Whole-genome analysis suggesting probiotic potential and safety properties of Pediococcus pentosaceus DSPZPP1, a promising LAB strain isolated from traditional fermented sausages of the Basilicata region (Southern Italy).

Introduction: Many lactic acid bacteria (LAB) strains are currently gaining attention in the food industry and various biological applications because of their harmless and functional properties. Given the growing consumer demand for safe food, further research into potential probiotic bacteria is beneficial. Therefore, we aimed to characterize Pediococcus pentosaceus DSPZPP1, a LAB strain isolated from traditional fermented sausages from the Basilicata region of Southern Italy. Methods: In this study, we analyzed the whole genome of the P. pentosaceus DSPZPP1 strain and performed in silico characterization to evaluate its applicability for probiotics and use in the food industry. Results and Discussion: The whole-genome assembly and functional annotations revealed many interesting characteristics of the DSPZPP1 strain. Sequencing raw reads were assembled into a draft genome of size 1,891,398 bp, with a G + C content of 37.3%. Functional annotation identified 1930 protein-encoding genes and 58 RNAs including tRNA, tmRNA, and 16S, 23S, and 5S rRNAs. The analysis shows the presence of genes that encode water-soluble B-group vitamins such as biotin, folate, coenzyme A, and riboflavin. Furthermore, the analysis revealed that the DSPZPP1 strain can synthesize class II bacteriocin, penocin A, adding importance to the food industry for bio-enriched food. The DSPZPP1 genome does not show the presence of plasmids, and no genes associated with antimicrobial resistance and virulence were found. In addition, two intact bacteriophages were identified. Importantly, the lowest probability value in pathogenicity analysis indicates that this strain is non-pathogenic to humans. 16 s rRNA-based phylogenetic analysis and comparative analysis based on ANI and Tetra reveal that the DSPZPP1 strain shares the closest evolutionary relationship with P. pentosaceus DSM 20336 and other Pediococcus strains. Analysis of carbohydrate active enzymes (CAZymes) identified glycosyl transferases (GT) as a main class of enzymes followed by glycoside hydrolases (GH). Our study shows several interesting characteristics of the isolated DSPZPP1 strain from fermented Italian sausages, suggesting its potential use as a promising probiotic candidate and making it more appropriate for selection as a future additive in biopreservation.

Alternative Ecological Products for Aphid Control on Plum.

Ecological farming is increasing worldwide, as more and more consumers prefer chemical-free fruits. As a result, ecological farming is becoming increasingly appealing to European farmers, including those in Romania. However, implementing an effective ecological phytosanitary program remains a challenge for farmers due to limited options and a lack of information about their effectiveness. Romania is a major producer of plums and ranks second in the world after China. Aphids are common pests of plum, and some species are vectors of the damaging Plum pox virus, and therefore require special attention regarding their control. Eight ecological products were tested both in the field and laboratory for a duration of three vegetative periods to determine their efficiency in aphid control. The effects of ecological products were compared with five chemical insecticides known to be effective against aphids. Observations were made 24 and 48 h after spraying. Two of the eight ecological products tested were proven to be efficient in aphid control, Ovipron Top and Prev-Am, with a mortality rate over 90%. The results indicate that these two ecological products are comparable in effectiveness to chemical insecticides and could be suitable candidates for both ecological and conventional treatment programs.

Antimicrobial and antioxidant properties and quantitative screening of phytochemicals of Fraxinus excelsior L. and Eschscholtzia californica Cham. mother tinctures.

The antioxidant and antimicrobial activities of Fraxinus excelsior L. and Eschscholtzia californica Cham. mother tinctures against a range of foodborne bacteria were investigated to determine the major components and to analyse the action spectrum and antimicrobial effectiveness of the extracts. Results demonstrated a significant antioxidant activity of Fraxinus excelsior L. and a lower activity of Eschscholtzia californica Cham. and a good chemical phenolic composition with the highest content of flavonoids. The Fraxinus excelsior L. and Eschscholtzia californica Cham. mother tinctures demonstrated a middle-high antimicrobial activity against, respectively, 66.67% and 43.33% of all tested bacteria. The inhibitory activity showed a moderate effect on the growth of the sensitive strains in presence of extracts minimum inhibitory concentration. The synergistic actions of bioactive compounds detected in the extracts might be on the basis of antioxidant and biological activities observed and should be used in pharmaceutical, food preservation, alternative medicine and natural therapies fields.

Effects of Below-Ground Microbial Biostimulant Trichoderma harzianum on Diseases, Insect Community, and Plant Performance in Cucurbita pepo L. under Open Field Conditions.

Agrochemicals are generally used in agriculture to maximize yields and product quality, but their overuse can cause environmental pollution and human health problems. To reduce the off-farm input of chemicals, numerous biostimulant products based on beneficial symbiont plant fungi are receiving a great deal of attention. The evolution of plant diseases and the performance of insects are influenced by plant chemical defences, both of which are, in turn, influenced by below-ground symbionts. Direct and indirect plant defences mediated by belowground symbionts against plant diseases and insect herbivores were demonstrated in greenhouses experiments. However, little attention has been paid to the use of Trichoderma under open field conditions, and no data are available for zucchini (Cucurbita pepo L.) plants in the field. To determine the effects of a commercial Trichoderma harzianum strain T22 on plant viruses, powdery mildew, the arthropod community, and on the agronomic performance associated with zucchini plants, an experiment was conducted in 2022 under open field conditions in South Italy. Our results indicate that T. harzianum T22 makes zucchini plants more attractive to aphids and to Hymenoptera parasitoid but failed to control zucchini pathogens. The complex plant-disease-arthropod-microorganism interactions that occurred in the field during the entire plant cycle are discussed to enrich our current information on the possibilities of using these microorganisms as a green alternative in agriculture.

Investigations on Fungi Isolated from Apple Trees with Die-Back Symptoms from Basilicata Region (Southern Italy).

Val d'Agri is an important orchard area located in the Basilicata Region (Southern Italy). A phenomenon affecting cv. "Golden Delicious" apples which lead to tree death has been observed in the past several years in this area. This phenomenon has already been detected in about 20 hectares and is rapidly expanding. The symptoms observed were "scaly bark" and extensive cankers, mainly located in the lower part of the trunk, associated with wood decay. Dead plants ranged from 20% to 80% and, in many cases, trees were removed by farmers. In order to identify the causes of this phenomenon, investigations were started in autumn/winter 2019. In order to determine the possible causal agents, fungal and bacterial isolations, from symptomatic tissues, were performed in laboratory. Bacterial isolations gave negative results, whereas pure fungal cultures (PFCs) were obtained after 3-4 passages on potato dextrose agar (PDA) media. Genetic material was extracted from each PFC and amplified by PCR using three pairs of primers: ITS5/4, Bt2a/Bt2b and ACT-512F/ACT-783R. The amplicons were directly sequenced, and nucleotide sequences were compared with those already present in the NCBI GenBank nucleotide database. All isolated fungi were identified based on morphological features and multilocus molecular analyses. Neofusicoccum parvum, Diaporthe eres and Trametes versicolor were most frequently isolated, while Pestalotiopsis funerea, Phomopsis spp. and Diaporthe foeniculina were less frequently isolated. All nucleotide sequences obtained in this study have been deposited into the EMBL database. Pathogenicity tests showed that N. parvum was the most pathogenic and aggressive fungus, while Phomopsis sp. was demonstrated to be the less virulent one. All the investigated fungi were repeatedly reisolated from artificially inoculated twigs of 2-year-old apple trees, cv. "Golden Delicious", and subsequently morphologically and molecularly identified. The role played by the above-mentioned fungi in the alterations observed in field is also discussed.

Effects of Trichoderma harzianum Strain T22 on the Arthropod Community Associated with Tomato Plants and on the Crop Performance in an Experimental Field.

Fungi belonging to the genus Trichoderma have received much attention in recent years due to their beneficial effects on crop health and their use as pest control agents. Trichoderma activates direct plant defenses against phytophagous arthropods and reinforces indirect plant defense through the attraction of predators. Although the plant defenses against insect herbivores were demonstrated in laboratory experiments, little attention has been paid to the use of Trichoderma spp. in open field conditions. In the present study, we investigated the effects of the inoculation of the commercial Trichoderma harzianum strain T22 on the arthropod community associated with tomato plants and on the crop performance in an experimental field located in South Italy. Our results showed that inoculation with T. harzianum could alter the arthropod community and reduce the abundance of specific pests under field conditions with respect to the sampling period. The present study also confirmed the beneficial effect of T. harzianum against plant pathogens and on tomato fruit. The complex tomato-arthropod-microorganism interactions that occurred in the field are discussed to enrich our current information on the possibilities of using Trichoderma as a green alternative agent in agriculture.

The Bio-Patina on a Hypogeum Wall of the Matera-Sassi Rupestrian Church "San Pietro Barisano" before and after Treatment with Glycoalkaloids.

The investigation focused on the deterioration of the walls in the hypogeum of "San Pietro Barisano" rupestrian church, located in the Matera-Sassi (Southern Italy), one of the UNESCO World Heritage sites. The study evaluated the biocide activity of a mixture of natural glycoalkaloids (GAs) extracted from the unripe fruit of Solanum nigrum and applied to clean a hypogeum wall surface in the church affected by bio-patinas. The analyzed bio-patina, collected before treatment and, at pre-established times, after treatment, showed changes in chemical composition detected by XPS, accompanied by visible discoloration and biological activity variation. The biocidal action of the glycoalkaloids mixture, directly employed on the wall surface, was effective after about four weeks for most bio-patina colonizers but not for the fungal species that can migrate and survive in the porosities of the calcarenite. Consequently, the cleaning procedure requires the integration of fungicidal actions, combined with the consolidation of the surfaces, to obtain complete bioremediation and avoid subsequent biological recolonization. SEM images and associated microanalysis of pretreated bio-patina have revealed the biocalcogenity of some autochthonous microorganisms, thus preluding to their eventual isolation and reintroduction on the wall surface to act as consolidants once the bio-cleaning phase has been completed.

Alternative Control of Phragmidium rubi-idaei Infecting Two Rubus Species.

Organic berry plantations have been gaining popularity among farmers during recent years. Even so, farmers experience serious challenges in disease control management, which is a concern in organic farming. Phragmidium rubi-idaei (DC) P. Karst is the pathogen responsible for blackberry and raspberry rust disease, one of the most present and active diseases in plantations. The antifungal certified products found on the organic farming market offer the opportunity for an efficient control strategy over plant pathogens in fruit shrub plantations. In this study, 5 natural based products-namely Altosan, Mimox, Canelys, Zitron, and Zeolite-were tested for their fungistatic effect over P. rubi-idaei. The experiments were carried out under laboratory conditions, performing observations over the impact of organic products, used at different concentration levels, on rust conidia germination. Moreover, field experiments were conducted in order to evaluate the efficiency of different treatments for rust control on raspberry ('Polka', 'Veten' and 'Heritage') and blackberry ('Thorn Free', 'Chester' and 'Loch Ness') varieties. Data analysis based on ANOVA tests showed significant differences between the tested variants and the control sample at p < 0.001. Furthermore, LSD test confirmed differences between all substances tested (p < 0.005). The natural products Canelys (formulated with cinnamon) and Zytron (based on citrus extract) have proven the highest inhibitory capacity for conidia germination during in vitro tests registering values of 80.42% and 78.34%, respectively. The same high inhibitory rates against rust pathogen were kept also in the field tests using the same two natural-based products mentioned earlier. In addition, outcomes from this study demonstrated that Zeolite is not recommended for raspberry or blackberry rust control.

An In Vitro Attempt for Controlling Severe Phytopathogens and Human Pathogens Using Essential Oils from Mediterranean Plants of Genus Schinus.

Growing concerns about food safety and environmental protection enhanced the need for new and safe plant disease control strategies. The chemical composition of the three essential oils (EOs) extracted from leaves and fruits of Schinus terebinthifolius and leaves of Schinus molle, growing in Tunisia, was studied by GC and GC-MS. In all, 12 compounds were identified. The oils were mainly composed of terpene compounds. α-Pinene, α-phellandrene, and D-limonene were the major constituents. The aim of the current study was to evaluate the in vitro antimicrobial effectiveness of three EOs derived from plants of genus Schinus and extracted from leaves and fruits of S. terebinthifolius and leaves of S. molle. Both antifungal and antibacterial activities of the EOs were examined. The antifungal activity of the studied EOs was investigated against Colletotrichum acutatum and Botrytis cinerea in comparison with the systemic fungicide azoxystrobin used at 0.8 µL mL(-1). The antibacterial activity was evaluated against three strains of Gram-positive (G+ve) bacteria (Bacillus megaterium, Bacillus mojavensis and Clavibacter michiganensis) and four strains of Gram-negative (G-ve) bacteria (Escherichia coli, Xanthomonas campestris, Pseudomonas savastanoi, and Pseudomonas syringae pv. phaseolicola) compared with the synthetic antibiotic tetracycline at a concentration of 1600 µg mL(-1). The minimum inhibitory concentration of the studied EOs has been evaluated against the above microorganisms using the 96-well microplate method. Tested microorganisms exhibited different levels of sensitivity to each tested EO. All investigated EOs reduced the fungal mycelial growth when used at low concentrations from 250 to 1000 ppm and from 2000 to 8000 ppm against C. acutatum and B. cinerea, respectively. Higher concentrations of the same EOs exhibited a fungicidal effect against both mitosporic fungi. The EO extracted from leaves of S. terebinthifolius significantly inhibited the growth of tested bacterial strains. Nevertheless, E. coli showed a weak resistance toward the same EO and a high resistance toward the other two tested EOs. Finally, P. savastanoi and P. syringae pv. phaseolicola showed a high resistance toward all tested EOs.