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The standard laboratory methods used to detect Salmonella are time-consuming, leading to unavoidable delays, which can be costly (e.g., food checks at customs) or even dangerous when quick alarms are needed (e.g., for potential outbreaks). To address this problem, we propose a rapid two-step colorimetric immunoassay. In the first step, gold nanoparticles interact with the sample, while in the second step, an optimized amount of saline solution is added to induce cluster formation among the unbound nanoparticles, providing a color change that inversely correlates with the bacteria concentration. The nanoparticles were functionalized using a technique based on UV activation of the antibodies, resulting in oriented antibodies with antigen-binding regions in the proximity of the surface. While a color change can be observed with the naked eye at relatively higher bacterial concentrations, a more quantitative approach based on UV–visible spectrum measurements allowed us to achieve a limit of detection of 1 cfu/mL in chicken samples. The test can be completed in 15 min and performed on-site, paving the way for widespread applications of our biosensor in the field of food safety. [ABSTRACT FROM AUTHOR]

Simple Summary: In dairy buffalo farms, the hygiene of the milking area represents a key task of internal biosecurity measures since it can directly influence the dairy animals' health and the microbiological quality of raw milk. The aim of this study was to evaluate the cleanliness and the bacterial contamination of milking parlours' surfaces and equipment of eight buffalo farms located in the Campania Region using an ATP-bioluminescence assay and bacteriological analysis during the year 2022. The findings of this study underline that an ATP-bioluminescence assay is a valid, complementary, cost-effective on-farm tool useful to quickly detect the contamination of milking parlours. Careful cleaning of a milking parlour and its equipment is fundamental to guarantee good raw milk quality and prevent the dissemination of bacteria and improve animal welfare. This study aimed to investigate, using an ATP-bioluminescence assay and bacteriological analysis, the bacterial contamination of milking parlours on milking parlour surfaces of buffalo farms in the Campania Region, evaluating the seasonal dynamics during the year 2022. Eight farms were selected by the Italian ClassyFarm system, which assesses the level of animal welfare and biosecurity according to risk analysis. Before sampling, all dairy farm owners filled out a questionnaire on milking management, animal hygiene, and health. The questionnaires evidenced similar cleaning procedures but an absence of a standardised cleaning protocol among the different farms. ATP bioluminescence results evidenced similar levels of contamination in all the selected buffalo farms, and the season comparison showed no significant differences. A variation in the percentages of bacterial isolates during the different seasons was observed, with a higher prevalence of Enterobacteriaceae (38%) in summer. A small number of samples exhibited an absence of bacterial growth. Identifying bacteria is crucial for understanding the microorganisms present in the milking parlour, yet employing ATP luminometry could offer broad and accurate applications in buffalo milking parlours. In conclusion, the use of ATP bioluminescence for evaluating the hygiene of a buffalo milking parlour could represent a further important advancement in dairy farming technology. [ABSTRACT FROM AUTHOR]

The challenges to which plants are exposed in urban environments represent, in miniature, the challenges plants face as a result of global environmental change. Hence, urban habitats provide a unique opportunity to assess whether processes of local adaptation are taking place despite the short temporal and geographical scales that characterize the Anthropocene.We quantified the ecological diversity of urban habitats hosting Arabidopsis thaliana populations. Using plant community indicators, we show that these patches differ in their levels of soil nutrient content and disturbance. Accordingly, plants in each patch displayed a range of flowering time, size and fitness.Using a deep sampling approach coupled with reduced genome sequencing, we demonstrate that most individuals can be assigned to a limited set of clonal lineages; the genetic diversity of these lineages is a sample of the diversity observed in western European populations of the species, indicating that established urban populations originate from a broad regional pool of lineages.We assessed the genetic and phenotypic diversity of these lineages in a set of common garden experiments. We report marked genetic differences in life‐history traits, including time of primary and secondary dormancy as well as of flowering. These genetic differences in life‐history traits are not randomly distributed but sorted out by ecological differences among sites of origin.Synthesis. Our study shows that the genetically diverse phenology of a regional Arabidopsis thaliana gene pool is not randomly distributed but filtered by heterogeneity in the urban environment. To our knowledge, this report is the first to show a pattern indicative of environmental filtering enhancing local genetic adaptation within urban environments. We conclude that environmental filtering helps maintain functional diversity within species. [ABSTRACT FROM AUTHOR]

Abstract Background Antimicrobial peptides (AMPs) are an ancient group of defense molecules. AMPs are widely distributed in nature (being present in mammals, birds, amphibians, insects, plants, and microorganisms). They display bactericidal as well as immunomodulatory properties. The aim of this study was to investigate the antimicrobial and anti-inflammatory activities of a combination of two AMPs (temporin B and the royal jellein I) against Staphylococcus epidermidis. Results The temporin B (TB-KK) and the royal jelleins I, II, III chemically modified at the C terminal (RJI-C, RJII-C, RJIII-C), were tested for their activity against 10 different Staphylococcus epidermidis strains, alone and in combination. Of the three royal jelleins, RJI-C showed the highest activity. Moreover, the combination of RJI-C and TB-KK (MIX) displayed synergistic activity. In vitro, the MIX displayed low hemolytic activity, no NO2- production and the ability to curb the synthesis of the pro-inflammatory cytokines TNF-α and IFN-γ to the same extent as acetylsalicylic acid. In vivo, the MIX sterilized mice infected with Staphylococcus epidermidis in eleven days and inhibited the expression of genes encoding the prostaglandin-endoperoxide synthase 2 (COX-2) and CD64, two important parameters of inflammation. Conclusion The study shows that the MIX – a combination of two naturally occurring peptides - displays both antimicrobial and anti-inflammatory activities.

Genomic variation in the model plant Arabidopsis thaliana has been extensively used to understand evolutionary processes in natural populations, mainly focusing on single-nucleotide polymorphisms. Conversely, structural variation has been largely ignored in spite of its potential to dramatically affect phenotype. Here, we identify 155,440 indels and structural variants ranging in size from 1 bp to 10 kb, including presence/absence variants (PAVs), inversions, and tandem duplications in 1,301 A. thaliana natural accessions from Morocco, Madeira, Europe, Asia, and North America. We show evidence for strong purifying selection on PAVs in genes, in particular for housekeeping genes and homeobox genes, and we find that PAVs are concentrated in defense-related genes (R-genes, secondary metabolites) and F-box genes. This implies the presence of a "core" genome underlying basic cellular processes and a "flexible" genome that includes genes that may be important in spatially or temporally varying selection. Further, we find an excess of intermediate frequency PAVs in defense response genes in nearly all populations studied, consistent with a history of balancing selection on this class of genes. Finally, we find that PAVs in genes involved in the cold requirement for flowering (vernalization) and drought response are strongly associated with temperature at the sites of origin. [ABSTRACT FROM AUTHOR]

Introduction: Pidotimod, a synthetic dipeptide molecule with biological and immunological activities, is used to reduce the number of exacerbations or pneumonitis in patients with inflammatory diseases. In the present study, we investigated whether Pidotimod modifies the metabolomic pathways measured in the exhaled breath condensate (EBC) of non-cystic fibrosis bronchiectatic patients (NCFB). Materials and Methods: We analyzed 40 adult patients affected by NCFB. They were randomly selected to receive Pidotimod 800 mg b/d for 21 consecutive days (3 weeks) per month for 6 months (20 patients, V1 group) or no drug (20 patients, V0 group), with a 1:1 criterion and then followed as outpatients. Results: EBC samples were collected from all patients at baseline and after 6 months. They were investigated by combined nuclear magnetic resonance (NMR) spectroscopy and multivariate statistical analysis to uncover metabolic differences between EBC from NCFB patients before and after therapy with Pidotimod. Pulmonary function test and pulmonary exacerbations were analyzed at baseline and at the end of Pidotimod therapy. The EBC metabolites were all identified, and through statistical evaluation, we were able to discriminate the two samples' classes, with acetate, acetoin, lactate, and citrate as statistically significant discriminatory metabolites. The model vas validated by using a blind set of 20 NCFB samples, not included in the primary analysis. No differences were observed in PFT after 6 months. At the end of the study, there was a significant decrease of exacerbation rate in V1 group as compared with V0 group, with a substantial reduction of the number of mild or severe exacerbations (p < 0.001). Discussion: Pidotimod modifies the respiratory metabolic phenotype ("metabotype") of NCFB patients and reduces the number of exacerbations. [ABSTRACT FROM AUTHOR]