Toxigenic Species Research Articles

Morphological and Molecular Diversity of Aspergillus From Corn Grain Used as Livestock Feed

The study of Aspergillus from corn grains used as livestock feed is important to ensure the safety of the grains as the occurrence of Aspergillus in the corn grain can give an indication of mycotoxin being produced. Morphological and molecular identifications were applied to identify Aspergillus isolated from corn grains used as livestock feed. Morphologically, six species were tentatively identified, namely Aspergillus niger (Groups I and II), Aspergillus flavus, Aspergillus oryzae, Aspergillus fumigatus, Aspergillus clavatus and Aspergillus terreus. Isolates of A. niger was divided into two groups based on slight differences of their colony appearances. Molecular identification using internal transcribed spacer and β-tubulin sequences supported morphological identification except isolates for A. niger Group II isolates, which were molecularly identified as Aspergillus tubingensis. Neighbour-joining phylogenetic tree showed that isolates from the same species were grouped in the same clade. The present study showed that diverse species of Aspergillus are prevalent in corn grain used as livestock feed. It is also necessary to correctly identify the Aspergillus species to employ correct treatment of contaminated corn grains. The occurrence of well-known toxigenic species such as A. niger, A. flavus and A. fumigatus suggested the possible risk of mycotoxin contamination of the corn grain.

Mycotoxins: An Underhand Food Problem.

Among the food safety issues, the occurrence of fungal species able to produce toxic metabolites on the agro-food products has acquired a general attention. These compounds, the mycotoxins, generally provided of low molecular weight, are the result of the secondary metabolism of the toxigenic fungi. They may have toxic activity toward the plants, but mostly represent a serious risk for human and animal health worldwide, since they can be accumulated on many final crop products and they have a broad range of toxic biological activities. In particular, mainly cereals are the most sensitive crops to the colonization of toxigenic fungal species which accumulate in the grains the related mycotoxins both in the field, until the harvest stage, and in the storage. According to a Food and Agriculture Organization study, approximately 25 % of the global food and feed output is contaminated by mycotoxins. Therefore, since a large proportion of the world's population consumes, as a staple food, the cereals, the consumption of mycotoxin-contaminated cereals is a main issue for health risk worldwide. Furthermore, mycotoxin contamination can have a huge economic and social impact, especially when mycotoxin occurrence on the food commodities is over the regulation limits established by different national and transnational institutions, implying that contaminated products must be discarded. Finally, the climate change due to the global warming can alter stages and rates of toxigenic fungi development and modify host-resistance and host-pathogen interactions, influencing deeply also the conditions for mycotoxin production that vary for each individual pathogen. New combinations of mycotoxins/host plants/geographical areas are arising to the attention of the scientific community and require new diagnostic tools and deeper knowledge of both biology and genetics of toxigenic fungi. Moreover, to spread awareness and knowledge at international level on both the hazard that mycotoxins represent for consumers and costs for stakeholders is of key importance for developing all possible measures aimed to control such dangerous contaminants worldwide.

Fusarium Species and Their Associated Mycotoxins.

The genus Fusarium includes numerous toxigenic species that are pathogenic to plants or humans, and are able to colonize a wide range of environments on earth. The genus comprises around 70 well-known species, identified by using a polyphasic approach, and as many as 300 putative species, according to phylogenetic species concepts; many putative species do not yet have formal names. Fusarium is one of the most economically important fungal genera because of yield loss due to plant pathogenic activity; mycotoxin contamination of food and feed products which often render them unaccep for marketing; and health impacts to humans and livestock, due to consumption of mycotoxins. Among the most important mycotoxins produced by species of Fusarium are the trichothecenes and the fumonisins. Fumonisins cause fatal livestock diseases and are considered potentially carcinogenic mycotoxins for humans, while trichothecenes are potent inhibitors of protein synthesis. This chapter summarizes the main aspects of morphology, pathology, and toxigenicity of the main Fusarium species that colonize different agricultural crops and environments worldwide, and cause mycotoxin contamination of food and feed.

Molecular diversity patterns among various phytoplankton size-fractions in West Greenland in late summer

Arctic regions have experienced pronounced biological and biophysical transformations as a result of global change processes over the last several decades. Current hypotheses propose an elevated impact of those environmental changes on the biodiversity, community composition and metabolic processes of species. The effects on ecosystem function and services, particularly when invasive or toxigenic harmful species become dominant, can be expressed over a wide range of temporal and spatial scales in plankton communities. Our study focused on the comparison of molecular biodiversity of three size-fractions (micro-, nano-, picoplankton) in the coastal pelagic zone of West Greenland and their association with environmental parameters. Molecular diversity was assessed via parallel amplicon sequencing the 28S rRNA hypervariable D1/D2 region. We showed that biodiversity distribution within the area of Uummannaq Fjord, Vaigat Strait and Disko Bay differed markedly within and among size-fractions. In general, we observed a higher diversity within the picoplankton size fraction compared to the nano- and microplankton. In multidimensional scaling analysis, community composition of all three size fractions correlated with cell size, silicate and phosphate, chlorophyll a (chl a) and dinophysistoxin (DTX). Individually, each size fraction community composition also correlated with other different environmental parameters, i.e. temperature and nitrate. We observed a more homogeneous community of the picoplankton across all stations compared to the larger size classes, despite different prevailing environmental conditions of the sampling areas. This suggests that habitat niche occupation for larger-celled species may lead to higher functional trait plasticity expressed as an enhanced range of phenotypes, whereas smaller organisms may compensate for lower potential plasticity with higher diversity. The presence of recently identified toxigenic harmful algal bloom (HAB) species (such as Alexandrium fundyense and A. ostenfeldii) in the area points out the potential risk for this vulnerable ecosystem in a changing world.

Контамінація та ушкодження мікроміцетами зерна та кормів

Контамінація та ушкодження мікроміцетами зерна та кормів

Evolution of fungal population and mycotoxins in sorghum silage

ABSTRACTSilage, one of the most important feed sources for cattle, is vulnerable to contamination by spoilage moulds and mycotoxins because ensilage materials are excellent substrates for fungal growth. The aim of this study was to identify the mycobiota of sorghum silages, to determine the presence of aflatoxins and fumonisins, and to correlate these results with physical parameters of the silage. A total of 275 samples of sorghum were collected from dairy farms in the south-west region of Uruguay were silage practices are developed. The presence of fungi was observed in all of the sorghum samples with values varying from 0.2 × 104 to 4085 × 104 UFC g−1. Significant difference were detected in the total number of fungi during the storage period; at six months there is a high risk of fungal spoilage. The most frequent genera isolated from sorghum samples were Penicillium (70%), Aspergillus (65%), Absidia (40%), Fusarium (35%), Paecilomyces (35%) and Alternaria, Cladosporium, Gliocadium and Mucor (30%). The toxigenic species most frequently found were Penicillium citrinum, Aspergillus flavus and Fusarium nygamai. Only two samples were contaminated by AFB1 with levels of 1 and 14 µg kg–1. Fumonisin was detected in 40% of freshly harvest samples with levels ranged from 533 µg kg–1 to 933 µg kg–1. The use of silo bags seems to be an effective tool to store sorghum. However, the presence of toxigenic fungi show that regular screening for mycotoxins levels in silages must be performed to avoid the exposure of animals to contaminated feed and the introduction of these compounds into the food chain.

Metagenomic analysis of medicinal Cannabis samples; pathogenic bacteria, toxigenic fungi, and beneficial microbes grow in culture-based yeast and mold tests

Background: The presence of bacteria and fungi in medicinal or recreational Cannabis poses a potential threat to consumers if those microbes include pathogenic or toxigenic species. This study evaluated two widely used culture-based platforms for total yeast and mold (TYM) testing marketed by 3M Corporation and Biomérieux, in comparison with a quantitative PCR (qPCR) approach marketed by Medicinal Genomics Corporation. Methods: A set of 15 medicinal Cannabis samples were analyzed using 3M and Biomérieux culture-based platforms and by qPCR to quantify microbial DNA. All samples were then subjected to next-generation sequencing and metagenomics analysis to enumerate the bacteria and fungi present before and after growth on culture-based media. Results: Several pathogenic or toxigenic bacterial and fungal species were identified in proportions of >5% of classified reads on the samples, including Acinetobacter baumannii, Escherichia coli, Pseudomonas aeruginosa, Ralstonia pickettii, Salmonella enterica, Stenotrophomonas maltophilia, Aspergillus ostianus, Aspergillus sydowii, Penicillium citrinum and Penicillium steckii. Samples subjected to culture showed substantial shifts in the number and diversity of species present, including the failure of Aspergillus species to grow well on either platform. Substantial growth of Clostridium botulinum and other bacteria were frequently observed on one or both of the culture-based TYM platforms. The presence of plant growth promoting (beneficial) fungal species further influenced the differential growth of species in the microbiome of each sample. Conclusions: These findings have important implications for the Cannabis and food safety testing industries.

Metagenomic analysis of medicinal Cannabis samples; pathogenic bacteria, toxigenic fungi, and beneficial microbes grow in culture-based yeast and mold tests.

Background: The presence of bacteria and fungi in medicinal or recreational Cannabis poses a potential threat to consumers if those microbes include pathogenic or toxigenic species. This study evaluated two widely used culture-based platforms for total yeast and mold (TYM) testing marketed by 3M Corporation and Biomérieux, in comparison with a quantitative PCR (qPCR) approach marketed by Medicinal Genomics Corporation. Methods: A set of 15 medicinal Cannabis samples were analyzed using 3M and Biomérieux culture-based platforms and by qPCR to quantify microbial DNA. All samples were then subjected to next-generation sequencing and metagenomics analysis to enumerate the bacteria and fungi present before and after growth on culture-based media. Results: Several pathogenic or toxigenic bacterial and fungal species were identified in proportions of >5% of classified reads on the samples, including Acinetobacter baumannii, Escherichia coli, Pseudomonas aeruginosa, Ralstonia pickettii, Salmonella enterica, Stenotrophomonas maltophilia, Aspergillus ostianus, Aspergillus sydowii, Penicillium citrinum and Penicillium steckii. Samples subjected to culture showed substantial shifts in the number and diversity of species present, including the failure of Aspergillus species to grow well on either platform. Substantial growth of Clostridium botulinum and other bacteria were frequently observed on one or both of the culture-based TYM platforms. The presence of plant growth promoting (beneficial) fungal species further influenced the differential growth of species in the microbiome of each sample. Conclusions: These findings have important implications for the Cannabis and food safety testing industries.

Inhibition of Fusarium Growth and Mycotoxin Production in Culture Medium and in Maize Kernels by Natural Phenolic Acids

The possible role of natural phenolic compounds in inhibiting fungal growth and toxin production has been of recent interest as an alternative strategy to the use of chemical fungicides for the maintenance of food safety. Fusarium is a worldwide fungal genus mainly associated with cereal crops. The most important Fusarium mycotoxins are trichothecenes, zearalenone, and fumonisins. This study was conducted to evaluate the potential of four natural phenolic acids (caffeic, ferulic, p-coumaric, and chlorogenic) for the control of mycelial growth and mycotoxin production by six toxigenic species of Fusarium . The addition of phenolic acids to corn meal agar had a marked inhibitory effect on the radial growth of all Fusarium species at levels of 2.5 to 10 mM in a dose-response pattern, causing total inhibition (100%) in all species except F. sporotrichioides and F. langsethiae . However, the effects of phenolic acids on mycotoxin production in maize kernels were less evident than the effects on growth. The fungal species differed in their responses to the phenolic acid treatments, and significant reductions in toxin concentrations were observed only for T-2 and HT-2 (90% reduction) and zearalenone (48 to 77% reduction). These results provide data that could be used for developing pre- and postharvest strategies for controlling Fusarium infection and subsequent toxin production in cereal grains.

Phycotoxins by Harmful Algal Blooms (HABS) and Human Poisoning: An Overview

Phycotoxins are potent natural toxins synthesized by certain marine algae and cyanobacteria species during quot Harmful Algal Blooms quot HABs often seen as water discoloration known as quot Red Tides quot quot Green Tide quot They are grouped by chemical structure mechanisms of action target tissues biological and health effects A constant threat to public health and economy environmental contaminants in aquatic ecosystems seafood drinking water requiring multidisciplinary action at the local and international level to manage their potentially harmful effects The bloom of toxigenic Pseudo nitzschia along the west coast of North America resulted in domoic acid contamination of crab and clams numerous harvesting closures and Consumer Warning issued by local public health authorities Whereas the massive cyanobacteria blooms reported in Florida in June lead to banning drinking water in some locations Public health vigilance monitoring and research need to be maintained and enhanced Despite scientific advancements phycotoxin research relating to human exposure and health consequences are sparse while blooms of toxigenic species have become more prevalent worldwide At present phycotoxins poisonings diagnosis and management largely based on the ability of health care provider to interpret presenting clinical symptoms collect exposure history identify and establish the exposure event Several phycotoxins are neurotoxic potentially lethal and or associated with chronic adverse health effects Human intoxications are often misdiagnosed under diagnosed and under reported to public health authorities hampering proper management and epidemiology data A regulatory standard alertness by public health organizations and primary health care providers in regions with a history of HABs helps to minimize and manage human health risks Nonetheless there are populations at higher risk of exposure due to cultural practices recreational shellfish pickers anglers aboriginals in coastal regions Human exposure and poisoning can occur worldwide after consuming contaminated seafood while traveling or products imported from world location with limited analytical facilities and regulatory implementation

Dietary mycotoxins binders: a strategy to reduce aflatoxin m1 residues and improve milk quality of lactating Beetal goats

Aflatoxin M1 (AFM1) is a metabolite of aflatoxin B1 (AFB1) and secreted in milk of animals that are kept on a moldy diet. AFB1 is produced by several toxigenic species of fungi. In this study, the comparative efficacy of two mycotoxin binders was evaluated in terms of reducing excretion of AFM1, total viable bacterial count and somatic cell count in milk obtained from goats fed with AFB1 contaminated diet. A total of 32 lactating Beetal goats were reared and divided into 4 equal groups: Animals of group A were kept as control, while those in groups B, C, and D were individually fed with AFB1 at 40 µg alone, 40 µg AFB1 along with mycotoxin binder Toxfin® at 3 g kg−1 of cotton seed cake, and 40 µg AFB1 along with mycotoxin binder Elitox® at a dose rate of 1 g kg−1 of cotton seed cake, respectively. Toxfin® significantly decreased the excretion of AFM1 in goats’ milk by 49.57 % while Elitox® non-significantly reduced the excretion of AFM1 in goats’ milk by 19.49 %. Total viable bacterial count in milk samples of group C and D, and somatic cell count in milk samples of group C did not reduce significantly. However, somatic cell count in milk samples of group D reduced significantly. We conclude that the addition of Toxfin® in the moldy diet significantly reduce the excretion of AFM1 in animals’ milk and minimize the risk of mycotoxin toxicity to public health.

Reprofiled anthelmintics abate hypervirulent stationary-phase Clostridium difficile.

Prolonged use of broad-spectrum antibiotics disrupts the indigenous gut microbiota, which consequently enables toxigenic Clostridium difficile species to proliferate and cause infection. The burden of C. difficile infections was exacerbated with the outbreak of hypervirulent strains that produce copious amounts of enterotoxins and spores. In recent past, membrane-active agents have generated a surge of interest due to their bactericidal property with a low propensity for resistance. In this study, we capitalized on the antimicrobial property and low oral bioavailability of salicylanilide anthelmintics (closantel, rafoxanide, niclosamide, oxyclozanide) to target the gut pathogen. By broth microdilution techniques, we determined the MIC values of the anthelmintics against 16 C. difficile isolates of defined PCR-ribotype. The anthelmintics broadly inhibited C. difficile growth in vitro via a membrane depolarization mechanism. Interestingly, the salicylanilides were bactericidal against logarithmic- and stationary-phase cultures of the BI/NAP1/027 strain 4118. The salicylanilides were poorly active against select gut commensals (Bacteroides, Bifidobacterium and Lactobacillus species), and were non-hemolytic and non-toxic to mammalian cell lines HepG2 and HEK 293T/17 within the range of their in vitro MICs and MBCs. The salicylanilide anthelmintics exhibit desirable properties for repositioning as anti-C. difficile agents.

Evaluation of an alternative spectroscopic approach for aflatoxin analysis: Comparative analysis of food and feed samples with UPLC–MS/MS

Increasing research has highlighted the effects of changing climates on the occurrence and prevalence of toxigenic Aspergillus species producing aflatoxins. There is concern of the toxicological effects to human health and animal productivity following acute and chronic exposure that may affect the future ability to provide safe and sufficient food globally. Considerable research has focused on the detection of these toxins, based on the physicochemical and biochemical properties of the aflatoxin compounds, in agricultural products for human and animal consumption. As improvements in food security continue more regulations for acceptable levels of aflatoxins have arisen globally; the most stringent in Europe. These regulations are important for developing countries as aflatoxin occurrence is high significantly effecting international trade and the economy. In developed countries analytical approaches have become highly sophisticated, capable of attaining results with high precision and accuracy, suitable for regulatory laboratories. Regrettably, many countries that are affected by aflatoxin contamination do not have resources for high tech HPLC and MS instrumentation and require more affordable, yet robust equally accurate alternatives that may be used by producers, processors and traders in emerging economies. It is especially important that those companies wishing to exploit the opportunities offered by lucrative but highly regulated markets in the developed world, have access to analytical methods that will ensure that their exports meet their customers quality and safety requirements.This work evaluates the ToxiMet system as an alternative approach to UPLC–MS/MS for the detection and determination of aflatoxins relative to current European regulatory standards. Four commodities: rice grain, maize cracked and flour, peanut paste and dried distillers grains were analysed for natural aflatoxin contamination. For B1 and total aflatoxins determination the qualitative correlation, above or below the regulatory limit, was good for all commodities with the exception of the dried distillers grain samples for B1 for which no calibration existed. For B1 the quantitative R2 correlations were 0.92, 0.92, 0.88 (<250μg/kg) and 0.7 for rice, maize, peanuts and dried distillers grain samples respectively whereas for total aflatoxins the quantitative correlation was 0.92, 0.94, 0.88 and 0.91. The ToxiMet system could be used as an alternative for aflatoxin analysis for current legislation but some consideration should be given to aflatoxin M1 regulatory levels for these commodities considering the high levels detected in this study especially for maize and peanuts.

MYCOTOXIN-PRODUCING PENICILLIUM AND OTHER FUNGI ISOLATED FROM SLOVAK WINE GRAPES AT VARIOUS STAGES OF MATURATION

The aim of the study was to monitor the mycobiota in grape samples at 3 maturation stages (pea berry, early veraison and ripe berry) and to test the ability of selected Penicillium strains to produce mycotoxins in year 2014. Grapes came from Nitra wine growing region, Šintava subregion and were analyzed by plating methods. From Devín grape variety the 339 strains were detected and identified. The most abundant genera found by descending order were Alternaria (42.8%), Cladosporium (24.2%) and Epicoccum (10.0%). Penicillium expansum was isolated from all 3 maturation stages but in low average relative density (1.2%). From Frankovka modrá (Frankish) grape variety the 562 strains were detected and identified. The most abundant genera found by descending order were Alternaria (30.6%), Cladosporium (21.3%) and Penicillium (20.3%). Totally were identified 8 species of the genus Penicillium. The most important species, on the basis of the isolation from all maturation stages and relative density, was P. expansum. Selected isolates of Penicillium species were tested for their toxigenity, according to thin layer chromatography method. In total 14 Penicillium strains representing 3 potentially toxigenic species were tested for their toxigenic ability, namely P. expansum, P. griseofulvum and P. chrysogenum. Out of 14 strains, 64% produced et least one mycotoxin as revealed by the method used here.

Polyphasic approach for detecting toxigenic Fusarium species collected from imported grain and seed commodities

Polyphasic approach for detecting toxigenic Fusarium species collected from imported grain and seed commodities

Influence of nitrogen availability on the expression of genes involved in the biosynthesis of saxitoxin and analogs in Cylindrospermopsis raciborskii

The development of cyanobacterial blooms in inland aquatic ecosystems is greatly promoted by nutrient availability, especially nitrogen and phosphorous. When blooms are dominated by toxigenic species the harmful effects of nutrient loading becomes particularly relevant. Among toxic species, Cylindrospermopsis raciborskii found in South American ecosystems is characterized by the production of saxitoxin and analogs (Paralytic Shellfish Poisoning, PSP), for which the factors that trigger their production have not been elucidated. In this study, the effect of nitrate availability on the relative transcript abundance of two genes (sxtU and sxtI), both involved in different steps of PSP biosynthetic pathway, was addressed in C. raciborskii MVCC19 by qPCR. The relative transcript abundance of both genes significantly increased from the beginning to the end of growth, independent of nitrate availability in the culture medium. Differences between the genes in terms of the levels of relative expression were also found, implying that during growth in nitrate-rich or nitrate-deprived conditions C. raciborskii MVCC19 has the ability to produce different kind of PSP molecules. The presence of nifH transcripts in the nitrogen-depleted treatment confirmed that in the absence of nitrate C. raciborskii fixed atmospheric N2. Moreover, after transferring filaments to nitrate-rich conditions the synthesis of nifH mRNA continued for few hours, suggesting that cell adjustments enabling the utilization of soluble nitrogen sources are not immediate. Our results show that biosynthesis of saxitoxin and analogs in C. raciborskii is not related to nitrate availability, but rather is linked to cyanobacteria growth rate.

Cannabis microbiome sequencing reveals several mycotoxic fungi native to dispensary grade Cannabis flowers

The Center for Disease Control estimates 128,000 people in the U.S. are hospitalized annually due to food borne illnesses. This has created a demand for food safety testing targeting the detection of pathogenic mold and bacteria on agricultural products. This risk extends to medical Cannabis and is of particular concern with inhaled, vaporized and even concentrated Cannabis products . As a result, third party microbial testing has become a regulatory requirement in the medical and recreational Cannabis markets, yet knowledge of the Cannabis microbiome is limited. Here we describe the first next generation sequencing survey of the fungal communities found in dispensary based Cannabis flowers by ITS2 sequencing, and demonstrate the sensitive detection of several toxigenic Penicillium and Aspergillus species, including P. citrinum and P. paxilli, that were not detected by one or more culture-based methods currently in use for safety testing.

The antagonistic effects of Candida parapsilosis on the growth of Fusarium species and fumonisin production.

Background and Purpose: Fusarium species are avid producers of secondary toxic and carcinogenic metabolites such as fumonisin. Contamination of food and feed products with fumonisin can be hazardous to the health of humans and animals and may lead to agricultural loss. Accordingly, in this study, we aimed to evaluate the effects of Candida parapsilosis on the growth and fumonisin production of Fusarium species. Materials and Methods:Mycelial growth rate of 26 Fusarium isolates, including F. verticillioides (n=6), F. proliferatum (n=18), F. solani (n=1), and F. oxysporum (n=1), in the presence of 42 C. parapsilosis strains was investigated by pour-plate method. The decline in fumonisin production was measured in co-cultured fungi in coarsely ground maize after four weeks of incubation in the dark at 22°C, using ELISA technique. For data analysis, paired t-test was performed, using SPSS version 20. Results:The mycelial growth and fumonisin production of Fusarium isolates significantly decreased in the presence of C. parapsilosis in comparison with the control cultures (P<0.05). The percentage of mycelial growth inhibition ranged from 56.36% to 74.54%. The minimum and maximum decline in total fumonisin production was 12% and 78%, respectively. F. oxysporum and F. solani were found to be minor fumonisin producers among the studied Fusarium species. On the other hand, a decline was reported in the growth of Fusarium species and fumonisin production in the presence of C. parapsilosis. Conclusion: C. parapsilosis showed notable inhibitory activities against Fusarium isolates. Therefore, this fungal species could be considered as a biocontrol agent against the growth and fumonisin production of toxigenic Fusarium species in the future.

Microbiological quality and sensory evaluation of new cured products obtained from sheep and goat meat

The present work aims to study the effect of species and seasoning time on the physicochemical, microbiological and sensory characteristics of cured legs of sheep and goats. Three cure periods were used: two for sheep and one for goat legs. Legs of lamb were cured for 7 and 8 months whereas legs of goat were cured for 8 months only. Samples were evaluated regarding pH, water activity and indicators of food microbial quality and safety. A trained panel carried out the sensory analysis, with aroma, texture, appearance and taste being the evaluated parameters. Significant differences were detected between the amount of aerobic mesophiles of the products cured during 7 months and the sheep legs cured during 8 months. Moulds and yeasts were between 1.81 × 106 ± 1.73 × 106 and 3.97 × 106 ± 5.45 × 106 colony-forming units/g, whereas total coliforms varied from 2.80 × 102 ± 4.13 × 102 to 1.31 × 104 ± 2.39 × 104. All samples were negative for toxigenic species. Concerning sensory analysis, hardness and taste persistence were the attributes that presented the highest and the lowest discriminative power, respectively. In general, the panel was able to characterise and distinguish the samples. The cured legs of goats were characterised as harder and as less succulent than those obtained from sheep. Sheep meat with larger time of cure was the brightest, whereas the one with a smaller time of cure was the most succulent. However, goat meat presented higher values of rancid and acid flavour. Sheep meat submitted to longer processing presented the most intense flavour and sheep meat with an inferior cure period presented the lowest intensity in all flavour attributes. This paper describes, for the first time in Portugal, the production and characterisation of cured legs of sheep and goats as a strategy to enhance economic value to good quality products obtained from animals of second category.

Effect of natural compounds on Fusarium graminearum complex.

A search is underway for new solutions to counter farm loss caused by fungal contamination of grains, since the active agents of fungicides can remain in the environment and contribute to the development of resistant and toxigenic species. In this study, the antifungal activity of natural compounds (γ-oryzanol, phenolic extract of neem seeds and of rice bran) was assessed on three toxigenic strains of Fusarium graminearum isolated from wheat, rice and barley. Their efficacy was compared to that of synthetic fungicides. The halo diameters were measured and the susceptible pathways were determined by the levels of structural compounds and activities of enzymes involved in the primary metabolism of the microorganisms. Moreover, mycotoxin production and gene expression were examined. Phenolic extracts were more effective at inhibiting F. graminearum than was γ-oryzanol, as evidenced by the minimum inhibitory concentration. This work contributed to the elucidation of the mechanism of action of natural antifungal agents. Natural antifungals effectively inhibited fungal growth, especially via the inactivation of the enzymatic systems of F. graminearum. Natural antifungals inhibited mycotoxin production by the fungi. A correlation between the levels of deoxynivalenol and the expression of Tri5 gene was observed, indicating that the natural compounds could be considered alternatives to synthetic antifungals. © 2015 Society of Chemical Industry.