Coregulated Genes Link Sulfide:Quinone Oxidoreductase and Arsenic Metabolism in Synechocystis sp. Strain PCC6803

Although the biogeochemistry of the two environmentally hazardous compounds arsenic and sulfide has been extensively investigated, the biological interference of these two toxic but potentially energy-rich compounds has only been hypothesized and indirectly proven. Here we provide direct evidence for the first time that in the photosynthetic model organism Synechocystis sp. strain PCC6803 the two metabolic pathways are linked by coregulated genes that are involved in arsenic transport, sulfide oxidation, and probably in sulfide-based alternative photosynthesis. Although Synechocystis sp. strain PCC6803 is an obligate photoautotrophic cyanobacterium that grows via oxygenic photosynthesis, we discovered that specific genes are activated in the presence of sulfide or arsenite to exploit the energy potentials of these chemicals. These genes form an operon that we termed suoRSCT, located on a transposable element of type IS4 on the plasmid pSYSM of the cyanobacterium. suoS (sll5036) encodes a light-dependent, type I sulfide:quinone oxidoreductase. The suoR (sll5035) gene downstream of suoS encodes a regulatory protein that belongs to the ArsR-type repressors that are normally involved in arsenic resistance. We found that this repressor has dual specificity, resulting in 200-fold induction of the operon upon either arsenite or sulfide exposure. The suoT gene encodes a transmembrane protein similar to chromate transporters but in fact functioning as an arsenite importer at permissive concentrations. We propose that the proteins encoded by the suoRSCT operon might have played an important role under anaerobic, reducing conditions on primordial Earth and that the operon was acquired by the cyanobacterium via horizontal gene transfer

Characterization of the activity of heavy metal-responsive promoters in the cyanobacterium Synechocystis PCC 6803

Aiming at developing cyanobacterial-based biosensors for heavy metal detection, expression of heavy metal inducible genes of the cyanobacterium Synechocystis PCC 6803 was investigated by quantitative RT-PCR upon 15 minutes exposure to biologically relevant concentrations of Co 2+ , Zn 2+ , Ni 2+ , Cd 2+ , Cr 6+ , As 3+ and As 5+ . The ziaA gene, which encodes a Zn 2+ -transporting P-type ATPase showed a marked- ly increased mRNA level after incubation with Cd 2+ and arsenic ions, besides the expected induction by Zn 2+ ions. The Co 2+ efflux system-encoding gene coaT was strongly induced by Co 2+ and Zn 2+ ions, mod- erately induced by As 3+ ions, and induced at a relatively low level by Cd 2+ and As 5+ ions. Expression of nrsB , which encodes a part of a putative Ni 2+ efflux system was highly induced by Ni 2+ salts and at a low extent by Co 2+ and Zn 2+ salts. The arsB gene, which encodes a putative arsenite-specific efflux pump was highly induced by As 3+ and As 5+ ions, while other metal salts provoked insignificant transcript level increase. The transcript of chrA , in spite of the high sequence similarity of its protein product with sev- eral bacterial chromate transporters, shows no induction upon Cr 6+ salt exposure. We conclude that due to the largely unspecific heavy metal response of the studied genes only nrsB and arsB are potential can- didates for biosensing applications for detection of Ni 2+ and arsenic pollutants, respectively

Prevalence of ESBL, AmpC and Carbapenemase-Producing Enterobacterales Isolated from Raw Vegetables Retailed in Romania

(1) Background: As β-lactamase-producing Enterobacterales are no longer exclusively associated with the health care system, investigating the potential risk they pose to the integrity of the environment and food safety has become of utmost importance. This study aimed to determine the prevalence of extended-spectrum β-lactamase (ESBL), AmpC, and carbapenemase-producing Enterobacterales isolates from retailed raw vegetables and to determine if household washing is an effective method of lowering bacterial load; (2) Methods: Seasonal vegetables (n = 165) were acquired from supermarkets (n = 2) and farmer markets (n = 2) in Romania. Following sample processing and isolation, identification of Enterobacterales was performed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF). Polymerase chain reaction (PCR) multiplex was used to ascertain the presence of the main ESBL, AmpC, and Carbapenemase genes. Phenotypic antibiotic resistance profiles of isolates were determined by extended antibiograms. Enterobacteriaceae colony-forming units (CFU) counts were compared between vegetable types; (3) Results: Beta-lactamase producing bacteria were observed on 7.9% of vegetables, with 5.5% displaying ESBL/AmpC phenotype and 2.4% identified as Carbapenemase producers. The most frequently detected β-lactamase genes were blaSHV (n = 4), followed by blaCTX-M and blaTEM (each with n = 3). Phenotypic antibiotic resistance analysis showed that 46% of isolates were multiple drug resistant, with aminoglycosides (38.5%) the most prevalent non-β-lactam resistance, followed by first-generation quinolones (38.5%). (4) Conclusions: The present study has described for the first time the presence of β-lactamase producing Enterobacterales in fresh produce retailed in Romania