Soil Microbial Communities in Lemon Orchards Affected by Citrus Mal Secco Disease.

Mal secco is a vascular disease of citrus caused by the mitosporic fungus Plenodomus tracheiphilus. Soil containing infected plant material constitutes an inoculum source for root infections. In this study, the soil bacterial and fungal communities of five lemon orchards located in Syracuse Province (Sicily, Italy) affected by mal secco were analyzed. Soil samples were collected under lemon tree canopies and subjected to total genomic DNA extraction. The fungal DNA was detected through qPCR in all orchards, with variable concentrations. Bacterial and fungal communities were profiled using 16S and ITS amplicon-based high-throughput sequencing, respectively. According to our results, the relative abundances of the most represented bacterial phyla (e.g., Proteobacteria, Actinobacteriota, Acidobacteriota) changed across the orchards, while in the fungal community, the phylum Ascomycota was dominant, with Basidiomycota and Mortierellomycota abundances fluctuating. On the whole, ß diversity analysis showed significant variation in the composition of the soil microbial communities across the orchards. This result was confirmed by the analysis of the core community (taxa present at ≥ 75% of total samples), where putative beneficial bacteria resulted in significantly enriched fungus-infected soil samples, suggesting complex microbial interactions. Our findings shed light on the composition and diversity of the soil microbiome in lemon orchards with the occurrence of mal secco infections.

Biocontrol efficacy of Pseudomonas mediterranea PVCT 3C against Plenodomus tracheiphilus: In vitro and in planta mechanisms at early disease stages.

In this study, we investigated the biocontrol activity of the P. mediterranea strain PVCT 3C against Mal secco, a severe disease of citrus caused by the vascular fungus Plenodomus tracheiphilus. In vitro, bacterial diffusible compounds, volatile organic compounds and culture filtrates produced by PVCT 3C reduced the mycelial growth and conidial germination of P. tracheiphilus, also affecting the mycelial pigmentation. The application of bacterial suspensions by leaf-spraying before the inoculation with the pathogen on plants of the highly susceptible species sour orange and lemon led to an overall reduction in incidence and disease index, above all during the early disease stage. PVCT 3C genome was subjected to whole-genome shotgun sequencing to study the molecular mechanisms of action of this strain. In silico annotation of biosynthetic gene clusters for secondary metabolites revealed the presence of numerous clusters encoding antimicrobial compounds (e.g. cyclic lipopeptides, hydrogen cyanide, siderophores) and candidate novel products. During the asymptomatic disease phase (seven days post-inoculation), bacterial treatments interfered with the expression of different fungal genes, as assessed with an NGS and de novo assembly RNA-seq approach. These results suggest that P. mediterranea PVCT 3C or its secondary metabolites may offer a potential effective and sustainable alternative to contain P. tracheiphilus infections via integrated management.

How the Electrical Conductivity of Water Fluids Affects Micro-EDM in the Short-Pulse Regime.

This work investigates micro-electro discharge machining (EDM) performance involving deionized and tap water. The chosen machining regime was semi-finishing, where open voltage (from 100 to 130 V) and current values (5-10 A) were applied using a 0.5 µs pulse-on time and a frequency of 150 kHz, i.e., a duty cycle of 25%. First, numerical analyses were performed via COMSOL Multiphysics and used to estimate the plasma channel distribution and melted material, varying the current, sparking gap, electrical conductivity, and permittivity of the two fluids. Then, experimentally, the micro-EDM of holes and channels in hardened thin steel plates were replicated three times for each considered fluid. The material removal rate (MRR), tool wear ratio (TWR), radius overcut, and surface roughness were plotted as a function of open voltage and electrical conductivity. The study proves that as voltage and current increase, the MRR and TWR decrease with electrical conductivity. Nonetheless, for higher electrical conductivity (tap water), the process did not proceed for lower open voltages and currents, and the radius overcut was reduced, contrary to what is commonly acknowledged. Finally, the crater morphology and size were evaluated using a confocal microscope and compared to simulated outcomes.

Replication Study of Molded Micro-Textured Samples Made of Ultra-High Molecular Weight Polyethylene for Medical Applications.

In articular joint implants, polymeric inserts are usually exploited for on-contact sliding surfaces to guarantee low friction and wear, a high load-bearing capacity, impact strength and stiffness, and biocompatibility. Surface micro-structuring can drastically reduce friction and wear by promoting hydrostatic friction due to synovial fluid. Ultra-High Molecular Weight Polyethylene (UHMWPE) is a suitable material for these applications due to its strong chemical resistance, excellent resistance to stress, cracking, abrasion, and wear, and self-lubricating property. However, surface micro-texturing of UHMWPE is hardly achievable with the currently available processes. The present study investigates UHMWPE's micro-textured surface replication capability via injection molding, comparing the results with the more easily processable High-Density Polyethylene (HDPE). Four different micro-texture cavities were designed and fabricated on a steel mold by micro-EDM milling, and used for the experimental campaign. Complete samples were fabricated with both materials. Then, the mold and samples were geometrically characterized, considering the dimensions of the features and the texture layout. The replication analysis showed that HDPE samples present geometrical errors that span from 1% to 9% resulting in an average error of 4.3%. In comparison, the UHMWPE samples display a higher variability, although still acceptable, with percentage errors ranging from 2% to 31% and an average error of 11.4%.

Editorial for the Special Issue on Micro-Electro Discharge Machining: Principles, Recent Advancements and Applications, Volume II.

The second volume of the Special Issue on "Micro-Electro Discharge Machining: Principles, Recent Advancements and Applications" confirms the growing interest in the micro-EDM technology as a suitable and efficient technology for machining novel, multi-material components, with demanding requirements in terms of precision, accuracy and productivity [...].

Micro Injection Molding of Thin Cavities Using Stereolithography for Mold Fabrication.

At the present time, there is a growing interest in additive manufacturing (AM) technologies and their integration into current process chains. In particular, the implementation of AM for tool production in micro injection molding (µ-IM), a well-established process, could introduce many advantages. First of all, AM could avoid the need for the time-consuming and expensive fabrication of molds for small series of customized products. In this work, the feasibility, quality, and reliability of an AM/µ-IM process chain were evaluated by designing and fabricating mold inserts for µ-IM by stereolithography (SLA) technology; the mold inserts were characterized and tested experimentally. The selected geometry is composed of four thin cavities: This particular feature represents an actual challenge for both the SLA and µ-IM perspective due to the large surface-to-volume ratio of the cavity. Two different materials were used for the mold fabrication, showing sharply different performance in terms of endurance limit and cavity degradation. The obtained results confirm that the µ-IM process, exploiting an SLA fabricated mold insert, is feasible but requires great accuracy in material choice, mold design, fabrication, and assembly.

Editorial for the Special Issue on Micro-Electro Discharge Machining: Principles, Recent Advancements and Applications.

Micro Electrical Discharge Machining (micro-EDM) is a thermo-electric and contactless process most suited for micro-manufacturing and high-precision machining, especially when difficult-to-cut materials, such as super alloys, composites, and electro conductive ceramics, are processed [...].

Pulse-Type Influence on the Micro-EDM Milling Machinability of Si3N4-TiN Workpieces.

In this paper, the effect of the micro-electro discharge machining (EDM) milling machinability of Si3N4-TiN workpieces was investigated. The material removal rate (MRR) and tool wear rate (TWR) were analyzed in relation to discharge pulse types in order to evaluate how the different pulse shapes impact on such micro-EDM performance indicators. Voltage and current pulse waveforms were acquired during micro-EDM trials, scheduled according to a Design of Experiment (DOE); then, a pulse discrimination algorithm was used to post-process the data off-line and discriminate the pulse types as short, arc, delayed, or normal. The analysis showed that, for the considered process parameter combinations, MRR was sensitive only to normal pulses, while the other pulse types had no remarkable effect on it. On the contrary, TWR was affected by normal pulses, but the occurrence of arcs and delayed pulses induced unexpected improvements in tool wear. Those results suggest that micro-EDM manufacturing of Si3N4-TiN workpiece is relevantly different from the micro-EDM process performed on metal workpieces such as steel. Additionally, the inspection of the Si3N4-TiN micro-EDM surface, performed by SEM and EDS analyses, showed the presence of re-solidified droplets and micro-cracks, which modified the chemical composition and the consequent surface quality of the machined micro-features.

Disposable Optical Stretcher Fabricated by Microinjection Moulding.

Microinjection moulding combined with the use of removable inserts is one of the most promising manufacturing processes for microfluidic devices, such as lab-on-chip, that have the potential to revolutionize the healthcare and diagnosis systems. In this work, we have designed, fabricated and tested a compact and disposable plastic optical stretcher. To produce the mould inserts, two micro manufacturing technologies have been used. Micro electro discharge machining (µEDM) was used to reproduce the inverse of the capillary tube connection characterized by elevated aspect ratio. The high accuracy of femtosecond laser micromachining (FLM) was exploited to manufacture the insert with perfectly aligned microfluidic channels and fibre slots, facilitating the final composition of the optical manipulation device. The optical stretcher operation was tested using microbeads and red blood cells solutions. The prototype presented in this work demonstrates the feasibility of this approach, which should guarantee real mass production of ready-to-use lab-on-chip devices.