Industrial scale bio-detoxification of raw olive mill wastewaters by the use of selected microbial yeast and bacterial strains to obtain a new source for fertigation.

The Olive Mill Wastewaters (OMWs) are one of the most important agro-industrial wastes of the Mediterranean Countries and the disposal by draining them onto land has been proved to be damaging for soils, plants and groundwater due to their polluting power. The present report describes a new method for bio-detoxification of undiluted fresh OMW based on the driven selection of aerobic yeasts and bacteria. The identified yeast Candida boidinii A5y and the bacterium Paenibacillus albidus R32b strains allowed the treatment of freshly produced raw OMW characterized by very high COD value and phenolic content, when applied as sequential inoculum. The treated OMW showed the absence of antimicrobial effects and a strongly reduction of phytotoxic activity on the germination of several plant seeds. The process was successfully validated on an industrial scale without any pre-treatment, dilution and/or supplementation of the raw waste. Bio-detoxified OMW produced by this sustainable and low-cost process would be suitable for new non-chemical fertigation or soilless applications. The described procedure represents a virtuous example of circular economy efficaciously applied for a depleting agri-food resource.

Expression of Lactobacillus brevis IOEB 9809 tyrosine decarboxylase and agmatine deiminase genes in wine correlates with substrate availability.

AIMS: Lactobacillus brevis IOEB 9809 is able to produce both tyramine and putrescine via tyrosine decarboxylase and agmatine deiminase enzymes, respectively, when cultured on synthetic media. The aims of this study were to assess the expression of L. brevis IOEB 9809 tdc and aguA1 genes, during wine fermentation and to evaluate the effect of substrate availability and pH on tdc and aguA1 expression, as well as on biogenic amine production and L. brevis viability. METHODS AND RESULTS: The relative expression of L. brevis IOEB 9809 tdc and aguA1 genes was analysed in wine by quantitative real-time RT-PCR (qRT-PCR) during a period of incubation of 30 days. Cell viability, pH values, putrescine and tyramine concentration were monitored throughout the experiments. CONCLUSIONS: The wine trials indicated that L. brevis IOEB 9809 is able to produce both tyramine and putrescine during wine fermentation. Increased cell viability was also observed in wine supplemented with tyrosine or agmatine. qRT-PCR analysis suggests a strong influence of substrate availability on the expression of genes coding for tyrosine decarboxylase and agmatine deiminase in L. brevis IOEB 9809. Less evident is the relationship between putrescine and tyramine production and tolerance to wine pH. SIGNIFICANCE AND IMPACT OF STUDY: To our knowledge, this study represents the first assessment of relative expression of L. brevis IOEB 9809 genes involved in biogenic amine production in wine. Furthermore, an effect of biogenic amine production on viability of L. brevis during wine fermentation was established.

Selection of non-conventional yeasts and their use in immobilized form for the bioremediation of olive oil mill wastewaters.

The yeast population dynamics in olive wastewaters (OMW), sampled in five mills from Salento (Apulia, Southern Italy), were investigated. Three hundred yeasts were isolated in five industrial mills and identified by molecular analysis. Strains belonging to Geotrichum, Saccharomyces, Pichia, Rhodotorula and Candida were detected. Five G. candidum strains were able to grow in OMW as the sole carbon source and to reduce phenolics, chemical oxygen demand (COD) and antimicrobial compounds. One G. candidum isolate was selected for whole-cell immobilization in calcium alginate gel. The COD and phenolic reduction obtained with immobilized cells showed a 2.2- and 2-fold increase compared to the removal obtained with free cells, respectively. The immobilization system enhanced yeast oxidative activity by avoiding the presence of microbial protease in treated OMW. To our knowledge, this is the first report on G. candidum whole-cell immobilization for OMW bioremediation.

Persistence of fenitrothion in oranges and clementines after treatment with emulsifiable concentrate and microencapsulate formulations.

The rate of decline of fenitrothion residues was investigated in oranges and clementines after treatment with two different kinds of commercial formulations: emulsifiable concentrate (Afidina M) and microencapsulate (Fenitrocap and IPM 400). The study was performed on the fruit and leaves over 131 and 161 days for oranges, and over 78 and 86 days for clementines, respectively. In fruit, the experimental data showed a similar behaviour of the active ingredient for both kinds of commercial formulations. High mean levels of fenitrothion (between about 0.4 and 0.8 mg kg(-1)) were persistent for at least 75 days after treatment in oranges and 50 days in clementines, with statistically significant declines observed only at days 110 and 78, respectively. A rapid decline of fenitrothion levels was observed in orange and clementine leaves during the starting phase followed by a slower decrease during the later stage; the decline was more pronounced with the treatment of emulsifiable concentrates. These findings are indicative of a poor degradability of fenitrothion in citrus fruits, and suggest that repeated or uneven applications of the pesticide should be avoided in order to exclude the risk of exceeding the maximum residue level permitted by the current regulations.

Detection of rice tungro bacilliform virus gene products in vivo.

To study the products of the open reading frames (ORFs) of rice tungro bacilliform virus in rice plants the sequences containing ORFs I (encoding a 24-kDa protein, P24) and IV (P46) and the protease and polymerase (reverse transcriptase+RNaseH) domains of ORF III were cloned into a pGEX expression vector. The proteins, which were C-terminal fusions to glutathione S-transferase, were expressed in Escherichia coli and antisera were raised against them which, together with an antiserum against virus particles, was used to probe blots of proteins from infected and uninoculated plants and from virus preparations. The P24 antiserum detected virus-specific proteins of 74, 60, and 52 kDa, which are much bigger than expected. These proteins were found in virus preparations and immunogold labeling suggested that they might be internal in the particles. Virus-specific proteins of 33, 37, 62, and > 150 kDa were revealed by antiserum to virus particles. The antiserum to the protease revealed proteins of 13.5, 37, and 68 kDa both in extracts from infected plants and in purified virus preparations. This antiserum decorated intact virus particles as did the particle antiserum. The polymerase domain antiserum reacted with products of 56, 65, and 68 kDa in extracts from infected plants but not in virus particles. The antiserum to the ORF IV product did not detect any bands in either infected plant extracts or virus preparations. The significance of these products is discussed.