Chemical Composition, Antibacterial, Antibiofilm and Synergistic Properties of Essential Oils from Eucalyptus globulus Labill. and Seven Mediterranean Aromatic Plants.

Essential oils (EOs) from Eucalyptus globulus Labill. ssp. globulus and from Mediterranean autochthonous aromatic plants - Thymus mastichina L., Mentha pulegium L., Rosmarinus officinalis L., Calamintha nepeta (L.) Savi ssp. nepeta, Cistus ladanifer L., Foeniculum vulgare L., Dittrichia viscosa (L.) Greuter ssp. viscosa - were extracted by hydrodistillation and characterized by GC-FID and NMR spectroscopy. EOs were evaluated for antimicrobial properties against several bacterial strains, using diverse methods, namely, the agar disc-diffusion method, the microdilution method, the crystal violet assay and the Live/Dead staining for assessment of biofilm formation. Potential synergy was assessed by a checkerboard method. EOs of R. officinalis and C. ladanifer showed a predominance in monoterpene hydrocarbons (> 60%); EOs of C. nepeta, M. pulegium, T. mastichina, E. globulus and F. vulgare were rich in oxygenated monoterpenes (62 - 96%) whereas EO of D. viscosa was mainly composed of oxygenated sesquiterpenes (54%). All EOs showed antimicrobial activity; M. pulegium and E. globulus generally had the strongest antimicrobial activity. EO of C. nepeta was the most promising in hampering the biofilm formation. The combinations D. viscosa/C. nepeta and E. globulus/T. mastichina were synergistic against Staphylococcus aureus. These results support the notion that EOs from the aromatic plants herein reported should be further explored as potential pharmaceuticals and/or food preservatives.

Follow-up of Patients with Hereditary Metabolic Diseases during the 3 Years of the Pandemic in the Reference Center for Hereditary Metabolism Diseases of the Centro Hospitalar Universitário Lisboa Norte.

INTRODUCTION: The adult unit of the Reference Center for Hereditary Metabolism Diseases of the Centro Hospitalar Universitário Lisboa Norte was created in 2002. The team that supports this unit is made up of several specialists, including 2 Internal Medicine Doctors and 2 Nutritionists. The unit carries out its activity mainly in an outpatient clinic on Fridays. Between March 2020 and December 2022, the team adapted the support given to this unit, transforming some external appointments into non-face-to-face appointments in order to maintain contact with patients and reduce the deslocation to the CHULN. METHODS: The analysis reported below reflects the activity in face-to-face and non-face-to-face appointment of the medical team and nutrition team in the critical period. RESULTS/CASE REPORT: A total of 220 patients were evaluated, 56.4% female and 43.6% male. Regarding the evaluated pathologies, these were divided into 3 groups: aminoacidopathies - 40%, organic aciduria - 6.5% and other metabolic diseases - 53.5%. During these three years a total of 580 nutrition appointments were scheduled, 457 face-to-face and 123 non-face-to-face. 32% of patients scheduled did not attend the appointments or it was not possible to make telephone contact. This percentage is divided into 90% face-to-face appointments and 10% nonface- to-face appointments. Regarding medical appointments, the total was 797, 667 of which were face-to-face and 130 non-face-to-face. 16.8% of absences were registered in face-to-face and non-face-to-face appointments. 98.5% of absences concern non-face-to-face appointments. CONCLUSION: Despite the troubled period worldwide, support for these patients was always ensured, both by the Medical team and the Nutrition team. Despite the solutions found to keep patients safe, in this specific period, the % of absences from the outpatient clinic is quite high, and one of the main justifications presented by the patients is related to the fact that they are afraid to go to the hospital. With regard to absences recorded in non-face-to-face appointments, there were several situations in which patients reported not being able to answer the phone during their working hours.

Anthracological study of a Chalcolithic funerary deposit from Perdigões (Alentejo, Portugal): A new analytical methodology to establish the wood burning temperature.

Anthracological analyses of charcoal samples retrieved from Pit 16 of Perdigões (Reguengos de Monsaraz, Portugal), a secondary deposition of cremated human remains dated back to the middle of the 3rd millennium BC, enabled the identification of 7 different taxa: Olea europaea, Quercus spp. (evergreen), Pinus pinaster, Fraxinus cf. angustifolia, Arbutus unedo, Cistus sp. and Fabaceae. All taxa are characteristic of both deciduous and evergreen Mediterranean vegetation, and this data might indicate that the gathering of woods employed for the human cremation/s occurred either on site, or in its vicinity. However, considering both the large distribution of the identified taxa and data about human mobility, it is not possible to conclusively determine the origin of the wood used in the cremation(s). Chemometric analysis were carried out to estimate the absolute burning temperature of woods employed for the human cremation/s. An in-lab charcoal reference collection was created by burning sound wood samples of the three main taxa identified from Pit 16, Olea europaea var. sylvestris, Quercus suber (evergreen type) and Pinus pinaster, at temperatures between 350 and 600 °C. The archaeological charcoal samples and the charcoal reference collection were chemically characterized by using mid-infrared (MIR) spectroscopy in the 1800-400 cm-1 range, and Partial Least Squares (PLS) regression method was used to build calibration models to predict the absolute combustion temperature of the archaeological woods. Results showed successful PLS forecasting of burn temperature for each taxon (significant (P <0.05) cross validation coefficients). The anthracological and chemometric analysis evidenced differences between the taxa coming from the two stratigraphic units within the Pit, SUs 72 and 74, suggesting that they may come from two different pyres or two different depositional moments.

Genetic diversity and population structure of Cynara cardunculus L. in southern Portugal.

Cynara cardunculus L. is a cardoon species native to the Mediterranean region, which is composed of three botanical taxa, each having distinct biological characteristics. The aim of this study was to examine wild populations of C. cardunculus established in Portugal, in order to determine their genetic diversity, geographic distribution, and population structure. Based on SSR markers, 121 individuals of C. cardunculus from 17 wild populations of the Portuguese Alentejo region were identified and analysed. Ten SSRs were found to be efficient markers in the genetic diversity analysis. The total number of alleles ranged from 9 to 17 per locus. The expected and observed means in heterozygosity, by population analysed, were 0.591 and 0.577, respectively. The wild population exhibited a high level of genetic diversity at the species level. The highest proportion of genetic variation was identified within a geographic group, while variation was lower among groups. Geographic areas having highest genetic diversity were identified in Alvito, Herdade da Abóboda, Herdade da Revilheira and Herdade de São Romão populations. Moreover, significant genetic differentiation existed between wild populations from North-Alentejo geographic locations (Arraiolos, Évora, Monte da Chaminé) and Centro Hortofrutícola, compared with other populations. This study reports genetic diversity among a representative number of wild populations and genotypes of C. cardunculus from Portugal. These results will provide valuable information towards future management of C. cardunculus germplasm.

Mediterranean Native Leguminous Plants: A Reservoir of Endophytic Bacteria with Potential to Enhance Chickpea Growth under Stress Conditions.

Bacterial endophytes, a subset of a plant's microbiota, can facilitate plant growth by a number of different mechanisms. The aims of this study were to assess the diversity and functionality of endophytic bacterial strains from internal root tissues of native legume species grown in two distinct sites in South of Portugal and to evaluate their ability to promote plant growth. Here, 122 endophytic bacterial isolates were obtained from 12 different native legume species. Most of these bacteria possess at least one of the plant growth-promoting features tested in vitro, with indole acetic acid production being the most common feature among the isolates followed by the production of siderophores and inorganic phosphate solubilization. The results of in planta experiments revealed that co-inoculation of chickpea plants with specific endophytic bacteria along with N2-fixing symbionts significantly improved the total biomass of chickpea plants, in particular when these plants were grown under saline conditions. Altogether, this study revealed that Mediterranean native legume species are a reservoir of plant growth-promoting bacteria, that are also tolerant to salinity and to toxic levels of Mn. Thus, these bacterial endophytes are well adapted to common constraints present in soils of this region which constitutes important factors to consider in the development of bacterial inoculants for stressful conditions in the Mediterranean region.

Dispersal mode and spatial extent influence distance-decay patterns in pond metacommunities.

Assuming that dispersal modes or abilities can explain the different responses of organisms to geographic or environmental distances, the distance-decay relationship is a useful tool to evaluate the relative role of local environmental structuring versus regional control in community composition. Based on continuing the current theoretical framework on metacommunity dynamics and based on the predictive effect of distance on community similarity, we proposed a new framework that includes the effect of spatial extent. In addition, we tested the validity of our proposal by studying the community similarity among three biotic groups with different dispersal modes (macrofaunal active and passive dispersers and plants) from two pond networks, where one network had a small spatial extent, and the other network had an extent that was 4 times larger. Both pond networks have similar environmental variability. Overall, we found that environmental distance had larger effects than geographical distances in both pond networks. Moreover, our results suggested that species sorting is the main type of metacommunity dynamics shaping all biotic groups when the spatial extent is larger. In contrast, when the spatial extent is smaller, the observed distance-decay patterns suggested that different biotic groups were mainly governed by different metacommunity dynamics. While the distance-decay patterns of active dispersers better fit the trend that was expected when mass effects govern a metacommunity, passive dispersers showed a pattern that was expected when species sorting prevails. Finally, in the case of plants, it is difficult to associate their distance-decay patterns with one type of metacommunity dynamics.

Field trial evaluating changes in prevalence and patterns of antimicrobial resistance among Escherichia coli and Enterococcus spp. isolated from growing broilers medicated with enrofloxacin, apramycin and amoxicillin.

The present study investigates, under field conditions, the influence of antimicrobial administration on prevalence and patterns of antimicrobial resistance among Escherichia coli and Enterococcus spp. isolated from growing broilers. For this purpose, a group of 16,000 commercial broiler chickens was treated with enrofloxacin from day 1 to day 3, gentamicin from day 19 to day 21, and ampicillin from day 26 to day 28. A control group of 16,000 broilers was placed in the same controlled environment poultry house. Fecal (from both groups) and feed samples were collected at regular intervals. Few E. coli isolates were obtained from either farm environment or poultry feed samples, while enterococci were found to be ubiquitous among these samples. The frequency of resistance against most antimicrobials tested was significantly higher (P<0.05) in E. coli isolated from broilers receiving intermittent antimicrobial pressure than that from non-medicated broilers, whereas in enterococci these differences were only observed among structurally related antimicrobial drugs and over a short period of time. By the time the broilers reached market age (33 days), several multi-resistant E. coli and enterococci were detected in the feces of the medicated group. Results suggest that antimicrobial resistance in E. coli was mainly medication-dependent, whereas among enterococci, changes observed over time were apparently influenced by factors apart from antimicrobial exposure, namely the resistance organisms previously present in farm environment and those present in feedstuffs.