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Background Sulforaphane (SFN) is a dietary isothiocyanate, derived from glucoraphanin, present in cruciferous vegetables belonging to the Brassica genus. It is a biologically active phytochemical that acts as a nuclear factor erythroid 2-related factor 2 (Nrf2) inducer. Thus, it has been reported to have multiple protective functions including anticancer responses and protection against a toxic agent's action. Purpose The present work systematically reviewed and synthesised the protective properties of sulforaphane against a toxic agent. This review reveals the mechanism of the action of SFN in each organ or system. Methods The PRISMA guideline was followed in this sequence: researched literature, organised retrieved documents, abstracted relevant information, assessed study quality and bias, synthesised data, and prepared a comprehensive report. Searches were conducted on Science Direct and PubMed using the keywords "Sulforaphane" AND (“protective effects” OR “protection against”). Results Reports showed that liver and the nervous system are the target organs on which attention was focused, and this might be due to the key role of oxidative stress in liver and neurodegenerative diseases. However, protective activities have also been demonstrated in the lungs, heart, immune system, kidneys, and endocrine system. SFN exerts its protective effects by activating the Nrf2 pathway, which enhances antioxidant defenses and reduces oxidative stress. It also suppresses inflammation by decreasing interleukin production. Moreover, SFN inhibits apoptosis by preventing caspase 3 cleavage and increasing Bcl2 levels. Overall, SFN demonstrates multifaceted mechanisms to counteract the adverse effects of toxic agents. Conclusion SFN has potential clinical applications as a chemoprotective agent. Nevertheless, more studies are necessary to set the safe doses of SFN in humans.

Anatoxin-a (ATX-a) is a potent neurotoxin produced by several species of cyanobacteria whose exposure can have direct consequences, including neurological disorders and death. The increasing prevalence of harmful cyanobacterial blooms makes the detection and reliable assessment of ATX-a levels essential to prevent the risk associated with public health. Therefore, the aim of this review is to compile the analytical methods developed to date for the detection and quantification of ATX-a levels alone and in mixtures with other cyanotoxins and their suitability. A classification of the analytical methods available is fundamental to make an appropriate choice according to the type of sample, the equipment available, and the required sensitivity and specificity for each specific purpose. The most widely used detection technique for the quantification of this toxin is liquid chromatography–tandem mass spectrometry (LC-MS/MS). The analytical methods reviewed herein focus mainly on water and cyanobacterial samples, so the need for validated analytical methods in more complex matrices (vegetables and fish) for the determination of ATX-a to assess dietary exposure to this toxin is evidenced. There is currently a trend towards the validation of multitoxin methods as opposed to single-ATX-a determination methods, which corresponds to the real situation of cyanotoxins’ confluence in nature.

Propyl-propane thiosulfonate (PTSO), an antioxidant organosulfur compound present in the genus Allium, has become a potential natural additive for food and feed, as well as a possible biopesticide for pest control in plants. A toxicological assessment is necessary to verify its safety for livestock, consumers, and the environment. As part of the risk assessment of PTSO, this study was designed to explore its potential reproductive toxicity in mice following the OECD 416 guideline. The investigation spans two generations to comprehensively evaluate potential reproductive, teratogenic, and hereditary effects. A total of 80 CD1 mice per sex and generation were subjected to PTSO exposure during three phases (premating, gestation, and lactation). This evaluation encompassed three dose levels: 14, 28, and 55 mg PTSO/kg b.w./day, administered through the feed. No clinical changes or mortality attributed to the administration of PTSO were observed in the study. Some changes in the body weight and food consumption were observed, but not related to sex or in a dose-dependent manner. The two parental generations (F0, F1) exhibited normal reproductive performance, and the offspring (F1 and F2) were born without any abnormalities. The serum sexual hormone levels (progesterone -P-, testosterone -T-, estradiol -E2-, follicular stimulating hormone -FSH-, and luteinizing hormone -LH-) were in a normal range. Although significant changes were observed in the sperm analysis in the case of F0 group, no variation was found for F1 group, and no alterations in fertility were recorded either. The absolute organ weights and relative organ weight/body weight and organ weight/brain weight ratios, and the complete histopathological study, showed no significant alterations in males and females for all the generations considered. Considering all the results obtained, PTSO is not considered a reproductive or developmental toxicant in mice under the assayed conditions. These results support the go

En la presente Tesis Doctoral, se investigó la toxicidad de diferentes materiales derivados de grafeno, como el óxido de grafeno (OG), el óxido de grafeno reducido (OGr) y sus derivados funcionalizados con dodecil amina (DA) y cloruro de [2-(metacriloiloxi) etil] trimetilamonio (MTAC), para su uso en la industria alimentaria en el área de envasado alimentario. Para garantizar la seguridad de estos materiales en contacto con alimentos, se llevaron a cabo extensos estudios toxicológicos. Como paso previo al estudio de su seguridad se realizó una exhaustiva revisión bibliográfica sobre la toxicidad de los materiales de grafeno en el marco del riesgo alimentario, publicados en la literatura científica hasta la actualidad. Con el fin de determinar las diferentes propiedades fisicoquímicas y así conocer la identidad de cada uno de los derivados de estudio, se llevó a cabo una caracterización fisicoquímica y morfológica de los derivados de grafeno elegidos. Los resultados revelaron diferente contenido atómico y diferente carga, además de la aglomeración de las partículas. La evaluación toxicológica se centró modelos posibles diana de una exposición oral. Se realizaron ensayos in vitro utilizando líneas celulares representativas del sistema digestivo (Caco-2) y el hígado (HepG2). Se observó la internalización de los materiales en estas células, y se detectó citotoxicidad y estrés oxidativo, especialmente a las concentraciones más altas. Se llevó a cabo una digestión gastrointestinal in vitro para simular las condiciones del tracto digestivo y evaluar cómo afectaba a la toxicidad de los materiales. Se encontraron cambios en la morfología de las partículas y en la toxicidad después de la digestión, con disminución de la toxicidad en el caso de OGr pero aumento en los derivados funcionalizados. Para la posible comercialización de estos compuestos y antes de su incorporación a los envases alimentarios, se debe llevar a cabo una evaluación del potencial mutagénico y genotóxico (

Anatoxin-a (ATX-a) is a cyanobacterial toxin whose occurrence has been reported worldwide and has attracted increasing scientific interest due to its toxicity. Moreover, in nature, ATX-a usually appears together with other cyanotoxins, such as cylindrospermopsin (CYN), so possible interaction phenomena could happen and should be considered for risk assessment purposes. For this reason, the aim of this work was to explore the potential mutagenicity and genotoxicity of pure ATX-a and an ATX-a/CYN mixture using a battery of in vitro assays, including the bacterial reverse-mutation assay in Salmonella typhimurium (OECD 471) and the micronucleus test (MN) (OECD 487) on L5178Y Tk+/− cells. The results showed that ATX-a was not mutagenic either alone or in combination with CYN under the conditions tested. Nevertheless, genotoxic effects were observed for both ATX-a and its mixture with CYN following the in vitro MN assay. The genotoxicity exhibited by ATX-a was only observed in the absence of S9 mix, whereas in the cyanotoxin mixture the concentration-dependent genotoxicity of ATX-a/CYN in vitro was observed only in the presence of S9. Thus, the toxicity induced by cyanotoxin mixtures may vary from that produced by toxins alone, and consequently more studies are necessary in order to perform more realistic risk assessments.

The " Scientific dissemination conference: Contributions of Toxicology in substance safety research and for society" were held on May 19th, 2023 at the Faculty of Pharmacy of the University of Seville, thanks to a financial support received by the VII Plan Propio of the University of Seville (VII PPIT-US) for scientific dissemination activities. The objective was to disseminate scientific knowledge among our students by highly qualified teaching and scientific personnel, and also to inform society in general of the high-level scientific research work that is being carried out in the University of Seville. The activity was aimed at students of the subjects of "Toxicology", "Chemoinformatics, Research and History of Pharmacy", " Laboratory of Pharmacy" (of the Degree in Pharmacy, Degree in Optics and Optometry, and Double Degree in Pharmacy and Optics and Optometry), and the subjects of "Toxicology of the Drugs of Abuse" or "Introduction to Forensic Sciences" (Degree in Criminology). During the Conference, which lasted 5.5 hours, 6 oral communications were given on graphene, cyanotoxins, alliaceous additives in human and animal feed, stilbenes in wines, or biopesticides. All of them were synthesized in informative triptychs distributed among the attendees for their dissemination in their social environment. Likewise, 2 invited presentations were given by experts in Forensic and Environmental Toxicology, respectively. The activity had 75 attendees and the results of the surveys generally showed a high degree of satisfaction with them, showing the Conference as an enriching outreach activity, due to the variety of topics covered, the new knowledge acquired and the approach to research and interest aroused among attendees., Las “Jornadas de divulgación científica: Aportaciones de la Toxicología en la investigación de la seguridad de las sustancias y para la sociedad” se celebraron el 19 de mayo de 2023 en la Facultad de Farmacia de la Universidad de Sevilla, gracias a una Ayuda recibida por el VII Plan Propio de la Universidad de Sevilla (VII PPIT-US) para actividades de divulgación científica. El objetivo fue divulgar entre nuestros alumnos los conocimientos científicos por parte de personal docente e investigador altamente cualificado, y también dar a conocer a la sociedad en general el trabajo de investigación científica de alto nivel que se está realizando en la US. La actividad iba dirigida a los alumnos de las asignaturas de “Toxicología”, “Quimioinformática, Investigación e Historia de la Farmacia”, “Laboratorio de Farmacia” (del Grado en Farmacia, Grado en Óptica y Optometría, y Doble Grado en Farmacia y Óptica y Optometría), y las asignaturas de “Toxicología de las Drogas de Abuso” o “Introducción a las Ciencias Forenses” (Grado en Criminología). Durante las Jornadas, que duraron 5,5 horas, se impartieron un total de 6 comunicaciones orales con diferente temática: el grafeno, las cianotoxinas, los aditivos aliáceos en alimentación humana y animal, los estilbenos en los vinos, o los bioplaguicidas; todas ellas sintetizadas en unos trípticos divulgativos repartidos entre los asistentes para su divulgación en su entorno social. Asimismo, se realizaron 2 ponencias invitadas por parte de expertos en Toxicología Forense y Ambiental, respectivamente. La actividad contó con casi un total de 75 asistentes y los resultados de las encuestas mostraron en general un alto grado de satisfacción con las mismas, mostrando las Jornadas como una enriquecedora actividad de divulgación científica, por la variedad de temas tratados, los nuevos conocimientos adquiridos y el acercamiento a la investigación e interés suscitado entre los asistentes.

Propyl-propane-thiosulfonate (PTSO) is an organosulfur compound found inAllium spp. Due to its antioxidant and antimicrobial activities, PTSO has been proposed for applications in the agri-food sector, such as feed additive. However, its use with commercial purposes depends on its toxicity evaluation. The present work aimed to perform a pilot-study of toxicokinetic profile of PTSO combining in silico and in vitro techniques, important steps in the risk assessment process. In silico ecotoxicity studies were also performed considering the importance of the environmental impact of the compound before its commercial use. First, an analytical method has been developed and validated to determine the original compound and its metabolites by ultra-performance liquid chromatography-tandem mass spectrometry. The phase I and II metabolism of PTSO was predicted using Meta-Pred Web Server. For the phase I metabolism, rat (male and female) and human liver microsomes were incubated with PTSO and NADPH regeneration system. Furthermore, in the phase II, microsomes were incubated with PTSO and glutathione or uridine 5′- diphosphoglucuronic acid. The analysis revealed the presence of propylpropane thiosulfinate (PTS) originated by redox reaction in phase I, and two conjugates from the phase II: S-propylmercaptoglutathione (GSSP) and S-propylmercaptocysteine (CSSP). Additionally, considering the environmental fate of PTSO and its metabolites, the ADME parameters and the potential ecotoxicity were also predicted using in silico softwares. The results of the ecotoxicity in silico study evidenced that the metabolism induced the formation of detoxified metabolites from the parent compound, except for dimercaprol and 3-mercaptopropane1,2-diol. Further in vivo assays are needed to confirm this prediction.

Las “Jornadas de divulgación científica: Aportaciones de la Toxicología en la investigación de la seguridad de las sustancias y para la sociedad” se celebraron el 19 de mayo de 2023 en la Facultad de Farmacia de la Universidad de Sevilla, gracias a una Ayuda recibida por el VII Plan Propio de la Universidad de Sevilla (VII PPIT-US) para actividades de divulgación científica. El objetivo fue divulgar entre nuestros alumnos los conocimientos científicos por parte de personal docente e investigador altamente cualificado, y también dar a conocer a la sociedad en general el trabajo de investigación científica de alto nivel que se está realizando en la US. La actividad iba dirigida a los alumnos de las asignaturas de “Toxicología”, “Quimioinformática, Investigación e Historia de la Farmacia”, “Laboratorio de Farmacia” (del Grado en Farmacia, Grado en Óptica y Optometría, y Doble Grado en Farmacia y Óptica y Optometría), y las asignaturas de “Toxicología de las Drogas de Abuso” o “Introducción a las Ciencias Forenses” (Grado en Criminología). Durante las Jornadas, que duraron 5,5 horas, se impartieron un total de 6 comunicaciones orales con diferente temática: el grafeno, las cianotoxinas, los aditivos aliáceos en alimentación humana y animal, los estilbenos en los vinos, o los bioplaguicidas; todas ellas sintetizadas en unos trípticos divulgativos repartidos entre los asistentes para su divulgación en su entorno social. Asimismo, se realizaron 2 ponencias invitadas por parte de expertos en Toxicología Forense y Ambiental, respectivamente. La actividad contó con casi un total de 75 asistentes y los resultados de las encuestas mostraron en general un alto grado de satisfacción con las mismas, mostrando las Jornadas como una enriquecedora actividad de divulgación científica, por la variedad de temas tratados, los nuevos conocimientos adquiridos y el acercamiento a la investigación e interés suscitado entre los asistentes., The " Scientific dissemination conference: Contributions of Toxicology in substance safety research and for society" were held on May 19th, 2023 at the Faculty of Pharmacy of the University of Seville, thanks to a financial support received by the VII Plan Propio of the University of Seville (VII PPIT-US) for scientific dissemination activities. The objective was to disseminate scientific knowledge among our students by highly qualified teaching and scientific personnel, and also to inform society in general of the high-level scientific research work that is being carried out in the University of Seville. The activity was aimed at students of the subjects of "Toxicology", "Chemoinformatics, Research and History of Pharmacy", " Laboratory of Pharmacy" (of the Degree in Pharmacy, Degree in Optics and Optometry, and Double Degree in Pharmacy and Optics and Optometry), and the subjects of "Toxicology of the Drugs of Abuse" or "Introduction to Forensic Sciences" (Degree in Criminology). During the Conference, which lasted 5.5 hours, 6 oral communications were given on graphene, cyanotoxins, alliaceous additives in human and animal feed, stilbenes in wines, or biopesticides. All of them were synthesized in informative triptychs distributed among the attendees for their dissemination in their social environment. Likewise, 2 invited presentations were given by experts in Forensic and Environmental Toxicology, respectively. The activity had 75 attendees and the results of the surveys generally showed a high degree of satisfaction with them, showing the Conference as an enriching outreach activity, due to the variety of topics covered, the new knowledge acquired and the approach to research and interest aroused among attendees.

The growing interest in graphene derivatives is a result of their variety of applications in many fields. Due to their use, the oral route could be a potential way of entrance for the general population. This work assesses the biotransformation of reduced graphene oxide (rGO) after an in vitro digestion procedure (mouth, gastric, intestinal, and colon digestion), and its toxic effects in different cell models (HepG2, Caco-2, and 3D intestinal model). The characterization of rGO digestas evidenced the agglomeration of samples during the in vitro gastrointestinal (g.i.) digestion. Internalization of rGO was only evident in Caco-2 cells exposed to the colonic phase and no cellular defects were observed. Digestas of rGO did not produce remarkable cytotoxicity in any of the experimental models employed at the tested concentrations (up to 200 µg/mL), neither an inflammatory response. Undigested rGO has shown cytotoxic effects in Caco-2 cells, therefore these results suggest that the digestion process could prevent the systemic toxic effects of rGO. However, additional studies are necessary to clarify the interaction of rGO with the g.i. tract and its biocompatibility profile.

Due to climate change and anthropogenic activities, the levels of pollution of aquatic and terrestrial environments have increased in the last decades. In this sense, the rise of cyanobacterial blooms, which release secondary metabolites with toxic properties, and the global use of pesticides for agricultural purposes have a negative impact on ecosystems. Thus, it would be interesting to study the concomitance of both types of toxicants in the same sample, since it is possible that they appear together. The aim of the present work was to state the effects of the interaction between the cyanotoxin cylindrospermopsin and the pesticide chlorpyrifos in differentiated SH-SY5Y neuronal cells to assess how they could affect the nervous system. To this end, cytotoxicity, morphological, and acetylcholinesterase activity studies were performed during 24 and 48 h. The results revealed a concentration-dependent decrease in viability and interaction between both toxicants, together with clear signs of apoptosis and necrosis induction. In this sense, different stages on the differentiation process would lead to differences in the toxicity exerted by the compounds both isolated as in combination, which it is not observed in non-differentiated cells. Additionally, the acetylcholinesterase activity appeared not to be affected, which is a clear difference compared to non-differentiated cells. These results show the importance of studying not only the toxicants themselves, but also in combination, to assess their possible effects in a more realistic scenario.

The incidence and interest of cyanobacteria are increasing nowadays because they are able to produce some toxic secondary metabolites known as cyanotoxins. Among them, the presence of cylindrospermopsin (CYN) is especially relevant, as it seems to cause damage at different levels in the organisms: the nervous system being the one most recently reported. Usually, the effects of the cyanotoxins are studied, but not those exerted by cyanobacterial biomass. The aim of the present study was to assess the cytotoxicity and oxidative stress generation of one cyanobacterial extract of R. raciborskii non-containing CYN (CYN−), and compare its effects with those exerted by a cyanobacterial extract of C. ovalisporum containing CYN (CYN+) in the human neuroblastoma SH-SY5Y cell line. Moreover, the analytical characterization of potential cyanotoxins and their metabolites that are present in both extracts of these cultures was also carried out using Ultrahigh Performance Liquid Chromatography-Mass Spectrometry, in tandem (UHPLC-MS/MS). The results show a reduction of cell viability concentration- and time-dependently after 24 and 48 h of exposure with CYN+ being five times more toxic than CYN−. Furthermore, the reactive oxygen species (ROS) increased with time (0–24 h) and CYN concentration (0–1.11 µg/mL). However, this rise was only obtained after the highest concentrations and times of exposure to CYN−, while this extract also caused a decrease in reduced glutathione (GSH) levels, which might be an indication of the compensation of the oxidative stress response. This study is the first one performed in vitro comparing the effect

The organosulfur compound propyl-propane thiosulfonate (PTSO), mainly found in Allium cepa, has a promising use in the agrifood industry. To confirm its safety for livestock, consumers, and environment, toxicological assessment is needed. In this regard, endocrine-disrupting chemicals (EDCs) are in the spotlight of research. Therefore, as part of the risk assessment of PTSO, in the present work, an in vivo study was performed in mice exposed to PTSO to investigate its potential reproductive toxicity considering fertility, genetic and endocrine endpoints. Five-weeks-old CD1 mice (80 males, 80 females) were exposed for 11 or 16 weeks (males or females, respectively) to different doses of PTSO (0, 14, 28 and 55 mg PTSO/kg b.w./day; 20 animals per group and sex) through the food pellets. No clinical observations or mortality and no changes in absolute organ weights and relative organ weights/body weight or brain ratios occurred during the study. The estrous cycle did not undergo any significant toxicologically relevant change. Most of the sex hormones displayed normal values. Some alterations in the expression of some genes related to reproduction is only observed in females, but they do not appear to have consequences in the development of sex organs. Docking results showed the impossibility of stable binding to estrogen and androgen receptors. Considering all the results obtained, the safe profile of PTSO can be confirmed for different agrifood applications at the conditions assayed.

Since the beginning of animal evolution, adaptation to the environmental factors has been key to survival in order to maintain the continuity of the spe cies. Therefore, numerous physiological mechanisms developed over the years to overcome adversities to the point where living organisms learnt to anticipate them through an internal biological clock: the circadian rhythm. Its discovery opened a new dimensi on of study in the scientific field with more and more researchers gaining interest about this subject up to the present day. It has been found that this internal rhythm is entrained by both internal and external factors and is responsible for the synchron ization of all physiological processes through peripheral clocks, where the main pacemaker, the suprachiasmatic nucleus (SCN), is located in the hypothalamus. The SCN releases neurotransmitters that orchestrate the rest of the system acting in different ta rget tissues, but recent studies showed that a deeper level of control governed the protein synthesis: the genetic molecular level. This means that our genetic material contains clock genes that regulate the biological clock. The goal of this work is to de scribe the main factors that negatively affect in different ways the correct function of the circadian rhythm, leading to health issues due to the chronodisruption and the consequent metabolic disorder. These can act in a wide range of situations, being cl assified as environmental, behavioural, intrinsic and iatrogenic factors. Finally, taken the fact that in modern life we’re constantly exposed to many of these factors, some ideas are exposed about the application of this knowledge into people’s lifestyles and into vital aspects such as pharmacological therapy, technological and chemical industry, etc which seek to protect and even promote human beings’ health.

Cylindrospermopsin (CYN) is a cyanotoxin whose incidence has been increasing in the last decades. Due to its capacity to exert damage at different levels of the organism, it is considered a cytotoxin. Although the main target organ is the liver, recent studies indicate that CYN has potential toxic effects on the nervous system, both in vitro and in vivo. Thus, the aim of the present work was to study the effects of this cyanotoxin on neuronal viability and synaptic integrity in murine primary cultures of neurons exposed to environmentally relevant concentrations (0–1 µg/mL CYN) for 12, 24, and 48 h. The results demonstrate a concentration-and time-dependent decrease in cell viability; no cytotoxicity was detected after exposure to the cyanotoxin for 12 h, while all of the concentrations assayed decreased this parameter after 48 h. Furthermore, CYN was also demonstrated to exert damage at the synaptic level in a murine primary neuronal culture in a concentration-and timedependent manner. These data highlight the importance of studying the neurotoxic properties of this cyanotoxin in different experimental models.

Cyanotoxins are secondary metabolites produced by different types of cyanobacteria. Among them, Cylindrospermopsin (CYN) and Microcystins (MCs) stand out due to their wide geographical distribution and toxicity in various organs, including the kidney, which is involved in their distribution and elimination. However, the renal toxicity caused by CYN and MCs has hardly been studied. The aim of this work was to assess the cytotoxicity effects caused by CYN and MC-LR in the renal cell line HEK293, and for the first time, the influence of CYN on the gene expression of selected genes in these cells by quantitative real-time PCR (qRT-PCR). CYN caused an upregulation in the gene expression after exposure to the highest concentration (5 µg/mL) and the longest time of exposure (24 h). Moreover, shotgun proteomic analysis was used to assess the molecular responses of HEK293 cells after exposure to the individuals and combinations of CYN + MC-LR. The simultaneous exposure to both cyanotoxins caused a greater number of alterations in protein expression compared to single toxins, causing changes in the cellular, lipid and protein metabolism and in protein synthesis and transport. Further studies are needed to complete the toxicity molecular mechanisms of both CYN and MC-LR at the renal level.

Microcystins (MCs) and cylindrospermopsin (CYN), although classified as hepatotoxins and cytotoxins, respectively, have been shown to also induce toxic effects in many other systems and organs. Among them, their potential endocrine disruption (ED) activity has been scarcely investigated. Considering the increasing relevance of ED on humans, mammals, and aquatic organisms, this work aimed to review the state-of-the-art regarding the toxic effects of MCs and CYN at this level. It has been evidenced that MCs have been more extensively investigated than CYN. Reported results are contradictory, with the presence or absence of effects, but experimental conditions also vary to a great extent. In general, both toxins have shown ED activity mediated by very different mechanisms, such as estrogenic responses via a binding estrogen receptor (ER), pathological changes in several organs and cells (testis, ovarian cells), and a decreased gonad-somatic index. Moreover, toxic effects mediated by reactive oxygen species (ROS), changes in transcriptional responses on several endocrine axes and steroidogenesis-related genes, and changes in hormone levels have also been reported. Further research is required in a risk assessment frame because official protocols for assessment of endocrine disrupters have not been used. Moreover, the use of advanced techniques would aid in deciphering cyanotoxins dose-response relationships in relation to their ED potential.

Organosulfur compounds (OSCs) are secondary metabolites produced by different Allium species which present important biological activities such as antimicrobial, antioxidant, anti-inflammatory antidiabetic, anticarcinogenic, antispasmodic, etc. In recent years, their use has been promoted in the agri-food industry as a substitute for synthetic preservatives, increasing potential accumulative exposure to consumers. Before their application in the food industry, it is necessary to pass a safety assessment as specified by the European Food Safety Authority (EFSA). This work reviews the scientific literature on OSCs regarding their in vitro toxicity evaluation following PRISMA guidelines for systematic reviews. Four electronic research databases were searched (Web of Science, Scopus, Science Database and PubMed) and a total of 43 works were selected according to predeterminate inclusion and exclusion criteria. Different data items and the risk of bias for each study were included. Currently, there are very few in vitro studies focused on investigating the potential toxicity of OSCs. Most research studies aimed to evaluate the cytotoxicity of OSCs to elucidate their antiproliferative effects focusing on their therapeutic aspects using cancer cell lines as the main experimental model. The results showed that diallyl disulfide (DADS) is the compound most studied, followed by diallyl trisulfide (DATS), diallyl sulfide (DAS), Allicin and Ajoene. Only 4 studies have been performed specifically to explore the safety of OSCs for agri-food applications, and genotoxicity studies are limited. More toxicity studies of OSCs are necessary to ensure consumers safety and should mainly be focused on the evaluation of genotoxicity and long-term toxicity effects.

Different applications have been suggested for graphene nanomaterials (GFNs) in the food and feed chain. However, it is necessary to perform a risk assessment before they become market-ready, and when consumer exposure is demonstrated. For this purpose, the European Food Safety Authority (EFSA) has published a guidance that has been recently updated. In this sense, the aim of this study is to identify and characterise toxicological hazards related to GFNs after oral exposure. Thus, existing scientific literature in relation to in vitro degradation studies, in vitro and in vivo genotoxicity, toxicokinetics data, in vivo oral studies, and other in-depth studies such as effects on the microbiome has been revised. The obtained results showed that the investigations performed up to now did not follow internationally agreed-upon test guidelines. Moreover, GFNs seemed to resist gastrointestinal digestion and were able to be absorbed, distributed, and excreted, inducing toxic effects at different levels, including genotoxicity. Also, dose has an important role as it has been reported that low doses are more toxic than high doses because GFNs tend to aggregate in the digestive system, changing the internal exposure scenario. Thus, further studies including a thorough toxicological evaluation are required to protect consumer's safety.

The presence of cyanotoxins and its bioaccumulation in the food chain is an increasingly common problem worldwide. Despite the toxic effects produced by Anatoxin-a (ATX-a), this neurotoxin has been less studied compared to microcystins (MCs) and cylindrospermopsin (CYN). Studies conducted under laboratory conditions are of particular interest because these provide information which are directly related to the effects produced by the toxin. Currently, the World Health Organization (WHO) considers the ATX-a toxicological database inadequate to support the publication of a formal guideline reference value. Therefore, the aim of the present work is to compile all of the in vitro and in vivo toxicological studies performed so far and to identify potential data gaps. Results show that the number of reports is increasing in recent years. However, more in vitro studies are needed, mainly in standardized neuronal cell lines. Regarding in vivo studies, very few of them reflect conditions occurring in nature and further studies with longer periods of oral exposure would be of interest. Moreover, additional toxicological aspects of great interest such as mutagenicity, genotoxicity, immunotoxicity and alteration of hormonal balance need to be studied in depth.

The organosulfur compounds (OSC) extracted from Allium spp. exhibit antibacterial, antifungal, and antioxidant properties. The agri-food industry is taking advantage of these properties by using them as natural feed and food additives. In the present work, an acute and a subchronic 90-days toxicity studies have been conducted for the first time to assess the safety of the OSC propyl-propane-thiosulfinate (PTS). Both studies were carried out following the Organization for Economic Co-operation and Development test guidelines (425 and 408, respectively). The acute study provided a maximum tolerated dose (MTD) of 175 mg/kg and the subchronic study established the Non Observed Adverse Effect Level (NOAEL) ≥ 55 mg/kg body weight (b.w.)/day in both sexes. In addition, the subchronic study performed on rats exposed to 14, 28 and 55 mg/kg b.w./day PTS, revealed no changes in any of the hematological parameters measured as well as no differences in body weight and water/food consumption. However, biochemical parameters were altered in some groups, although they were not biologically significant (Ca2+ in female rats, and the thyroids hormones T3 and T4 in rat males). Furthermore, the histopathological assessment evidenced no abnormality on the gastrointestinal, respiratory, lymphoid, urinary, circulatory, nervous, musculoskeletal, and reproductive systems.

The occurrence of toxic cyanobacteria and their cyanotoxins production are increasing phenomena reported worldwide. Cyanotoxins, secondary metabolites of different cyanobacterial species, are considered a potential hazard that can become a risk for human and environmental health after exposure. Global changes (rising temperatures, eutrophication due to human activities, etc.) favour their appearance leading to increased exposures and, consequently, risks. In order to avoid their negative impacts, the World Health Organization has established provisional drinking-water and recreational-water guideline values, and different countries have implemented limits in their regulations. However, these limits are not enough taking into account the increasing number of known cyanotoxins. Also, the establishment of these legal limits is hampered by the gaps in the knowledge of their toxicological profiles. Thus, this work aimed to establish the state of the art on the human toxicity of cyanotoxins (mainly microcystins, cylindrospermopsin and anaxotin-A) and to shed light on aspects not completely elucidated, such as target organ toxicity, immunotoxicity or genotoxicity.

Recientemente, el interés por el óxido de grafeno reducido (OGr) se ha visto incrementado debido a sus numerosas propiedades. Considerando que el uso de estos materiales ha aumentado en los últimos años, esto puede suponer un mayor riesgo para la salud humana. La vía oral es una de las principales rutas de exposición, por lo que es importante determinar cuáles son los principales efectos tóxicos sobre el sistema hepato-gastrointestinal. En este sentido, el objetivo de este estudio es revisar los posibles efectos tóxicos in vitro del OGr en las principales líneas celulares de los sistemas intestinal y hepático. Los resultados muestran que el OGr podría ser tóxico en modelos in vitro; sin embargo, son necesarios un mayor número de investigaciones para determinar los posibles mecanismos de toxicidad., Recently, the interest in reduced graphene oxide (rGO) has increased due to its myriad of properties. Considering the higher use of this material in the last years, it is important to evaluate its potential risk on human health. The oral pathway is one of the main exposure routes, therefore, it is important to assess its toxic effects on the hepato- gastrointestinal system. In this sense, the aim of this study is to review the potential in vitro toxic effects of rGO on gastrointestinal and liver cell lines. The results obtained showed that rGO could be toxic in in vitro models; however, further toxicological studies are required to define its mechanisms of toxicity.

En la actualidad, como consecuencia del cambio climático, la eutrofización de las aguas y el desarrollo de actividades antropogénicas, se está produciendo un aumento en la frecuencia y presencia de floraciones de cianobacterias productoras de cianotoxinas a nivel mundial. El hombre puede estar expuesto a estas toxinas mediante diferentes vías de exposición, siendo la oral a través del consumo de agua y alimentos contaminados la principal. Por ello, es necesario realizar una correcta caracterización del perfil toxicológico de estas toxinas, y en concreto, de las mezclas de cianotoxinas, al ser el escenario de exposición más posible, al encontrarse de manera simultánea en la naturaleza. Conocer más detalladamente la toxicidad de las combinaciones de cianotoxinas permitiría realizar una evaluación de riesgos más realista y fiable. Uno de los aspectos principales para poder llevar a cabo una correcta caracterización del peligro de las cianotoxinas es determinar su presencia, su capacidad de acumulación y su toxicidad. Por ello, con el fin de establecer y describir el estado del arte sobre las cianotoxinas, y en concreto de las Microcistinas (MCs), nos pareció necesario revisar la información existente en la literatura científica sobre la presencia, concentración y toxicidad de los congéneres de MCs diferentes de MC-LR, ya que aunque esta variante es la más estudiada por su toxicidad, ubicuidad y prevalencia, existen otras muchas que van cobrando cada vez más importancia. En primer lugar, nos centramos en el análisis de la bibliografía publicada sobre MC-RR, al ser el congénere más estudiado tras MC-LR. MC-RR es la variante más detectada en aguas del continente asiático, y está presente en alimentos de consumo humano (peces, bivalvos, vegetales, etc.), y su toxicidad ha sido observada tanto in vitro como in vivo. Sin embargo, son escasos los estudios que evalúan de modo específico sus propiedades toxicológicas, siendo necesarios para determinar los riesgos asociados a su, At present, as a consequence of climate change, eutrophication of waters and anthropogenic activities, there is an increase in the frequency and presence of cyanobacterial blooms producers of cyanotoxins worldwide. Humans can be exposed to these toxins through different exposure routes, and the oral intake through consumption of contaminated food and water is the main one. For this reason, it is necessary to carry out a correct characterization of the toxicological profile of these toxins, and in particular, of the cyanotoxin mixtures, as this is the most likely exposure scenario, since they are found simultaneously in nature. A more detailed understanding of the toxicity of cyanotoxins combinations would allow a more realistic and reliable risk assessment. One of the main aspects to perform a correct characterization of the risk of cyanotoxins is to determine their presence, accumulation capacity and toxicity. Therefore, in order to establish and describe the state of the art on cyanotoxins, and specifically Microcystins (MCs), it was considered necessary to review the existing information in the scientific literature on the presence, concentration and toxicity of MCs congeners different from MC-LR, since although this variant is the most studied due to its toxicity, ubiquity and prevalence, there are many others that are gaining more and more importance. First, we focus on the analysis of the published literature on MC-RR, as it is the most studied congener after MC-LR. MC-RR is the variant most detected in waters of the Asian continent, and it is present in food for human consumption (fish, bivalves, vegetables, etc.), and its toxicity has been observed both in vitro and in vivo. However, there are few studies that specifically evaluate its toxicological properties, being necessary to determine the risks associated with its exposure, both in human and in the environment. Afterwards, a review of the scientific literature on minority MCs congeners was carried out.

Propyl-propanethiosulfinate (PTS) is a component of Allium essential oils. This organosulfur molecule can be used as a feed additive to decrease the appearance of bacterial resistances caused by the residues of antibiotics. In previous in vitro genotoxicity studies, contradictory results were reported for PTS. In this work, the in vivo genotoxicity of PTS in male and female rats was assessed for the first time, following OECD (Organisation for Economic Co-operation and Development) guidelines. After oral administration (doses: 5.5, 17.4, and 55.0 mg/kg PTS body weight), a combination of the micronucleus (MN) assay (OECD 474) in bone marrow and the standard and enzyme-modified comet assay (OECD 489) was performed. After necropsy, histopathological studies were also carried out. The results did not show the in vivo genotoxicity of PTS at any doses assayed, revealed by the absence of increased MN, and DNA strand breaks or oxidative DNA damage in the standard and enzyme-modified comet assays. The histopathological study revealed that only the highest dose tested (55.0 mg/kg) in the liver and all dose groups in the stomach presented minimal pathological lesions in the organs studied. Consequently, the present work confirms that PTS is not genotoxic at the doses assayed, and it is a promising natural alternative to synthetic preservatives and antibiotics in animal feed.

Since 2013, Xylella fastidiosa Wells et al. has been reported to infect several hosts and to be present in different areas of Europe. The main damage has been inflicted on the olive orchards of southern Apulia (Italy), where a severe disease associated with X. fastidiosa subspecies pauca strain De Donno has led to the death of millions of trees. This dramatic and continuously evolving situation has led to European and national (Italian and Spanish) measures being implemented to reduce the spread of the pathogen and the associated olive quick decline syndrome (OQDS). Research has been also carried out to find solutions to better and directly fight the bacterium and its main insect vector, Philaenus spumarius L. In the course of this frantic effort, several treatments based on chemical or biological substances have been tested, in addition to plant breeding techniques and integrated pest management approaches. This review aims to summarize the attempts made so far and describe the prospects for better management of this serious threat, which poses alarming questions for the future of olive cultivation in the Mediterranean basin and beyond.

Propyl-propane-thiosulfonate (PTSO) and Propyl-propane-thiosulfinate (PTS) are organosulfur compounds used to supplement the diet of livestock because of their beneficial effects on feed palatability, their antibacterial, anti-inflammatory, and antimethanogenic activities. Besides, antibiotic residues in the environment can be reduced by using these natural bioactive compounds. The objective of this study was to optimize the extraction parameters for the analysis of PTSO and PTS in feed matrices by performing a solid-liquid extraction and quantification by Ultra performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS). Optimization was performed using the Response Surface Methodology on a Box–Behnken experimental design, optimizing the following parameters: solvent:sample ratios and evaporation temperature set for the rotary evaporator. The method was validated for 3 concentration levels for both PTSO (100, 500, 1000 ng g−1) and PTS (500, 1150, 2300 ng g−1). The highest recoveries of PTSO and PTS were obtained using 12.5 mL of 100% acetonitrile, stirring for 15 min, and an evaporation temperature of 20 °C. The validated method was further applied to detect and quantify these compounds in different feed matrices. In conclusion, this is the first study to simultaneously analyze PTSO and PTS at low concentrations, employing a sensitive technique such as UPLC-MS/MS.

Cyanotoxin occurrence is gaining importance due to anthropogenic activities, climate change and eutrophication. Among them, Microcystins (MCs) and Cylindrospermopsin (CYN) are the most frequently studied due to their ubiquity and toxicity. Although MCs are primary classified as hepatotoxins and CYN as a cytotoxin, they have been shown to induce deleterious effects in a wide range of organs. However, their effects on the immune system are as yet scarcely investigated. Thus, to know the impact of cyanotoxins on the immune system, due to its importance in organisms’ homeostasis, is considered of interest. A review of the scientific literature dealing with the immunotoxicity of MCs and CYN has been performed, and both in vitro and in vivo studies have been considered. Results have confirmed the scarcity of reports on the topic, particularly for CYN. Decreased cell viability, apoptosis or altered functions of immune cells, and changed levels and mRNA expression of cytokines are among the most common effects reported. Underlying mechanisms, however, are still not yet fully elucidated. Further research is needed in order to have a full picture of cyanotoxin immunotoxicity.

Las cianobacterias son bacterias Gram negativas fotoautótrofas presentes en ecosistemas acuáticos, que suponen una amenaza para la salud humana y ambiental debido a la producción de floraciones. Éstas, a su vez, pueden producir también cianotoxinas, las cuales pueden actuar como hepatotoxinas, neurotoxinas, citotoxinas o dermatotoxinas. Entre las cianotoxinas de mayor interés se encuentran las microcistinas (MC) y la cilindrospermopsina (CYN) por su amplia distribución y toxicidad. La CYN puede causar daños en distintos órganos, tales como hígado o riñón, siendo este último objeto del presente estudio, es decir, el objetivo principal de esta revisión bibliográfica se centra en poner de manifiesto las alteraciones que produce la CYN a nivel renal. Tras la realización de una búsqueda bibliográfica, se han seleccionado un total de veintiocho artículos en relación con la nefrotoxicidad de la CYN. Se evidencian efectos producidos como resultado de una exposición a CYN por diferentes vías tanto en modelos experimentales in vivo como in vitro, sin embargo, la nefrotoxicidad de CYN sigue siendo un tema aún poco estudiado. Los principales resultados observados incluyeron cambios histopatológicos, acompañados de variaciones en niveles de biomarcadores séricos tanto en organismos acuáticos como en mamíferos. Por tanto, se considera que la CYN presenta un alto potencial nefrotóxico, siendo de interés abordar una investigación más profunda sobre su mecanismo de acción tóxica en este órgano.

Toxic cyanobacterial blooms are a major contaminant in inland aquatic ecosystems. Furthermore, toxic blooms are carried downstream by rivers and waterways to estuarine and coastal ecosystems. Concerning marine and estuarine animal species, very little is known about how these species are affected by the exposure to freshwater cyanobacteria and cyanotoxins. So far, most of the knowledge has been gathered from freshwater bivalve molluscs. This work aimed to infer the sensitivity of the marine mussel Mytilus galloprovincialis to single as well as mixed toxic cyanobacterial cultures and the underlying molecular responses mediated by toxic cyanobacteria. For this purpose, a mussel exposure experiment was outlined with two toxic cyanobacteria species, Microcystis aeruginosa and Chrysosporum ovalisporum at 1 × 105 cells/mL, resembling a natural cyanobacteria bloom. The estimated amount of toxins produced by M. aeruginosa and C. ovalisporum were respectively 0.023 pg/cell of microcystin-LR (MC-LR) and 7.854 pg/cell of cylindrospermopsin (CYN). After 15 days of exposure to single and mixed cyanobacteria, a depuration phase followed, during which mussels were fed only non-toxic microalga Parachlorella kessleri. The results showed that the marine mussel is able to filter toxic cyanobacteria at a rate equal or higher than the non-toxic microalga P. kessleri. Filtration rates observed after 15 days of feeding toxic microalgae were 1773.04 mL/ind.h (for M. aeruginosa), 2151.83 mL/ind.h (for C. ovalisporum), 1673.29 mL/ind.h (for the mixture of the 2 cyanobacteria) and 2539.25 mL/ind.h (for the non-toxic P. kessleri). Filtering toxic microalgae in combination resulted in the accumulation of 14.17 ng/g dw MC-LR and 92.08 ng/g dw CYN. Other physiological and biochemical endpoints (dry weight, byssus production, total protein and glycogen) measured in this work did not change significantly in the groups exposed to toxic cyanobacteria with regard to control group, suggesting that muss

Cylindrospermopsin (CYN) and microcystins (MC) are cyanotoxins that can occur simultaneously in contaminated water and food. CYN/MC-LR mixtures previously investigated in vitro showed an induction of micronucleus (MN) formation only in the presence of the metabolic fraction S9. When this is the case, the European Food Safety Authority recommends a follow up to in vivo testing. Thus, rats were orally exposed to 7.5 + 75, 23.7 + 237, and 75 + 750 μg CYN/MC-LR/kg body weight (b.w.). The MN test in bone marrow was performed, and the standard and modified comet assays were carried out to measure DNA strand breaks or oxidative DNA damage in stomach, liver, and blood cells. The results revealed an increase in MN formation in bone marrow, at all the assayed doses. However, no DNA strand breaks nor oxidative DNA damage were induced, as shown in the comet assays. The histopathological study indicated alterations only in the highest dose group. Liver was the target organ showing fatty degeneration and necrotic hepatocytes in centrilobular areas, as well as a light mononuclear inflammatory periportal infiltrate. Additionally, the stomach had flaking epithelium and mild necrosis of epithelial cells. Therefore, the combined exposure to cyanotoxins may induce genotoxic and histopathological damage in vivo.