Improving self-defense in plants. Martial arts for vegetables*

Abstract: From the dawn of agriculture there has been an ever-intensifying human effort to improve yields by having crops with enhanced biological similarity (i.e., characteristics of product, maturation time, height, color, etc.). The ultimate stage is to plant a crop where all individuals behave in exactly the same way, being clones of each other. This very intensive approach leads to loss of intrapopulation biodiversity and to unstable systems, prone to disastrous losses should anything go wrong. Biological evolutionary success is usually derived from high adaptability to everchanging external conditions. Highly specialized plants (such as certain orchids) or animals survive by correctly performing a high-wire act of enormous risk. External disbalances have catastrophic results on these species. Nature excels and corrects imbalances increased biodiversity within natural populations. Given this situation, we should study the defensive systems used by plants and improve on those natural systems

A first approximation to the adjustment of pesticide use and theoretical MRL's and ADI accomplishment in fruits and vegetables in Uruguay

Pesticide residues in fruits and vegetables (F&V) are of primary concern as these foods can be usually consumed as such. Little if any processing is done before their consumption apart from peeling and washing, depending on the type of F&V considered. Because of that, monitoring programs are performed all over the world to ensure legal MRL's accomplishment. The MRLs pursue two main objectives: to enforce Good Agricultural Practices accomplishment and protect consumers' health. In Uruguay, the number of legally allowed pesticides dropped from 453 to 285 active principles during the past decade. For instance, no pesticide of toxicological Level I is permitted in the country. In this work, thorough research on the active principles employed in Uruguay in F&V was performed. Of 233 pesticides registered for the 35 most cultured and consumed F&V in the country, 72 are insecticides, 60 are fungicides, and 101 herbicides. Among the insecticides, 20 were OPs, 10 carbamates, 11 pyrethroids, 6 neonicotinoids, and 25 belong to other chemical classes (matrine, azadirachtin, spinosad, abamectin, chlorantraniliprole a.o.). Most of the 60 fungicides active principles belong to the dithiocarbamates, Cyt-P450 inhibitors, and strobilurins chemical classes. A deterministic approach for evaluating chronic dietary risk for pesticide intake was performed following the recommended procedures by the World Health Organization (WHO) for chlorpyrifos, an insecticide from the organophosphate class, and the two most employed dithiocarbamates in Uruguay: Mancozeb and Ziram. The National Maximum Theoretical Daily Intake (IDTMN) was calculated using consumption data of fruits and vegetables of the National Survey of Household Expenditure and Income (ENGIHS), the Accepted Daily Intake (ADI), and the Maximum Residue Limits (MRL). As a result of the deterministic analysis, chlorpyrifos theoretical intake represented 92% of the IDA value below the safety limits. Nevertheless, as chlorpyrifos is allowed to be used in other highly consumed commodities such as cereals and grains, the maximum value of IDA for it can be easily reachable. In the case of dithiocarbamates, the results varied depending on the studied compound. The MRL for the whole chemical class is expressed as mg of CS 2 /kg sample, and correction factors for each specific compound must be applied to assess dietary risk assessment. Within this context, Mancozeb represented 48% of the IDA and Ziram 427% IDA. The results of a monitoring program on a seasonal basis of pesticide residues in fruits and vegetables will be presented to refine the values obtained with the theoretical calculations; aiming to verify the adjustment of the established MRLs to the Uruguayan diet will be presented and discussed.Agencia Nacional de Investigación e Innovació

Seasonal surveillance of pesticide residues and preliminary dietary risk assessment in commonly consumed F&V in Uruguay

Fruits and vegetables are essential foods for a balanced diet. They have the particularity that they are mostly consumed as such, without further processing. Therefore, the concentration of residues from the application of phytosanitary products for crop protection must be controlled. On average, Uruguay consumes less than half of the F&V recommended by the WHO. For that reason, a consumption incentive program called "Smart Basket or Smart List” has been developed in the last few years. In this work, seasonal monitoring of fruits and vegetables is being carried out to know the positive findings in the different matrices, their concentrations, and their relationship regarding the maximum residue limits. The monitoring results since the summer of 2020 for matrices such as onion, tomato, citrus, apple, pumpkin, carrot, eggplant, and grapes are presented. Each analytical methodology for the selected F&V was validated using a QuEChERS AOAC 2007.01 scheme, working with the Codex Alimentarius groups and contrasting the figures of merit required by the current SANTE document to ensure the quality of the results obtained. The method was adjusted for the GC-MS/MS determination of approximately 80 analytes, depending on the matrix under study. For most of the matrices studied, at least one pesticide residue was found within the method’s scope, belonging to the technological package used for each matrix. In turn, almost all the concentrations for the cases studied were below the Codex MRL values. Regarding the identities of the findings, 22 compounds were the most detected, mainly insecticides and fungicides. The generation of empirical data will allow us to refine the calculations presented in previous communications and go from working with theoretical concentrations or Maximum Limits to distributions of total concentrations. This work aims to obtain a phased or gradual approach to an intake risk assessment and a global overview of the compliance of the Codex MRLs with the findings for the Uruguayan situation.Agencia Nacional de Investigación e Innovació

Pesticide residues in onion varieties marketed in Uruguay

Onion is one of the most consumed vegetables all over the world, and Uruguay is not the exception. As a part of a global study of the pesticide intake of the Uruguayan population, we present the results of the evaluation of pesticide residues content in three onion varieties marketed in Montevideo. Samples of spring onion (Allium fistulosum), white and red onion (Allium cepa) were analyzed for 57 pesticide residues through gas chromatography coupled to tandem mass spectrometry. The compounds studied included the pesticides approved to be applied to onion plus the banned organochlorines. As onion is a bulb, the food contamination by non-approved and no longer used organochlorines cannot be dismissed because they are very persistent compounds that are still found in soils of old farms, where horticultural activities have been carried out for decades. The method will be useful for the monitoring of pesticde residues in onions markted in Uruguay, aiming to perfrmo risk assesment studies on pesticde exposure of the uruguayan population.Agencia Nacional de Investigación e Innovació

Flora alimenticia disponible para Apis mellifera (Himenoptera: Apidae) en un área de bosque serrano en Uruguay

Título alternativo : Flora alimentar disponível para Apis mellifera (Himenoptera: Apidae) em uma área de floresta de altitude no UruguaiKnowing the blooms is useful to predict the behavior of Apis mellifera (honeybee) colonies. The objective was to determine the floristic characteristics of a mountain forest in Uruguay from the beekeeping point of view. We made eight visits in every three sites surrounding the apiary, at 10-20 m, 500 m and 1500 m distance. The first visit determined the species, families, origin (native or exotic), and frequency of each taxon. In subsequent visits, we censed which ones remained in bloom (presence or absence). We assigned each species a value of one to five for abundance (A) and the duration of flowering (P), during two years. Using the Shannon Index the floristic diversity was established. A generalized linear model was employed to analyze flowering through the response variables: presence of flowering in winter, spring, summer and autumn, as well as flowering duration. We calculated a Shannon Index of 3.6, indicating high biodiversity. Then we correlated the findings of the survey and literature reports on the occurrence of each species in melissopalynological studies in mature and immature honey. The expression “Convertible Flora” was coined to identify the floral resources whose pollen is stored when no flower is available. These were Lithraea brasiliensis, Baccharis articulata, Baccharis trimera, Blepharocalyx salicifolius, Eugenia uniflora, Colletia Paradoxa, Oxalis sp., Scutia buxifolia, Jodina rhombifolia, Aloysia gratissima for the spring and autumn seasons (P <0.05). The expression “Support Flora” (P <0.05) is used to classify those resources that are found in immature honey and are almost absent in mature honey, and are used as nutrients during the colony expansion: Schinus engleri, Maytenus ilicifolia, Eryngium pandanifolium, Baccharis punctulata, Abutilon pauciflorum, Daphnopsis racemosa, Allophylus edulis, and Celtis tala. This flora classification allows planning the expected developments and yields of honey from a region to establish technological useful and efficient packages of honey harvests.El conocimiento de las floraciones es útil para predecir el comportamiento de las colonias de Apis mellifera (abeja de la miel). El objetivo fue determinar las características florísticas desde el punto de vista apícola de un bosque serrano en Uruguay. Realizamos ocho visitas en tres sitios aledaños a la colmena: a 10-20 m, 500 m y 1500 m de distancia. En la primera visita determinamos especies, familias, origen (nativo o exótico) y frecuencia de cada taxón. En visitas posteriores, censamos cuáles estaban florecidas (presencia o ausencia). Asignamos a cada especie un valor de uno a cinco para la abundancia (A) y la duración de la floración (P), durante dos años. Utilizamos el índice de Shannon para medir la diversidad florística. Se utilizó un modelo lineal generalizado para analizar la floración a través de las variables respuesta: presencia de floración en invierno, primavera, verano y otoño, así como duración de la floración. Calculamos un índice de Shannon de 3,6, lo que indica una alta biodiversidad. Luego correlacionamos los hallazgos de la prospección con datos de la literatura sobre la ocurrencia de cada especie en estudios melisopalinológicos en mieles maduras e inmaduras. Por último, definimos el término flora convertible para identificar los recursos florales cuyo polen se almacena cuando no hay ninguna flora disponible. Estas fueron Lithraea brasiliensis, Baccharis articulata, Baccharis trimera, Blepharocalyx salicifolius, Eugenia uniflora, Colletia Paradoxa, Oxalis sp., Scutia buxifolia, Jodina rhombifolia, Aloysia gratissima, para las temporadas de primavera y otoño (P<0.05). El término flora de soporte (P<0.05) lo definimos para clasificar aquellos recursos que se encuentran en la miel inmadura y están casi ausentes en la miel madura, y se utilizan como nutrientes durante la expansión de la colonia: Schinus engleri, Maytenus ilicifolia, Eryngium pandanifolium, Baccharis punctulata, Abutilon pauciflorum, Daphnopsis racemosa, Allophylus edulis y Celtis tala. Esta clasificación de la flora permite planificar los desarrollos y rendimientos de la miel esperados de una región, y establecer paquetes tecnológicos útiles y eficientes de cosechas de miel.O conhecimento das flores é útil para prever o comportamento de colônias de Apis mellifera (abelha do mel). O objetivo foi determinar as características florísticas de uma floresta montanhosa no Uruguai do ponto de vista apícola. Fizemos oito visitas em três locais no entorno do apiário: a 10-20m, 500m e 1500m de distância dele. A primeira visita determinou as espécies, famílias, origem (nativa ou exótica) e frequência de cada táxon. Nas visitas subsequentes, censuramos quais permaneceram em flor (presença ou ausência). Atribuímos a cada espécie um valor de um a cinco para abundância (A) e duração da floração (P), durante dois anos. Usando o Índice de Shannon, a diversidade florística foi estabelecida. Um modelo linear generalizado foi empregado para analisar a floração através das variáveis ​​resposta: presença de floração no inverno, primavera, verão e outono, bem como a duração da floração. Calculamos um Índice de Shannon de 3,6, indicando alta biodiversidade. Após correlacionar os achados do levantamento e relatos da literatura sobre a ocorrência de cada espécie em estudos melisopalinológicos em mel maduro e imaturo. O termo “Flora Conversível” foi cunhado para identificar os recursos florais cujo pólen é armazenado quando nenhuma flor está disponível. Estes foram Lithraea brasiliensis, Baccharis articulata, Baccharis trimera, Blepharocalyx salicifolius, Eugenia uniflora, Colletia Paradoxa, Oxalis sp., Scutia buxifolia, Jodina rhombifolia, Aloysia gratissima para as estações de primavera e outono (P <0,05). O termo “Flora de Apoio” (P <0,05) é usado para classificar os recursos encontrados no mel imaturo e quase ausentes no mel maduro e são usados ​​como nutrientes durante a expansão da colônia: Schinus engleri, Maytenus ilicifolia, Eryngium pandanifolium, Baccharis punctulata, Abutilon pauciflorum, Daphnopsis racemosa, Allophylus edulis and Celtis tala. Esta classificação da flora permite planejar os desenvolvimentos e rendimentos esperados do mel de uma região para estabelecer embalagens tecnológicas úteis e eficientes de colheita de mel

Monitoreo estacional de residuos de plaguicidas en hortalizas en la búsqueda de fijar criterios para asegurar la inocuidad alimentaria. El caso de las cucurbitáceas

Como contribución al consumo seguro de F&H por parte de nuestra población, se ha llevado a cabo un monitoreo estacional piloto de frutas y verduras seleccionadas durante dos años para determinar la naturaleza y los niveles de residuos de plaguicidas en los diferentes productos básicos y su relación con el Límite Máximo legal establecido para ellos. En el presente trabajo presentamos los hallazgos en vegetales de la familia de las cucurbitaceas.Agencia Nacional de Investigación e Innovació

Desarrollo y validación de metodología para la determinación de residuos de plaguicidas en garbanzos y lentejas mediante GC-QqQ-MS

Últimamente ha aumentado la población que no consume alimentos de origen animal, por lo cual el consumo de legumbres aumentó considerablemente. Es probable que a estos cultivos se les administre plaguicidas, siendo posible encontrar residuos de estos afectando de esta manera la seguridad alimentaria. Teniendo en cuenta esto se plantea como objetivo el desarrollo de metodologías para determinar residuos de plaguicidas en lentejas y garbanzos. Debido a que las matrices tienen diferente composición se desarrolló una metodología para cada una. Luego de varias pruebas y diversos cambios se partió de 5 g, se hidrataron con H2O y se extrajeron con ACN con 1% de HAc. Para favorecer la separación de fases se agregó NaCl y luego MgSO 4 con AcONa. Luego se realizó limpieza del extracto con PSA y MgSO 4. Se seleccionaron 157 plaguicidas según su compatibilidad con GC y LC, teniendo en cuenta además las prácticas agrícolas para estos cultivos. De ellos 101 se analizaron mediante GC-QqQ-MS y 56 mediante LC-MS/MS. Al realizar la validación de ambos métodos se demostró veracidad en los niveles de trabajo (10, 20, 50 y 100μg kg -1 ) con recuperaciones promedio (70-123%) y RSD inferiores al 20%. Se evaluó la linealidad de las curvas de calibración en solvente y matriz, así como límites de cuantificación y el efecto de matriz según la guía SANTE 1 vigente. Estos métodos multicontaminante resultan útiles para asegurar la inocuidad de estos alimentos. Se analizaron 5 muestras reales de cada legumbre, no habiendo positivos cuantificables en ninguna de ellas.Agencia Nacional de Investigación e Innovació

Evaluación de la dinámica de residuos de plaguicidas durante la cocción de lentejas y garbanzos por LC-MS/MS

En nuestro país la población que no consume alimentos de origen animal crece día a día, por lo cual el consumo de legumbres ha aumentado considerablemente, siendo las semillas de leguminosas una fuente económica de proteínas, carbohidratos, vitaminas y minerales dietéticos. A estos cultivos se les administre plaguicidas, siendo posible encontrar residuos de éstos que afecten la seguridad alimentaria. Teniendo en cuenta esto es importante contar con metodologías analíticas para la determinación de residuos de plaguicidas que puedan estar presentes en las legumbres, y por otro lado, conocer, que sucede con ellos cuando el alimento está listo para su consumo. Como respuesta a esta situación, el presente trabajo plantea desarrollar un método de preparación de muestra empleando lentejas y garbanzos cocidos y sus respectivas aguas de cocción como matriz de estudio para determinar la proporción en la cual los plaguicidas existentes en las legumbres secas se retienen en las cocidas y transfieren a su agua de cocción. Debido a que las matrices tienen diferente composición se desarrollaron metodologías específicas para cada una de ellas. Luego de varias pruebas, una modificación del método AOAC 2007.01 fue utilizada. Se partió de 5 g de legumbre cocida, se extrajeron con ácido acético al 1% en acetonitrilo. Para favorecer la separación de fases se agregó cloruro de sodio, sulfato de magnesio (MgSO4) y acetato de sodio y se realizó limpieza del extracto con Amina Primaria y Secundaria y MgSO4. Se centrifugó, filtró e inyectó en HPLC-MS/MS. Respecto al agua de remojo y de cocción, se tomaron 25mL, se extrajeron con ácido acético al 1% en acetonitrilo. Para favorecer la separación se agregaron 8g de MgSO4. Se agitó manualmente y luego se centrifugó. Se filtró la fase orgánica y se inyectó en HPLC-MS/MS. Se seleccionaron 58 plaguicidas teniendo en cuenta las prácticas agrícolas para estos cultivos y además la compatibilidad de estos con el equipo. Se validaron las metodologías desarrolladas evaluando veracidad en los niveles de 20, 50 y 100 μg.kg-1, obteniéndose recuperaciones promedio (70-123%) y RSD inferiores al 20%. Se evaluó la linealidad de las curvas de calibración en solvente y matriz en el rango de 10-200 μg.kg-1, así como límites de cuantificación y el efecto de matriz según la guía SANTE [1] vigente. Con las metodologías validadas y las previamente validadas para legumbres secas fue posible determinar, para los residuos de plaguicidas encontrados, el factor de procesamiento de los mismos en el material seco a las legumbres cocidas. A 20 μg.kg-1, producto del proceso de cocción, el 10% de los plaguicidas no se detectaron, y para los restantes se observó un muy bajo contenido residual (<30%). En el agua de cocción solo un 45% de los pesticidas fueron detectados. Se discuten los factores de procesamiento obtenidos con las propiedades fisicoquímicas, como la solubilidad en agua (Ws), el coeficiente octanol-agua (Kow), la constante de Henry (KH) y su estabilidad térmica. La cocción, previo remojo de la legumbre es una manera de asegurar su inocuidad al consumirlas.Agencia Nacional de Investigación e Innovació

Study of the matrix effect of table and wine grapes using GC-QqQ-MS

The matrix effect in analytics is known as the variation in the response of the analytical system, induced by the presence of some components of the matrix (coextractives) in which the analyte is found. This effect occurs throughout the entire analytical determination. In separative techniques, the most common case is the coelution of the analyte with the matrix components. In other cases, the characteristics of the analytical system also participate in this effect. In gas chromatography, these two aspects are involved: how the matrix influences the degradation of compounds at the injection port and their ability throughout the chromatographic run. The study of it is contemplated in the validation of analytical methodologies. There are different ways of approaching the study of the matrix effect, ranging from statistical methods to methods based on instrumental determinations. According to the SANTE document, the matrix effect can be studied by analyzing the analytical sensitivity through the quotient of the slopes of the calibration curves prepared in the matrix and solvent, respectively. Using the equation below, complementary information is obtained when its sign and module are analyzed, classifying the effect as low, moderate, and high and, in turn, signal suppression/increase. EM (%) = ((matrix matched calibration slope)/(solvent calibration slope)-1) ×100 The present work studies the matrix effect that occurs using gas chromatography coupled with tandem mass spectrometry taking a table and wine grapes as a case study. As expected, the differences between the two cases not only lay in their morphological characteristics, such as size, the shape of the bunches, and thickness of the skin, nor in their cultivation method but also the profile of co-extractives for each case.,When the information of the matrix effect is not taken into account or the correspondence between the matrix selected as the target and the one being analyzed, the consequence is thus being able to In this way, alter changes in the result of quantification of thea pesticide residues either, under or over quantifying it. , when the information of the matrix effect is not taken into account or the correspondence between the matrix selected as the target and the one being analyzed. Different calibration curves with table grapes (muscatel and white) and wine grapes (Chardonnay, Tannat, Merlot, and Albariño) were prepared, and the responses were compared for a representative group of compounds analyzable by GC-MS/MS, which mainly comprised organophosphates, pyrethroids, organochlorines, azoles, and strobilurins. The matrix effect was generally high in most cases, regardless of the type of grape, but the analyte quantification varied from matrix to matrix . In turn, the coextractives profile was evaluated by gas chromatography (Q3-Scan mode) and thin layer chromatography using UV absorption and universal developer to relate the information qualitatively, detecting significant differences in the coextractives profile. Depending on whether they are wine or table grapes, white or red.Agencia Nacional de Investigación e Innovació