Browsing by Author "Rivas, Lina"
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Item A Multicenter Study To Evaluate Ceftaroline Breakpoints: Performance in an Area with High Prevalence of Methicillin-Resistant Staphylococcus aureus Sequence Type 5 Lineage(2019) Khan, Ayesha; Rivas, Lina; Spencer, María; Martínez, Rodrigo; lam, Marusella; Rojas, Pamela; Porte, Lorena; Silva, Francisco; Braun, Estefanía; Valdivieso, Francisca; Lhauser, Margareta; Lafourcade, Mónica; Miller, Guillermo; García, Patricia; Arias, César; Munita, JoséCeftaroline (CPT) is a broad-spectrum agent with potent activity against methicillin-resistant Staphylococcus aureus (MRSA). The sequence type 5 (ST5) Chilean-Cordobés clone, associated with CPT nonsusceptibility, is dominant in Chile, a region with high rates of MRSA infections. Here, we assessed the in vitro activity of CPT against a collection of MRSA isolates collected between 1999 and 2018 from nine hospitals (n = 320) and community settings (n = 41) in Santiago, Chile, and evaluated performance across testing methodologies. We found that our hospital-associated isolates exhibited higher CPT MIC distributions (MIC50 and MIC90 of 2 mg/liter) than the community isolates (MIC50 and MIC90 of 0.5 mg/liter), a finding that was consistent across time and independent of the culture source. High proportions (64%) of isolates were CPT nonsusceptible despite the absence of CPT use in Chile. Across methodologies, the Etest underestimated the MIC relative to the gold standard broth microdilution (BMD) test (MIC50 and MIC90 of 1 and 1.5 mg/liter, respectively). There was low (∼51%) categorical agreement (CA) between Etest and BMD results across CLSI and EUCAST breakpoints. The recent revision of CLSI guidelines abolished "very major error" (VME) from the previous guidelines (81%), which perform similarly to the EUCAST guidelines. The level of concordance between CLSI and EUCAST for BMD testing and Etest was >95%. Disk diffusion performed poorly relative to BMD under CLSI (CA, 55%) and EUCAST (CA, 36%) guidelines. Comparison of EUCAST to CLSI for disk diffusion (with EUCAST used as the reference) showed low agreement (CA, 25%; VME, 70%). In summary, CPT-nonsusceptible MRSA are dominant in clinical settings in Chile. Our results provide data to support the reevaluation of CPT breakpoints and to improve agreement across methodologies and agencies.Item A multispecies outbreak of carbapenem-resistant bacteria harboring the blaKPC gene in a non-classical transposon element(2021) Wozniak, Aniela; Figueroa, Cristian; Moya-Flores, Francisco; Guggiana, Piero; Castillo, Claudia; Rivas, Lina; Munita, José; García, Patricia C.Klebsiella pneumoniae is the most frequent KPC-producing bacteria. The blaKPC gene is frequently embedded in Tn4401 transposon, and less frequently in non-Tn4401 elements (NTEKPC) variants I-III. The first case of KPC in the UC-CHRISTUS Clinical Hospital was detected in Pseudomonas aeruginosa. Soon after this event, KPC was detected in 2 additional Pseudomonas aeruginosa, 3 Escherichia coli, 3 Enterobacter cloacae, 3 Klebsiella pneumoniae, and 1 Citrobacter freundii, isolated from 6 different patients. We aimed to elucidate the possible mechanisms of genetic transfer and dissemination of the blaKPC gene among isolates of this multispecies outbreak. A molecular epidemiology analysis of the above mentioned clinical isolates (n = 13) through Multi-Locus Sequence Typing, plasmid analysis, Pulsed-Field Gel-Electrophoresis, and Whole-genome sequencing (WGS) was performed.Item A Natural Deep Eutectic Solvent Formulated to Stabilize β-Lactam Antibiotics(2018) Olivares, Belén; Martínez, Fabián; Rivas, Lina; Calderon, Cristian; Munita, José; Campodónico, Paolaβ-lactam antibiotics, such as penicillin share a very unstable chemical structure. In water-based solutions, such as those used for clinical applications, the β-lactam ring is readily opened due to a nucleophilic or electrophilic attack, leading to the loss of antimicrobial activity. Since the achievement and maintenance of optimum therapeutic levels of β-lactam antibiotics is critical for the resolution of many infectious clinical situations, and to avoid antibiotic resistance generation, the design of new non-aqueous dosage forms is urgent. Recently, natural deep eutectic solvents (NADES) have emerged as alternative non-toxic and non-aqueous solvents for different biomedical applications. In this work, we formulated and characterized a NADES composed by betaine and urea (BU). Using this solvent, we evaluated the stability of clavulanic acid (CLV) and imipenem (IMP) and characterized their antimicrobial activities calculating the minimal inhibitory concentration. Characterization of BU solvent by infrared spectroscopy (IR) and nuclear magnetic resonance spectroscopy (NMR) indicated that the obtained solvent has a microstructure mainly based on hydrogen bonding interactions and water addition strongly affects its dynamic. The stability of β-lactam antibiotic IMP and CLV using this solvent was increased by 7 fold and 2.5 fold respectively compared to water when analysed seven days after being dissolved. Microbiological assays showed that antibacterial activity at day seven was significantly decreased for both CLV and IMP when dissolved in water, while no change in their antibacterial properties was observed when antibiotics were dissolved in BU. The increased stability of IMP and CLV in BU may be related to the inert behaviour of the solvent and the higher dynamic restriction that helps antibiotics to maintain a more stable conformation. These data suggest the potential use of BU as a solvent to prevent degradation of β-lactam antibiotics.Publication Dynamics of the MRSA Population in a Chilean Hospital: a Phylogenomic Analysis (2000-2016)(2023) Martínez, José; Planet, Paul; Spencer, Maria; Rivas, Lina; Díaz, Lorena; Moustafa, Ahmed; Riquelme, Roberto; Alcalde, Manuel; Hanson, Blake; Carvajal, Lina; Rincón, Sandra; Reyes, Jinnethe; Lam, Marusella; Calderon, Juan; Araos, Rafael; García, Patricia; Arias, César; Munita, Jose M.The global dissemination of methicillin-resistant Staphylococcus aureus (MRSA) is associated with the emergence and establishment of clones in specific geographic areas. The Chilean-Cordobes clone (ChC) (ST5-SCCmecI) has been the predominant MRSA clone in Chile since its first description in 1998, despite the report of other emerging MRSA clones in recent years. Here, we characterize the evolutionary history of MRSA from 2000 to 2016 in a Chilean tertiary health care center using phylogenomic analyses. We sequenced 469 MRSA isolates collected between 2000 and 2016. We evaluated the temporal trends of the circulating clones and performed a phylogenomic reconstruction to characterize the clonal dynamics. We found a significant increase in the diversity and richness of sequence types (STs; Spearman r = 0.8748, P < 0.0001) with a Shannon diversity index increasing from 0.221 in the year 2000 to 1.33 in 2016, and an effective diversity (Hill number; q = 2) increasing from 1.12 to 2.71. The temporal trend analysis revealed that in the period 2000 to 2003 most of the isolates (94.2%; n = 98) belonged to the ChC clone. However, since then, the frequency of the ChC clone has decreased over time, accounting for 52% of the collection in the 2013 to 2016 period. This decline was accompanied by the rise of two emerging MRSA lineages, ST105-SCCmecII and ST72-SCCmecVI. In conclusion, the ChC clone remains the most frequent MRSA lineage, but this lineage is gradually being replaced by several emerging clones, the most important of which is clone ST105-SCCmecII. To the best of our knowledge, this is the largest study of MRSA clonal dynamics performed in South America. IMPORTANCE Methicillin-resistant Staphylococcus aureus (MRSA) is a major public health pathogen that disseminates through the emergence of successful dominant clones in specific geographic regions. Knowledge of the dissemination and molecular epidemiology of MRSA in Latin America is scarce and is largely based on small studies or more limited typing techniques that lack the resolution to represent an accurate description of the genomic landscape. We used whole-genome sequencing to study 469 MRSA isolates collected between 2000 and 2016 in Chile providing the largest and most detailed study of clonal dynamics of MRSA in South America to date. We found a significant increase in the diversity of MRSA clones circulating over the 17-year study period. Additionally, we describe the emergence of two novel clones (ST105-SCCmecII and ST72-SCCmecVI), which have been gradually increasing in frequency over time. Our results drastically improve our understanding of the dissemination and update our knowledge about MRSA in Latin America.Item ESBL-Producing Escherichia coli Carrying CTX-M Genes Circulating among Livestock, Dogs, and Wild Mammals in Small-Scale Farms of Central Chile(2021) Benavides, Julio A.; Salgado-Caxito, Marília; Opazo-Capurro, Andrés; González Muñoz, Paulina; Piñeiro, Ana; Otto Medina, Macarena; Rivas, Lina; Munita, José; Millán, JavierAntibiotic-resistant bacteria of critical importance for global health such as extended-spectrum beta-lactamases-producing (ESBL)-Escherichia coli have been detected in livestock, dogs, and wildlife worldwide. However, the dynamics of ESBL-E. coli between these animals remains poorly understood, particularly in small-scale farms of low and middle-income countries where contact between species can be frequent. We compared the prevalence of fecal carriage of ESBL-E. coli among 332 livestock (207 cows, 15 pigs, 60 horses, 40 sheep, 6 goats, 4 chickens), 82 dogs, and wildlife including 131 European rabbits, 30 rodents, and 12 Andean foxes sharing territory in peri-urban localities of central Chile. The prevalence was lower in livestock (3.0%) and wildlife (0.5%) compared to dogs (24%). Among 47 ESBL-E. coli isolates recovered, CTX-M-group 1 was the main ESBL genotype identified, followed by CTX-M-groups 2, 9, 8, and 25. ERIC-PCR showed no cluster of E. coli clones by either host species nor locality. To our knowledge, this is the first report of ESBL-E. coli among sheep, cattle, dogs, and rodents of Chile, confirming their fecal carriage among domestic and wild animals in small-scale farms. The high prevalence of ESBL-E. coli in dogs encourages further investigation on their role as potential reservoirs of this bacteria in agricultural settingsItem Higher Prevalence of Extended-Spectrum Cephalosporin-Resistant Enterobacterales in Dogs Attended for Enteric Viruses in Brazil Before and After Treatment with Cephalosporins(2021) Salgado-Caxito, Marília; Moreno-Switt, Andrea I.; Paes, Antonio Carlos; Shiva, Carlos; Munita, José; Rivas, Lina; Benavides, Julio A.The extensive use of antibiotics is a leading cause for the emergence and spread of antimicrobial resistance (AMR) among dogs. However, the impact of using antibiotics to treat viral infections on AMR remains unknown. In this study, we compared the prevalence of extended-spectrum cephalosporin-resistant Enterobacterales (ESCR-E) between dogs with a suspected infection of canine parvovirus (CPV) and canine distemper (CDV) before and after treatment with third-generation cephalosporins. We found a higher prevalence of ESCR-E faecal carriage in dogs suspected of CPV (37%) and CDV (15%) compared to dogs with noninfectious pathologies (9%) even prior to the start of their treatment. A 7-day course of ceftriaxone or ceftiofur administrated to CPV and CDV-suspected dogs substantially increased their ESCR-E faecal carriage during treatment (85% for CPV and 57% for CDV), and 4 weeks after the treatment ended (89% for CPV and 60% for CDV) when dogs were back in their households. Most of the observed resistance was carried by ESCR-E. coli carrying blaCTX-M genes. Our results suggest the need to optimize prophylactic antibiotic therapy in dogs treated for a suspected viral infection to prevent ESCR-E emergence and spread in the communityItem Isolation of Ciprofloxacin and Ceftazidime-Resistant Enterobacterales From Vegetables and River Water Is Strongly Associated With the Season and the Sample Type(2021) Díaz-Gavidia, Constanza; Rivas, Lina; Barría, Carla; García, Patricia; Alvarez, Francisca P.; González-Rocha, Gerardo; Opazo-Capurro, Andrés; Araos, Rafael; Munita, José; Cortes, Sandra; Olivares-Pacheco, Jorge; Adell, Aiko; Moreno-Switt, AndreaThe dissemination of antibiotic-resistant bacteria (ARB) from water used for crop irrigation to vegetables is poorly studied. During a year, five farmer markets in a city in Central Chile were visited, and 478 vegetable samples (parsleys, corianders, celeries, lettuces, chards, and beets) were collected. Simultaneously, 32 water samples were collected from two rivers which are used to irrigate the vegetables produced in the area. Resistant Enterobacterales were isolated and identified. Colistin resistance gene mcr-1 and extended spectrum β-lactamases (ESBL) were molecularly detected. The association of environmental factors was evaluated, with the outcomes being the presence of Enterobacterales resistant to four antibiotic families and the presence of multidrug resistance (MDR) phenotypes. Parsley, coriander, and celery showed the highest prevalence of resistant Enterobacterales (41.9% for ciprofloxacin and 18.5% for ceftazidime). A total of 155 isolates were obtained, including Escherichia coli (n=109), Citrobacter sp. (n=20), Enterobacter cloacae complex (n=8), Klebsiella pneumoniae (n=8), and Klebsiella aerogenes (n=1). Resistance to ampicillin (63.2%) and ciprofloxacin (74.2%) was most frequently found; 34.5% of the isolates showed resistance to third-generation cephalosporins, and the MDR phenotype represented 51.6% of the isolates. In two E. coli isolates (1.29%), the gene mcr-1 was found and ESBL genes were found in 23/62 isolates (37%), with bla CTX-M being the most frequently found in 20 isolates (32%). Resistant Enterobacterales isolated during the rainy season were less likely to be MDR as compared to the dry season. Understanding environmental associations represent the first step toward an improved understanding of the public health impact of ARB in vegetables and water.Item Mechanical and Antimicrobial Polyethylene Composites with CaO Nanoparticles(2020) Silva, Cristián; Bobillier, Felipe; Canales, Daniel; Sepúlveda, Francesca Antonella; Cament, Alejandro; Amigo, Nicolás; Rivas, Lina; Ulloa, María T.; Reyes, Pablo; Ortiz, J. Andrés; Gómez, Tatiana; Loyo, Carlos; Zapata, Paula ALow-density polyethylene composites containing different sizes of calcium oxide (CaO) nanoparticles were obtained by melt mixing. The CaO nanoparticles were synthesized by either the sol-gel or sonication methods, obtaining two different sizes: ca. 55 nm and 25 nm. These nanoparticles were used either as-synthesized or were modified organically on the surface with oleic acid (Mod-CaO), at concentrations of 3, 5, and 10 wt% in the polymer. The Mod-CaO nanoparticles of 25 nm can act as nucleating agents, increasing the polymer’s crystallinity. The Young’s Modulus increased with the Mod-CaO nanoparticles, rendering higher reinforcement effects with an increase as high as 36%. The reduction in Escherichia coli bacteria in the nanocomposites increased with the amount of CaO nanoparticles, the size reduction, and the surface modification. The highest antimicrobial behavior was found in the composites with a Mod-CaO of 25 nm, presenting a reduction of 99.99%. This strong antimicrobial effect can be associated with the release of the Ca2+ from the composites, as studied for the composite with 10 wt% nanoparticles. The ion release was dependent on the size of the nanoparticles and their surface modification. These findings show that CaO nanoparticles are an excellent alternative as an antimicrobial filler in polymer nanocomposites to be applied for food packaging or medical devices.Publication Mechanical and antimicrobial properties of low-density-polyethylene/MgO nanocomposites(2022) Cament, Alejandro; Moreno-Serna, Viviana; Loyo, Carlos; Quintana, Pabla; Seura, Pablo; Vallejos Baier, Raúl; Benavente, Rosario; Ulloa, María Teresa; Rivas, Lina; Pino, Eduardo; Gómez, Tatiana; Zapata, Paula A.Low-density polyethylene (LDPE) nanocomposites containing magnesium oxide (MgO) nanoparticles are obtained by melt mixing. MgO nanoparticles ca. 29 ± 2 nm are synthesized by sol–gel and organically modified on the surface with oleic acid (Mod-MgO), whose final concentration in the polymers is 3, 5, and 10 wt%. The presence of these nanoparticles did not affect the crystallization process of LDPE. However, Young's modulus increases with 10 wt% of Mod-MgO nanoparticles, rendering higher reinforcement effects with an increase as high as 48%. This affects the elongation at break, which decreases ca. 57% compared to neat LDPE. The storage and loss modulus of the LDPE/MgO and LDPE/Mod-MgO nanocomposites increases at room temperature and low temperatures ( 150 to 50C) compared to neat LDPE. SEM analysis showed that the organic modification of MgO nanoparticles improved their dispersion within the polymer matrix. Nanocomposites present antimicrobial properties against Escherichia coli, reaching an efficiency ca. 53%.Publication PLA/CaO nanocomposites with antimicrobial and photodegradation properties(2022) Loyo, Carlos; Moreno-Serna, Viviana; Fuentes, Jairo; Amigo, Nicolás; Sepúlveda, Francesca Antonella; Ortiz, Andrés; Rivas, Lina; Ulloa, María Teresa; Benavente, Rosario; Zapata , Paula A.CaO nanoparticles sized ca. 26 nm were organically modified with oleic acid (Ol-CaO), and both were incorporated into PLA at concentrations of 5 and 8 wt.% by a melting process. Modification of nanoparticles improved the distribution into PLA, as seen by transmission electron microscopy (TEM). Thermal analysis revealed that the presence of Ol-CaO in the PLA matrix promoted a decrease ca. 13% in the glass transition temperature (Tg). The thermal stability of the PLA/Ol-CaO decreased ca. 23% compared to the neat PLA due to the catalytic activity from nanoparticles, while Vickers Microhardness (HV) for nanocomposites PLA/Ol-CaO increased ca. 9%, compared with the neat PLA, due to the good dispersion of modified-surface Ol-CaO nanoparticles in PLA. PLA/Ol-CaO nanocomposites reached 99.9% of antimicrobial effectiveness against E. coli for nanoparticles content above 8 wt.%. From photodegradation tests under irradiation during five days, it was verified that the presence of CaO nanoparticles accelerated the photodegradation of the polymer matrix nanoparticles into PLA promoted a decreasing ca. 13% of Tg and an increase in the degree of crystallinity (Xc) (ca. 7%), compared to PLA/CaO without irradiation. Besides, the viscosity molecular weight (M¯ v) of PLA/CaO showed a higher decrease than neat PLA after irradiation, and SEM analysis showed that the nanocomposites presented cavities around the nanoparticles after irradiation. Our results showed that incorporating CaO nanoparticles into the PLA polymer matrix allows future development of more sustainable materials as nanocomposites for food packaging or medical devices.Item Real-world Performance of Susceptibility Testing for Ceftolozane/Tazobactam Against Non-Carbapenemase-Producing Carbapenem-Resistant Pseudomonas aeruginosa(2021) Rivas, Lina; Alcalde-Rico, Manuel; Martínez, José R.; Moreno, Victoria; Rojas, Pamela; Wozniak, Aniela; García, Patricia; Olivares, Jorge; Miller, William R.; Arias, Cesar A.; Khan, Ayesha; Munita, JoséCeftolozane/tazbactam (C/T) is a potent anti-pseudomonal agent that has clinical utility against infections caused by non-carbapenemase producing carbapenem-resistant P. aeruginosa (non-CP-CR-PA). Accurate, precise and reliable antimicrobial susceptibility testing (AST) is crucial to guide clinical decisions. However, studies assessing the performance of different AST methods against non-CP-CR-PA- (the main clinical niche for C/T), are lacking. Here, we evaluated performance of gradient strips (Etest and MIC test strip (MTS), and disk diffusion (DD) using CLSI breakpoints. Additionally, we assessed the performance of DD using EUCAST breakpoints. For all susceptibility tests, we used a collection of 97 non-CP-CR-PA clinical isolates recovered from 11 Chilean hospitals. Both gradient strips and DD had acceptable performance when using CLSI breakpoints, yielding a categorical agreement (CA) of >90% and 92%, respectively. In contrast, DD using EUCAST breakpoints performed sub-optimally (CA 81%). MTS yielded a higher essential agreement (EA, >90%) than Etest (84%). Importantly, the performance of all methods varied significantly when the isolates were stratified by their degree of susceptibility to other anti-pseudomonal β-lactams. All methods had 100% CA when testing isolates that were pan-susceptible to all β-lactams (Pan-β-S). However, the CA markedly decreased when testing isolates resistant to all β-lactams (Pan-β-R). Indeed, the CA was 81% for Etest (6 errors), 78% for MTS (7 errors) and 78% and 56% for DD when using CLSI (7 errors) or EUCAST breakpoints (14 errors), respectively. Our results suggest that all manual AST methods have strikingly decreased performance in the context of Pan-β-R P. aeruginosa with potentially major clinical implications.Item Risk factors associated with faecal carriage of extended-spectrum cephalosporin-resistant Escherichia coli among dogs in Southeast Brazil(2021) Salgado-Caxito, Marília; Benavides, Julio A.; Munita, José; Rivas, Lina; García, Patricia; Listoni, Fernando J. P.; Paes, Antonio C.; Moreno-Switt, Andrea I.Faecal carriage of extended-spectrum cephalosporin-resistant Escherichia coli (ESC-R E. coli) in dogs has been reported worldwide and can reduce the effectiveness of treatments against bacterial infections. However, the drivers that influence faecal carriage of ESC-R E. coli in dogs are poorly understood. The aims of this study were to estimate the prevalence of ESC-R E. coli among dogs prior to their admission to a veterinary teaching hospital and to identify risk factors associated with the faecal carriage of ESC-R E. coli. Rectal swabs (n = 130) were collected from dogs and screened for ESC-R E. coli using MacConkey agar supplemented with cefotaxime (2 μg/mL). E. coli species was confirmed by MALDI-TOF and screening of extended-spectrum beta-lactamase (ESBL) genes was conducted by multiplex PCR. Questionnaires were completed by each dog's owner to test several human and dog characteristics associated with ESC-R E. coli. The prevalence of faecal carriage of ESC-R E. coli was 9.2 % and 67 % of ESC-R E. coli isolates harboured ESBL genes including CTX-M alone or in combination with TEM. All ESC-R E. coli isolates were resistant to ceftriaxone, cefpodoxime, and cefotaxime and were susceptible to cefoxitin and carbapenems. The likelihood of carrying ESC-R E. coli was 15 times higher (OR = 14.41 [95 % CI: 1.80-38.02], p < 0.01) if the dog was treated with antibiotics 3-12 months prior to sampling and 8 times higher (OR = 7.96 [95 % CI: 2.96-92.07], p < 0.01) if the dog had direct contact with livestock, but 15 times lower (OR = 0.07 [95 % CI: 0.01-0.32], p < 0.01) if the dog was dewormed during the previous year. Our findings confirm the faecal carriage of ESC-R E. coli in subclinical dogs and call for further investigation regarding the impact of deworming on antibiotic-resistant bacteria in companion animals.