Incidence and Genomic Background of Antibiotic Resistance in Food-Borne and Clinical Isolates of Serovar Derby from Spain.

serovar Derby (. Derby) ranks fifth among nontyphoidal serovars causing human infections in the European Union. .

Spread of and Other Clinically Relevant Resistance Genes, Such as and , Driven by IncHI2-ST1 Plasmids in Clinical Isolates of Monophasic Serovar Typhimurium ST34.

The monophasic 4,[5],12:i:-variant of serovar Typhimurium with sequence type ST34 has become one of the most prevalent non-typhoidal salmonellae worldwide. In the present study, we thoroughly characterized seven isolates of this variant detected in a Spanish hospital and selected based on cefotaxime resistance and cefoxitin susceptibility, mediated by . For this, conventional microbiological techniques, together with whole genome sequencing performed with the Illumina platform, were applied.

Genomic Analysis of Two MDR Isolates of Serovar Infantis from a Spanish Hospital Bearing the Gene with or without in pESI-like Plasmids.

serovar Infantis (. Infantis) is a broiler-associated pathogen which ranks in the fourth position as a cause of human salmonellosis in the European Union. Here, we report a comparative genomic analysis of two clinical .

Detection of the Gene Among Polyclonal Linezolid-Susceptible Isolates of Recovered from Community Patients.

Dispersion of transferable oxazolidinone resistance genes among enterococci poses a serious problem to human health. Prompt detection of bacteria carrying these genes is crucial to avoid their spread to multidrug-resistant bacteria. The aim of the study was to describe the presence of -positive isolates among enterococci in a Spanish hospital, and to determine their genetic context and location through whole genome sequencing.

Plasmid-Mediated Quinolone Resistance (PMQR) in Two Clinical Strains of Serovar Corvallis.

Non-typhoid serovars of are one of the main causes of bacterial food-borne infections worldwide. For the treatment of severe cases of salmonellosis in adults, fluoroquinolones are amongst the drugs of choice. They are categorized by the World Health Organization (WHO) as "critically important with highest priority in human medicine".

Colistin Resistance in Monophasic Isolates of ST34 Collected From Meat-Derived Products in Spain, With or Without CMY-2 Co-production.

Colistin is a last-resort antibiotic in fighting severe infections caused by multidrug resistant Gram negative pathogens in hospitals. Zoonotic bacteria acquire colistin resistance in animal reservoirs and mediate its spread along the food chain. This is the case of non-typhoid serovars of .

Genetic and Physiological Characterization of Fructose-1,6-Bisphosphate Aldolase and Glyceraldehyde-3-Phosphate Dehydrogenase in the Crabtree-Negative Yeast .

The milk yeast degrades glucose through glycolysis and the pentose phosphate pathway and follows a mainly respiratory metabolism. Here, we investigated the role of two reactions which are required for the final steps of glucose degradation from both pathways, as well as for gluconeogenesis, namely fructose-1,6-bisphosphate aldolase (FBA) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). In silico analyses identified one gene encoding the former (), and three genes encoding isoforms of the latter (, , ).

Genomic Analysis of Ciprofloxacin-Resistant Serovar Kentucky ST198 From Spanish Hospitals.

serovar Kentucky (. Kentucky) with sequence type (ST) 198 and highly resistant to ciprofloxacin (ST198-Cip ) has emerged as a global MDR clone, posing a threat to public health. In the present study, whole genome sequencing (WGS) was applied to characterize all Cip Kentucky detected in five Spanish hospitals during 2009-2018.

Genomic analysis and phylogenetic position of the complex IncC plasmid found in the Spanish monophasic clone of Salmonella enterica serovar Typhimurium.

pUO-STmRV1 is an IncC plasmid discovered in the Spanish clone of the emergent monophasic variant of Salmonella enterica serovar Typhimurium, which has probably contributed to its epidemiological success. The sequence of the entire plasmid determined herein revealed a largely degenerated backbone with accessory DNA incorporated at four different locations. The acquired DNA constitutes more than two-thirds of the pUO-STmRV1 genome and originates from plasmids of different incompatibility groups, including IncF (such as R100 and pSLT, the virulence plasmid specific of S.

A Plasmid-Encoded FetMP-Fls Iron Uptake System Confers Selective Advantages to Serovar Typhimurium in Growth under Iron-Restricted Conditions and for Infection of Mammalian Host Cells.

The resistance plasmid pUO-StVR2, derived from virulence plasmid pSLT, is widespread in clinical isolates of serovar Typhimurium recovered in Spain and other European countries. pUO-StVR2 carries several genes encoding a FetMP-Fls system, which could be involved in iron uptake. We therefore analyzed .

Analysis of the Degradation of Broad-Spectrum Cephalosporins by OXA-48-Producing Using MALDI-TOF MS.

The objective of the study was to evaluate the activity of OXA-48 against different broad-spectrum cephalosporins and to identify the reaction products by MALDI-TOF MS. The action of OXA-48 on cefotaxime, ceftazidime, and ceftriaxone was assessed by this method, using an J53 transconjugant carrying only the ~62 Kb IncL plasmid containing the gene, and the same strain without any plasmid was included as a negative control. In addition, a collection of 17 clinical OXA-48-producing , which were susceptible to broad-spectrum cephalosporins, was evaluated.

Lack of the NAD-dependent glycerol 3-phosphate dehydrogenase impairs the function of transcription factors Sip4 and Cat8 required for ethanol utilization in Kluyveromyces lactis.

The NAD-dependent glycerol 3-phosphate dehydrogenase (KlGpd1) is an important enzyme for maintenance of the cytosolic redox balance in the milk yeast Kluyveromyces lactis. The enzyme is localized in peroxisomes and in the cytosol, indicating its requirement for the oxidation of NADH in both compartments. Klgpd1 mutants grow more slowly on glucose than wild-type cells and do not grow on ethanol as a sole carbon source.

Protein kinase C in fungi-more than just cell wall integrity.

Human protein kinase C (PKC) isoforms have been implicated in diseases such as Alzheimer's, diabetes and cancers. In contrast to mammals, which have at least nine genes, fungi have only one or two. The yeast Saccharomyces cerevisiae produces only a single Pkc1 and is employed in the study of specific human isozymes, including their susceptibility to pharmacological drugs.

Cell wall synthesis and central carbohydrate metabolism are interconnected by the SNF1/Mig1 pathway in Kluyveromyces lactis.

The trimeric AMP-activated kinase complex (AMPK) is conserved from yeast to humans and is best known for its role in balancing energy metabolism. Additional functions, including the regulation of cell wall biosynthesis, have been proposed for the SNF1 complex, the baker's yeast homolog of AMPK. We here demonstrate that this function is conserved in the Crabtree-negative milk yeast Kluyveromyces lactis.

Incidence and Genetic Bases of Nitrofurantoin Resistance in Clinical Isolates of Two Successful Multidrug-Resistant Clones of Salmonella enterica Serovar Typhimurium: Pandemic "DT 104" and pUO-StVR2.

In this study, the incidence and genetic bases of nitrofurantoin resistance were established for clinical isolates of two successful clones of Salmonella enterica serovar Typhimurium, the pandemic "DT 104" and the pUO-StVR2 clone. A total of 61 "DT 104" and 40 pUO-StVR2 isolates recovered from clinical samples during 2008-2014 and assigned to different phage types, were tested for nitrofurantoin susceptibility. As previously shown for older isolates, all newly tested pUO-StVR2 isolates were highly resistant to nitrofurantoin (minimal inhibitory concentration [MIC] of 128 μg/ml), while 42.

The role of IS26 in evolution of a derivative of the virulence plasmid of Salmonella enterica serovar Enteritidis which confers multiple drug resistance.

pUO-SeVR1 is a resistance derivative of pSEV, the virulence plasmid specific of Salmonella enterica serovar Enteritidis. It was first detected in a Spanish isolate involved in gastroenteritis, but closely related plasmids are widespread in highly invasive isolates originating from Africa. According to its nucleotide sequence, pUO-SeVR1 consists of 110,982bp with a GC content of 53.

Transposition and homologous recombination drive evolution of pUO-StVR2, a multidrug resistance derivative of pSLT, the virulence plasmid specific of Salmonella enterica serovar Typhimurium.

Five variants of a resistant derivative of pSLT (termed pUO-StVR2) were detected in clinical isolates of Salmonella enterica serovar Typhimurium recovered in Spain. The structure of these variants revealed the involvement of IS1, IS26 and Tn21-like transposition, as well as homologous recombination in the generation of deletions, inversions and insertions which, depending on the variant, affected an orf of unknown function, genes encoding a possible iron acquisition system, and/or resistance properties. These variants, which appeared at a relatively low frequency, can be used as a model to understand the co-selection mechanisms that are helping to maintain multidrug resistance in bacterial pathogens, despite the structural instability of the responsible DNA.

Mutations in SNF1 complex genes affect yeast cell wall strength.

The trimeric SNF1 complex from Saccharomyces cerevisiae, a homolog of mammalian AMP-activated kinase, has been primarily implicated in signaling for the utilization of alternative carbon sources to glucose. We here find that snf1 deletion mutants are hypersensitive to different cell wall stresses, such as the presence of Calcofluor white, Congo red, Zymolyase or the glucan synthase inhibitor Caspofungin in the growth medium. They also have a thinner cell wall.

Yeast on the milky way: genetics, physiology and biotechnology of Kluyveromyces lactis.

The milk yeast Kluyveromyces lactis has a life cycle similar to that of Saccharomyces cerevisiae and can be employed as a model eukaryote using classical genetics, such as the combination of desired traits, by crossing and tetrad analysis. Likewise, a growing set of vectors, marker cassettes and tags for fluorescence microscopy are available for manipulation by genetic engineering and investigating its basic cell biology. We here summarize these applications, as well as the current knowledge regarding its central metabolism, glucose and extracellular stress signalling pathways.

Efficient mobilization of a resistance derivative of pSLT, the virulence plasmid specific of Salmonella enterica serovar Typhimurium, by an IncI1 plasmid.

pUO-StVR2 is a derivative of pSLT, the virulence plasmid specific of Salmonella enterica serovar Typhimurium, which confers multidrug resistance. This plasmid is widespread among closely related isolates of S. Typhimurium, and often coexists with other plasmids like pStR12.

Milk and sugar: regulation of cell wall synthesis in the milk yeast Kluyveromyces lactis.

The milk yeast Kluyveromyces lactis is an alternative model yeast to the well established Saccharomyces cerevisiae. The cell wall of these fungi consists of polysaccharides (i.e.

Together we are strong--cell wall integrity sensors in yeasts.

The integrity of the fungal cell wall is ensured by a signal transduction pathway, the so-called CWI pathway, which has best been studied in the model yeast Saccharomyces cerevisiae. In this context, environmental stress and other perturbations at the cell surface are detected by a small set of plasma membrane-spanning sensors, viz. Wsc1, Wsc2, Wsc3, Mid2 and Mtl1.

A tool kit for molecular genetics of Kluyveromyces lactis comprising a congenic strain series and a set of versatile vectors.

A set of different marker deletions starting with a ura3 derivative of the Kluyveromyces lactis type strain CBS2359 was constructed. After a first cross to obtain a strain with the opposite mating type that also carried a leu2 allele, continuous back-crosses were used to obtain a congenic strain series with different marker combinations, including deletions in KlHIS3, KlADE2 and KlLAC4. Enzymes involved in carbohydrate metabolism were shown to behave very similarly to the original type strain and other K.

Molecular basis and evolution of multiple drug resistance in the foodborne pathogen Salmonella enterica serovar Ohio.

The molecular basis and evolution of multidrug resistance were established for 54 isolates of Salmonella enterica serovar Ohio, recorded between 1991 and 2005 in Asturias, a northern region of Spain. All isolates were closely related, as shown by cluster analysis of XbaI-BlnI combined profiles. Of these, 33.