Clonal dissemination of Acinetobacter radioresistens among Humboldt penguins (Spheniscus humboldti) inhabiting a barren northern Peruvian island.

Acinetobacter spp. are often isolated from natural sources, but knowledge about their presence in wild animals is fragmented and uncomplete. The present study aimed to characterize a series of Acinetobacter radioresistens isolated from Humboldt penguins (Spheniscus humboldti). Fifteen Humboldt penguins from an inhabited northern Peruvian island were sampled. Microorganisms were identified by MALDI-TOF MS. Antibiotic susceptibility to 12 antimicrobial agents was established, and clonal relationships were determined. A representative isolate was selected for whole genome sequencing (WGS). A. radioresistens were isolated from the feces of 12 (80%) Humboldt penguins, being susceptible to all the antimicrobial agents tested, except eight cefotaxime-intermediate isolates. All A. radioresistens were clonally related. WGS showed that the isolate belonged to ST1972, the presence of two chromosomal encoded carbapenemases (blaOXA-23 and a putative subclass B3 metallo-ß-lactamase), and a series of point mutations in antibiotic-resistance related chromosomal genes, which were considered as polymorphisms. In addition, a few virulence factors, including a capsule-encoding operon, superoxide dismutases, catalases, phospholipases and a siderophore receptor were identified. The present results suggest that A. radioresistens may be a common member of the gut microbiota of Humboldt penguins, but further studies in other geographical areas are needed to establish this finding.

Cytokine Profile Response of Human Peripheral Blood Mononuclear Cells Stimulated by Bartonella bacilliformis.

Carrion's disease is a neglected endemic disease found in remote Andean areas. As an overlooked disease, knowledge of innate immune responses to Bartonella bacilliformis, the etiological agent, is scarce. This study aimed to evaluate the cytokine response to B. bacilliformis using in vitro human peripheral blood mononuclear cells (PBMCs) stimulations. PBMCs from naive adults were isolated by gradient centrifugation and cocultured with heat-inactivated (HI) B. bacilliformis at different incubation times (3, 6, 12, 24, and 36 h). Cytokines, chemokines, and growth factors were determined in culture supernatants by multiplex fluorescent bead-based quantitative suspension array technology. During the first 36 h, a proinflammatory response was observed, including tumor necrosis factor-α, interleukin (IL)-1α, IL-1ß, interferon-α2, and IL-6, followed by an anti-inflammatory response mainly related to IL-1RA. Moreover, high expression levels of chemokines IL-8, monocyte chemoattractant protein-1α, and macrophage inflammatory protein (MIP)-1ß were detected from 3 h poststimulation and MIP-1α was detected at 24 h. Some growth factors, mainly granulocyte macrophage colony-stimulating factor and granulocyte colony-stimulating factor, and in minor concentrations vascular endothelial growth factor, epidermal growth factor, and eotaxin, were also detected. Innate response to HI B. bacilliformis stimulation consists of a rapid and strong proinflammatory response characterized by a wide range of cytokines and chemokines followed by an anti-inflammatory response and increased specific growth factors.

Dengue and COVID-19 Co-Circulation in the Peruvian Amazon: A Population-Based Study.

The COVID-19 pandemic affected the main Amazon cities dramatically, with Iquitos City reporting the highest seroprevalence of anti-SARS-CoV-2 antibodies during the first COVID-19 wave worldwide. This phenomenon raised many questions about the possibility of a co-circulation of dengue and COVID-19 and its consequences. We carried out a population-based cohort study in Iquitos, Peru. We obtained a venous blood sample from a subset of 326 adults from the Iquitos COVID-19 cohort (August 13-18, 2020) to estimate the seroprevalence of anti-dengue virus (DENV) and anti-SARS-CoV-2 antibodies. We tested each serum sample for anti-DENV IgG (serotypes 1, 2, 3, and 4) and SARS-CoV-2 antibodies anti-spike IgG and IgM by ELISA. We estimated an anti-SARS-CoV-2 seroprevalence of 78.0% (95% CI, 73.0-82.0) and an anti-DENV seroprevalence of 88.0% (95% CI, 84.0-91.6), signifying a high seroprevalence of both diseases during the first wave of COVID-19 transmission in the city. The San Juan District had a lower anti-DENV antibody seroprevalence than the Belen District (prevalence ratio, 0.90; 95% CI, 0.82-0.98). However, we did not observe these differences in anti-SARS-CoV-2 antibody seroprevalence. Iquitos City presented one of the highest seroprevalence rates of anti-DENV and anti-SARS-CoV-2 antibodies worldwide, but with no correlation between their antibody levels.

KPC-2-NDM-1-producing Serratia marcescens: first description in Peru.

Two clonal extensively-resistant S. marcescens were isolated in Peru. The presence and transferability of extended-spectrum ß-lactamases and IMP, KPC, NDM, OXA-48 and VIM were determined. The concomitant presence of KPC-2 and NDM-1 was confirmed in both isolates, with KPC-2 being able to be transferred.

Zonulin, a marker of gut permeability, is associated with mortality in a cohort of hospitalised peruvian COVID-19 patients.

Zonulin has previously been related to intestinal permeability in various inflammatory diseases, and more recently to the physiopathology of severe COVID-19 infections. We analysed serum samples from a previous study of a Peruvian cohort of hospitalised COVID-19 patients, for the quantification of zonulin by sandwich ELISA. Comparisons with clinical data, haematological and biochemical parameters and cytokine/chemokine levels were made. We found higher baseline zonulin levels in deceased patients, and zonulin was associated with fatal outcome in multivariable analyses, even after adjustment for age, gender, and obesity. There were also positive correlations between zonulin, creatinine, D-dimer values and prothrombin time, while inverse correlations were found for Sa/FiO2 ratio and CCL5 (RANTES). Further longitudinal studies are recommended to analyse the variation of zonulin levels over time as well as their relationship with long-COVID.

Worrying levels of antimicrobial resistance in Gram-negative bacteria isolated from cell phones and uniforms of Peruvian intensive care unit workers.

BACKGROUND: Healthcare worker (HCW) uniforms and cell phones are involved in pathogen transmission. This study aimed to characterize pathogenic microorganism isolates from HCW uniforms and cell phones. METHODS: Gram-negative microorganisms were recovered from HCW uniforms and cell phones. Antimicrobial susceptibility and the presence of extended-spectrum ß-lactamases (ESBL) and carbapenemases were determined. RESULTS: Escherichia coli was the most prevalent microorganism. Overall, high levels of resistance to cephalosporins, quinolones, co-trimoxazole and colistin were found. ESBL were mainly related to blaCTX-M-15 and blaSHV- genes. Carbapenem-resistant isolates presented as blaKPC or blaNDM. CONCLUSIONS: High levels of antimicrobial resistance, including colistin, were detected. Therefore, strategies are urgently needed to prevent bacterial dissemination.

Corrigendum: Zonulin, a marker of gut permeability, is associated with mortality in a cohort of hospitalised peruvian COVID-19 patients.

[This corrects the article DOI: 10.3389/fcimb.2022.1000291.].

Cytokine Profiles Associated With Worse Prognosis in a Hospitalized Peruvian COVID-19 Cohort.

Cytokines, chemokines and growth factors present different expression profiles related to the prognosis of COVID-19. We analyzed clinical parameters and assessed the expression of these biomarkers in patients with different disease severity in a hospitalized Peruvian cohort to determine those associated with worse prognosis. We measured anti-spike IgG antibodies by ELISA and 30 cytokines by quantitative suspension array technology in 123 sera samples. We analyzed differences between patients with moderate, severe and fatal COVID-19 by logistic regression at baseline and in longitudinal samples. Significant differences were found among the clinical parameters: hemoglobin, neutrophils, lymphocytes and C-reactive protein (CRP), creatinine and D-dimer levels. Higher anti-spike IgG antibody concentrations were associated to fatal patient outcomes. At hospitalization, IL-10, IL-6, MIP-1α, GM-CSF, MCP-1, IL-15, IL-5, IL1RA, TNFα and IL-8 levels were already increased in fatal patients´ group. Meanwhile, multivariable analysis revealed that increased GM-CSF, MCP-1, IL-15, and IL-8 values were associated with fatal outcomes. Moreover, longitudinal analysis identified IL-6 and MCP-1 as the main risk factors related to mortality in hospitalized COVID-19 patients. In this Peruvian cohort we identified and validated biomarkers related to COVID-19 outcomes. Further studies are needed to identify novel criteria for stratification of SARS-CoV-2 infected patients at hospital entry. Background: In the most severe forms of SARS-CoV-2 infection, large numbers of innate and adaptive immune cells become activated and begin to produce pro-inflammatory cytokines, establishing an exacerbated feedback loop of inflammation. Methods: A total of 55 patients with laboratory-confirmed COVID-19 admitted to the Hospital Nacional Guillermo Almenara Irigoyen in Lima, Peru were enrolled during August-October 2020. Of these, 21 had moderate disease, 24 severe diseases and 10 died. We measured 30 cytokines and chemokines by quantitative suspension array technology and anti-spike IgG antibodies using a commercial ELISA. We evaluated these parameters in peripheral blood every 2-5 days until patient discharge or death. Patient information and clinical parameters related were obtained from the respective clinical histories. Results: The frequency of obesity differed among the 3 groups, being most frequent in patients who died. There were also significant differences in clinical parameters: hemoglobin, segmented neutrophils, lymphocytes,C-reactive protein, creatinine and D-dimer levels. Greater anti-spike IgG antibody concentrations were associated to fatal outcomes. In univariate analyses, higher baseline concentrations of IL-6, MIP-1α, GM-CSF, MCP-1, IL-15, IL-5, IL1RA, TNFα, IL-8 and IL-12p70 correlated with severity, while multivariable analysis showed that increased concentrations in 4 biomarkers (GM-CSF, MCP-1, IL-15, IL-8) were associated with fatal outcomes. Longitudinal analysis showed IL-6 (hazard ratio [HR] 6.81, 95% confidence interval [CI] 1.6-28.7) and MCP-1 (HR 4.61, 95%CI 1.1-19.1) to be related to mortality in hospitalized COVID-19 patients. Conclusions: Cytokine, chemokine and growth factor profiles were identified and validated related to severity and outcomes of COVID-19. Our findings may be useful to identify novel criteria for COVID-19 patient stratification at hospital entry.

Tolerance to disinfectants (chlorhexidine and isopropanol) and its association with antibiotic resistance in clinically-related Klebsiella pneumoniae isolates.

Disinfectants play an essential role in controlling the dissemination of bacteria in health care settings, but it may also contribute to the selection of antibiotic resistance bacteria. This study looked at Klebsiella pneumoniae isolates collected from three hospitals in Lima, Peru, in order to evaluate: their susceptibility to chlorhexidine [CHG] and isopropanol [ISP]), and their association with antimicrobial susceptibility. We analyzed 59 K. pneumoniae isolates and assessed their CHG and ISP susceptibility by minimum inhibitory concentrations (MICs). Additionally, we performed a regression analysis to assess the association between disinfectant tolerance and antibiotic resistance (measured by the disc diffusion method), colistin resistance (by microdilution), carbapenemases presence (by polymerase chain reaction [PCR]), and clonal relationships (by pulsed-field gel electrophoresis [PFGE]). Eleven K. pneumoniae strains were isolated from fomites, and 48 strains from clinical samples. The MIC range of these isolates was 8-128 µg/ml for CHG and 16-256 mg/ml for ISP. We found that resistance to trimethoprim/sulfamethoxazole (TMP/SMX) was the main factor associated with CHG log2 MIC (ß = 0.65; 95%CI: 0.03, 1.27; R2 = 0.07). In the case of ISP, the log2(MIC) was associated with the institution of origin, showing lower ISP log2(MIC) in fomites compared to clinical samples(ß = -0.77; 95%CI: -1.54, -0.01; R2 = 0.08). Resistance to CHG and ISP among K. pneumoniae isolates found in Peruvian hospitals seems to be elevated and highly variable. Further studies are needed to confirm our results and implement actionable interventions if necessary.

Resistencia a la colistina en cepas de Klebsiella pneumoniae multidrogorresistente del período 2015-2018 en un instituto materno perinatal de Lima, Perú / Colistin resistance in multidrug-resistant Klebsiella pneumoniae strains at a perinatal maternal institute in Lima, Peru, 2015-2018

RESUMEN El objetivo del estudio fue evaluar los niveles y mecanismos de resistencia a la colistina y a los carbapenémicos en cepas de Klebsiella pneumoniae multidrogorresistente aisladas durante el periodo 2015-2018 en el Instituto Materno Perinatal de Lima. Se analizó la sensibilidad mediante difusión en disco y microdilución. La presencia de genes de resistencia a los carbapenémicos y a la colistina se determinó por reacción en cadena de la polimerasa (PCR, por sus siglas en inglés) y se la relacionó con la clonalidad. Se analizaron 36 cepas de K. pneumoniae, cinco (13,8%) fueron resistentes a la colistina, pertenecían a diferentes grupos clonales. Se encontraron dos cepas con carbapenemasas (bla KPC y bla NDM) y no se detectaron genes plasmídicos para la colistina. Los niveles de resistencia al resto de antimicrobianos testados fueron elevados, a excepción de amikacina (13,9%). Los resultados destacan la presencia de cepas resistentes a la colistina (33,3% en 2018), situación preocupante por ser esta parte de las últimas alternativas de tratamiento para las infecciones causadas por patógenos multirresistentes.

ABSTRACT The objective of this study was to evaluate the presence of colistin- and carbapenemic-resistant genes in multidrug-resistant Klebsiella pneumoniae strains isolated at the Instituto Materno Perinatal de Lima (2015-2018). Susceptibility levels were analyzed by disk diffusion and microdilution. The presence of colistin- and carbapenemic-resistant genes was determined by polymerase chain reaction (PCR) and was related to clonality. A total of 36 K. pneumoniae strains were analyzed, 5 (13.8%) were resistant to colistin and belonged to different clonal groups. Only 2 strains were found with carbapenemases (bla KPC and bla NDM), and no colistin plasmid genes were detected. High resistance levels to the other tested antimicrobials were observed, except for amikacin (13.9%). The results highlight the presence of colistin-resistant strains (33.3% in 2018), a worrying situation as they are part of the latest treatment alternatives for infections caused by multiresistant pathogens.

Spread of ST348 Klebsiella pneumoniae Producing NDM-1 in a Peruvian Hospital.

The aim of this study was to characterize carbapenem-resistant Klebsiella pneumoniae (CR-Kp) isolates recovered from adults and children with severe bacteremia in a Peruvian Hospital in June 2018. Antimicrobial susceptibility was determined by disc/gradient diffusion and broth microdilution when necessary. Antibiotic resistance mechanisms were evaluated by PCR and DNA sequencing. Clonal relatedness was assessed using pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Plasmid typing was performed with a PCR-based method. Thirty CR-Kp isolates were recovered in June 2018. All isolates were non-susceptible to all ß-lactams, ciprofloxacin, gentamicin and trimethoprim-sulfamethoxazole, while mostly remaining susceptible to colistin, tigecycline, levofloxacin and amikacin. All isolates carried the blaNDM-1 gene and were extended spectrum ß-lactamase (ESBL) producers. PFGE showed four different pulsotypes although all isolates but two belonged to the ST348 sequence type, previously reported in Portugal. blaNDM-1 was located in an IncFIB-M conjugative plasmid. To our knowledge, this is the first report of an New Delhi metallo-ß-lactamase (NDM)-producing K. pneumoniae recovered from both children and adults in Lima, Peru, as well as the first time that the outbreak strain ST348 is reported in Peru and is associated with NDM. Studies providing epidemiological and molecular data on CR-Kp in Peru are essential to monitor their dissemination and prevent further spread.

Colistin resistance in multidrug-resistant Klebsiella pneumoniae strains at a perinatal maternal institute in Lima, Peru, 2015-2018. / Resistencia a la colistina en cepas de Klebsiella pneumoniae multidrogorresistente del período 2015-2018 en un instituto materno perinatal de Lima, Perú.

The objective of this study was to evaluate the presence of colistin- and carbapenemic-resistant genes in multidrug-resistant Klebsiella pneumoniae strains isolated at the Instituto Materno Perinatal de Lima (2015-2018). Susceptibility levels were analyzed by disk diffusion and microdilution. The presence of colistin- and carbapenemic-resistant genes was determined by polymerase chain reaction (PCR) and was related to clonality. A total of 36 K. pneumoniae strains were analyzed, 5 (13.8%) were resistant to colistin and belonged to different clonal groups. Only 2 strains were found with carbapenemases (bla KPC and bla NDM), and no colistin plasmid genes were detected. High resistance levels to the other tested antimicrobials were observed, except for amikacin (13.9%). The results highlight the presence of colistin-resistant strains (33.3% in 2018), a worrying situation as they are part of the latest treatment alternatives for infections caused by multiresistant pathogens.

El objetivo del estudio fue evaluar los niveles y mecanismos de resistencia a la colistina y a los carbapenémicos en cepas de Klebsiella pneumoniae multidrogorresistente aisladas durante el periodo 2015-2018 en el Instituto Materno Perinatal de Lima. Se analizó la sensibilidad mediante difusión en disco y microdilución. La presencia de genes de resistencia a los carbapenémicos y a la colistina se determinó por reacción en cadena de la polimerasa (PCR, por sus siglas en inglés) y se la relacionó con la clonalidad. Se analizaron 36 cepas de K. pneumoniae, cinco (13,8%) fueron resistentes a la colistina, pertenecían a diferentes grupos clonales. Se encontraron dos cepas con carbapenemasas (bla KPC y bla NDM) y no se detectaron genes plasmídicos para la colistina. Los niveles de resistencia al resto de antimicrobianos testados fueron elevados, a excepción de amikacina (13,9%). Los resultados destacan la presencia de cepas resistentes a la colistina (33,3% en 2018), situación preocupante por ser esta parte de las últimas alternativas de tratamiento para las infecciones causadas por patógenos multirresistentes.