Surveillance of COVID-19 vaccines: A comprehensive analysis of the first immunization drive in Ecuador.

The initial phase of the COVID-19 vaccination in Ecuador occurred between April and November 2021. Initially, it focused on priority populations, including health professionals and other front-line workers. During this period, there was limited knowledge about the vaccine's adverse effects. A non-probability, observational study was conducted among university staff in Guayaquil, Ecuador, who received the AstraZeneca vaccine (n = 423) between April and November 2021. This study aimed to compare the acute adverse reactions by doses and to report the incidence of long-term adverse reactions within the AstraZeneca group. As a result, comparing acute adverse reactions between doses, the odds ratio for local pain, headache, muscle pain, fever, and chills are statistically higher after the first dose than the second dose. Survival curves indicated these symptoms appeared mainly within the first 6 h post-vaccination. This is the first pharmacovigilance study from Ecuador that analyzes survival probabilities for the AstraZeneca vaccine's adverse effects.

Alignment of multiple metabolomics LC-MS datasets from disparate diseases to reveal fever-associated metabolites.

Acute febrile illnesses are still a major cause of mortality and morbidity globally, particularly in low to middle income countries. The aim of this study was to determine any possible metabolic commonalities of patients infected with disparate pathogens that cause fever. Three liquid chromatography-mass spectrometry (LC-MS) datasets investigating the metabolic effects of malaria, leishmaniasis and Zika virus infection were used. The retention time (RT) drift between the datasets was determined using landmarks obtained from the internal standards generally used in the quality control of the LC-MS experiments. Fitted Gaussian Process models (GPs) were used to perform a high level correction of the RT drift between the experiments, which was followed by standard peakset alignment between the samples with corrected RTs of the three LC-MS datasets. Statistical analysis, annotation and pathway analysis of the integrated peaksets were subsequently performed. Metabolic dysregulation patterns common across the datasets were identified, with kynurenine pathway being the most affected pathway between all three fever-associated datasets.

A report on SARS-CoV-2 first wave in Ecuador: drug consumption dynamics.

Introduction: The first COVID-19 wave in Ecuador started in March 2020 and extended until November. Several types of drugs have been proposed as a potential treatment during this period, and some affected people have self-medicated. Method: A retrospective study was conducted with 10,175 individuals who underwent RT-PCR tests for SARS-CoV-2 from July to November 2020. We compared the number of positive and negative cases in Ecuador with symptoms and drug consumption. The Chi-square test of independence compared clinical and demographic data and PCR test results. Odds ratios analyzed drug consumption dynamics. Results: Of 10,175 cases, 570 were positive for COVID-19, while 9,605 were negative. In positive cases, there was no association between the RT-PCR result and sex, age, or comorbidities. When considering demographic data, Cotopaxi and Napo had the highest rates of positive cases (25.7% and 18.8%, respectively). Manabí, Santa Elena, and Guayas regions had fewer than 10% positive cases. The Drug consumption dynamic analysis showed that negative COVID-19 cases presented higher drug consumption than positive cases. In both groups, the most consumed medication was acetaminophen. Acetaminophen and Antihistamines had higher odds of consumption in positive PCR cases than in negative. Symptoms like fever and cough were more related to positive RT-PCR results. Conclusion: The first COVID-19 wave in Ecuador has affected the provinces differently. At a national level, the consumption of drugs has been highly associated with self-medication.

Antarctic fungi with antibiotic potential isolated from Fort William Point, Antarctica.

The Antarctic continent is one of the most inhospitable places on earth, where living creatures, mostly represented by microorganisms, have specific physiological characteristics that allow them to adapt to the extreme environmental conditions. These physiological adaptations can result in the production of unique secondary metabolites with potential biotechnological applications. The current study presents a genetic and antibacterial characterization of four Antarctic fungi isolated from soil samples collected in Pedro Vicente Maldonado Scientific Station, at Fort William Point, Greenwich Island, Antarctica. Based on the sequences of the internal transcribed spacer (ITS) region, the fungi were identified as Antarctomyces sp., Thelebolus sp., Penicillium sp., and Cryptococcus gilvescens. The antibacterial activity was assessed against four clinical bacterial strains: Escherichia coli, Klebsiella pneumoniae, Enterococcus faecalis, and Staphylococcus aureus, by a modified bacterial growth inhibition assay on agar plates. Results showed that C. gilvescens and Penicillium sp. have potential antibiotic activity against all bacterial strains. Interestingly, Thelebolus sp. showed potential antibiotic activity only against E. coli. In contrast, Antarctomyces sp. did not show antibiotic activity against any of the bacteria tested under our experimental conditions. This study highlights the importance of conservation of Antarctica as a source of metabolites with important biomedical applications.

Gold Nanoshells-Based Lateral Flow Assay for the Detection of Chagas Disease at the Point-of-Care.

Chagas disease is a neglected parasitic infection and a major public health problem in the Americas. It remains underdiagnosed in the United States and internationally due to the lack of affordable testing and disparities in healthcare, particularly for those most at risk. We describe a proof-of-concept lateral flow immunoassay employing a recombinant Chagas multiantigen conjugated to gold nanoshells (AuNS) to detect circulating human anti-Chagas IgG antibodies. This is one of the first lateral flow immunoassays to capitalize on the larger surface area of AuNS compared with nanoparticles that can help amplify low-magnitude signals. Results were compared with 42 positive and negative Chagas serum samples, of which a subset of 27 samples was validated against an ELISA (Hemagen®). The sensitivity and specificity of our assay were 83% and 95%, respectively. These results suggest that an AuNS-based rapid testing for Chagas disease could facilitate in-field screening/diagnosis with a performance comparable to commercial methods.

High sensitivity-low cost detection of SARS-CoV-2 by two steps end point RT-PCR with agarose gel electrophoresis visualization.

More than one year since Coronavirus disease 2019 (COVID-19) pandemic outbreak, the gold standard technique for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) detection is still the RT-qPCR. This is a limitation to increase testing capacities, particularly at developing countries, as expensive reagents and equipment are required. We developed a two steps end point RT-PCR reaction with SARS-CoV-2 Nucleocapsid (N) gene and Ribonuclease P (RNase P) specific primers where viral amplicons were verified by agarose gel electrophoresis. We carried out a clinical performance and analytical sensitivity evaluation for this two-steps end point RT-PCR method with 242 nasopharyngeal samples using the CDC RT-qPCR protocol as a gold standard technique. With a specificity of 95.8%, a sensitivity of 95.1%, and a limit of detection of 20 viral RNA copies/uL, this two steps end point RT-PCR assay is an affordable and reliable method for SARS-CoV-2 detection. This protocol would allow to extend COVID-19 diagnosis to basic molecular biology laboratories with a potential positive impact in surveillance programs at developing countries.

Vitamin A status, inflammation adjustment, and immunologic response in the context of acute febrile illness: A pilot cohort study among pediatric patients.

BACKGROUND: Vitamin A is necessary for an adequate immune response to infections. Infection also alters vitamin A biomarkers, which interferes with assessment of vitamin A deficiency and thus impairs clinical management. Here we apply multiple strategies to adjust vitamin A biomarkers for inflammation during acute infection and evaluate associations between adjusted vitamin A status and immunologic response markers. METHODS: We measured biomarkers in pediatric patients presenting with acute febrile illness in Guayaquil, Ecuador at paired acute and convalescent visits. Four adjustment strategies were applied to retinol-binding protein (RBP) concentrations: Thurnham correction factor (TCF), BRINDA regression correction (BRC), CRP-only adjustment factor (CRP), and proof-of-concept for a proposed interleukin 6 regression model (IL-6 RM). Adjusted RBP concentrations were compared between visits using the paired Wilcoxon signed-rank test. Multivariate regression analysis was used to assess associations between adjusted vitamin A status and immunologic response markers. RESULTS: A sample of 57 participants completed the acute visit 1, and 18 of these individuals completed the convalescent visit 2. The IL-6 RM was the only strategy resulting in adjusted RBP concentrations that were not significantly different between paired visits (p = 0.20). Following RBP adjustment, 0.0% of participants were classified as vitamin A deficient (RBP ≤ 0.70 µmol/L) and 14.0% were classified as vitamin A insufficient (RBP ≤ 1.05 µmol/L). Adjusted vitamin A insufficiency was associated with an increase in macrophage inflammatory protein 1-alpha (MIP-1α, p = 0.03) and a pro-inflammatory immune response profile (p = 0.03) during the acute visit. CONCLUSIONS: We introduce a strategy for adjusting vitamin A in the context of clinical illness based on IL-6 concentrations that will need to be validated in larger studies. Assessment of vitamin A during infection allows for further understanding of how vitamin A status modulates immunopathology and enables targeted strategies for vitamin A supplementation in the context of infection among children in settings with high burdens of undernutrition and infectious diseases.

A Comparison of Deep Learning Models for Detecting COVID-19 in Chest X-ray Images.

COVID-19 has spread around the world rapidly causing a pandemic. In this research, a set of Deep Learning architectures, for diagnosing the presence or not of the disease have been designed and compared; such as, a CNN with 4 incremental convolutional blocks; a VGG-19 architecture; an Inception network; and, a compact CNN model known as MobileNet. For the analysis and comparison, transfer learning techniques were used in forty-five different experiments. All four models were designed to perform binary classification, reaching an accuracy above 95%. A set of different scores were implemented to compare the performance of all models, showing that the VGG-19 and Inception configurations performed the best.

Current state of the art in rapid diagnostics for antimicrobial resistance.

Antimicrobial resistance (AMR) is a fundamental global concern analogous to climate change threatening both public health and global development progress. Infections caused by antimicrobial-resistant pathogens pose serious threats to healthcare and human capital. If the increasing rate of AMR is left uncontrolled, it is estimated that it will lead to 10 million deaths annually by 2050. This global epidemic of AMR necessitates radical interdisciplinary solutions to better detect antimicrobial susceptibility and manage infections. Rapid diagnostics that can identify antimicrobial-resistant pathogens to assist clinicians and health workers in initiating appropriate treatment are critical for antimicrobial stewardship. In this review, we summarize different technologies applied for the development of rapid diagnostics for AMR and antimicrobial susceptibility testing (AST). We briefly describe the single-cell technologies that were developed to hasten the AST of infectious pathogens. Then, the different types of genotypic and phenotypic techniques and the commercially available rapid diagnostics for AMR are discussed in detail. We conclude by addressing the potential of current rapid diagnostic systems being developed as point-of-care (POC) diagnostic tools and the challenges to adapt them at the POC level. Overall, this review provides an insight into the current status of rapid and POC diagnostic systems for AMR.

Micronutrients, Immunological Parameters, and Dengue Virus Infection in Coastal Ecuador: A Nested Case-Control Study in an Infectious Disease Surveillance Program.

BACKGROUND: Micronutrients are known to modulate host immunity, and there is limited literature on this association in the context of dengue virus infection (DENV). METHODS: Using a nested case-control design in a surveillance program, we measured the following: anthropometry; nutritional biomarkers including serum ferritin, soluble transferrin receptor, retinol-binding protein (RBP), 25-hydroxy vitamin D, folate, and vitamin B12; and a panel of immune response markers. We then compared these measures across 4 illness categories: healthy control, nonfebrile DENV, other febrile illness (OFI), and apparent DENV using multivariate polytomous logistic regression models. RESULTS: Among 142 participants, serum ferritin (ng/mL) was associated with apparent DENV compared to healthy controls (odds ratio [OR], 2.66; confidence interval [CI], 1.53-4.62; P = .001), and RBP concentrations (µmol/L) were associated with apparent DENV (OR, 0.03; CI, 0.00-0.30; P = .003) and OFI (OR, 0.02; CI, 0.00-0.24; P = .003). In a subset of 71 participants, interleukin-15 levels (median fluorescent intensity) were positively associated with apparent DENV (OR, 1.09; CI, 1.03-1.14; P = .001) and negatively associated with nonfebrile DENV (OR, 0.89; CI, 0.80-0.99; P = .03) compared to healthy controls. CONCLUSIONS: After adjusting for the acute-phase response, serum ferritin and RBP concentrations were associated with apparent DENV and may represent biomarkers of clinical importance in the context of dengue illness.

A two-colour multiplexed lateral flow immunoassay system to differentially detect human malaria species on a single test line.

BACKGROUND: Malaria continues to impose a tremendous burden in terms of global morbidity and mortality, yet even today, a large number of diagnoses are presumptive resulting in lack of or inappropriate treatment. METHODS: In this work, a two-colour lateral flow immunoassay (LFA) system was developed to identify infections by Plasmodium spp. and differentiate Plasmodium falciparum infection from the other three human malaria species (Plasmodium vivax, Plasmodium ovale, Plasmodium malariae). To achieve this goal, red and blue colours were encoded to two markers on a single test line of strips, for simultaneous detection of PfHRP2 (red), a marker specific for P. falciparum infection, and pLDH (blue), a pan-specific marker for infections by all species of Plasmodium. The assay performance was first optimized and evaluated with recombinant malarial proteins spiked in washing buffer at various concentrations from 0 to 1000 ng mL-1. The colour profiles developed on the single test line were discriminated and quantified: colour types corresponded to malaria protein species; colour intensities represented protein concentration levels. RESULTS: The limit of detection (the lowest concentrations of malaria antigens that can be distinguished from blank samples) and the limit of colour discrimination (the limit to differentiate pLDH from PfHRP2) were defined for the two-colour assay from the spiked buffer test, and the two limits were 31.2 ng mL-1 and 7.8 ng mL-1, respectively. To further validate the efficacy of the assay, 25 human whole blood frozen samples were tested and successfully validated against ELISA and microscopy results: 15 samples showed malaria negative; 5 samples showed P. falciparum positive; 5 samples showed P. falciparum negative, but contained other malaria species. CONCLUSIONS: The assay provides a simple method to quickly identify and differentiate infection by different malarial parasites at the point-of-need and overcome the physical limitations of traditional LFAs, improving the multiplexing potential for simultaneous detection of various biomarkers.

First report of fibropapillomatosis in an olive ridley turtle Lepidochelys olivacea from the southeastern Pacific.

An adult olive ridley turtle Lepidochelys olivacea with lesions suggestive of fibropapillomatosis was rescued on the coast of San Antonio, central Chile. Histopathologic analysis showed an exophytic and pedunculated mass formed by epidermal papillary projections supported by fibrovascular cores, epidermal hyperplasia and marked orthokeratotic hyperkeratosis. ChHV5 unique long genes UL27, UL28 and UL30 were amplified from tumor lesions and sequenced for phylogeny. Phylogenetic reconstruction showed the Chilean sequences clustering with the Eastern Pacific group. This is the first case of fibropapillomatosis in an olive ridley turtle diagnosed in Chile and in the southeastern Pacific region. Our results suggest a regional grouping of ChHV5 variants independent of the marine turtle's species.

Rapid Diagnostic Platform for Colorimetric Differential Detection of Dengue and Chikungunya Viral Infections.

In this work, we demonstrate a rapid diagnostic platform with potential to transform clinical diagnosis of acute febrile illnesses in resource-limited settings. Acute febrile illnesses such as dengue and chikungunya, which pose high burdens of disease in tropical regions, share many nonspecific symptoms and are difficult to diagnose based on clinical history alone in the absence of accessible laboratory diagnostics. Through a unique color-mixing encoding and readout strategy, our platform enabled consistent and accurate multiplexed detection of dengue and chikungunya IgM/IgG antibodies in human clinical samples within 30 min. Our multiplex assay offers several advantages over conventional rapid diagnostic tests deployed in resource-limited settings, including a low sample volume requirement and the ability to concurrently detect four analytes. Our platform is a step toward multiplexed diagnostics that will be transformative for disease management in resource-limited settings by enabling informed treatment decisions through accessible evidence-based diagnosis.

Fibropapillomatosis in a Green Sea Turtle ( Chelonia mydas) from the Southeastern Pacific.

Fibropapillomatosis is a neoplastic disease that afflicts sea turtles. Although it is disseminated worldwide, cases of the disease have not been reported in the southeastern Pacific region. We describe a case of fibropapillomatosis in a green sea turtle ( Chelonia mydas) during its rehabilitation at the Machalilla National Park Rehabilitation Center, Ecuador. Viral presence was confirmed by PCR, targeting fragments of the chelonid alphaherpesvirus 5 (ChHV5) unique long (UL) genes, UL27, UL28, and UL30. The amplicons were sequenced and included in a global phylogenetic analysis of the virus with other reported sequences from GenBank. Results showed that the available viral sequences segregated into five phylogeographic groups: western Atlantic and eastern Caribbean, central Pacific, western Pacific, Atlantic, and eastern Pacific groups. The concatenated ChHV5 sequences from Ecuador clustered with the eastern Pacific sequences.

The Social and Spatial Ecology of Dengue Presence and Burden during an Outbreak in Guayaquil, Ecuador, 2012.

Dengue fever, a mosquito-borne arbovirus, is a major public health concern in Ecuador. In this study, we aimed to describe the spatial distribution of dengue risk and identify local social-ecological factors associated with an outbreak of dengue fever in the city of Guayaquil, Ecuador. We examined georeferenced dengue cases (n = 4248) and block-level census data variables to identify social-ecological risk factors associated with the presence/absence and burden of dengue in Guayaquil in 2012. Local Indicators of Spatial Association (LISA), specifically Anselin’s Local Moran’s I, and Moran’s I tests were used to locate hotspots of dengue transmission, and multimodel selection was used to identify covariates associated with dengue presence and burden at the census block level. We identified significant dengue transmission hotspots near the North Central and Southern portions of Guayaquil. Significant risk factors for presence of dengue included poor housing conditions, access to paved roads, and receipt of remittances. Counterintuitive positive correlations with dengue presence were observed with several municipal services such as garbage collection and access to piped water. Risk factors for increased burden of dengue included poor housing conditions, garbage collection, receipt of remittances, and sharing a property with more than one household. Social factors such as education and household demographics were negatively correlated with increased dengue burden. These findings elucidate underlying differences with dengue presence versus burden, and suggest that vulnerability and risk maps could be developed to inform dengue prevention and control; this is information that is also relevant for emerging epidemics of chikungunya and Zika viruses.

Spatiotemporal Variation in Environmental Vibrio cholerae in an Estuary in Southern Coastal Ecuador.

Cholera emergence is strongly linked to local environmental and ecological context. The 1991-2004 pandemic emerged in Perú and spread north into Ecuador's El Oro province, making this a key site for potential re-emergence. Machala, El Oro, is a port city of 250,000 inhabitants, near the Peruvian border. Many livelihoods depend on the estuarine system, from fishing for subsistence and trade, to domestic water use. In 2014, we conducted biweekly sampling for 10 months in five estuarine locations, across a gradient of human use, and ranging from inland to ocean. We measured water-specific environmental variables implicated in cholera growth and persistence: pH, temperature, salinity, and algal concentration, and evaluated samples in five months for pathogenic and non-pathogenic Vibrio cholerae, by polymerase chain reaction (PCR). We found environmental persistence of pandemic strains O1 and O139, but no evidence for toxigenic strains. Vibrio cholerae presence was coupled to algal and salinity concentration, and sites exhibited considerable seasonal and spatial heterogeneity. This study indicates that environmental conditions in Machala are optimal for cholera re-emergence, with risk peaking during September, and higher risk near urban periphery low-income communities. This highlights a need for surveillance of this coupled cholera-estuarine system to anticipate potential future cholera outbreaks.

The Burden of Dengue Fever and Chikungunya in Southern Coastal Ecuador: Epidemiology, Clinical Presentation, and Phylogenetics from the First Two Years of a Prospective Study.

Here, we report the findings from the first 2 years (2014-2015) of an arbovirus surveillance study conducted in Machala, Ecuador, a dengue-endemic region. Patients with suspected dengue virus (DENV) infections (index cases, N = 324) were referred from five Ministry of Health clinical sites. A subset of DENV-positive index cases (N = 44) were selected, and individuals from the index household and four neighboring homes within 200 m were recruited (N = 400). Individuals who entered the study, other than the index cases, are referred to as associates. In 2014, 70.9% of index cases and 35.6% of associates had acute or recent DENV infections. In 2015, 28.3% of index cases and 12.8% of associates had acute or recent DENV infections. For every DENV infection captured by passive surveillance, we detected an additional three acute or recent DENV infections in associates. Of associates with acute DENV infections, 68% reported dengue-like symptoms, with the highest prevalence of symptomatic acute infections in children aged less than 10 years. The first chikungunya virus (CHIKV) infections were detected on epidemiological week 12 in 2015; 43.1% of index cases and 3.5% of associates had acute CHIKV infections. No Zika virus infections were detected. Phylogenetic analyses of isolates of DENV from 2014 revealed genetic relatedness and shared ancestry of DENV1, DENV2, and DENV4 genomes from Ecuador with those from Venezuela and Colombia, indicating the presence of viral flow between Ecuador and surrounding countries. Enhanced surveillance studies, such as this, provide high-resolution data on symptomatic and inapparent infections across the population.

Micronutrients and Leptospirosis: A Review of the Current Evidence.

BACKGROUND: Leptospirosis is one of the most widespread zoonoses and represents a major threat to human health. Due to the high burden of disease, limitations in diagnostics, and limited coverage and availability of effective human and veterinary vaccines, leptospirosis remains an important neglected zoonotic disease. Improved surveillance and identification of modifiable risk factors for leptospirosis are urgently needed to inform preventive interventions and reduce the risk and severity of Leptospira infection. METHODOLOGY/PRINCIPAL FINDINGS: This review was conducted to examine the evidence that links micronutrient status and Leptospira infection. A total of 56 studies were included in this review: 28 in vitro, 17 animal, and 11 observational human studies. Findings indicated that Leptospira infection is associated with higher iron and calcium concentrations and hypomagnesemia. CONCLUSIONS/SIGNIFICANCE: Few prospective studies and no randomized trials have been conducted to date to examine the potential role of micronutrients in Leptospira infection. The limited literature in this area constrains our ability to make specific recommendations; however, the roles of iron, calcium, and magnesium in leptospirosis represent important areas for future research. The role of micronutrients in leptospirosis risk and severity needs to be elucidated in larger prospective human studies to inform public health interventions.

Rabies Epidemiology and Control in Ecuador.

OBJECTIVE: Describe the epidemiology and the control effort for rabies in Ecuador. METHODS: This observational study included data from the Ecuadorian National Institute of Census and Statistics (INEC), and mortality and morbidity data reported by the Ministry of Public Health and the National Institute for Social Security. We conducted a phylogeny analyses to compare the N gene from the Challenge Virus Standard (CVS) vaccine strain used in Ecuador with published Cosmopolitan, Asian and Sylvatic strains. Descriptive and inferential statistics were used to determine the significance of the data. RESULTS: In 1996 Ecuador suffered the highest rate of rabies per capita in the Americas, with an incidence rate of 0.56 cases per 100 000 people per year. Human and canine rabies showed a sharp decline until 2012. Between 1994 and 2014, we found a correlation of 0.925 (p<0.01) between annual cases of dog and human rabies. In 2011, there was an epidemic of sylvatic rabies transmitted to people by vampire bats (Desmodus rotundus) in the Amazon region, specifically in Morona Santiago, leading to 11 fatalities. Phylogenetic analyses of the CVS vaccine N gene showed an association with urban canine rabies strains (the Cosmopolitan lineage and Asian strains), whereas sylvatic rabies, like those reported in the Amazon region, were found to be grouped in a different clade represented mainly by bat-derived strains. CONCLUSIONS: This study presents the first compilation of epidemiological data on rabies in Ecuador. The incidence of human and canine rabies, also known as urban rabies, has clearly decreased due to massive canine vaccination campaigns. Phylogenetic analysis of the prevailing vaccine used in the country showed a clear separation from bat-derived rabies, the source of recent rabies outbreaks. Efforts are ongoing to develop rabies vaccines that are highly specific to the rabies virus genotype circulating in the region, including sylvatic rabies. These efforts include the implementation of reverse genetics to generate recombinant virus coding for the prevailing glycoprotein gene.

Micronutrients and dengue.

Dengue virus infection is the most widespread mosquito-borne viral infection in humans and has emerged as a serious global health challenge. In the absence of effective treatment and vaccine, host factors including nutritional status, which may alter disease progression, need investigation. The interplay between nutrition and other infections is well-established, and modulation of nutritional status often presents a simple low-cost method of interrupting transmission, reducing susceptibility, and/or ameliorating disease severity. This review examines the evidence on the role of micronutrients in dengue virus infection. We found critical issues and often inconsistent results across studies; this finding along with the lack of sufficient literature in this field have limited our ability to make any recommendations. However, vitamins D and E have shown promise in small supplementation trials. In summary, the role of micronutrients in dengue virus infection is an exciting research area and needs to be examined in well-designed studies with larger samples.