Emerging Target-Directed Approaches for the Treatment and Diagnosis of Microbial Infections.

With the rising levels of drug resistance, developing efficient antimicrobial therapies has become a priority. A promising strategy is the conjugation of antibiotics with relevant moieties that can potentiate their activity by target-directing. The conjugation of siderophores with antibiotics allows them to act as Trojan horses by hijacking the microorganisms' highly developed iron transport systems and using them to carry the antibiotic into the cell. Through the analysis of relevant examples of the past decade, this Perspective aims to reveal the potential of siderophore-antibiotic Trojan horses for the treatment of infections and the role of siderophores in diagnostic techniques. Other conjugated molecules will be the subject of discussion, namely those involving vitamin B12, carbohydrates, and amino acids, as well as conjugated compounds targeting protein degradation and ß-lactamase activated prodrugs.

Tryptophan derived natural marine alkaloids and synthetic derivatives as promising antimicrobial agents.

Antimicrobial resistance has become a major threat to public health worldwide, as pathogenic microorganisms are finding ways to evade all known antimicrobials. Therefore, the demand for new and effective antimicrobial agents is also increasing. Natural products have always played an important role in drug discovery, either by themselves or as inspiration for synthetic compounds. The marine environment is a rich source of bioactive metabolites, and among them, tryptophan-derived alkaloids stand out for their abundance and by displaying a variety of biological activities, with antimicrobial properties being among the most significant. This review aims to reveal the potential of marine alkaloids derived from tryptophan as antimicrobial agents. Relevant examples of these compounds and their synthetic analogues reported in the last decades are presented and discussed in detail, with their mechanism of action and synthetic approaches whenever relevant. Several tryptophan-derived marine alkaloids have shown potent and promising antimicrobial activities, whether against bacteria, fungi, or virus. Synthetic approaches to many of the compounds have been developed and recent methodologies are proving to be efficient. Even though most of the studies regarding the antimicrobial activity are still preliminary, this class of compounds has proven to be worth of further investigation and may provide useful lead compounds for the development of antimicrobial agents. Overall, marine alkaloids derived from tryptophan are revealed as a valuable class of antimicrobials and molecular modifications in order to reduce the toxicity of these compounds and additional studies regarding their mechanism of action are interesting topics to explore in the future.

Veterinary Expert Opinion on Potential Drivers and Opportunities for Changing Antimicrobial Usage Practices in Livestock in Denmark, Portugal, and Switzerland.

Reducing antimicrobial use (AMU) in livestock is requested by Public Health authorities. Ideally, this should be achieved without jeopardizing production output or animal health and welfare. Thus, efficient measures must be identified and developed to target drivers of AMU. Veterinarians play a central role in the identification and implementation of such efficient interventions. Sixty-seven veterinarians with expertise in livestock production in Denmark, Portugal, and Switzerland participated in an expert opinion study aimed at investigating experiences and opinions of veterinarians about the driving forces and practices related to AMU in the main livestock sectors (broiler, dairy cattle, fattening/veal calf, and pig industry) of the aforementioned countries. Opinions on potential factors influencing the choice of antimicrobials and opportunities to reduce AMU were collected. Antibiograms are seldom used, mainly due to the time lag between testing and obtaining the results. The perceived percentage of treatment failures varied between countries and livestock sectors; however, little changes were reported over time (2005-2015). The animal health problems of each livestock sector most frequently leading to AMU did not vary substantially between countries. Mandatory official interventions (i.e., binding measures applied by national or international authorities) were highlighted as having the biggest impact on AMU. There was a variation in the experts' opinion regarding feasibility and impact of interventions both between countries and livestock sectors. Nevertheless, improved biosecurity and education of veterinarians frequently received high scores. Most veterinarians believed that AMU can be reduced. The median potential reduction estimates varied from 1% in Swiss broilers to 50% in Portuguese broilers and veal/fattening calves in all countries. We hypothesize that the differences in views could be related to disease epidemiology, animal husbandry, and socio-economic factors. A profound investigation of these disparities would provide the required knowledge for developing targeted strategies to tackle AMU and consequently resistance development. However, experts also agreed that mandatory official interventions could have the greatest impact on antimicrobial consumption. Furthermore, improvement of biosecurity and education of veterinarians, the use of zinc oxide (in pigs), improving vaccination strategies, and the creation of treatment plans were the measures considered to have the largest potential to reduce AMU. This paper can inform policymakers in Europe and countries with a similar animal production regarding their AMU policy.

Exposure assessment of extended-spectrum beta-lactamases/AmpC beta-lactamases-producing Escherichia coli in meat in Denmark.

INTRODUCTION: Extended-spectrum beta-lactamases (ESBL) and AmpC beta-lactamases (AmpC) are of concern for veterinary and public health because of their ability to cause treatment failure due to antimicrobial resistance in Enterobacteriaceae. The main objective was to assess the relative contribution (RC) of different types of meat to the exposure of consumers to ESBL/AmpC and their potential importance for human infections in Denmark. MATERIAL AND METHODS: The prevalence of each genotype of ESBL/AmpC-producing E. coli in imported and nationally produced broiler meat, pork and beef was weighted by the meat consumption patterns. Data originated from the Danish surveillance program for antibiotic use and antibiotic resistance (DANMAP) from 2009 to 2011. DANMAP also provided data about human ESBL/AmpC cases in 2011, which were used to assess a possible genotype overlap. Uncertainty about the occurrence of ESBL/AmpC-producing E. coli in meat was assessed by inspecting beta distributions given the available data of the genotypes in each type of meat. RESULTS AND DISCUSSION: Broiler meat represented the largest part (83.8%) of the estimated ESBL/AmpC-contaminated pool of meat compared to pork (12.5%) and beef (3.7%). CMY-2 was the genotype with the highest RC to human exposure (58.3%). However, this genotype is rarely found in human infections in Denmark. CONCLUSION: The overlap between ESBL/AmpC genotypes in meat and human E. coli infections was limited. This suggests that meat might constitute a less important source of ESBL/AmpC exposure to humans in Denmark than previously thought - maybe because the use of cephalosporins is restricted in cattle and banned in poultry and pigs. Nonetheless, more detailed surveillance data are required to determine the contribution of meat compared to other sources, such as travelling, pets, water resources, community and hospitals in the pursuit of a full source attribution model.

Antibiotic resistance of Enterococcus spp. isolated from wastewater and sludge of poultry slaughterhouses.

Antibiotimicrobial resistance was investigated in 537 Enterococcus spp. isolates recovered from 22 samples of crude inflow, treated effluent and sludge collected in wastewater treatment plants of eight poultry slaughterhouses of Portugal. No significant differences (P > 0.05) were found in the resistance to each antimicrobial agent with regards to the origin of the sample (inflow, sludge and effluent). Many of the isolates displayed resistance to tetracycline (85.7%), erythromycin (45.7%), nitrofurantoin (34.0%) and rifampicin (17.8%). Resistance was also observed, but to a lesser extent, to ciprofloxacin (10.2%), ampicillin (8.0%), chloramphenicol (4.6%), vancomycin (0.9%) and gentamicin (0.4%). Resistance to three or more antimicrobial classes was present in 37.1% of the isolates. Wastewater treatment resulted in viable enterococci decrease between less than 1 log and 4 log; nevertheless, more than 4.4 x 10(5) colony forming units (CFU)/100 mL were present in the outflow of the plants and thus resistant enterococci are not prevented from reaching the general environment.

The role of a basic surgical skills laboratory as viewed by medical students (6th year).

In the last academic year, a training period in a surgical skills laboratory, using plastic models, was included as a part of the 'Cirurgia 3' ('Surgery 3', 6th year) programme of Faculdade Medicina Lisboa. To evaluate the training period, a structured, anonymous questionnaire was administered at the end, seeking students' responses on their satisfaction in attending the course. A global view of the answers points out students' favourable opinion of the laboratory-training period: concerning students' perceived goals for attending the course, a high degree of satisfaction was reported (globally 73% of computed scores were rated as 'good' or 'very good'); the teaching/learning environment and the delivered content got 90% 'very good' and 'good' ratings; teaching staff performance got the highest percentage of rating scores 'very good' and 'good' (95%). These ratings were dependent on variables that are unlikely to be a reflection only of glamour and seduction (clear explanations and clear demonstration of each component of the skill) and were less dependent on other variables such as 'enthusiasm for teaching' and 'relationship with students', which could be expected to be influenced by the charisma of the tutors. Helping students learn and train in surgical skills in a laboratory is challenging and our results affirm that the environment and the content were valuable for the understanding of the subject, while sessions planning and appropriate teaching technique are essential when practical skills are to be taught and learned.