatins, ochratoxin A, zearalenone (100 nM each), individually and combined, and pumpkin extract (500 nM). Reactive oxygen species were measured by fluorescence using the dichlorofluorescein diacetate probe at 0 h, 2 h and 4 h. Intracellular ROS generation reported was condition dependent. RNA extraction was performed and gene expression was analyzed by qPCR after 2 h exposure. The selected genes were related to the Electron Transport Chain (ETC) and mitochondrial activity. Gene expression reported upregulation for exposures including mycotoxins plus pumpkin extract versus individual mycotoxins. Beauvericin and Beauvericin-Enniatins exposure significantly downregulated Complex I and pumpkin addition reverted the effect upregulating Complex I. Complex IV was the most downregulated structure of the ETC. Thioredoxin Interacting Protein was the most upregulated gene. These data confirm that mitochondrial processes in the BBB could be compromised by mycotoxin exposure and damage could be modulated by dietary antioxidants like carotenoids.

Proteomics evaluation of enniatins acute toxicity in rat liver.

Enniatins (ENs) are emerging mycotoxins produced by Fusarium fungi which are cytotoxic also at low concentrations due to its ionophoric properties. The aim of this study was to evaluate the hepatic toxicity of ENs exposure at different concentrations in Wistar rats through a proteomic approach. Animals were intoxicated by oral gavage with medium (EN A 256, ENA1 353, ENB 540, ENB1 296 µg/mL) and high concentrations (ENA 513, ENA1 706, ENB 1021, ENB1 593 µg/mL) of an ENs mixture and sacrificed after 8 h. Protein extraction was performed using powdered liver. Peptides were analyzed using a liquid chromatography coupled with a quadrupole time-of-flight mass spectrometer. Proteins were filtered by abundance using Mass Professional Profiler software (Agilent Technologies) and 57 were differentially expressed when compared to the control. In terms of abundance, the liver biomarker Carboamoyl-phosphate synthase showed the highest levels in all conditions employed while actin-1 had the lowest. Bioinformatic analysis using DAVID platform reported acetylation, nucleotide phosphate-binding region:NAD and catalytic activity as the most represented terms. Furthermore, metabolism was the most significant and enriched pathway in Reactome overrepresentation. In conclusion, ENs acute exposure caused protein expression changes related to major cellular processes in rats, hinting its involvement in liver disturbance.

In vitro exposure to pumpkin extract induces a protective transcriptomic profile in blood brain barrier electron transport chain

C. maxima (var. Delica) is a variety of pumpkin known for its beneficial effects and its high content in carotenoids (violaxanthin, astaxanthin, antheraxanthin, zeaxanthin, lutein, lycopene and β-carotene), which are natural antioxidants bioavailable to humans through food consumption. Numerous biological effects have been attributed to carotenoids due to their antioxidant activity: improved immune response, anti-inflammatory and anti-tumor properties and reduced risk of cardiovascular and chronic degenerative diseases. They are capable of accumulating in the brain after crossing the blood-brain barrier. Therefore, the aim of this study is to analyze changes in mitochondrial gene expression using an in vitro cell model (ECV304) of the blood brain barrier, after exposure to pumpkin extract. Cells were treated during 24 h at 5 different β-carotene concentrations, as reference extract compound: 1.72×10-4 – 1.72×10-3– 1.72×10-2 – 0.172 – 1.72 nM in DMSO 0.5%. The extracted RNA was used to perform qPCR analysis on 15 mitochondrial related genes: MT-ND2, MT-ND3, MT-ND4, MT-ND4L, MT-ND5, MT-CO1, MT-CO3, MT-ATP6, MT-ATP8, MT-RNR2, MRPL12, OSGIN1, SRXN1, TXNIP, UCP2, and S18 as reference gene. Results demonstrate that dietary carotenoids act at transcriptional level, especially on the genes belonging to the electron transport chain, reporting an overall protective pattern. The findings show a dose dependent differential gene expression pattern by carotenoids exposure, even at low concentrations. (AU)

Una actualización sobre las manifestaciones clínicas y el diagnóstico de la enfermedad celiaca / An update on the clinical manifestations and diagnosis of celiac disease

La enfermedad celíaca (EC) es un trastorno sistémico, crónico y de origen autoinmune que afecta a individuos genéticamente susceptibles. Se presenta con una gran variedad de signos, síntomas y enfermedades asociadas, incluso también puede cursar de manera asintomática, lo que hace difícil su diagnóstico. La EC se diagnostica con mayor frecuencia en las mujeres y a edades más tempranas que a los hombres. Debido a la gran heterogeneidad en la forma de presentación de la EC, se ha llevado a cabo una revisión sistemática, donde estudios recientes, muestran una variación del espectro clínico de la enfermedad en función de la edad. En niños pequeños predomina la forma clásica con síntomas como distensión abdominal, pérdida del apetito, diarrea y pérdida de peso; esta forma de presentación típica es más intensa en niños menores de dos años. En niños mayores son frecuentes las manifestaciones atípicas como el dolor abdominal, reflujo, vómitos, fatiga y talla baja, entre otros. En adultos se observa una reducción de la forma de presentación clásica e incremento de la no clásica o atípica. El incremento en la prevalencia en los últimos años y la variación del espectro clínico hacia una forma atípica con síntomas extradigestivos o incluso asintomática, causa retraso en su diagnóstico, por lo que deben reconocerse para que se detecte de manera más precoz. El diagnóstico de la EC ha mejorado mucho en los últimos años debido a la creciente incidencia de esta patología y a la precisión de los nuevos métodos de detección. Se ha realizado una búsqueda bibliográfica de artículos sobre grupos de riesgo y métodos de diagnóstico en bases de datos como: Pubmed, Scopus, Web of Science de los últimos 6 años (2014-2019), se realizaron dos búsquedas en cada una de las bases de datos consultadas, con las siguientes palabras clave: 1) celiac disease AND sign AND symptom AND prevalence; 2) celiac disease AND sign AND symptom AND autoimmune disease. (AU)

Celiac disease (CD) is a systemic, chronic and autoimmune disorder affecting genetically susceptible individuals. It presents a wide variety of signs, symptoms and associated diseases, it can even be asymptomatic, difficulting their diagnosis. CD is diagnosed more often in women and at a younger age than in men. Due to the great hererogeneity in the presentation of CD, a systematic review has been carried out, where recent studies show a variation in the clinical spectrum of the disease as an age function. The classic form predominates in young children with symptoms such as abdominal distension, loss of appetite, diarrhea and weight loss; this typical presentation is more intense in children under two years of age. Atypical manifestations such as abdominal pain, reflux, vomiting, fatigue and short stature, among others, are frequent in older children. In adults, a reduction in the classic presentation and an increase in the non-classic or atypical form was shown. The increase in prevalence in recent years and the variation of the clinical spectrum towards an atypical form with extradigestive or even asymptomatic symptoms, causes a delay in the diagnosis, so they must be recognized to get an early detection. The diagnosis of CD has improved greatly in recent years due to the increasing incidence of this pathology and the precision of new detection methods. A bibliographic search has been carried out for articles on risk groups and diagnostic methods in databases such as Pubmed, Scopus and Web of Science for the last 6 years (2014-2019), two searches were carried out in each consulted database, with the following keywords: 1) celiac disease AND sign AND sym ptom AND prevalence; 2) celiac disease AND sign AND symptom AND autoimmune disease. (AU)

Association between small-bowel mucosal damage and related diseases: Observational study in celiac patients / Asociación entre lesiones en la mucosa del intestino delgado y otras enfermedades: Estudio observacional en pacientes celiacos

Celiac disease is a systemic, chronic and autoimmune disorder that affects genetically susceptible individuals. Due to the increasing incidence of this pathology and the precision of new detection methods, celiac disease diagnosis has improved dramatically in recent years. Hereby, a study was performed to evaluate celiac disease’s prevalence, attending to associated diseases as well as clinical determinants. A convenience sample of 254 patients diagnosed between 2007 and 2017 in the South of Spain was selected: 212 were confirmed for celiac disease, 18 remained with suspected celiac disease and 24 were considered silent patients. Multivariate logistic regression models were applied to patients’ data. 95.3% of the subjects obtained a positive result in the genetic-molecular diagnosis, with prevalence of female patients’ group (58.7%). Moreover, females were associated with diarrhea and abdominal pain to a greater extent (54.3% and 66.2%, respectively). Youngsters had accused villi atrophy and larger concentrations of anti-tTG antibodies compared to adults, but had more adhesion to treatment and recovered better than the older group. Deficit in Fe and multimorbidity were also factors associated with villi atrophy. The multivariate analysis adjusted for sex and age showed a direct association between intestinal lesion and Fe deficit, the presence of vomiting and the number of diseases associated with celiac disease. Novel results of the present study refer to the association between the level of intestinal injury and the multimorbidity associated with celiac disease. (AU)

La enfermedad celíaca es un trastorno sistémico, crónico y autoinmune que afecta a individuos genéticamente susceptibles. Debido al incremento en la incidencia de esta patología y la precisión de los nuevos métodos de detección, el diagnóstico de la enfermedad celíaca ha mejorado drásticamente en los últimos años. De esta manera, se ha realizado un estudio para evaluar la prevalencia de la enfermedad celíaca, atendiendo tanto a las enfermedades asociadas como a los determinantes clínicos. Se seleccionó una muestra de conveniencia de 254 pacientes diagnosticados entre 2007 y 2017 en el sur de España: 212 fueron confirmados de enfermedad celíaca, 18 permanecieron con sospecha de enfermedad celíaca y 24 fueron considerados pacientes silentes. Se aplicaron modelos de regresión logística multivariante a los datos de los pacientes. El 95,3% de los sujetos obtuvo un resultado positivo en el diagnóstico genético-molecular, con predominio del grupo de pacientes del sexo femenino (58,7%). Además, las mujeres se asociaron en mayor medida con diarrea y dolor abdominal (54,3% y 66,2%, respectivamente). Los jóvenes se asociaron a una mayor atrofia de las vellosidades y concentraciones de anticuerpos-tTG superiors en comparación con los adultos, pero mostraron una mayor adherencia al tratamiento y se recuperaron mejor que el grupo de mayor edad. El déficit de Fe y la multimorbilidad también fueron factores asociados con la atrofia de las vellosidades. El análisis multivariante ajustado por sexo y edad mostró una asociación directa entre la lesión intestinal y el déficit de Fe, la presencia de vómitos y el número de enfermedades asociadas a la enfermedad celíaca. Los nuevos resultados del presente estudio se refieren a la asociación entre el nivel de lesión intestinal y la multimorbilidad asociada a la enfermedad celíaca. (AU)

Mitochondrial proteomics profile points oxidative phosphorylation as main target for beauvericin and enniatin B mixture.

Beauvericin (BEA) and enniatin B (EN B) are non-legislated Fusarium mycotoxins usually found in cereal and cereal-based products all around the world. By the proteomic analysis of mitochondria enriched extracts from Jurkat cells exposed for 24 h to three concentrations of BEA:EN B (0.01-0.1-0.5 µM), a number of 1821 proteins (202 mitochondrial) were identified and relatively quantified. 340 proteins (59 mitochondrial) were statistically significant altered in our samples (Anova p-value ≤ 0.05 and fold change (FC) ≥1.5). The protein mitochondrial translational release factor 1 like (MTRF1L) was the most abundant protein in the three mycotoxin exposures studied. The mycotoxins mixture exposure induced concentration dependent changes at mitochondrial proteins levels that mainly involve inner and outer membrane complexes, Electron Transport Chain (ETC) and ribosomes. These results showed alteration of proteins levels related to oxidative phosphorylation, metabolic and neurodegenerative diseases related pathways.

Toxicity of mycotoxins in vivo on vertebrate organisms: A review.

Mycotoxins are considered to be a major risk factor affecting human and animal health as they are one of the most dangerous contaminants of food and feed. This review aims to compile the research developed up to date on the toxicological effects that mycotoxins can induce on human health, through the examination of a selected number of studies in vivo. AFB1 shows to be currently the most studied mycotoxin in vivo, followed by DON, ZEA and OTA. Scarce data was found for FBs, PAT, CIT, AOH and Fusarium emerging mycotoxins. The majority of them concerned the investigation of immunotoxicity, whereas the rest consisted in the study of genotoxicity, oxidative stress, hepatotoxicity, cytotoxicity, teratogenicity and neurotoxicity. In order to assess the risk, a wide range of different techniques have been employed across the reviewed studies: qPCR, ELISA, IHC, WB, LC-MS/MS, microscopy, enzymatic assays, microarray and RNA-Seq. In the last decade, the attention has been drawn to immunologic and transcriptomic aspects of mycotoxins' action, confirming their toxicity at molecular level. Even though, more in vivo studies are needed to further investigate their mechanism of action on human health.

Mitochondrial transcriptional study of the effect of aflatoxins, enniatins and carotenoids in vitro in a blood brain barrier model.

C. maxima (var. Delica), a variety of pumpkin, is well known for its high concentration on carotenoids, possessing dietary benefits and antioxidant properties. Aflatoxins and enniatins are common mycotoxins present in food and feed with an extended toxicity profile in humans and animals. Both types of substances reach a wide range of tissues and organs and have the capability to penetrate the blood brain barrier. Since carotenoids and mycotoxins have been reported to modify diverse mitochondrial processes individually, transcriptional in vitro studies on human epithelial cells ECV 304 were conducted to analyze the relative expression of 13 mitochondria related genes. ECV 304 cells were differentiated for 9 days and treated for 2 h with: a) pumpkin (500 nM); b) aflatoxins (100 nM); c) enniatins (100 nM); d) aflatoxins (100 nM) and pumpkin (500 nM); e) enniatins (100 nM) and pumpkin (500 nM). Even at low concentrations, dietary carotenoids activity on mitochondrial genes expression reported a beneficial effect and, for most of the genes studied across the Electron Transport Chain (ETC), developed a protective effect when mixed with aflatoxins (AFs) or enniatins (ENs).

Defensa Alimentaria:revisión de herramientas y estrategias / Food Defense: tools review and strategies

La defensa alimentaria se refiere principalmente a la contaminación intencional del suministro de alimentos, amenazando de esta manera la seguridad de la cadena agroalimentaria lo que puede significar grandes riesgos e inmensurables impactos. La prevención de la contaminación de los alimentos y la reducción de la probabilidad de la contaminación deliberada y el conocimiento de los peligros son los primeros pasos para construir unas estrategias efectivas de food defense y requieren un esfuerzo recurrente en toda la cadena de suministro de alimentos. La naturaleza intencional de estos riesgos ofensivos que se pueden originar en el entorno interno o externo de la industria alimentaria, significa que se deben considerar diferentes factores y se requieren diferentes tácticas para avanzar más allá de los enfoques de seguridad alimentaria comunes. Sin embargo, el conocimiento sobre técnicas básicas de defensa alimentaria sigue siendo limitado. En el presente estudio, se exploran conceptos básicos de food defense para delimitar la vulnerabilidad alimentaria a nivel del fabricante, los proveedores y los clientes de la industria alimentaria así como las bases de un sistema de vigilancia sensible y programas de capacitación en comunicación

Food defense refers mainly to the intentional contamination of the food supply, thus threatening the safety of the agri-food chain, which can mean great risks and huge impacts. The prevention of food contamination and the reduction of the likelihood of deliberate contamination and knowledge of hazards are the first steps to build effective food defense strategies and require a recurring effort throughout the food supply chain. The intentional nature of these offensive risks that may originate in the internal or external environment of the food industry means that different factors must be considered and different tactics are required to move beyond common food security approaches. However, knowledge about basic food defense techniques remains limited. In the present study, basic food defense concepts are explored to delimit food vulnerability at the level of the manufacturer, suppliers and customers of the food industry, as well as the foundations of a sensitive surveillance system and communication training programs

FoodSimplex - a public health tool to improve restaurants cleaning and sanitation status / FoodSimplex - una herramienta de salud pública para mejorar el estado de sanitario de los restaurantes

Foodborne diseases have a negative impact on health, and little consideration has been given to such because the symptoms are often moderate and self-limiting. The microbiological contamination of meals by catering systems is one of the main issues, and it must be assessed to ensure safer meals. The aim of this study is to evaluate the hygienic quality in portuguese micro, small and medium restaurants through audit data and microbial sampling through the application of new food safety methodology - FoodSimplex. This four-year longitudinal study revealed maintenance or an improvement in the compliance with hygiene requirements according to the FoodSimplex checklist, and with statistical significance evolution in food handlers hands microbial analyses (p=0,003), which confirms that FoodSimplex contributed for a general food safety status uplift

Las enfermedades transmitidas por los alimentos tienen un impacto negativo en la salud y se les ha prestado poca atención porque los síntomas suelen ser moderados y autolimitados. La contaminación microbiológica de las comidas por los sistemas de restauración es uno de los principales problemas, y debe evaluarse para garantizar comidas más seguras. El objetivo de este estudio es evaluar la calidad higiénica en micro, pequeños y medianos restaurantes portugueses a través de datos de auditoría y muestreo microbiano mediante la aplicación de una nueva metodología de seguridad alimentaria: FoodSimplex. Este estudio longitudinal de cuatro años reveló un mantenimiento o una mejora en el cumplimiento de los requisitos de higiene de acuerdo con la lista de verificación de FoodSimplex, y con una evolución estadísticamente significativa en los análisis microbianos (p = 0,003), que confirma que FoodSimplex contribuyó para aumentar la seguridad general de los alimentos

Enniatin B induces expression changes in the electron transport chain pathway related genes in lymphoblastic T-cell line.

Enniatin B is a ionophoric and lipophilic mycotoxin which reaches the bloodstream and has the ability to penetrate into cellular membranes. The purpose of this study was to reveal changes in the gene expression profile caused by enniatin B in human Jurkat lymphoblastic T-cells after 24 h of exposure at 1.5, 3 and 5 µM by next generation sequencing. It was found that up to 27% of human genome expression levels were significantly altered (5750 genes for both down-regulation and up-regulation). In the three enniatin B concentrations studied 245 differentially expressed genes were found to be overlapped, 83 were down and 162 up-regulated. ConsensusPathDB analysis of over-representation of differentially expressed genes provided a list of gene ontology terms in which several biological processes related to nucleoside monophosphate metabolic process, respiratory chain complex, electron transport chain, oxidative phosphorylation and cellular respiration were the most altered. Also, an interesting correlation was found between enniatin B toxicity and the up-regulation of the UCP protein complex. In summary, the transcriptomic analysis revealed that mitochondria are the organelles showing more related differentially expressed genes. Consequently, differentially expressed genes involved in biological processes, molecular functions and pathways related to mitochondrial metabolism and respiration were significantly changed.

Beauvericin and enniatin B effects on a human lymphoblastoid Jurkat T-cell model.

Several mycotoxins exert their effect on the immunological system; some are classified as immunotoxic. Jurkat T-cells were used to study toxic effects of beauvericin (BEA) and enniatin B (ENN B). Both are not legislated mycotoxins with increasing presence in feed and food. Concentrations studied were from 1 to 15 µM at 24, 48 and 72 h. Cell death by increasing the percentage of apoptotic/necrotic cells was: BEA > ENN B. IC50 values ranged from 3 to 7.5 µM for BEA. ENN B 15 µM decreased viability (21-29%). The percentage of apoptotic/necrotic cells was BEA > ENN B at 24 h but not at 48 h. Caspase-3&7 activation profile varied, although both mycotoxins increased this activation. No difference in ROS production for any mycotoxin was observed. Arrest in S phase for both mycotoxins was obtained. BEA increased the percentage of DNA in the tail (18% and 20%) with respect to the control, whereas not for ENN B. In summary, cytotoxicity of BEA involved mitochondrial alterations; while ENN B only at highest concentrations and time assayed. BEA had cell cycle disturbances and apoptotic and apoptotic/necrotic cells increased; for ENN B these were not evident. Different toxic responses in Jurkat T-cells may be involved in BEA and ENN B toxicity.

Transcriptomic study of the toxic mechanism triggered by beauvericin in Jurkat cells.

Beauvericin (BEA), an ionophoric cyclic hexadepsipeptide mycotoxin, is able to increase oxidative stress by altering membrane ion permeability and uncoupling oxidative phosphorylation. A toxicogenomic study was performed to investigate gene expression changes triggered by BEA exposure (1.5, 3 and 5 µM; 24 h) in Jurkat cells through RNA-sequencing and differential gene expression analysis. Perturbed gene expression was observed in a concentration dependent manner, with 43 differentially expressed genes (DEGs) overlapped in the three studied concentrations. Gene ontology (GO) analysis showed several biological processes related to electron transport chain, oxidative phosphorylation, and cellular respiration significantly altered. Molecular functions linked to mitochondrial respiratory chain and oxidoreductase activity were over-represented (q-value < 0.01). Pathway analysis revealed oxidative phosphorylation and electron transport chain as the most significantly altered pathways in all studied doses (z-score > 1.96; adj p-value < 0.05). 77 genes involved in the respiratory chain were significantly down-regulated at least at one dose. Moreover, 21 genes related to apoptosis and programmed cell death, and 12 genes related to caspase activity were significantly altered, mainly affecting initiator caspases 8, 9 and 10. The results demonstrated BEA-induced mitochondrial damage affecting the respiratory chain, and pointing to apoptosis through the caspase cascade in human lymphoblastic T cells.

Análisis de los inhibidores de las alfa-amilasa y la tripsina contenidos en el trigo y otros cereales: potenciales promotores de la inflamación intestinal / Analysis of Amylase/Trypsin's Inhibitors contained in wheat and other cereals: potential promoters of intestinal inflammation

Las plantas han desarrollado un sistema de resistencia a insectos y plagas mediante la síntesis de compuestos como los inhibidores de las alfa-amilasas y/o las proteasas digestivas, proteínas que actúan también contra las enzimas digestivas de mamíferos. Diversos estudios han demostrado su implicación en la activación del sistema inmune y posiblemente, en la sintomatología de patologías como la Sensibilidad al Gluten no Celíaca, siendo esta reacción proporcional al contenido en el cereal. El objetivo de este trabajo fue realizar una revisión de las diferentes estrategias para la extracción, purificación, detección y cuantificación de estas proteínas en el trigo y en otros cereales de la dieta actual. Esta revisión incluyó los métodos de análisis con espectrometría de masas del periodo 2000 - 2017, identificándose 114 artículos relevantes de los que se seleccionaron 26. Actualmente no existe un método estandarizado de control que permita determinar estas proteínas de forma clara, concisa y fiable

Plants have developed a system of resistance to insects and pests through the synthesis of certain compounds such as alpha-amylase inhibitors and/or digestive proteases, proteins that can also act against digestive enzymes of mammals. Studies have shown their involvement in the activation of the immune system and possibly in the symptomatology of non-coeliac Gluten Sensitivity, being the reaction proportional to cereal content. The objective of this study was to review the different strategies used for extraction, purification, detection and quantification of these inhibitors in wheat and other cereals of the current diet. A review of the literature published between 2000-2017 on methods of analysis by mass spectrometry was carried out. 114 relevant articles were identified from which 26 were selected for review. At present, there is no standardized control method for the clear, concise and reliable determination of these proteins

A decade of Food Safety Management System based on ISO 22000: a global overview / Una década del Sistema de gestión de la seguridad alimentaria basado en ISO 22000: visión general

The worldwide implementation and certification of food safety management systems (FSMS) have significantly increased during the last decade, reflecting the importance of assuming these standards in activity sectors involved in the food chain. Companies of all types and sizes worldwide have made several efforts for implementing a FSMS based on International Organization for Standardization (ISO) 22000. Certification system and quality management tools development in international context, are analyzed in food processing and food service sectors. Studies of food safety, performance of ISO 22000 implemented in food companies and hurdles of context characteristics wherein operate were reviewed. To summarize, improved product quality and safety has been identified as the major benefit of implementing ISO 22000. Other benefits highlighted include enhanced employee skills, improved company image, increased product sales, increased market share and access to new markets (AU)

La implementación y certificación mundial de los sistemas de gestión de la seguridad alimentaria (SGSA) se han incrementado significativamente durante la última década, lo que refleja la importancia de asumir estos estándares en los sectores de actividad involucrados en la cadena alimentaria. Empresas de todos los tipos y tamaños han realizado esfuerzos para implementar un SGSA basado en la Organización Internacional de Normalización (ISO) 22000. El sistema de certificación y el desarrollo de herramientas de gestión de calidad en el contexto internacional se analizan en los sectores de procesamiento de alimentos y servicios de alimentos. Se revisaron los estudios de inocuidad de los alimentos, el desarrollo de ISO 22000 implementado en las empresas de alimentos y los problemas según las características del contexto en donde opera. En resumen, la mejora en la calidad y seguridad del producto ha sido identificada como el principal beneficio de implementar ISO 22000. Otros beneficios destacados incluyen mejoras en las habilidades de los empleados, en la imagen de la compañía, mayores ventas de productos, mayor participación de mercado y acceso a nuevos mercados (AU)