Cytokine expression and mortality risk among COVID-19 hospitalized patients over 60 years of age in a referral hospital in Cartagena, Colombia.

Background: Cytokine dysregulation in COVID-19 patients aged over 60 has been associated to adverse outcomes. While serum levels have been studied, cellular expression, particularly in Afro-Colombians, remains understudied. This research aims to describe cytokine expression in peripheral blood leukocytes and its association with adverse outcomes in COVID-19 patients aged over 60 at Cartagena's referral hospital. Methods: A cohort study was conducted, encompassing severe and critical cases of COVID-19 between November 2021 and February 2022. At baseline, the cellular expression level of cytokines IL-2, IL-4, IL-6, IL-10, TNF-α and IFN-γ was assessed using flow cytometry. Additionally, various biochemical, hematological, and coagulation markers were evaluated. The main outcome was time to death. Results: Among the 50 enrolled participants, the median age was 76.5 years, 60% were male, 60% were admitted to the ICU, and 42% died. Lactate dehydrogenase and hemoglobin were the only markers that differed between fatal and surviving cases. Regarding cytokines, the level of IL-6 expression was associated with an increased risk of death. Specifically, a one percent increase in the expression was associated with a 7.3% increase in the risk of death. Stratifying the analysis by death and ICU admission, the median expression level remained high in fatal cases who were admitted to the ICU. Conclusions: Our findings revealed a significant association between high cellular expression levels of IL-6 and an increased risk of mortality. These results provide valuable scientific insights that could inform the prioritization of case management, providing especially advantageous for the vulnerable Afro-Colombian group.

Sampling is decisive to determination of Leishmania (Viannia) species.

BACKGROUND: Accuracy of molecular tools for the identification of parasites that cause human cutaneous leishmaniasis (CL) could largely depend on the sampling method. Non-invasive or less-invasive sampling methods such as filter paper imprints and cotton swabs are preferred over punch biopsies and lancet scrapings for detection methods of Leishmania based on polymerase chain reaction (PCR) because they are painless, simple, and inexpensive, and of benefit to military and civilian patients to ensure timely treatment. However, different types of samples can generate false negatives and there is a clear need to demonstrate which sample is more proper for molecular assays. METHODOLOGY: Here, we compared the sensitivity of molecular identification of different Leishmania (Viannia) species from Peru, using three types of sampling: punch biopsy, filter paper imprint and lancet scraping. Different composite reference standards and latent class models allowed to evaluate the accuracy of the molecular tools. Additionally, a quantitative PCR assessed variations in the results and parasite load in each type of sample. PRINCIPAL FINDINGS: Different composite reference standards and latent class models determined higher sensitivity when lancet scrapings were used for sampling in the identification and determination of Leishmania (Viannia) species through PCR-based assays. This was consistent for genus identification through kinetoplastid DNA-PCR and for the determination of species using FRET probes-based Nested Real-Time PCR. Lack of species identification in some samples correlated with the low intensity of the PCR electrophoretic band, which reflects the low parasite load in samples. CONCLUSIONS: The type of clinical sample can directly influence the detection and identification of Leishmania (Viannia) species. Here, we demonstrated that lancet scraping samples consistently allowed the identification of more leishmaniasis cases compared to filter paper imprints or biopsies. This procedure is inexpensive, painless, and easy to implement at the point of care and avoids the need for anesthesia, surgery, and hospitalization and therefore could be used in resource limited settings for both military and civilian populations.

Molecular characterization of carbapenemase-producing Enterobacterales in a tertiary hospital in Lima, Peru.

Carbapenemase-producing Enterobacterales (CPE) are a growing threat to global health and the economy. Understanding the interactions between resistance and virulence mechanisms of CPE is crucial for managing difficult-to-treat infections and informing outbreak prevention and control programs. Here, we report the characterization of 21 consecutive, unique clinical isolates of CPE collected in 2018 at a tertiary hospital in Lima, Peru. Isolates were characterized by phenotypic antimicrobial susceptibility testing and whole-genome sequencing to identify resistance determinants and virulence factors. Seven Klebsiella pneumoniae isolates were classified as extensively drug-resistant. The remaining Klebsiella, Enterobacter hormaechei, and Escherichia coli isolates were multidrug-resistant. Eighteen strains carried the metallo-ß-lactamase NDM-1, two the serine-carbapenemase KPC-2, and one isolate had both carbapenemases. The blaNDM-1 gene was located in the truncated ΔISAba125 element, and the blaKPC-2 gene was in the Tn4401a transposon. ST147 was the most frequent sequence type among K. pneumoniae isolates. Our findings highlight the urgent need to address the emergence of CPE and strengthen control measures and antibiotic stewardship programs in low- and middle-income settings.IMPORTANCEGenomic surveillance of antimicrobial resistance contributes to monitoring the spread of resistance and informs treatment and prevention strategies. We characterized 21 carbapenemase-producing Enterobacterales collected at a Peruvian tertiary hospital in 2018, which exhibited very high levels of resistance and carried numerous resistance genes. We detected the coexistence of carbapenemase-encoding genes (blaNDM-1 and blaKPC-2) in a Klebsiella pneumoniae isolate that also had the PmrB(R256G) mutation associated with colistin resistance. The blaKPC-2 genes were located in Tn4401a transposons, while the blaNDM-1 genes were in the genetic structure Tn125 (ΔISAba125). The presence of high-risk clones among Klebsiella pneumoniae (ST11 and ST147) and Escherichia coli (ST410) isolates is also reported. The study reveals the emergence of highly resistant bacteria in a Peruvian hospital, which could compromise the effectiveness of current treatments and control.

Prediction models of COVID-19 fatality in nine Peruvian provinces: A secondary analysis of the national epidemiological surveillance system.

There are initiatives to promote the creation of predictive COVID-19 fatality models to assist decision-makers. The study aimed to develop prediction models for COVID-19 fatality using population data recorded in the national epidemiological surveillance system of Peru. A retrospective cohort study was conducted (March to September of 2020). The study population consisted of confirmed COVID-19 cases reported in the surveillance system of nine provinces of Lima, Peru. A random sample of 80% of the study population was selected, and four prediction models were constructed using four different strategies to select variables: 1) previously analyzed variables in machine learning models; 2) based on the LASSO method; 3) based on significance; and 4) based on a post-hoc approach with variables consistently included in the three previous strategies. The internal validation was performed with the remaining 20% of the population. Four prediction models were successfully created and validate using data from 22,098 cases. All models performed adequately and similarly; however, we selected models derived from strategy 1 (AUC 0.89, CI95% 0.87-0.91) and strategy 4 (AUC 0.88, CI95% 0.86-0.90). The performance of both models was robust in validation and sensitivity analyses. This study offers insights into estimating COVID-19 fatality within the Peruvian population. Our findings contribute to the advancement of prediction models for COVID-19 fatality and may aid in identifying individuals at increased risk, enabling targeted interventions to mitigate the disease. Future studies should confirm the performance and validate the usefulness of the models described here under real-world conditions and settings.

Development and validation of a vero cell-based suspension method for the detection of Zika virus. / Desarrollo y validación de un método de suspensión basado en células Vero para la detección del virus Zika.

OBJECTIVE.: To develop and validate a cell suspension method using Vero 76 cells for culturing Zika virus (ZIKV) based on infection of detached freshly seeded cells. MATERIAL AND METHODS.: Three different multiplicities of infection of ZIKV were used to develop and compare this novel method to the standard confluent cell monolayer method. In addition, we preliminary validated the cell suspension method using well-characterized ZIKV positive and negative clinical samples. The standard confluent cell monolayer method was used as the reference method, and viral isolation was confirmed by a ZIKV-specific RT-PCR. The sensitivity and its 95% confidence intervals for the cell suspension method were estimated. Also, a technical comparison of the cell suspension method against the cell monolayer method was performed. RESULTS.: Our findings suggested that both the viral load and replication of ZIKV were comparable between both monolayer- and suspension-infection methods. Although both methods were suitable for culturing and isolating ZIKV, the cell suspension method was easier, cheaper, and quicker as well as a sensitive isolation technique. The cell suspension method was significantly more sensitive in detecting Zika in inconclusive cases by RT-PCR, with a fourfold increase compared to the confluent cell monolayer method. CONCLUSION.: The cell suspension method has the potential to be an effective method for cultivating and isolating ZIKV and its application is potentially useful in both research and clinical settings.

OBJETIVO: . Desarrollar y validar un método de suspensión celular utilizando células Vero 76 para el cultivo del virus Zika (ZIKV) basado en la infección de células recién sembradas no adheridas. MATERIAL Y MÉTODOS: . Se utilizaron tres multiplicidades de infección diferentes del ZIKV para desarrollar y comparar este novedoso método con el método estándar de monocapa de células confluentes. Además, validamos preliminarmente el método de suspensión utilizando muestras clínicas caracterizadas como positivas o negativas para el ZIKV. El método estándar de monocapa se utilizó como método de referencia, y el aislamiento viral se confirmó mediante un RT-PCR específico del ZIKV. Se estimó la sensibilidad e intervalos de confianza del 95% para el método de suspensión. Asimismo, se realizó una comparación técnica del método de suspensión contra el método de monocapa. RESULTADOS: . Nuestros hallazgos sugieren que tanto la carga viral como la replicación del ZIKV fueron comparables entre los métodos de infección en monocapa y en suspensión. Aunque ambos métodos fueron adecuados para cultivar y aislar el ZIKV, el método de suspensión se caracterizó por ser más fácil, barato y rápido, así como una técnica de aislamiento sensible. En comparación con el método de monocapa, el método de suspensión fue cuatro veces más sensible en la detección del ZIKV en casos inconclusos por RT-PCR. CONCLUSIONES: . El método de suspensión tiene el potencial de ser un método eficaz para cultivar y aislar el ZIKV y su uso es potencialmente útil tanto en la investigación como en entornos clínicos.

Desarrollo y validación de un método de suspensión basado en células Vero para la detección del virus Zika / Development and validation of a vero cell-based suspension method for the detection of Zika virus

Objetivo . Desarrollar y validar un método de suspensión celular utilizando células Vero 76 para el cultivo del virus Zika (ZIKV) basado en la infección de células recién sembradas no adheridas. Material y métodos . Se utilizaron tres multiplicidades de infección diferentes del ZIKV para desarrollar y comparar este novedoso método con el método estándar de monocapa de células confluentes. Además, validamos preliminarmente el método de suspensión utilizando muestras clínicas caracterizadas como positivas o negativas para el ZIKV. El método estándar de monocapa se utilizó como método de referencia, y el aislamiento viral se confirmó mediante un RT-PCR específico del ZIKV. Se estimó la sensibilidad e intervalos de confianza del 95% para el método de suspensión. Asimismo, se realizó una comparación técnica del método de suspensión contra el método de monocapa. Resultados . Nuestros hallazgos sugieren que tanto la carga viral como la replicación del ZIKV fueron comparables entre los métodos de infección en monocapa y en suspensión. Aunque ambos métodos fueron adecuados para cultivar y aislar el ZIKV, el método de suspensión se caracterizó por ser más fácil, barato y rápido, así como una técnica de aislamiento sensible. En comparación con el método de monocapa, el método de suspensión fue cuatro veces más sensible en la detección del ZIKV en casos inconclusos por RT-PCR. Conclusiones . El método de suspensión tiene el potencial de ser un método eficaz para cultivar y aislar el ZIKV y su uso es potencialmente útil tanto en la investigación como en entornos clínicos.

Objective. To develop and validate a cell suspension method using Vero 76 cells for culturing Zika virus (ZIKV) based on infection of detached freshly seeded cells. Material and methods. Three different multiplicities of infection of ZIKV were used to develop and compare this novel method to the standard confluent cell monolayer method. In addition, we preliminary validated the cell suspension method using well-characterized ZIKV positive and negative clinical samples. The standard confluent cell monolayer method was used as the reference method, and viral isolation was confirmed by a ZIKV-specific RT-PCR. The sensitivity and its 95% confidence intervals for the cell suspension method were estimated. Also, a technical comparison of the cell suspension method against the cell monolayer method was performed. Results. Our findings suggested that both the viral load and replication of ZIKV were comparable between both monolayer- and suspension-infection methods. Although both methods were suitable for culturing and isolating ZIKV, the cell suspension method was easier, cheaper, and quicker as well as a sensitive isolation technique. The cell suspension method was significantly more sensitive in detecting Zika in inconclusive cases by RT-PCR, with a fourfold increase compared to the confluent cell monolayer method. Conclusion. The cell suspension method has the potential to be an effective method for cultivating and isolating ZIKV and its application is potentially useful in both research and clinical settings.

Factors Associated with COVID-19 Death in a High-Altitude Peruvian Setting during the First 14 Months of the Pandemic: A Retrospective Multicenter Cohort Study in Hospitalized Patients.

Risk factors for COVID-19 death in high-altitude populations have been scarcely described. This study aimed to describe risk factors for COVID-19 death in three referral hospitals located at 3399 m in Cusco, Peru, during the first 14 months of the pandemic. A retrospective multicenter cohort study was conducted. A random sample of ~50% (1225/2674) of adult hospitalized patients who died between 1 March 2020 and 30 June 2021 was identified. Of those, 977 individuals met the definition of death by COVID-19. Demographic characteristics, intensive care unit (ICU) admission, invasive respiratory support (IRS), disease severity, comorbidities, and clinical manifestation at hospital admission were assessed as risk factors using Cox proportional-hazard models. In multivariable models adjusted by age, sex, and pandemic periods, critical disease (vs. moderate) was associated with a greater risk of death (aHR: 1.27; 95%CI: 1.14-1.142), whereas ICU admission (aHR: 0.39; 95%CI: 0.27-0.56), IRS (aHR: 0.37; 95%CI: 0.26-0.54), the ratio of oxygen saturation (ROX) index ≥ 5.3 (aHR: 0.87; 95%CI: 0.80-0.94), and the ratio of SatO2/FiO2 ≥ 122.6 (aHR: 0.96; 95%CI: 0.93-0.98) were associated with a lower risk of death. The risk factors described here may be useful in assisting decision making and resource allocation.

Epidemiological, Clinical, and Virological Investigation of the First Four Cases of Monkeypox in Cartagena during the 2022 Outbreak.

Since early May 2022, numerous cases of Monkeypox (Mpox) have been reported globally in non-endemic areas. However, despite numerous reports worldwide, the epidemiological and genomic information related to the 2022 multi-country outbreak remains scarce in South America. By late June 2022, the first Mpox cases were detected in Colombia. Cartagena is a Colombian Caribbean city with high domestic and international connectivity, and, therefore, is vulnerable to the introduction of the Monkeypox virus (MPXV). This report provides an in-depth description of the epidemiological, clinical, and virological characteristics of the first four cases detected in Cartagena including three cases with no history of recent travel and one imported case. Using various laboratory tools based on PCR, next-generation sequencing, and viral isolation and quantification methods, the MPXV clade IIB was detected and isolated. Importantly, infectious viral particles were identified in lesion swabs collected from all cases and in oropharyngeal swabs collected from two cases. Blood samples tested negative using PCR and isolation. In summary, our work contributes complete genomic, clinical, and epidemiological information that will be useful for a number of studies going forward, and it also documents local information that contributes to our understanding of Mpox at the local level.

Detection of Zika Virus in Oropharyngeal Swabs from Patients with Acute Febrile Illness.

The isolation of Zika virus (ZIKV) from serum of suspected human cases for diagnostic purposes can be challenging due to infrastructure constraints of laboratory testing technology. Therefore, as an alternative method, the objective of this study was to evaluate a random sample of oropharyngeal swabs for the diagnosis of ZIKV infection among patients with symptoms of arboviral and respiratory illness. The results revealed that ZIKV RNA could be detected by a reverse transcriptase polymerase chain reaction (RT-PCR) assay and isolated from oropharyngeal swabs from five of 38 samples, but serum samples from the same patients were negative for ZIKV by a variety of laboratory diagnostic approaches including RT-PCR and viral isolation followed by immunofluorescence assays. The findings suggested that the molecular detection and isolation of ZIKV in oropharyngeal swab warrants further study for consideration as an improved diagnostic procedure.

Climatic factors and the incidence of dengue in Cartagena, Colombian Caribbean Region.

BACKGROUND: The influence of climate on the epidemiology of dengue has scarcely been studied in Cartagena. METHODS: The relationship between dengue cases and climatic and macroclimatic factors was explored using an ecological design and bivariate and time-series analyses during lag and non-lag months. Data from 2008-2017 was obtained from the national surveillance system and meteorological stations. RESULTS: Cases correlated only with climatic variables during lag and non-lag months. Decreases in precipitation and humidity and increases in temperature were correlated with an increase in cases. CONCLUSIONS: Our findings provide useful information for establishing and strengthening dengue prevention and control strategies.

Actividad antimicrobiana in vitro del extracto de hojas de Bixa orellana L. contra bacterias anaerobias asociadas a la vaginosis bacteriana y Lactobacillus spp / In vitro antimicrobial activity of Bixa orellana L. leaves extract against anaerobic bacteria associated to bacterial vaginosis and Lactobacillus spp.

Objetivos . Describir la actividad antimicrobiana in vitro del extracto metanólico de las hojas de Bixa orellana L. contra las bacterias anaerobias asociadas a la vaginosis bacteriana y Lactobacillus spp. Materiales y métodos . Se incluyeron en el estudio ocho cepas de referencia ATCC; Gardnerella vaginalis, Prevotella bivia, Peptococcus niger, Peptostreptococcus anaerobius, Mobiluncus curtisii, Atopobium vaginae, Veillonella parvula y Lactobacillus crispatus, y 22 aislamientos clínicos; once aislados de Gardnerella vaginalis y once aislados de Lactobacillus. La susceptibilidad antimicrobiana se determinó mediante el método de difusión en agar. La concentración mínima inhibitoria (CMI) y la concentración bactericida mínima (CBM) fueron determinadas utilizando el método de dilución en agar y un método de dilución modificado, respectivamente. Resultados . Todas las cepas de referencia ATCC tuvieron un alto nivel de susceptibilidad al extracto, con excepción de P. vibia, V. parvula y L. crispatus. Interesantemente, los aislamientos clínicos de G. vaginalis y la cepa ATCC de G. vaginalis fueron los más susceptibles al extracto dados los bajos valores de CMI (1,0 - 2,0 mg/mL) y CBM (1,0 - 4,0 mg/mL), mientras que, los aislamientos clínicos de Lactobacillus spp. y la cepa ATCC de L. crispatus fueron los menos susceptibles debido a los altos valores de CMI (32,0 mg/mL) y CBM (≥ 32,0 mg/mL). Conclusiones . Los experimentos in vitro sugieren que el extracto posee propiedades antibacterianas selectivas dada su alta actividad contra bacterias anaerobias asociadas a vaginosis bacteriana y baja actividad contra especies de Lactobacillus.

Objective. To describe the in vitro antimicrobial activity of the methanolic extract of Bixa orellana L. leaves against anaerobic bacteria associated to bacterial vaginosis and Lactobacillus spp. Materials and methods. Eight ATCC reference strains; Gardnerella vaginalis, Prevotella bivia, Peptococcus niger, Peptostreptococcus anaerobius, Mobiluncus curtisii, Atopobium vaginae, Veillonella parvula, and Lactobacillus crispatus, and twenty-two clinical isolates; eleven Gardnerella vaginalis and eleven Lactobacillus strains, were included in the study. The antimicrobial susceptibility was determined by the agar diffusion method. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined by using agar dilution and a modified dilution plating method, respectively. Results. All ATCC reference strains showed high levels of susceptibility to the extract, except P. vibia, V. parvula and L. crispatus. Interestingly, all G. vaginalis clinical isolates and the G. vaginalis ATTC strain were the most susceptible to the extract, given their low MIC (1.0 - 2.0 mg/mL) and MBC (1.0 - 4.0 mg/mL) values, whereas, the Lactobacillus spp. clinical isolates and the L. crispatus ATCC strain were the least susceptible bacteria given their high MIC (32.0 mg/mL) and MBC (≥ 32.0 mg/mL) values. Conclusions. In vitro experiments suggest that the extract possesses selective antimicrobial properties given its high activity against bacterial vaginosis-associated anaerobic bacteria and low activity against Lactobacillus species.

Epidemiology of COVID-19 in Individuals under 18 Years Old in Cartagena, Colombia: An Ecological Study of the First 14 Months of the Pandemic.

The epidemiology of the coronavirus disease (COVID-19) has been scarcely described in individuals under 18 years old, particularly during the first months of the pandemic. The study aimed to describe the COVID-19 epidemiology in the Colombian department of Bolívar from March 2020 to April 2021 among individuals under 18 years. Furthermore, we explored whether the use of data generated by a Bolívar reference laboratory captures the departmental epidemiology. Two information sources were used; the national COVID-19 surveillance system and the Bolívar COVID-19 reference laboratory. In using a population-based ecological approach and information from confirmed symptomatic cases, epidemic curves and heat maps were constructed to assess the COVID-19 dynamics and patterns by sex, age, and residence (Cartagena vs. 45 other municipalities). The COVID-19 incidence was comparable between males and females but varied by age group, being higher in children aged 10 years and older. Cartagena had a significantly higher number of cases and experienced early epidemic peaks. Our analyses suggest that information generated by the reference laboratory does not capture the COVID-19 departmental epidemiology, despite conducting population-based surveillance across Bolívar. The study provides a retrospective characterization of the COVID-19 epidemiology in an understudied population and information that may be useful for future evidence-based responses.

Molecular Characterization by Multilocus Sequence Typing and Diversity Analysis of Rickettsia asembonensis in Peru.

Despite several reports worldwide documenting the presence of Rickettsia asembonensis in samples derived from ectoparasites, animals and more recently humans, genomic information of these specimens remains scarce, and when available, is usually limited to small genomic fragments of limited value. We generated complete sequences for two conserved (17-kDa antigen gene and gltA) and three variable (sca4, ompB and ompA) genes in five R. asembonensis DNA samples detected in cat and dog fleas in Peru. Complete gene sequences were used to conduct multi-locus sequence typing and phylogenetic analyses to assess diversity and infer relationships among strains and other reference sequences. The 17-kDa antigen gene was highly conserved across Rickettsia species. Of the variable genes ompB was the most variable, but this diversity was not captured through phylogenetics alone even when efforts were made to maximize potential diversity in terms of flea species, animal host and location. Through a combination of de novo and reference-based genome assembly we identified a 75 bp insertion in ompA that encodes a 25 aa repetitive motif found in other Rickettsia species, but not present in the original prototype strain from Kenya. R. asembonensis has only recently been shown to be a bona-fide human pathogen. As such, and compounded by a lack of available genomic information, it remains understudied. Our work directly addresses the lack of genomic information available worldwide for the study of these novel Rickettsia species and specifically contributes to our understanding of the diversity and molecular epidemiology of R. asembonensis in Peru.

High titers of anti-A1 and anti-B antibodies among Peruvian group O platelet donors.

Critical antibody titers have been described as factors associated with hemolysis in ABO plasma-incompatible platelet (PLT) transfusions. This study was carried out to describe the frequency of high-titers anti-A and antiB IgM and IgG antibodies in group O apheresis platelet donors, and to explore differences according to the donor characteristics. A cross-sectional study was carried out at the Blood Bank of a National Hospital in Peru from January to March 2019. IgM and IgG antibodies against A1 and B antigens were quantified in 339 platelet donors using the direct hemagglutination technique and the solid-phase adherence technique, respectively. For analysis purposes, two cut-off points; ≥128 and ≥64, were used to define a critical titer for IgM due to a lack of consensus. An IgG titer of ≥256 was also defined as critical. Of the donors, 22.1 % had critical IgM titers when the cut-off point was defined as ≥128. However, when the IgM cut-off was ≥64, the frequency of platelet donors with critical titers increased to 54.0 %. The frequency of donors with critical IgG titers was 23.5 %. Higher IgG titers were associated with female donors while higher IgM titers were negative associated with age. One in two or three platelet donors, depending on the cutoff point used to define a critical IgM titer, had at least one critical titer of anti-A or anti-B antibodies. Early identification of platelet donors with critical antibody titers could prevent passive transfusion of ABO antibodies to non-isogroup recipients.

In vitro antimicrobial activity of Bixa orellana L . Leaves extract against anaerobic bacteria associated to bacterial vaginosis and Lactobacillus spp. / Actividad antimicrobiana in vitro del extracto de hojas de Bixa orellana L. contra bacterias anaerobias asociadas a la vaginosis bacteriana y Lactobacillus spp.

OBJECTIVE.: Motivation for the study: bacterial vaginosis is a bacterial infection that frequently affects women of reproductive age. The treatment is based on synthetic antimicrobials. Bixa orellana L. possesses antimicrobial properties and could represent a potential non-synthetic therapeutic alternative. Main findings: in vitro results suggest that, methanolic extract of Bixa orellana L. leaves possesses potential antimicrobial properties against bacteria associated to bacterial vaginosis. Implications: to identify new sources with therapeutic potential, and to promote research, discovery, and characterization of non-synthetic antimicrobials. To describe the in vitro antimicrobial activity of the methanolic extract of Bixa orellana L. leaves against anaerobic bacteria associated to bacterial vaginosis and Lactobacillus spp. MATERIALS AND METHODS.: Eight ATCC reference strains; Gardnerella vaginalis, Prevotella bivia, Peptococcus niger, Peptostreptococcus anaerobius, Mobiluncus curtisii, Atopobium vaginae, Veillonella parvula, and Lactobacillus crispatus, and twenty-two clinical isolates; eleven Gardnerella vaginalis and eleven Lactobacillus strains, were included in the study. The antimicrobial susceptibility was determined by the agar diffusion method. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined by using agar dilution and a modified dilution plating method, respectively. RESULTS.: All ATCC reference strains showed high levels of susceptibility to the extract, except P. vibia, V. parvula and L. crispatus. Interestingly, all G. vaginalis clinical isolates and the G. vaginalis ATTC strain were the most susceptible to the extract, given their low MIC (1.0 - 2.0 mg/mL) and MBC (1.0 - 4.0 mg/mL) values, whereas, the Lactobacillus spp. clinical isolates and the L. crispatus ATCC strain were the least susceptible bacteria given their high MIC (32.0 mg/mL) and MBC (≥ 32.0 mg/mL) values. CONCLUSIONS.: In vitro experiments suggest that the extract possesses selective antimicrobial properties given its high activity against bacterial vaginosis-associated anaerobic bacteria and low activity against Lactobacillus species.

OBJETIVOS: Motivación para realizar el estudio: la vaginosis bacteriana es una infección bacteriana que afecta de forma frecuente a las mujeres en edad reproductiva. El tratamiento se basa en antimicrobianos sintéticos. Bixa orellana L. posee propiedades antimicrobianas y podría representar una potencial alternativa terapéutica no sintética. Principales hallazgos: los resultados in vitro sugieren que el extracto metanólico de las hojas de Bixa orellana L. posee potenciales propiedades antimicrobianas contra las bacterias asociadas a vaginosis bacteriana. Implicancias: identificar nuevas fuentes con potencial terapéutico, y promover la investigación, descubrimiento y caracterización de antimicrobianos no sintéticos. Describir la actividad antimicrobiana in vitro del extracto metanólico de las hojas de Bixa orellana L. contra las bacterias anaerobias asociadas a la vaginosis bacteriana y Lactobacillus spp. MATERIALES Y MÉTODOS: . Se incluyeron en el estudio ocho cepas de referencia ATCC; Gardnerella vaginalis, Prevotella bivia, Peptococcus niger, Peptostreptococcus anaerobius, Mobiluncus curtisii, Atopobium vaginae, Veillonella parvula y Lactobacillus crispatus, y 22 aislamientos clínicos; once aislados de Gardnerella vaginalis y once aislados de Lactobacillus. La susceptibilidad antimicrobiana se determinó mediante el método de difusión en agar. La concentración mínima inhibitoria (CMI) y la concentración bactericida mínima (CBM) fueron determinadas utilizando el método de dilución en agar y un método de dilución modificado, respectivamente. RESULTADOS: . Todas las cepas de referencia ATCC tuvieron un alto nivel de susceptibilidad al extracto, con excepción de P. vibia, V. parvula y L. crispatus. Interesantemente, los aislamientos clínicos de G. vaginalis y la cepa ATCC de G. vaginalis fueron los más susceptibles al extracto dados los bajos valores de CMI (1,0 - 2,0 mg/mL) y CBM (1,0 - 4,0 mg/mL), mientras que, los aislamientos clínicos de Lactobacillus spp. y la cepa ATCC de L. crispatus fueron los menos susceptibles debido a los altos valores de CMI (32,0 mg/mL) y CBM (≥ 32,0 mg/mL). CONCLUSIONES: . Los experimentos in vitro sugieren que el extracto posee propiedades antibacterianas selectivas dada su alta actividad contra bacterias anaerobias asociadas a vaginosis bacteriana y baja actividad contra especies de Lactobacillus.

Climatic factors and the incidence of dengue in Cartagena, Colombian Caribbean Region

ABSTRACT Background: The influence of climate on the epidemiology of dengue has scarcely been studied in Cartagena. Methods: The relationship between dengue cases and climatic and macroclimatic factors was explored using an ecological design and bivariate and time-series analyses during lag and non-lag months. Data from 2008-2017 was obtained from the national surveillance system and meteorological stations. Results: Cases correlated only with climatic variables during lag and non-lag months. Decreases in precipitation and humidity and increases in temperature were correlated with an increase in cases. Conclusions: Our findings provide useful information for establishing and strengthening dengue prevention and control strategies.

Climatic Variability and Human Leptospirosis Cases in Cartagena, Colombia: A 10-Year Ecological Study.

Leptospirosis is an acute febrile disease that mainly affects developing countries with tropical climates. The complexity and magnitude of this disease is attributed to socioeconomic, climatic, and environmental conditions. In this study, in a 10-year period from 2008 to 2017, the relationship between human leptospirosis cases and climatic factors in Cartagena de Indias, Colombia were evaluated. Monthly leptospirosis cases, climatic variables, and macroclimatic phenomena (El Niño and La Niña) were obtained from public datasets. Local climatic factors included temperature (maximum, average, and minimum), relative humidity, precipitation, and the number of precipitation days. Time series graphs were drawn and correlations between cases of leptospirosis and climatic variables considering lags from 0 to 10 months were examined. A total of 360 cases of leptospirosis were reported in Cartagena during the study period, of which 192 (53.3%) were systematically notified between October and December. Several correlations were detected between the number of cases, local climatic variables, and macroclimatic phenomena. Mainly, the increase of cases correlated with increased precipitation and humidity during the La Niña periods. Herein, seasonal patterns and correlations suggest that the climate in Cartagena could favor the incidence of leptospirosis. Our findings suggest that prevention and control of human leptospirosis in Cartagena should be promoted and strengthened, especially in the last quarter of the year.

Evaluation of Two Serological Assays for Diagnosing Zika Virus Infection.

Zika virus (ZIKV) emerged and spread rapidly in South American countries during 2015. Efforts to diagnose ZIKV infection using serological tools were challenging in dengue-endemic areas because of antigenic similarities between both viruses. Here, we assessed the performance of an in-house developed IgM antibody capture enzyme-linked immunosorbent assay (MAC-ELISA) and the plaque reduction neutralization test (PRNT) to diagnose ZIKV infection. Acute and convalescent paired serum samples from 51 patients who presented with clinical symptoms suggestive of an arbovirus illness in dengue-endemic areas of Honduras, Venezuela, Colombia and Peru were used in the assessment. Samples were tested for ZIKV, dengue and chikungunya virus using a variety of laboratory techniques. The results for the ZIKV-RNA screening and seroconversion detected by the microneutralization test were used to construct a composite reference standard. The overall sensitivity and specificity for the MAC-ELISA were 93.5% and 100.0%, respectively. Contrastingly, the overall sensitivity and specificity for the PRNT were 96.8% and 95.0%, respectively. Restricting the analysis according to IgM or neutralizing antibodies against dengue, the performances of both serological assays were adequate. The findings of this study reveal that the MAC-ELISA and PRNT would provide initial reliable laboratory diagnostic assays for ZIKV infection in dengue-endemic areas.

Antimicrobial Resistance Patterns and Dynamics of Extended-Spectrum ß-Lactamase-Producing Uropathogenic Escherichia coli in Cusco, Peru.

Urinary tract infections (UTIs) are a common human infection. Antibiotic resistance in extended-spectrum ß-lactamase (ESBL)-producing uropathogenic E. coli (UPEC) is a major therapeutic challenge due to limited treatment alternatives. The aim was to characterize the antimicrobial resistance (AMR) and dynamics of ESBL-producing UPEC isolates from UTI cases seen at a local hospital in Cusco, Peru. Ninety-nine isolates from respective patients were characterized against 18 different antibiotics. Latent class analysis (LCA) was used to evaluate the dynamics across the study time according to resistance patterns. The median age of patients was 51 years old, and nearly half were women. ESBL-producing UPEC isolates were slightly more frequent in outpatient services than emergency rooms, and there were higher resistance rates in males compared to females. Half of the ESBL producers were resistant to aminoglycosides and nitrofurantoin. Cefoxitin and fosfomycin resistance was 29.3% and 14.1%, respectively. Resistance to carbapenems was not observed. All isolates were multidrug-resistant bacteria, and 16.2% (16/99) were also classified as extensively drug-resistant bacteria. The resistance patterns varied across the study time and differed regarding sex and healthcare service. The study revealed high levels of AMR to commonly used antimicrobials and a dynamic circulation of ESBL-producing UPEC isolates with varying resistance patterns.