Mariana Papalia

Biodegradability and resistance from indigenous bacterial communities to dyes were tested using samples from both polluted and unpolluted surface waters in Buenos Aires. Five dyes were selected for the study: Acid Black 210, Direct Orange 39, Malachite Green, Gentian Violet, and Alizarin Red. Water quality was assessed by measuring chemical oxygen demand, biochemical oxygen demand, and both Escherichia coli and enterococci counts. Biodegradability was tested using a respirometric method, while resistance was assessed by determining the minimum inhibitory concentration (MIC). No bacterial strains capable of degrading the dyes as the sole carbon source were isolated from the respirometric tests. However, from the MIC tests, 28 strains capable of dye discolouration were identified, using nutrient broth as a supplement. Two of them were able to degrade Malachite Green and Acid Black 210 at a concentration of 50 mg L−1 in less than 24 h and with an efficiency greater than 87%. These strains were identified as Aeromonas sp. and Shewanella sp. through MALDI-TOF/MS and 16S rRNA gene sequencing. The determination of biodegradability and resistance can be used to enhance the characterization of watercourses. Furthermore, this methodology provides a means to isolate biodegrading bacteria that could be applied in effluent treatment processes.

Achromobacter spp. are intrinsically resistant to multiple antibiotics and can also acquire resistance to those commonly used for the treatment of respiratory infections, especially in patients with cystic fibrosis. The aim of this study was to perform the genetic and biochemical characterization of AXC-2 from A. ruhlandii and to analyze all available AXC variants. Steady-state kinetic parameters were determined on a purified AXC-2 enzyme. It exhibited higher catalytic efficiencies towards amino-penicillins and older cephalosporins, while carbapenems behaved as poor substrates. Phylogenetic analysis of all bla_AXC variants available in the NCBI was conducted. AXC was encoded in almost all A. ruhlandii genomes, whereas it was only found in 30% of A. xylosoxidans. AXC-1 was prevalent among A. xylosoxidans. AXC variants were clustered in two main groups, correlating with the Achromobacter species. No association could be established between the presence of bla_AXC variants and a specific lineage of A. xylosoxidans; however, a proportion of AXC-1-producing isolates corresponded to ST 182 and ST 447. In conclusion, this study provides valuable insights into the genetic context and kinetic properties of AXC-2, identified in A. ruhlandii. It also provides a thorough description of all AXC variants and their association with Achromobacter species and various lineages.

Burkholderia contaminans is a member of the Burkholderia cepacia complex (Bcc), a pathogen with increasing prevalence among cystic fibrosis (CF) patients and the cause of numerous outbreaks due to the use of contaminated commercial products. The antibiotic resistance determinants, particularly β-lactamases, have been poorly studied in this species. In this work, we explored the whole genome sequence (WGS) of a B. contaminans isolate (FFH 2055) and detected four putative β-lactamase-encoding genes. In general, these genes have more than 93% identity with β-lactamase genes found in other Bcc species. Two β-lactamases, a class A (Pen-like, suggested name PenO) and a class D (OXA-like), were further analyzed and characterized. Amino acid sequence comparison showed that Pen-like has 82% and 67% identity with B. multivorans PenA and B. pseudomallei PenI, respectively, while OXA-like displayed strong homology with class D enzymes within the Bcc, but only 22–44% identity with available structures from the OXA family. PCR reactions designed to study the presence of these two genes revealed a heterogeneous distribution among clinical and industrial B. contaminans isolates. Lastly, bla_PenO gene was cloned and expressed into E. coli to investigate the antibiotic resistance profile and confers an extended-spectrum β-lactamase (ESBL) phenotype. These results provide insight into the presence of β-lactamases in B. contaminans, suggesting they play a role in antibiotic resistance of these bacteria.

Purpose. The aim of this work was to investigate the presence of selected potential virulence factors, susceptibility and clonal relatedness among 63 Stenotrophomonas maltophilia isolates recovered from patients exposed to invasive devices in a university hospital in Argentina between January 2004 and August 2012. Methodology. Genetic relatedness was assessed by enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) and pulsed-field gel electrophoresis (PFGE). Isolates were characterized by antimicrobial resistance, the presence and/or expression of potential virulence determinants, and virulence in the Galleria mellonella model. Results/Key findings. ERIC-PCR generated 52 fingerprints, and PFGE added another pattern. Resistance to trimethoprim–sulfamethoxazole (6.35 %), levofloxacin (9.52 %) and ciprofloxacin (23.80 %) was detected. All isolates were susceptible to minocycline. All isolates were lipase, protease and siderophore producers, while all but Sm61 formed biofilms. However, 11/63 isolates did not amplify the major extracellular protease-coding gene (stmPr1). Sm61 is an stmPr1-negative isolate, and showed (as did Sm13 and the reference strain K279a) strong proteolysis and siderophore production, and high resistance to hydrogen peroxide. The three isolates were virulent in the G. mellonella model, while Sm10, a low-resistance hydrogen peroxide stmPr1-negative isolate, and weak proteolysis and siderophore producer, was not virulent. Conclusion. This is the first epidemiological study of the clonal relatedness of S. maltophilia clinical isolates in Argentina. Great genomic diversity was observed, and only two small clusters of related S. maltophilia types were found. Minocycline and trimethoprim–sulfamethoxazole were the most active agents. S. maltophilia virulence in the G. mellonella model is multifactorial, and further studies are needed to elucidate the role of each potential virulence factor.

Water from six points from the Riachuelo-Matanza basin was analyzed in order to assess ibuprofen biodegradability. In four of them biodegradation of ibuprofen was proved and degrading bacterial communities were isolated. Biodegradation in each point could not be correlated with sewage pollution. The indigenous bacterial community isolated from the point localized in the La Noria Bridge showed the highest degradative capacity and was selected to perform batch and continuous degradation assays. The partial 16S rRNA gene sequence showed that the community consisted of Comamonas aquatica and Bacillus sp. In batch assays the community was capable of degrading 100 mg L⁻¹ of ibuprofen in 33 h, with a specific growth rate (μ) of 0.21 h⁻¹. The removal of the compound, as determined by High performance liquid chromatography (HPLC), exceeded 99% of the initial concentration, with a 92.3% removal of Chemical Oxygen Demand (COD). In a down-flow fixed-bed continuous reactor, the community shows a removal efficiency of 95.9% of ibuprofen and 92.3% of COD for an average inlet concentration of 110.4 mg. The reactor was kept in operation for 70 days. The maximal removal rate for the compound was 17.4 g m⁻³ d⁻¹. Scanning electron microscopy was employed to observe biofilm development in the reactor. The ability of the isolated indigenous community can be exploited to improve the treatment of wastewaters containing ibuprofen.