Drosophila Model for Gut-Mediated Horizontal Transfer of Narrow- and Broad-Host-Range Plasmids

Microbial evolution in the gut of animals due to horizontal gene transfer (HGT) is of significant interest for microbial evolution as well as within the context of human and animal health. Microbial populations evolve within the host, and factors from the bacteria and host interact to regulate this evolution.

Carbapenemase Producing Enterobacterales in Romania: Investigating the Origins

Carbapenemase-producing Enterobacterales (CPE) have emerged and spread in Romania since 2010. According to the reports of the EuSPACE (European survey of carbapenemase-producing Enterobacteriaceae) the epidemio-logical stage of the CPE expansion in Romania has shifted from sporadic occurrence in 2013 directly to inter-regional spread in 2014-2015. In this study we aimed to provide data from the timeframe when the dissemination of the carbapenemase genes in Romania began, by retrospectively analyzing CPE strains in a tertiary care university hospital. During the period of November 2012 – October 2013 we found 107 CPE (8.78%) out of 1219 non-duplicate Enterobacterales strains. 26 isolates of various Enterobacterales species carried blaNDM-1, 83 Klebsiella pneumoniae strains were positive for blaOXA-48-like and 2 of these co-harboured blaNDM-1. The increased incidence of OXA-48 producing K. pneumoniae was linked to a two-peaked hospital outbreak during February and May 2013. The percentage of 24.3% of NDM-1 producers was alarming due to the diversity of involved species and the higher resistance levels to carbapenems compared with blaOXA-48-like gene carriers. Plasmid replicon typing revealed a great diversity of plasmids in NDM-1-positive strains, belonging to incompatibility groups A/C, FII, FIIk, HI2, L and M. The strong connection between certain plasmid groups and host species suggests that the transfer of broad host-range plasmids through conjugation does not play the main role in the successful spread of blaNDM-1 among Enterobacterales species.

Proma Plasmids are Instrumental in the Dissemination of Linuron Catabolic Genes between Different Genera

ABSTRACTPromA plasmids are broad host range plasmids, which are often cryptic and hence have an uncertain ecological role. We present three novel PromA γ plasmids which carry genes associated with degradation of the phenylurea herbicide linuron, two (pPBL-H3-2 and pBPS33-2) of which originate from unrelated Hydrogenophaga hosts isolated from different environments, and one (pEN1) which was exogenously captured from an on-farm biopurification system. Both Hydrogenophaga plasmids carry all three necessary gene clusters determining the three main steps for conversion of linuron to Krebs cycle intermediates, while pEN1 only determines the initial linuron hydrolysis step. Linuron catabolic gene clusters that determine the same step were identical on all plasmids, encompassed in differently arranged constellations and characterized by the presence of multiple IS1071 elements. In all plasmids except pEN1, the insertion spot of the catabolic genes in the PromA γ plasmids was the same. Highly similar PromA plasmids carrying the linuron degrading gene cargo at the same insertion spot were were previously identified in linuron degrading Variovorax sp. Interestingly, in both Hydrogenophaga populations not every PromA plasmid copy carries catabolic genes. The results indicate that PromA plasmids are important vehicles of linuron catabolic gene dissemination, rather than being cryptic and only important for the mobilization of other plasmids.

Broad-Host-Range Plasmids for Constitutive and Inducible Gene Expression in the Absence of Antibiotic Selection

Plasmid vectors are a valuable research tool for characterizing bacterial gene function, but there is a limited range of plasmids that are functional in nonmodel bacterial species. Described here is a set of broad-host-range plasmids modified for stability in the absence of antibiotic selection and for gene expression manipulation.

Emergence of plasmid-mediated oxazolidinone resistance gene poxtA from CC17 Enterococcus faecium of pig origin

ObjectivesTo characterize the oxazolidinone resistance gene poxtA on broad-host-range Inc18 plasmids from CC17 Enterococcus faecium of pig origin.MethodsOxazolidinone-resistant E. faecium isolates were screened for the presence of poxtA. The poxtA-carrying isolates were characterized by antimicrobial susceptibility testing, conjugation, S1-PFGE and hybridization. The poxtA-carrying plasmids were completely sequenced and their instability was verified.ResultsTwo individual CC17 E. faecium strains were positive for poxtA. S1-PFGE and hybridization revealed the presence of a poxtA-carrying plasmid of ∼62 kb in both WZ27-2 and the transconjugant, while poxtA-carrying plasmids of different sizes were observed in QF25-1 and the transconjugant. The two poxtA-carrying plasmids, pC25-1 and pC27-2, belonged to the broad-host-range plasmids of the Inc18 family and carried dfrG, aadE, Δsat4, aph(3′)-III, erm(B), tet(M), tet(L) and fexB. Plasmid pC27-2 was virtually identical to pC25-1, with minor differences. The calculated transfer frequency was ∼0.87 × 10−8 and ∼1.03 × 10−7 per recipient to plasmids pC25-1 and pC27-2, respectively. Instability assays of the region with four adjacent IS1216Es, which forms three IS1216E translocatable units, revealed the formation of a series of mosaic circular intermediates.ConclusionsWe report the emergence of the plasmid-mediated oxazolidinone resistance gene poxtA in E. faecium from different farms in China. Comparison of the poxtA genetic context suggests that IS1216E elements play an important role in the dissemination of poxtA. The co-occurrence of poxtA with other antimicrobial and heavy metal resistance genes on the broad-host-range plasmids of the Inc18 family may lead to the co-selection of poxtA, contributing to its persistence and accelerating its dissemination.