Expanding the Repertoire of Carbapenem-Hydrolyzing Metallo-ß-Lactamases by Functional Metagenomic Analysis of Soil Microbiota

Carbapenemases are bacterial enzymes that hydrolyze carbapenems, a group of last-resort β-lactam antibiotics used for treatment of severe bacterial infections. They belong to three β-lactamase classes based amino acid sequence (A, B, and D). The aim of this study was to elucidate occurrence, diversity and functionality of carbapenemase-encoding genes in soil microbiota by functional metagenomics. Ten plasmid libraries were generated by cloning metagenomic DNA from agricultural (n = 6) and grassland (n = 4) soil into Escherichia coli. The libraries were cultured on amoxicillin-containing agar and up to 100 colonies per library were screened for carbapenemase production by CarbaNP test. Presumptive carbapenemases were characterized with regard to DNA sequence, minimum inhibitory concentration (MIC) of β-lactams, and imipenem hydrolysis. Nine distinct class B carbapenemases, also known as metallo-beta-lactamases (MBLs), were identified in six soil samples, including two subclass B1 (GRD23-1 and SPN79-1) and seven subclass B3 (CRD3-1, PEDO-1, GRD33-1, ESP-2, ALG6-1, ALG11-1, and DHT2-1). Except PEDO-1 and ESP-2, these enzymes were distantly related to any previously described MBLs (33 to 59% identity). RAIphy analysis indicated that six enzymes (CRD3-1, GRD23-1, DHT2-1, SPN79-1, ALG6-1, and ALG11-1) originated from Proteobacteria, two (PEDO-1 and ESP-2) from Bacteroidetes and one (GRD33-1) from Gemmatimonadetes. All MBLs detected in soil microbiota were functional when expressed in E. coli, resulting in detectable imipenem-hydrolyzing activity and significantly increased MICs of clinically relevant β-lactams. Interestingly, the MBLs yielded by functional metagenomics generally differed from those detected in the same soil samples by antibiotic selective culture, showing that the two approaches targeted different subpopulations in soil microbiota. © 2016 Gudeta, Bortolaia, Pollini, Docquier, Rossolini, Amos, Wellington and Guardabassi.Grant HEALTH-F3-2011-282004(EvoTAR

Greening crowdfunding campaigns: an investigation of message framing and effective communication strategies for funding success

Purpose: Recognising the growing importance of environmental and sustainable activities and the role of communication strategies in soliciting their financing, this work investigates the influence of message framing, green emphasis and quantitative information on the probability of green crowdfunding campaigns' success. Design/methodology/approach: This analysis is based on crowdfunding campaigns published between 2015 and 2020 on the Indiegogo platform in the category “Community projects – Environment”. The study develops an in-depth qualitative content analysis of the projects before performing an empirical examination to determine funding causes. Findings: Communication strategies (message framing, green emphasis and quantitative goals) affect funding success. However, project category moderates the impact of message framing and green emphasis on campaign success. While positive framing increases agri-food campaign success, negative framing is more effective for clean energy and climate preservation projects. Moreover, indication of a quantitative goal and a marked green emphasis in a project's presentation increase campaign success, but a too marked green emphasis is only effective for agri-food projects. Practical implications: Green entrepreneurs and campaign managers must work carefully on their projects' communication, accounting for the type of product proposed, emphasising green components in its description and utilising quantitative information to present future goals. These strategies maximise backers' responses and enable entrepreneurs to obtain funding. The authors’ findings may be extended to other contexts, including the banking sector, to craft effective communication strategies for green financial products. Originality/value: By applying framing theory in a new context (i.e. the online financing of green entrepreneurs), this study identifies new campaign success determinants and provides evidence for the moderating role of project category. Furthermore, the study highlights the need to develop different communication strategies for social and environmental-oriented projects

Genome-based study of a spatio-temporal cluster of invasive meningococcal disease due to Neisseria meningitidis serogroup C, clonal complex 11

SummaryObjectivesTo describe a spatio-temporal cluster of invasive meningococcal disease (IMD) due to serogroup C meningococci, occurred in a restricted area of Tuscany between January and October 2015, and the results of whole genome sequencing (WGS).MethodsSurveillance activities and public health measures were implemented in the Region. Bacterial isolates from IMD cases were characterized by the National Reference Laboratory of the Istituto Superiore di Sanità (ISS), and WGS was performed on available strains. The kSNP software was used to identify core genome SNPs.ResultsOverall, 28 IMD cases due to meningococcus C were identified up to 31st October, 2015. Of them, 26 were due to meningococcus C:P1.5-1,10-8: F3-6:ST-11 (cc11) and 2 to C:P1.5-1,10-8: F3-6:ST-2780 (cc11). WGS of 13 meningococci isolated during the outbreak occurred in Tuscany in 2015 showed higher similarity when compared with those of 47 C: P1.5-1,10-8: F3-6:ST-11 (cc11) invasive strains from sporadic cases previously detected in Italy.ConclusionsA highly aggressive meningococcal C strain was involved in the cluster of severe IMD occurred in Tuscany, a Region with high vaccine coverage among children. Whether this was due to low herd immunity related to the short duration of vaccine protection needs further investigation