Desert soil hosts many microorganisms, whose activities are essential from an ecological viewpoint. Moreover, they are of great anthropic interest. The knowledge of extreme environments microbiomes may be beneficial for agriculture, technology, and human health. In this study, 11 Arthrobacter strains from topsoil samples collected from the Great Gobi A Strictly Protected Area in the Gobi Desert, were characterized by a combination of different techniques. The phylogenetic analysis, performed using their 16S rDNA sequences and the most similar Arthrobacter sequences found in databases, revealed that most of them were close to A. crystallopoietes, while others joined a sister group to the clade formed by A. humicola, A. pascens, and A. oryzae. The resistance of each strain to different antibiotics, heavy-metals, and NaCl was also tested as well as the inhibitory potential against human pathogens (i.e., Burkholderia ssp., Klebsiella pneumoniae, Pseudomonas aeruginosa, and Staphylococcus ssp.) via cross-streaking, to check the production of metabolites with antimicrobial activity. Data obtained revealed that all strains were resistant to heavy metals and were able to strongly interfere with the growth of many of the human pathogens tested. The volatile organic compounds (VOCs) profile of the 11 Arthrobacter strains was also analyzed. A total of 16 different metabolites were found, some of which were already known for having an inhibitory action against different Gram-positive and Gram-negative bacteria. Isolate MS-3A13, producing the highest quantity of VOCs, is the most efficient against Burkholderia cepacia complex (Bcc), K. pneumoniae, and coagulase-negative Staphylococci (CoNS) strains. This work highlights the importance of understanding microbial populations' phenotypical characteristics and dynamics in extreme environments to uncover the antimicrobial potential of new species and strains.
Understanding how microbial communities survive in extreme environmental pressure is critical for interpreting ecological patterns and microbial diversity. Great Gobi A Strictly Protected Area represents an intriguing model for studying the bacterial community since it is a protected and intact wild area of the Mongolian desert. In this work, the composition of a bacterial community of the soil from four oases was characterized by extracting total DNA and sequencing through the Illumina NovaSeq platform. In addition, the soil’s chemical and physical properties were determined, and their influence on shaping the microbial communities was evaluated. The results showed a high variability of bacterial composition among oases. Moreover, combining specific chemical and physical parameters significantly shapes the bacterial community among oases. Data obtained suggested that the oases were highly variable in physiochemical parameters and bacterial communities despite the similar extreme climate conditions. Moreover, core functional microbiome were constituted by aerobic chemoheterotrophy and chemoheterotrophy, mainly contributed by the most abundant bacteria, such as Actinobacteriota, Pseudomonadota, and Firmicutes. This result supposes a metabolic flexibility for sustaining life in deserts. Furthermore, as the inhabitants of the extreme regions are likely to produce new chemical compounds, isolation of key taxa is thus encouraged.
The emergence of new multidrug-resistant (MDR) pathogens brought attention to the need for new antimicrobial compounds. One of the advances in addressing the global issue of antibiotic resistance is the discovery that microorganisms residing inside medicinal plants (i.e., the endophytes) may contribute to the production of metabolites of pharmaceutical interest. In this work, the composition of the essential oil (EO) and the cultivable bacterial communities isolated from three different species belonging to the genus Origanum, i.e., O. vulgare ssp. vulgare, O. vulgare ssp. hirtum and O. heracleoticum were analyzed, to check the possible nexus existing between them. The three species exhibited different EO aroma profiles, revealing two trans-sabinene hydrate chemotypes and a carvacrol chemotype. The composition of the bacterial endophytic cultivable microbiome isolated from flowers, leaves, and stems and the bacterial communities isolated from bulk soil were characterized at the genus, species, and/or strain level. The analyses revealed a high degree of biodiversity and a low number of strains shared between the three plant species and compartments of the same plant, suggesting the existence of one or more forces responsible for the distribution of the endophytes in the different microenvironments of the plant. Data obtained also suggested that the composition of the EO might be involved in the structuring of the microbial community and that bacteria might be able to colonize the plant since they are resistant to the antimicrobial activity exhibited by the EO itself and/or able to utilize some compounds as a carbon and energy source. Lastly, the composition of microbial communities might enhance or influence the EO composition.
The Italian-native Mediterranean brown trout (Salmo ghigii) is a seriously threatened freshwater fish, especially by anthropogenic hybridisation with the domestic strains of Atlantic origin that have been repeatedly released into the wild for angling. A PCR-restriction fragment length polymorphism (RFLP) assay of the diagnostic lactate dehydrogenase chain-1 (LDH-C1) gene sequences has been routinely applied to distinguish exotic from native brown trout lineages and detect Atlantic introgression signals in the Mediterranean wild populations. Here, we used dermal swab DNA obtained from 28 wild trout to improve laboratory procedures to genetically characterise trout samples at the LDH-C1gene through (1) a capillary electrophoresis analysis of the RFLP fragments and (2) the optimisation of a diagnostic single nucleotide polymorphism analysable through mini-sequencing approaches. The developed methods were fully consistent with those obtained through the traditional approach, but their analytical process is almost entirely automated and digitalised, thus improving result readability and accuracy in the detection of alien introgressed traces in wild Mediterranean brown trout populations.
Seed-associated microbiota are believed to play a crucial role in seed germination, seedling establishment, and plant growth and fitness stimulation, due to the vertical transmission of a core microbiota from seeds to the next generations. It might be hypothesized that medicinal and aromatic plants could use the seeds as vectors to vertically transfer beneficial endophytes, providing plants with metabolic pathways that could influence phytochemicals production. Here, we investigated the localization, the structure and the composition of the bacterial endophytic population that resides in Origanum heracleoticum L. seeds. Endocellular bacteria, surrounded by a wall, were localized close to the aleurone layer when using light and transmission electron microscopy. From surface-sterilized seeds, cultivable endophytes were isolated and characterized through RAPD analysis and 16S RNA gene sequencing, which revealed the existence of a high degree of biodiversity at the strain level and the predominance of the genus Pseudomonas. Most of the isolates grew in the presence of six selected antibiotics and were able to inhibit the growth of clinical and environmental strains that belong to the Burkholderia cepacia complex. The endophytes production of antimicrobial compounds could suggest their involvement in plant secondary metabolites production and might pave the way to endophytes exploitation in the pharmaceutical field.
Multidrug-resistant pathogens represent a serious threat to human health. The inefficacy of traditional antibiotic drugs could be surmounted through the exploitation of natural bioactive compounds of which medicinal plants are a great reservoir. The finding that bacteria living inside plant tissues, (i.e., the endophytic bacterial microbiome) can influence the synthesis of the aforementioned compounds leads to the necessity of unraveling the mechanisms involved in the determination of this symbiotic relationship. Here, we report the genome sequence of four endophytic bacterial strains isolated from the medicinal plant Origanum vulgare L. and able to antagonize the growth of opportunistic pathogens of cystic fibrosis patients. The in silico analysis revealed the presence of gene clusters involved in the production of antimicrobial compounds, such as paeninodin, paenilarvins, polymyxin, and paenicidin A. Endophytes' adaptation to the plant microenvironment was evaluated through the analysis of the presence of antibiotic resistance genes in the four genomes. The diesel fuel degrading potential was also tested. Strains grew in minimum media supplemented with diesel fuel, but no n-alkanes degradation genes were found in their genomes, suggesting that diesel fuel degradation might occur through other steps involving enzymes catalyzing the oxidation of aromatic compounds.
Strigiformes are affected by a substantial decline mainly caused by habitat loss and destruction, poaching, and trapping. Moreover, the increasing trend in bird trade and the growing interest in wild-caught rather than captive-bred birds are expected to encourage illegal trade. The biomolecular investigation represents a valuable tool to track illegal trade and to explore the genetic variability to preserving biodiversity. Microsatellite loci (STRs) are the most used markers to study genetic variability. Despite the availability of species-specific microsatellite loci in Strigiformes, a unique panel permitting the description of the genetic variability across species has not been identified yet. We tested 32 highly polymorphic microsatellite markers to evaluate the reliability of a unique microsatellite panel in different species of Strigiformes and its use for conservation and forensic purposes. We included in the study 84 individuals belonging to 28 parental groups and 11 species of Strigiformes. After screening polymorphic microsatellite loci, the description of genetic variability, and the kinship assessment, we characterized a final panel of 12 microsatellite loci able to identify individuals in 9 Strigiformes species. This STR panel might support the authorities in the forensic investigation for suspected smugglers and false parental claims; moreover, it can be useful to evaluate relatedness among individuals in captive-bred populations and to implement research projects finalized to the description of the genetic variability in wild populations.
This study aimed to characterise the gut microbiome composition of European hares (Lepus europaeus) and its potential changes after a short-term diet modification. The high sensitivity of European hare to habitat changes makes this species a good model to analyse possible alterations in gut microbiome after the introduction of additional nourishment into the diet. In total, 20 pairs were chosen for the experiments; 10 pairs formed the control group and were fed with standard fodder. The other 10 pairs represented the experimental group, whose diet was integrated with apples and carrots. The DNA from fresh faecal pellets collected after 4 days from the start of the experiment was extracted and the V3-V4 hypervariable regions were amplified and sequenced using the Illumina MiSeq® platform. The obtained amplicon sequence variants were classified into 735 bacterial genera belonging to 285 families and 36 phyla. The control and the experimental groups appeared to have a homogenous dispersion for the two taxonomic levels analysed with the most abundant phyla represented by Bacteroidetes and Firmicutes. No difference between control and experimental samples was detected, suggesting that the short-term variation in food availability did not alter the hares’ gut microbiome. Further research is needed to estimate significant time threshold.
Health technology assessment has long been considered the most comprehensive and suitable approach to support health policy decision-making for specific technologies, pharmaceuticals included, and Europe is no exception. The expected outcomes of this multidisciplinary approach – also acknowledged by the World Health Organization are recommendations bridging evidence-based medicine and economic evaluation and including the social, legal and ethical implications. The first move for coordination of health technology assessment in Europe was a project funded by the European Commission in the mid-1990s, followed by two other projects which led to the establishment in 2006 of EUnetHTA, a large network of organisations designed to back health technology assessment as a decision-making tool for the European Union and its Member States. Here we first summarise the health technology assessment experience on pharmaceuticals in the European Union in the very last years. Then, we raise an alternative proposal to that recently issued by the European Commission.
Pharmacy has been historically regarded as a discipline between health and chemistry devoted to drug development, production, and compounding. These tasks have been almost lost with the industrial manufacturing, and dispensing remains the main activity of pharmacists. Hospital pharmacists are usually employees in their workplace, while the professional framework of community pharmacists is very different, being pharmacies predominantly private shops in almost all European countries. In the last years pharmacists have strongly advocated that the focus of their services should switch from 'product' to 'patient'. Clinical pharmacy and pharmaceutical care are the two most cited concepts to support this shift. Clinical pharmacy was originally defined as the area of pharmacy concerned with the science and practice of rational medication use, pharmaceutical care as the responsible provision of drug therapies to achieve definite outcomes. The practice of clinical pharmacy should embrace the philosophy of pharmaceutical care. The new wave of pharmacists' patient-centered care in Europe still seems to be a reaction against the loss of their traditional professional role after the drug manufacturing revolution. To depict a realistic scenario for progress, it is worth differentiating between hospital and community. Hospital pharmacists should strengthen their pivotal role of medication gatekeepers to improve among clinicians the appropriateness of drug prescriptions and generate savings in expenditures. Any proposal for clinical services provided by community pharmacists is inevitably affected by the issue of their potential remuneration, especially in countries where the remuneration for reimbursable drugs is still a proportion of the retail price.
The Editorial "Defensive medicine in Europe: a 'full circle'?"
Individuals may have conflicts of interest (CoI) when they choose between the duties of their jobs and their own private interests. In medicine, CoI are potentially ubiquitous and their disclosure has now become the most frequent strategy to address them in professional lives. In the medical literature, CoI are classified into two different types-financial and non-financial. Financial CoI are easy to identify and can bias any kind of results in research; so, their disclosure is very important. The unsolvable dilemma is where to set the lowest limit for sums received from industry. Non-financial CoI are a very large category intrinsically related to the individuals concerned, ranging from family relationships to religious beliefs, and the mere disclosure of many of them can raise privacy and ethical issues. Two opposite narratives characterize the debate on financial CoI caused by pharmaceutical industry. The critical side argues that, because the primary goal of pharmaceutical industry is inevitably to promote its products, the best strategy is to stay away from financial CoI. On the other hand, the defensive side claims that financial CoI are boosted by ideology but meaningless in real practice, since any kind of interest can raise a potential conflict. A missing point in the debate on financial CoI is that health care is a classical example of 'market failure' in the economic theory. Since health cannot be considered a 'consumer good', the economic paradigm of 'free market' does not fit for healthcare products. To conclude, even though transparency on financial CoI cannot itself deter the risk of bias, rejecting it would be an even bigger mistake. At variance, mandatory disclosure of non-financial CoI risks to be confusing and questionable in many cases, paradoxically distracting attention from the potential bias created by financial CoI.
Defensive medicine (DM) refers to all medical care provided by physicians mainly for preventing the risk of litigation. Redundant DM practices induced by the threat of medical liability are expected to increase total healthcare expenditures.
Following the letter by Colombo and Parisi [ [1] Colombo A. Parisi G. Seeking identity in primary care. A survey on GPs trainees in Lombardy. Eur J Intern Med. 2019; ([in press]) Abstract Full Text Full Text PDF PubMed Scopus (6) Google Scholar ], first of all we thank the authors for their positive comments on our earlier editorial [ [2] Garattini L. Padula A. English and Italian national health services: time for more patient-centered primary care?. Eur J Intern Med. 2018; 57: 19-21 Abstract Full Text Full Text PDF PubMed Scopus (9) Google Scholar ]. Actually, their letter adds interesting information from Lombardy (by far the largest Italian Region), collected through a survey on trainers and trainees in general practice. The results highlighted a major difference in the educational aims between tutors and their younger colleagues, the former being more focussed on the importance of developing relational skills with patients, and the latter on their clinical and technical skills (e.g. information technology). Although these results may partly reflect a difference in the generational approach to labour, as happens in the majority of fields, they do confirm the urgent need to introduce a national academic specialty for general practice to plan at best the education of future Italian General Practitioners (GPs), instead of the present regional courses open to widely differing approaches. This change should also help cancel out the arguable difference of pay between training specialists and GPs reported in the letter, which certainly penalizes the latter.