Biofilm and Pathogenesis-Related Proteins in the Foodborne P. fluorescens ITEM 17298 With Distinctive Phenotypes During Cold Storage

In food chain, Pseudomonas spp. cause spoilage by reducing shelf life of fresh products, especially during cold storage, with a high economic burden for industries. However, recent studies have shed new light on health risks occurring when they colonize immunocompromised patient tissues. Likewise to P. aeruginosa, they exhibit antibiotic resistance and biofilm formation, responsible for their spread and persistence in the environment. Biofilm formation might be induced by environmental stresses, such as temperature fluctuations causing physiological and metabolic changes exacerbating food spoilage (by protease and pigment synthesis), and the production of adhesion molecules, chemotactic or underestimated virulence factors. In order to provide a new insight into phenotypic biodiversity of Pseudomonas spoilers isolated from cold stored cheese, in this work 19 Pseudomonas spp. were investigated for biofilm, pigments, exopolysaccharide production and motility at low temperature. Only nine strains showed these phenotypic traits and the blue pigmenting cheese strain P. fluorescens ITEM 17298 was the most distinctive. In addition, this strain decreased the survival probability of infected Galleria mellonella larvae, showing, for the first time, a pathogenic potential. Genomic and proteomic analyses performed on the ITEM 17298 planktonic cells treated or not with lactoferrin derived antibiofilm peptides allowed to reveal specific biofilm related-pathways as well as proteins involved in pathogenesis. Indeed, several genes were found related to signaling system by cGMP-dependent protein kinases, cellulose, rhamnolipid and alginate synthesis, antibiotic resistance, adhesion and virulence factors. The proteome of the untreated ITEM 17298, growing at low temperature, showed that most of the proteins associated with biofilm regulation, pigmentation motility, antibiotic resistance and pathogenecity were repressed, or decreased their levels in comparison to that of the untreated cultures. Thus, the results of this work shed light on the complex pathways network allowing psychrotrophic pseudomonads to adapt themselves to food-refrigerated conditions and enhance their spoilage. In addition, the discovery of virulence factors and antibiotic resistance determinants raises some questions about the need to deeper investigate these underestimated bacteria in order to increase awareness and provide input to update legislation on their detection limits in foods

Molecular characteristics of a strain (Salento-1) of Xylella fastidiosa isolated in Apulia (Italy) from an olive plant with the quick decline syndrome

DNA-based approaches were used to characterize a strain (Salento-1) of Xylella fastidiosa obtained from an olive plant suffering from the syndrome of quick decline in Apulia (South Italy). Salento-1 was indistinguishable from strain CoDiRO previously isolated from olive in Apulia and assigned to  X. fastidiosa subsp. pauca. Based on our results and comparative analysis with reported data, the subspecies pauca, multiplex, and fastidiosa may invade olive throughout the world (California, Italy, Argentina and Brazil). The strain Salento-1 has been deposited in the National Collection of Plant Pathogenic Bacteria (NCPPB), England, and in the Belgian Coordinated Collections of Microorganisms (BCCM), Belgium

AFLP variability, toxin production, and pathogenicity of Alternaria species from Argentinean tomato fruits and puree

Large amounts of tomato fruits and derived products are produced in Argentina and may be contaminated by Alternaria toxins. Limited information is available on the genetic variability, toxigenicity, and pathogenicity of Alternaria strains occurring on tomato. We analyzed 65 Alternaria strains isolated in Argentina from tomato fruits affected by black mould and from tomato puree, using amplified fragment length polymorphisms (AFLPs) technique. AFLP analysis resolved the set of strains in 3 main clusters (DICE similarity values of 58 and 60%) corresponding to A. alternata/tenuissima (44 strains), A. arborescens (15 strains) and to an unknown group (6 strains). Most of the representative strains, belonging to each AFLP cluster, when cultured on rice, produced tenuazonic acid (up to 46,760. mg/kg), alternariol monomethyl ether (AME, up to 1860. mg/kg), and alternariol (up to 70. mg/kg). The toxin profile related to the strains was not related to any AFLP cluster, except for AME which was produced at lower level by A. arborescens. Most of strains were pathogenic on two types of commonly cultivated tomato fruits. These findings provide new information on the variability within the Alternaria species complex associated with tomato disease.Fil: Somma, Stefania. Institute of Sciences of Food Production; ItaliaFil: Pose, Graciela Noemi. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Pardo, Alejandro Guillermo. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Mulè, Giuseppina. Institute of Sciences of Food Production; ItaliaFil: Fernández Pinto, Virginia Elena. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Orgánica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Moretti, Antonio. Institute of Sciences of Food Production; ItaliaFil: Logrieco, Antonio Francesco. Institute of Sciences of Food Production; Itali

Molecular characteristics of a strain (Salento-1) of Xylella fastidiosa isolated in Apulia (Italy) from an olive plant with the quick decline syndrome

DNA-based approaches were used to characterize a strain (Salento-1) of Xylella fastidiosa obtained from an olive plant suffering from the syndrome of quick decline in Apulia (South Italy). Salento-1 was indistinguishable from strain CoDiRO previously isolated from olive in Apulia and assigned to  X. fastidiosa subsp. pauca. Based on our results and comparative analysis with reported data, the subspecies pauca, multiplex, and fastidiosa may invade olive throughout the world (California, Italy, Argentina and Brazil). The strain Salento-1 has been deposited in the National Collection of Plant Pathogenic Bacteria (NCPPB), England, and in the Belgian Coordinated Collections of Microorganisms (BCCM), Belgium

“Ectomosphere”: Insects and Microorganism Interactions

This study focuses on interacting with insects and their ectosymbiont (lato sensu) microorganisms for environmentally safe plant production and protection. Some cases help compare insect-bearing, -driving, or -spreading relevant ectosymbiont microorganisms to endosymbionts’ behaviour. Ectosymbiotic bacteria can interact with insects by allowing them to improve the value of their pabula. In addition, some bacteria are essential for creating ecological niches that can host the development of pests. Insect-borne plant pathogens include bacteria, viruses, and fungi. These pathogens interact with their vectors to enhance reciprocal fitness. Knowing vector-phoront interaction could considerably increase chances for outbreak management, notably when sustained by quarantine vector ectosymbiont pathogens, such as the actual Xylella fastidiosa Mediterranean invasion episode. Insect pathogenic viruses have a close evolutionary relationship with their hosts, also being highly specific and obligate parasites. Sixteen virus families have been reported to infect insects and may be involved in the biological control of specific pests, including some economic weevils. Insects and fungi are among the most widespread organisms in nature and interact with each other, establishing symbiotic relationships ranging from mutualism to antagonism. The associations can influence the extent to which interacting organisms can exert their effects on plants and the proper management practices. Sustainable pest management also relies on entomopathogenic fungi; research on these species starts from their isolation from insect carcasses, followed by identification using conventional light or electron microscopy techniques. Thanks to the development of omics sciences, it is possible to identify entomopathogenic fungi with evolutionary histories that are less-shared with the target insect and can be proposed as pest antagonists. Many interesting omics can help detect the presence of entomopathogens in different natural matrices, such as soil or plants. The same techniques will help localize ectosymbionts, localization of recesses, or specialized morphological adaptation, greatly supporting the robust interpretation of the symbiont role. The manipulation and modulation of ectosymbionts could be a more promising way to counteract pests and borne pathogens, mitigating the impact of formulates and reducing food insecurity due to the lesser impact of direct damage and diseases. The promise has a preventive intent for more manageable and broader implications for pests, comparing what we can obtain using simpler, less-specific techniques and a less comprehensive approach to Integrated Pest Management (IPM).The present work acknowledges the support from: European Union’s Horizon 2020 research and innovation programme under Grant Agreements No. 635646-POnTE “Pest Organisms Threatening Europe”, No. 727987-XF-ACTORS “Xylella Fastidiosa Active Containment Through a multidisciplinary-Oriented Research Strategy”, Grant number 952337-MycoTWIN “Enhancing Research and Innovation Capacity of Tubitak MAM Food Institute on Management of Mycotoxigenic Fungi and Mycotoxins”, and CURE-Xf, H2020-Marie Sklodowska-Curie Actions—Research and Innovation Staff Exchange. Reference number: 634353, coordinated by CIHEAM Bari. The EU Funding Agency is not responsible for any use that may be made of the information it contains. European Union’s StopMedWaste “Innovative Sustainable technologies TO extend the shelf-life of Perishable MEDiterranean fresh fruit, vegetables and aromatic plants and to reduce WASTE” a PRIMA project ID: 1556. European Union’s Euphresco BasicS “Basic substances as an environmentally friendly alternative to synthetic pesticides for plant protection” project ID: 2020-C-353. The work was partially carried out in the framework of the National Projects: RIGENERA, granted by MASAF n. 207631, 9 May 2022, and GENFORAGRIS, granted by MASAF n. 207631, 9 May 2022; and regional projects “Laboratory network for the selection, characterisation and conservation of germplasm and for preventing the spread of economically-relevant and quarantine pests (SELGE) No. 14”, founded by the Apulia Region, PO FESR 2007–2013—Axis I, Line of intervention 1.2., Action 1.2.1; Research for Innovation (REFIN) POR Puglia 2014–2020 Project: 8C6E699D, and PON AIM, COD. AIM 1809249-Attività 1 Linea 1

Biodiversity of Aspergillus Section Flavi in Europe in relation to the management of aflatoxin risk

Aflatoxins and the producing fungi Aspergillus Section Flavi are widely known as the most serious and dangerous mycotoxin issue in agricultural products. In Europe, before the outbreak of aflatoxins on maize (2003-2004) due to new climatic conditions, their contamination was confined to imported foods. Little information is available on molecular biodiversity and population structure of Aspergillus Section Flavi in Europe. Preliminary reports evidenced the massive presence of A. flavus L-morphotype as the predominant species in maize field, no evidence of the highly toxigenic S-morphotype and of other aflatoxigenic species are reported. The risk of a shift in traditional occurrence areas for aflatoxins is expected in the world and in particular in South East of Europe due to the increasing average temperatures. Biological control of aflatoxin risk in the field by atoxigenic strains of A. flavus starts to be widely used in Africa and USA. Studies are necessary on the variation of aflatoxin production in populations of A. flavus to characterize stable atoxigenic A. flavus strains. The aim of present article is to give an overview on biodiversity and genetic variation of Aspergillus Sect. Flavi in Europe in relation to the management of aflatoxins risk in the field

In Vitro and in Field Response of Different Fungicides against Aspergillus flavus and Fusarium Species Causing Ear Rot Disease of Maize

Aspergillus flavus, the main aflatoxin B1 producing fungal species, Fusarium graminearum, a deoxynivalenol producer, and the fumonisin-producing species F. proliferatum and F. verticillioides are the main toxigenic fungi (TF) that colonize maize. Several strategies are available to control TF and related mycotoxins, such as chemical control. However, there is poor knowledge on the efficacy of fungicides on maize plants since few molecules are registered. The sensitivity of F. graminearum, F. proliferatum, F. verticillioides, and A. flavus to eleven fungicides, selected based on their different modes of action, was evaluated in both in vitro assays and, after selection, in the field. In vitro, demethylation inhibitors (DMI) showed excellent performances, followed by thiophanate-methyl and folpet. Among the succinate dehydrogenase inhibitors (SDHI), isopyrazam showed a higher effectiveness against Fusarium species than boscalid, which was ineffective against Fusarium, like the phenyl-pyrrole fludioxonil. Furthermore, both SDHIs and fludioxonil were more active against A. flavus than Fusarium species. In field trials, prothioconazole and thiophanate-methyl were confirmed to be effective to reduce F. graminearum (52% and 48%) and F. proliferatum contamination (44% and 27%). On the other hand, prothioconazole and boscalid could reduce A. flavus contamination at values of 75% and 56%, respectively

Boron Biofortification of Portulaca oleracea L. through Soilless Cultivation for a New Tailored Crop

Purslane (Portulaca oleracea L.) is a wild edible plant, traditionally consumed in the Mediterranean area and recently proposed as a new ready-to-eat vegetable; it is also called the “vegetable for long life” because of the high contents of several healthy compounds. Although boron (B) is not considered to be essential for humans, a daily intake of about 2 mg to obtain positive effects on aging in adult men and women has been suggested. In this study, two genotypes of purslane (wild collected and commercial variety) are grown by using a hydroponic system with three boron (B) levels in the nutrient solution (NS) (0.3 mg/L—control, 3 mg/L—low level of biofortification, and 6 mg/L—high level of biofortification) in order to increase the B content in the edible parts of the plant. The crop yield, color traits, and content of glucose, fructose, total phenols, chlorophylls, carotenoids, mineral elements (Al, B, Ca, Cr, Fe, K, Mg, Mn, Na, and Zn), nitrate, and oxalate are analyzed. Independent of the genotype, the B content in edible purslane was successfully increased in comparison with the control, obtaining 1.8- to 10.7-fold higher values of B tissue concentrations by using, respectively, 3 and 6 mg/L of B in the NS without affecting crop performances. From a nutritional point of view, the average daily intake of B could be satisfied by consuming about 75 or 48 g of purslane, grown by using 3 and 6 mg/L B level in the NS, respectively. Apart from B and Fe, the content of mineral elements in edible parts of purslane was not strongly influenced by different B levels in the NS but it was affected by genotypes. A lower sugar content was found in wild purslane grown with the highest B level. A higher content of both chlorophylls and carotenoids was found in the control but only for the commercial genotype. No differences in oxalate content were observed among B levels in the NS, while only in the case of wild genotype, we found a lower nitrate content when a B concentration of 3 mg/L was used in the NS. In conclusion, we demonstrated the possibility of using the floating hydroponic system, combined with specific B concentrations in the NS composition, as a method to calibrate the B uptake in edible parts of purslane

Hydroponic Production of Reduced-Potassium Swiss Chard and Spinach: A Feasible Agronomic Approach to Tailoring Vegetables for Chronic Kidney Disease Patients

Tailored foods are specifically suitable for target groups of people with particular nutritional needs. Although most research on tailored foods has been focused on increasing the nutrient content in plant tissues (biofortification), in populations with specific physiological conditions, it is recommended to reduce the uptake of specific nutrients in order to improve their health. People affected by chronic kidney disease (CKD) must limit their consumption of vegetables because of the generally high potassium (K) content in the edible parts. This study aimed to define an appropriate production technique for two baby leaf vegetables, spinach (Spinacia oleracea L.) and Swiss chard (Beta vulgaris L. ssp. vulgaris), with reduced K tissue content, minimizing the negative effects on their crop performance and overall nutritional quality. Plants were grown in a hydroponic floating system. The K concentration in the nutrient solution (NS) was reduced from 200 mg/L (K-200, the concentration usually used for growing baby leaf vegetables in hydroponic conditions) to 50 mg/L over the entire growing cycle (K-50) or only during the seven days before harvest (K50-7d). The reduction of K in the NS resulted in a significant decrease of K tissue content in both species (32% for K-50 and 10% for K50-7d, on average), while it did not, in general, compromise the crop performance and quality traits or the bioaccessibility of K, magnesium, and calcium. The production of reduced-potassium leafy vegetables is a feasible tailored nutrition approach for CKD patients in order to take advantage of the positive effects of vegetable consumption on health without excessively increasing potassium intake

Boron Biofortification of Portulaca oleracea L. through Soilless Cultivation for a New Tailored Crop

Purslane (Portulaca oleraceaL.) is a wild edible plant, traditionally consumed in the Mediterranean area and recently proposed as a new ready-to-eat vegetable; it is also called the "vegetable for long life" because of the high contents of several healthy compounds. Although boron (B) is not considered to be essential for humans, a daily intake of about 2 mg to obtain positive effects on aging in adult men and women has been suggested. In this study, two genotypes of purslane (wild collected and commercial variety) are grown by using a hydroponic system with three boron (B) levels in the nutrient solution (NS) (0.3 mg/L-control, 3 mg/L-low level of biofortification, and 6 mg/L-high level of biofortification) in order to increase the B content in the edible parts of the plant. The crop yield, color traits, and content of glucose, fructose, total phenols, chlorophylls, carotenoids, mineral elements (Al, B, Ca, Cr, Fe, K, Mg, Mn, Na, and Zn), nitrate, and oxalate are analyzed. Independent of the genotype, the B content in edible purslane was successfully increased in comparison with the control, obtaining 1.8- to 10.7-fold higher values of B tissue concentrations by using, respectively, 3 and 6 mg/L of B in the NS without affecting crop performances. From a nutritional point of view, the average daily intake of B could be satisfied by consuming about 75 or 48 g of purslane, grown by using 3 and 6 mg/L B level in the NS, respectively. Apart from B and Fe, the content of mineral elements in edible parts of purslane was not strongly influenced by different B levels in the NS but it was affected by genotypes. A lower sugar content was found in wild purslane grown with the highest B level. A higher content of both chlorophylls and carotenoids was found in the control but only for the commercial genotype. No differences in oxalate content were observed among B levels in the NS, while only in the case of wild genotype, we found a lower nitrate content when a B concentration of 3 mg/L was used in the NS. In conclusion, we demonstrated the possibility of using the floating hydroponic system, combined with specific B concentrations in the NS composition, as a method to calibrate the B uptake in edible parts of purslane