The chemical memory of smoking tobacco.

The concentration in urine of N-acetyl-hydroxy-propyl-cisteine (3HPMA), an acrolein metabolite, has been employed as a marker of the risk of illness of smokers and the relative concentration of creatinine has been evaluated to verify the effect of moving from the practice of burning tobacco to nicotine vaping. From the results concerning the urine samples of 38 subjects, collected from 2021 to 2023 and analyzed by LC-MS/MS, corresponding to 5 active smokers, 13 previously heavy smokers who replaced traditional tobacco by vaping, and 20 non-smokers, a dramatic reduction was found in 3HPMA/creatinine in urine. 3HPMA varied from values of 2150-3100 µg gcreatinine-1 to levels of 225-625 µg gcreatinine-1 found for non-smokers, with the time decay described by the equation y = 0.3661x2 - 94.359x + 6246.4 (R2: 0.757), providing a time of approximately 10 years for tobacco memory after the cessation of the consumption of burned tobacco.

Biomarkers of exposure in urine of active smokers, non-smokers, and vapers.

The exposure to smoking related products has been evaluated through urine illness risk marker determination through the analysis of urine samples of smokers and vapers. Biomarkers and their metabolites such as N-acetyl-S-(2-cyanoethyl)-L-cysteine (CEMA), N-acetyl-S-(3,4-dihydroxybutyl)-L-cysteine (DHBMA), N-acetyl-S-[1-(hydroxymethyl)-2-propen-1-yl)-L-cysteine (MHBMA), N-acetyl-S-(3-hydroxypropyl)-L-cysteine (3HPMA), 2R-N-acetyl-S-(4-hydroxybutan-2-yl)-L-cysteine (HMPMA), and N-acetyl-S-(3-carboxy-2-propyl)-L-cysteine (CMEMA) together with nicotine and cotinine were identified and quantified by LC-HRMS and LC-MS/MS, and data found normalized to the creatinine level. One hundred two urine samples were collected from smokers, non-smokers, and vapers, spanning an age range from 16 to 79 years. Results obtained showed that CEMA was only detected in urine samples from smokers and MHBMA was in the same order of magnitude in all the urine samples analyzed. HMPMA was found in the urine of vapers at the same order of concentration as in non-smokers. 3HPMA in vapers was lower than in the urine of smokers, presenting an intermediate situation between smokers and non-smokers. On the other hand, DHBMA in vapers can reach similar values to those found for smokers, while CMEMA shows concentrations in the urine of vapers higher than in the case of non-smokers and traditional smokers, requiring new research to link this metabolite to the use of electronic cigarettes and possible alternative metabolomic routes. In general, this study seems to verify that traditional smoking practice constitutes a major source of carcinogenic chemicals compared with substitutive practices, although those practices are not free of potential harm.

Tools for Nutrition Assessment of Adults with Cerebral Palsy: Development of a Gold Standard.

PURPOSE OF REVIEW: Cerebral palsy (CP) is a group of disorders caused by non-stabilized cerebral lesions. Individuals with this disorder are at a higher risk of suffering from malnutrition and other related detrimental effects to their quality of life. For this reason, accurate methods of nutritional assessment are vital for people suffering from this condition. While assessment of nutritional status in children with CP has been extensively studied, very few studies have been carried out on adults. These limitations are due to the great anatomical-functional variability characteristic of this syndrome. Difficulties that derive from this variability in adult patients with CP mean that there remains an urgent need for certain standards of nutritional assessment for this population. The objective of this review is to compile the latest trends in nutritional assessment in adults with CP to guide the development of a conceptual framework for future research. RECENT FINDINGS: With this aim, relevant studies have been identified. The most commonly used technique to evaluate nutritional status is the BMI because of its ease-of-use. However, its well-known limitations fail to adequately estimate the nutritional status in this population, with measurements of patients with CP yielding results that are much less accurate than those that already exist in the general population. Although more studies are needed, kinanthropometry is considered one of the most reliable techniques; nevertheless, the anatomical limitation characteristic of CP plays a limiting factor.

Presence and significance of intestinal unicellular parasites in a morbidly obese population.

BACKGROUND: Obesity is a chronic disease whose pathogenesis has been related to changes in the intestinal microbiota. Yet, the role of protozoa and other unicellular eukaryotic parasites in this microenvironment is still largely unknown. Their presence within the gut ecosystem in obese subjects warrants further study, as well as their influence on the host metabolism and comorbidities. METHODS: Herein, a single center, cross-sectional study of 104 obese individuals was performed to assess the presence of six intestinal unicellular parasites in stool using a commercially available kit, and to evaluate its relationship with the presence of abdominal symptoms, metabolic comorbidities, variations in body composition and nutritional deficiencies. RESULTS: The overall parasitic colonization rate was 51%, with Blastocystis sp., identified as the most frequent (44.2%), followed by Dientamoeba fragilis (11.5%) and Giardia intestinalis (8.7%), and significantly related to the consumption of ecological fruits and vegetables. Contrary to what previous studies pointed out, colonization with parasites species was significantly associated with fewer abdominal symptoms and depositions per day. The presence of parasites did not correlate with any nutritional deficiencies nor differences in body composition, while it did with significant lower HOMA-IR levels and a lower trend towards metabolic syndrome. CONCLUSION: Obese subjects frequently harbor unicellular enteric parasites, apparently without clinical nor nutritional harm. This evidence suggests that carrying these microorganisms, from an endocrinological perspective, has a beneficial effect, especially on insulin resistance and possibly on the development of related comorbidities.

Monitoring serologic response to single in ovo vaccination with an immune complex vaccine against infectious bursal disease in broilers.

The infectious bursal disease (IBD) virus is one of the most resistant and prevalent virus worldwide in the poultry industry, being vaccination the main tool to control the disease. For this reason, consistent and uniform immunization of broiler flocks against IBD is necessary to avoid the disease spreading. The aim of this study was to apply and assess an epidemiologic mapping tool focused on the immunization by in ovo single broiler vaccination using an immune complex IBD vaccine. With this regard, 7,576 serum samples were collected from 603 broiler flocks raised in 354 Spanish farms. To do so, blood samples were randomly collected from birds with ages between 35 to 51 d, and the serum was analyzed by ELISA. The results obtained from this study suggested a high uniform immunization against IBDV and a protective immunization between 35 and 51 d of age, with mean titer values ranging between 6,331 and 7,426. In addition, seroprevalence titer data of this large-scale monitoring study fitted a polynomial equation with a R2 value of 0.77, helping to explain and predict the humoral response to IBD vaccination. This seroprevalence map was applied to broiler production and was based on business intelligence tool that incorporates newly developed mapping tool to cover the need of having real-time information of humoral response to IBD vaccination and could be an effective tool for veterinary services to control and prevent IBD.

The use of dynamic prevalence maps to assess the prevalence of Salmonella in broiler flocks in the Valencian Community, Spain.

The aim of this study was to design and implement a dynamic prevalence map for managing the data collected within the framework of the Salmonella spp. National control plan in broiler flocks, using a data set from the Valencian Community, Spain. Such a map would be a useful tool for veterinarians seeking to define control options. The development of the mapping tool for this study involved three main phases: data collection, data analysis and data representation. The data used were obtained from the results of the National Control Programme (based on European Commission [EC] Regulation No. 2160/2003). The Salmonella prevalence data were represented on a map of the Valencian Community, which included geographical information on flock locations to facilitate the interpretation of the results from monitoring. The average prevalence of Salmonella spp. was 2.74% from 2009 to 2013. Moreover, the proportion of broiler flocks that remained positive for the target serovars according to EC Regulation No. 2160/2003 (S. Enteritidis and/orS. Typhimurium [including monophasic S. Typhimurium]) was less than 1%. Over the five-year period of the study (2009-2013), the area of Bajo Maestrazgo had the highest level of Salmonella spp. prevalence every year, thereby validating the usefulness of this mapping tool. The aim of the study is to detect geographical 'highpressure' Salmonella zones over time and make suggestions on where efforts should be focused in this region to reduce future Salmonella spp. prevalence. Local Veterinary Services could, using the map, identify the infection pressure of Salmonella in the geographical areas where farms are located. Proximity to higher prevalences may imply an increased risk of flock contamination. Although annual data are represented, data can be presented in real time with regular frequency (daily, weekly, monthly or quarterly) or even over periods of several years. Therefore, the dynamic prevalence map provides extremely useful epidemiological information to veterinarians on the prevention of Salmonella.

L'étude présentée par les auteurs visait à concevoir et mettre en application une carte dynamique de la prévalence afin de gérer les données collectées dans le cadre du plan national de contrôle des Salmonella spp. dans les cheptels de poulets de chair à partir des informations transmises par la Communauté valencienne (Espagne). L'objectif attendu était que cette carte puisse aider les vétérinaires à définir la meilleure stratégie de lutte. La conception du logiciel de cartographie s'est faite en trois étapes, portant respectivement sur la collecte, l'analyse et la représentation des données. Les données utilisées provenaient des résultats du Programme national de lutte (basé sur le Règlement [CE] n° 2160/2003 du Parlement européen et du Conseil). Les taux de prévalence de Salmonella ont été reportés sur une carte de la Communauté valencienne, qui contenait également des informations géographiques relatives à la localisation des élevages afin de faciliter l'interprétation des résultats du suivi. De 2009 à 2013, la prévalence moyenne de Salmonella spp. était de 2,74 %. En outre, le pourcentage de cheptels de poulets de chair restant positifs au regard des sérovars visés par le Règlement CE n° 2160/2003 (S. Enteritidis et/ou S. Typhimurium [y compris la souche monophasique de S. Typhimurium]) était inférieur à 1 %. Sur les cinq années de l'étude (2009­2013), la prévalence annuelle la plus élevée de Salmonella spp. restait concentrée dans la région de Bajo Maestrazgo, ce qui confirme l'utilité de cet outil cartographique. Le but de l'étude est de pouvoir détecter les zones géographiques à « forte pression ¼ de Salmonella dans le temps afin d'orienter en conséquence les efforts de lutte dans cette région et de réduire ainsi la prévalence des Salmonella spp. à l'avenir. Grâce à cette carte, les Services vétérinaires locaux pourront connaître la pression exercée par Salmonella dans les zones géographiques où se situent les élevages. La proximité des zones à forte prévalence peut représenter un risque accru de contamination des cheptels. L'outil propose par défaut une représentation annuelle des données mais celles-ci peuvent également être présentées en temps réel à intervalles réguliers (fréquence quotidienne, hebdomadaire, mensuelle ou trimestrielle), voire même sur des périodes couvrant plusieurs années. Ainsi, la carte dynamique des prévalences fournit aux vétérinaires des informations épidémiologiques extrêmement utiles pour la prévention de Salmonella.

Los autores describen un estudio encaminado a concebir y realizar un mapa dinámico de prevalencia para explotar los datos obtenidos como parte del plan nacional de lucha contra las salmonelas en bandadas de pollos asaderos, utilizando para ello un conjunto de datos correspondientes a la Comunidad Valenciana (España). Semejante mapa ofrecería a los veterinarios una herramienta útil para definir las fórmulas idóneas de lucha. La elaboración del dispositivo de cartografía para este estudio entrañaba tres grandes fases: la de obtención, la de análisis y la de representación de los datos. Los datos utilizados derivan de los resultados obtenidos con el Programa Nacional de Control (basado en el Reglamento de la Comisión Europea [CE] Nº 2160/2003). Para facilitar la interpretación de los resultados obtenidos con la labor de vigilancia, los datos de prevalencia de salmonelas fueron representados en un mapa de la Comunidad Valenciana que contenía información geográfica sobre la localización de las bandadas. De 2009 a 2013, la prevalencia media de Salmonella spp. fue del 2,74%. Por otra parte, la proporción de bandadas de pollos asaderos que seguían siendo positivas para las serovariantes estudiadas con arreglo al Reglamento (CE)Nº 2160/2003 (S. Enteritidis y/o S. Typhimurium [incluida S. Typhimurium monofásica]) era inferior al 1%. En el quinquenio cubierto por el estudio (2009­2013), la zona del Bajo Maestrazgo registró cada año la mayor prevalencia de salmonelas, hecho que confirmó la utilidad de esta herramienta cartográfica. El estudio tiene por objetivo determinar a lo largo del tiempo las zonas geográficas de 'alta presión' de salmonelas y hacer propuestas sobre las áreas de esa región en las que convendría centrar el trabajo para reducir en el futuro la prevalencia de salmonelas. Empleando el mapa, los Servicios Veterinarios podrían determinar la presión infecciosa de salmonelas en las zonas geográficas en que se encuentran las explotaciones. Cuanto más cerca esté una explotación de zonas con niveles altos de prevalencia, mayor puede ser el riesgo de contaminación de las bandadas. Aunque en el mapa se representan datos anuales, también es posible presentarlos en tiempo real con determinada periodicidad (diaria, semanal, mensual o trimestral) o incluso cubrir periodos de varios años. El mapa dinámico de prevalencia proporciona así a los veterinarios información epidemiológica de gran utilidad para la prevención de las salmonelosis.

Prevalence of bacteria and absence of anisakid parasites in raw and prepared fish and seafood dishes in Spanish restaurants.

This study evaluated the presence of bacteria and anisakid parasites in 45 samples of raw anchovies in vinegar, a dish widely eaten in Spain, and in 227 samples of cooked fish and cephalopods served in Spanish food service establishments. Our analysis showed that, according to European and Spanish regulation, 14 to 30% of the prepared fish and cephalopod dishes exceeded the maximum allowable level for mesophilic aerobic counts, and 10 to 40% of these samples exceeded the allowable levels for Enterobacteriaceae. None of the studied samples showed evidence of anisakid parasites, Escherichia coli, Staphylococcus aureus, Salmonella, or Listeria monocytogenes. These results indicate that application of hazard analysis and critical control points, food safety training courses, and routine inspections in compliance with current European and Spanish legislation help protect consumer health.

Curcumin and trans-resveratrol exert cell cycle-dependent radioprotective or radiosensitizing effects as elucidated by the PCC and G2-assay.

Curcumin and trans-resveratrol are well-known antioxidant polyphenols with radiomodulatory properties, radioprotecting non-cancerous cells while radiosensitizing tumor cells. This dual action may be the result of their radical scavenging properties and their effects on cell-cycle checkpoints that are activated in response to radiation-induced chromosomal damage. It could be also caused by their effect on regulatory pathways with impact on detoxification enzymes, the up-regulation of endogenous protective systems, and cell-cycle-dependent processes of DNA damage. This work aims to elucidate the mechanisms underlying the dual action of these polyphenols and investigates under which conditions they exhibit radioprotecting or radiosensitizing properties. The peripheral blood lymphocyte test system was used, applying concentrations ranging from 1.4 to 140µM curcumin and 2.2 to 220µM trans-resveratrol. The experimental design focuses first on their radioprotective effects in non-cycling lymphocytes, as uniquely visualized using cell fusion-mediated premature chromosome condensation, excluding, thus, cell-cycle interference to repair processes and activation of checkpoints. Second, the radiosensitizing potential of these chemicals on the induction of chromatid breaks in cultured lymphocytes following G2-phase irradiation was evaluated by a standardized G2-chromosomal radiosensitivity predictive assay. This assay uses caffeine for G2-checkpoint abrogation and it was applied to obtain an internal control for radiosensitivity testing, which simulates conditions similar to those of the highly radiosensitive lymphocytes of AT patients. The results demonstrate for the first time the cell-cycle-dependent action of these polyphenols. When non-cycling cells are irradiated, the radioprotective properties of curcumin and trans-resveratrol are more prominent. However, when cycling cells are irradiated during G2-phase, the radiosensitizing features of these compounds are more pronounced. This observation offers a new biological basis for the mechanisms underlying the action of these polyphenols in cancer radiotherapy.

Characterization of heat-labile toxin-subunit B from Escherichia coli by liquid chromatography-electrospray ionization-mass spectrometry and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

The possibilities of characterizing the heat-labile enterotoxin (LT) of enterotoxigenic Escherichia coli (ETEC) by liquid chromatography electrospray mass spectrometry (LC/ESI-MS) and matrix-assisted laser desorption with time-of-flight mass spectrometry (MALDI-TOF-MS) were investigated. The B subunit from recombinant E. coli (expression in Pichia pastoris) can be detected by LC/ESI-MS expressed in P. pastoris and the charge envelope signals can be observed; LC/ESI-MS and MALDI-TOF-MS analysis allowed the acquisition of labile toxin subunit B (LTB) molecular weight and preliminary structural characterization of LTB toxin. MALDI-TOF analysis after reduction and alkylation of the protein evidenced the presence of one disulfide bond in the structure of the protein. Confirmatory analysis was carried out by detection of most of the tryptic fragments of the B subunit by MALDI-TOF-MS, obtaining total coverage of the protein sequence. Possible biovariations in the toxin can mostly be determined by sequencing, where an increase of molecular mass in the N-terminal side of the protein was identified. This modification may be due to an O-GlcNAc-1-phosphorylation.

Report of toxic shock syndrome toxin 1 (TSST-1) from Staphylococcus aureus isolated in food handlers and surfaces from foodservice establishments.

A set of 53 Staphylococcus aureus isolates collected from food handlers and foodservice establishments in Spain was analyzed for toxic shock syndrome toxin (TSST-1) production. S. aureus strains were isolated from 908 samples collected from different surfaces such as dish towels, workers' hands, cutting boards, stainless steel tables and slicers, but they were not detected neither in clean plates nor in kitchen knives. Only one food worker hand has been reported to be contaminated by TSST-1 in a restaurant. Despite this, proper hygiene practices should be respected for the surfaces of contact with food, as well as for the hands of the manipulators This is the first article, in Spain, that reports the detection of TSST-1 in a restaurant worker hand.

Assessment of microbial quality of commercial and home-made tiger-nut beverages.

AIMS: We aimed to assess the microbiological quality of Spanish commercial tiger-nut beverages as well as home-made samples collected from supermarkets, street vendors, juice bars and ice-cream parlours located in Valencia. METHODS AND RESULTS: Microbiological analysis of 44 tiger-nut beverages samples were carried out according to International Standard Organization (ISO) norms and published works which included the total viable count, Enterobacteriaceae, Escherichia coli, Staphylococcus aureus, Salmonella, Bacillus cereus, yeasts, moulds, Yersinia enterocolitca, Clostridium perfringens, Vibrio spp. and Listeria monocytogenes. The obtained results indicated that all the commercial samples were below the detection limit for the viable microorganisms. Results of analysis of those home-made tiger-nut samples revealed that 67% (16 samples) harboured total plate counts while the rest of samples were free from these microorganisms. Enterobacteriaceae were detected in 62% (15 samples). E. coli were found in only one sample (4%), yeasts and moulds were detected in 62% (15 samples) each, Shigella was found in 21% (five samples); however, all samples were free from S. aureus, Salmonella, Y. enterocolotica, C. perfringens, Vibrio spp. and L. monocytogenes. CONCLUSIONS: These results reflected that there exists a rather high contamination level in home-made tiger-nut beverages indicating the need to apply correct and strict HACCP system(s) during manufacturing and storage of these food products. SIGNIFICANCE AND IMPACT OF STUDY: This study demonstrates the great need to carry out microbiological tests frequently in these products and even more the need to apply correct HACCP system (s). Tiger-nut beverages are especially well-known products in Spain, hence it is extremely important to ensure an adequate microbiological quality to guarantee consumers health.

Assessment in vitro of cytogenetic and genotoxic effects of propolis on human lymphocytes.

We evaluated the genetic damage by ethanolic extract of propolis (EEP) induced to human lymphocytes which were exposed to increasing concentrations (0-2000µgml(-1)). The results indicated that EEP reduced significantly the mitotic index (MI) and proliferation index (PI) when high concentrations of EEP were used. Sister chromatid exchange (SCE) rates indicated that EEP could have genotoxic effects at high concentrations. Exposure of the cells to the amount of ethanol used as solvent did not alter either the MI and cell proliferation kinetics (CPK), or the rate of SCE. The results showed: (a) statistical increase in the percentage the cells with CAs and in the frequency of SCE at the highest concentrations, (b) a decrease in MI and in the CPK values was observed, (c) no effect was noticed in negative controls. In conclusion, it can be assumed that high concentrations of EEP have a cyto and genotoxic effect, in vitro, for human peripheral lymphocytes.

Assessment of Salmonella spp. in feces, cloacal swabs, and eggs (eggshell and content separately) from a laying hen farm.

Microbial pathogens of the genus Salmonella are among the leading causes of foodborne illness in the world. The present study was done on a laying hen farm with a Salmonella enterica serovar Enteritidis-positive result according to the testing specified by European regulation 2160/2003. The aim of this study was to compare the Salmonella contamination on a laying hen farm with the Salmonella presence in the hen eggs. The strains were isolated by ISO method 6579:2002 (standard method for the detection of Salmonella spp. in the European regulation for food and animal feeding stuffs, animal feces, and environmental samples from the primary production stage, including poultry farms) and were confirmed as Salmonella Enteritidis by the Kauffmann-White method. In addition, strains were compared with genomic macrorestriction followed by pulsed-field gel electrophoresis. Four types of samples, namely, feces (n = 50), cloacal swabs (n = 150), eggshells (n = 50), and egg contents (n = 50), were taken from each of 50 randomly selected battery cages. Results demonstrated that feces (92%) were the most positive sample, followed by eggshells (34%) and cloacal swabs (4%). No Salmonella spp. were detected in the egg contents. Our results show that a Salmonella Enteritidis-positive result on a laying hen farm, according to the testing specified by European regulation 2160/2003, did not imply the presence of the pathogen in the egg contents. Additionally, XbaI-digested genomic DNA of Salmonella Enteritidis strains isolated from several samples resulted in the same pattern, so were probably of the same origin.

Concentration-Dependent Protection by Ethanol Extract of Propolis against γ-Ray-Induced Chromosome Damage in Human Blood Lymphocytes.

Radioprotection with natural products may be relevant to the mitigation of ionizing radiation-induced damage in mammalian systems; in this sense, propolis extracts have shown effects such as antioxidant, antitumoral, anti-inflammatory, and immunostimulant. We report for the first time a cytogenetic study to evaluate the radioprotective effect, in vitro, of propolis against radiation-induced chromosomal damage. Lymphocytes were cultured with increasing concentrations of ethanol extract of propolis (EEP), including 20, 40, 120, 250, 500, 750, 1000, and 2000 µg mL(-1) and then exposed to 2 Gy γ-rays. A significant and concentration-dependent decrease is observed in the frequency of chromosome aberrations in samples treated with EEP. The protection against the formation of dicentrics was concentration-dependent, with a maximum protection at 120 µg mL(-1) of EEP. The observed frequency of dicentrics is described as negative exponential function, indicating that the maximum protectible fraction of dicentrics is approximately 44%. Free radical scavenging and antioxidant activities are the mechanisms that these substances use to protect cells from ionizing radiation.

One-year monitoring of aflatoxins and ochratoxin A in tiger-nuts and their beverages.

A sensitive and selective liquid chromatography-triple quadrupole-tandem mass spectrometry (LC-ESI-MS-MS) method was developed for the routine analysis of aflatoxins (AFB(1), AFB(2), AFG(1) and AFG(2)) and ochratoxin A (OTA) in tiger nuts and tiger-nut beverage (horchata). A matrix solid phase dispersion was adapted to eliminate lipidic interferences. The solid support was C(18), while the elution solvent was acetonitrile. Mean recoveries obtained at two fortification levels were 72-83% and 71-81% for horchata and tiger nut respectively with relative standard deviations (RSDs) <13% and 15% respectively. The LC-MS-MS method allowed quantification and identification at low levels in two matrices. The method was applied for the routine analysis of tiger-nuts and horchata samples collected from different supermarkets of Valencia (Spain) during one year (March 2009-March 2010). A total of 238 samples were analysed and 32 samples were found positives for OTA, AFB(1), AFB(2) and AFG(2).

Intestinal parasitic infections in hosted Saharawi children.

Literatures on intestinal parasitic infections in Saharawi children were scarce and distributed in non parasitological journals. This was the first article that specifically highlighted on the prevalence of these infections in 270 Saharawi children aged from 6 to 12 years hosted in Spain. Six different intestinal parasites were identified in this study and 78, 46, 40, 24, 13 and 5 were positive for Giardia lamblia (29%), Entamoeba coli (17%), Blastocystis hominis (15%), Endolimax nana (9%), Hymenolepis nana (5%) and Enterobius vermicularis (2%), respectively. Mixed intestinal parasitic infections were seen in 12 (4.4%) studied children. Six (2.2%) double infections for G. lamblia and B. hominis were seen in these children while in four (1.5%) had G. lamblia and H. nana. Triple intestinal parasitic infections of G. lamblia, B. hominis and H. nana were observed in two (0.7%) of the children studied. In the other hand, about 14.8% of the studied children had a mild anaemia and 15.5 and 16.6% had iron deficiency and eosinophilia, respectively.

Antifungal effects of the bioactive compounds enniatins A, A(1), B, B(1).

To produce enniatin (ENs), Fusarium tricinctum CECT 20150 was grown in a liquid medium of potato (PDB), being mycotoxin purified by high performance liquid chromatography (HPLC) with a reverse phase semipreparative column using a mobile phase of acetonitrile/water using gradient condition. The purity of the ENs fractions was verified by analytical HPLC and LC/MS-MS. The pure fractions of ENs were utilized to study the biological activity on several mycotoxigenic moulds as Fusarium verticilloides, Fusarium sporotrichioides, Fusarium tricinctum, Fusarium poae, Fusarium oxysporum, Fusarium proliferatum, Beauveria bassiana, Trichoderma harzianum, Aspergillus flavus, Aspergillus parasiticus, Aspergillus fumigatus, Aspergillus ochraceus and Penicillium expansum. The results obtained demonstrated that in several antibiograms, ENs induced the inhibition of the grown microorganisms tested.

Isolation and purification of enniatins A, A(1), B, B(1), produced by Fusarium tricinctum in solid culture, and cytotoxicity effects on Caco-2 cells.

Enniatins (ENs) are antibiotic compounds of hexadepsipeptidic structure produced by several strains of Fusarium spp. The ENs A, A(1), B, B(1) were purified from extracts of Fusarium tricinctum grown on a solid medium of corn, by a low pressure liquid chromatography (LPLC) on reverse phase of Amberlite XAD-7 followed by semipreparative LC. The purity and the structure of the isolated compounds were confirmed by LC-MS/MS. The technique of the purification of the fungal extract enabled complete separation of the ENs A, A(1), B, B(1) with a mean purity of 97% for all the compounds. The cytoxicity of the ENs was tested in the cell lines of human origin (epithelial colorectal adenocarcinoma cells, Caco-2) by MTT assays. Only EN A(1) and B(1) evoked toxicity at the tested concentrations. The inhibitory concentration (IC(50)) for EN A(1) on Caco-2 cells was 12.3 microM, whereas the IC(50) produced by the EN B(1) was 19.5 microM. This study indicates that ENs, fungal metabolites that are commonly found in corn and in general in product composed by corn, may have a toxic potential for human health.

Antibacterial effect of the bioactive compound beauvericin produced by Fusarium proliferatum on solid medium of wheat.

To obtain the bioactive compound beauvericin (BEA), Fusarium proliferatum CECT 20569 was grown on a solid medium of wheat, utilizing the technique of the solid state fermentation (SSF), being this mycotoxin purified by high performance liquid chromatography (HPLC) with a reverse phase semi-preparative column using as the mobile phase acetonitrile/water in gradient condition. The purity of the BEA was verified by analytical HPLC and liquid chromatography tandem mass spectrometry (LC/MS-MS). The pure fractions of BEA were utilized to determinate the antibiotic effects on several bacterial strains that are considered normally pathogens of the intestinal tract as: Escherichia coli, Enterococcus faecium, Salmonella enterica, Shigella dysenteriae, Listeria monocytogenes, Yersinia enterocolitica, Clostridium perfringens, Pseudomonas aeruginosa and Staphylococcus aureus.

Glucose influence on the production of T-2 toxin by Fusarium sporotrichioides.

Toxigenic isolate of Fusarium sporotrichioides was tested for the T-2 toxin production on PDA plates during 10 days under various glucose concentrations. T-2 toxin was determined by LC-MS and confirmed with LC-MS/MS. This analytical method has been applied, for the first time, to an extensive study of T-2 accumulation. Results showed that the production of this mycotoxin is directly correlated to the concentration of glucose present in the medium. Concentrations of T-2 toxin produced by the strain of F. sporotrichioides ranged from 0 to 1.45 mg/kg. The better T-2 production was evidenced in the fermentation operated with 20% of glucose.