In order to identify antifungal compounds from natural sources to be used as ingredients in the bakery industry, water/salt-soluble extracts (WSE) from different legume flour hydrolysates obtained by the use of a fungal protease were assayed against Penicillium roqueforti DPPMAF1. The agar diffusion assays allowed the selection of the pea (Pisum sativum) hydrolysate as the most active. As shown by the hyphal radial growth rate, the WSE had inhibitory activity towards several fungi isolated from bakeries. The MIC of the WSE was 9.0 mg/ml. Fungal inhibition was slightly affected by heating and variations in pH. The antifungal activity was attributed to three native proteins (pea defensins 1 and 2 and a nonspecific lipid transfer protein [nsLTP]) and a mixture of peptides released during hydrolysis. The three proteins have been reported previously as components of the defense system of the plant. Five peptides were purified from WSE and were identified as sequences encrypted in leginsulin A, vicilin, provicilin, and the nsLTP. To confirm antifungal activity, the peptides were chemically synthesized and tested. Freeze-dried WSE were used as ingredients in leavened baked goods. In particular, breads made by the addition of 1.6% (wt/wt) of the extract and fermented by baker's yeast or sourdough were characterized for their main chemical, structural, and sensory features, packed in polyethylene bags, stored at room temperature, and compared to controls prepared without pea hydrolysate. Artificially inoculated slices of a bread containing the WSE did not show contamination by fungi until at least 21 days of storage and behaved like the bread prepared with calcium propionate (0.3%, wt/wt). Contamination by fungi is the most common cause of microbial spoilage and a costly problem for bakeries. In many cases, it is the major factor governing shelf life. In addition to the repellent sight of visible growth, fungi may be responsible for off-flavors and may synthesize mycotoxins and allergenic compounds (1, 2).Chemical preservatives, novel ingredients with fungus-inhibiting properties, and packaging techniques are all technology options that, together with hygiene during processing, may contribute to decreasing the growth of fungi in baked goods (3). Routinely, salts of propionic, sorbic, and benzoic acids, as well as ethanol, are used as chemical preservatives. Nevertheless, consumers require high-quality, preservative-free, safe but mildly processed foods with extended shelf life and preserved sensory properties. These requirements have led to a search for new preservatives derived mainly from natural sources. In nature, antifungal compounds from plants comprise proteins and peptides that are thought to play an important role in plant disease resistance, serving as protective agents against fungal invasion (4). In particular, leguminous plants represent an important reservoir to be investigated owing to their abundance of proteins with important biological activities (5). Overall, leguminous proteins are classified as prote...

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The adoption of sewage sludge as an agricultural management strategy to improve soil properties and crop production is attracting great interest. Despite many positive effects on soil inorganic and organic components reported for different soil types, little information is available on sewage sludge application on Mediterranean soils, as well as on its use at different dose rates. The objectives of the present research was to evaluate, through an integrated approach, the effects of sewage sludge compost from urban wastewater on physicochemical, hydrological, biochemical parameters, and microbiota composition in soil pots under a three-year crop rotation system. Four different doses of sewage sludge compost (C3, C6, C9, C12) from municipal wastewater and a dose of them in combination with mineral fertilizer (C6N) were used. We have used 3-6-9-12 Mg/ha of sewage sludge compost for the treatments C3, C6, C9 and C12, respectively, and 6 Mg/ha of sewage sludge compost in combination with 60 kg/ha of ammonium nitrate for the treatment C6N.The effects were compared to non-fertilized (C0) and mineral fertilized (Min) sets of controls. The electrical conductivity, soil pH, stability of soil aggregates, percent of moisture of the dry soil both at the field capacity and at the wilting point, available P, and exchangeable K were all positively affected by increasing the amounts of composted sludge. The organic carbon and total N increased up to 66% and 39%, respectively. Increased enzymatic activities and microbial biomass were also observed in soil after the application of sewage sludge compost when compared to un-amended control. A higher richness and evenness among the soil plots amended with sewage sludge compost was observed, with no significant differences among the application dose rates, when compared to the un-amended soil control and soil treated with a mineral fertilizer. A three-year amendment was able to separate soil plots amended with high doses of sewage sludge compost from the low dose amended and control samples. Among the microbial groups responsible for such marked separation, bacteria belonging to Actinobacteria, Acidobacteria, Cyanobacteria and Bacteroidetes contribute the most, with a shift from oligotrophic to copiotrophic taxa. Significant changes in bacterial composition and taxonomic structure should be considered in order to properly balance agronomic and economic advantages with environmental concerns. After all, our results have evidenced the effects of sewage sludge amendment on different soil properties, microbial activity, and composition already after a short period of application. The findings are particularly relevant in semiarid soils, where an immediate restoration of soil fertility by short-term compost application is needed.

Coagulase Negative Staphylococci (CoNS) are becoming increasingly recognized as an important cause of human and animal infections. Notwithstanding their clinical relevance, annotation of genes potentially involved in pathogenicity and/or antibiotic resistance in the CoNS species Staphylococcus arlettae (SAR) is currently very limited. In the current work we describe the genome of a novel methicillin resistant isolate of SAR, which we named Bari, and present a comprehensive analysis of predicted antibiotic resistance profiles and virulence determinants for all the 22 currently available SAR genomes. By comparing predicted antibiotic resistance and virulence-associated genes with those obtained from a manual selection of 148 bacterial strains belonging to 14 different species of staphylococci and to two “outgroup” species, Bacillus subtilis (BS) and Macrococcus caseoliticus (MC), we derived some interesting observations concerning the types and number of antibiotic resistance-related and virulence-like genes in SAR. Interestingly, almost 50% of the putative antibiotic resistance determinants identified in this work, which include the clinically relevant mec, van, and cls genes, were shared among all the SAR strains herein considered (Bari included). Moreover, comparison of predicted antibiotic resistance profiles suggest that SAR is closely related to well-known pathogenic Staphylococcus species, such as Staphylococcus aureus (SA) and Staphylococcus epidermidis (SE). A similar analysis of predicted virulence factors, revealed that several genes associated with pathogenesis (including, for example, ica, nuc, and ssp), which are commonly found in the genomes of pathogenic staphylococci such as Staphylococcus haemolyticus (SH) and Staphylococcus saprophyticus (SS), are observed also in the SAR strains for which a genomic sequence is available. All in all, we believe that the analyses presented in the current study, by providing a consistent and comprehensive annotation of virulence and antibiotic resistance-related genes in SAR, can constitute a valuable resource for the study of molecular mechanisms of opportunistic pathogenicity in this species.

The genome sequences of three new strains of Staphylococcus arlettae named Bari1, Bari2, and Bari3 are presented. The strains exhibited tolerance to hexavalent chromium ions.