Continued waves, new variants, and limited vaccine deployment mean that SARS-CoV-2 tests remain vital to constrain the coronavirus disease 2019 (COVID-19) pandemic. Affordable, point-of-care (PoC) tests allow rapid screening in non-medical settings. Reverse-transcription loop-mediated isothermal amplification (RT-LAMP) is an appealing approach. A crucial step is to optimize testing in low/medium resource settings. Here, we optimized RT-LAMP for SARS-CoV-2 and human β-actin, and tested clinical samples in multiple countries. “TTTT” linker primers did not improve performance, and while guanidine hydrochloride, betaine and/or Igepal-CA-630 enhanced detection of synthetic RNA, only the latter two improved direct assays on nasopharygeal samples. With extracted clinical RNA, a 20 min RT-LAMP assay was essentially as sensitive as RT-PCR. With raw Canadian nasopharygeal samples, sensitivity was 100% (95% CI: 67.6% - 100%) for those with RT-qPCR Ct values ≤ 25, and 80% (95% CI: 58.4% - 91.9%) for those with 25 < Ct ≤ 27.2. Highly infectious, high titer cases were also detected in Colombian and Ecuadorian labs. We further demonstrate the utility of replacing thermocyclers with a portable PoC device (FluoroPLUM). These combined PoC molecular and hardware tools may help to limit community transmission of SARS-CoV-2.

Aedes mosquitoes, as vectors of medically important arthropod-borne viruses (arboviruses), constitute a major public health threat that requires entomological and epidemiological surveillance to guide vector control programs to prevent and reduce disease transmission. In this study, we present the collaborative effort of 1 year of Aedes ageypti (Linnaeus, 1762) mosquito-based arbovirus surveillance in 2 geographically distinct regions of Latin America (Nicaragua and Ecuador). Adult female mosquitoes were collected using backpack aspirators in over 2,800 randomly selected households (Nicaragua, Ecuador) and 100 key sites (Nicaragua) from 8 distinct communities (Nicaragua: 2, Ecuador: 6). A total of 1,358 mosquito female pools were processed for RNA extraction and viral RNA detection using real-time reverse transcription-polymerase chain reaction. Ten positive dengue virus (DENV) pools were detected (3 in Nicaragua and 7 in Ecuador), all of which were found during the rainy season and matched the serotypes found in humans (Nicaragua: DENV-1 and DENV-4; Ecuador: DENV-2). Infection rates ranged from 1.13 to 23.13, with the Nicaraguan communities having the lowest infection rates. Our results demonstrate the feasibility of detecting DENV-infected Aedes mosquitoes in low-resource settings and underscore the need for targeted mosquito arbovirus sampling and testing, providing valuable insights for future surveillance programs in the Latin American region.

Background/Objectives: There is limited information on human milk (HM) microbiome composition and function in Latin America. Also, interactions between HM constituents and its microbiome have received partial attention. Objective: To characterize the HM microbiota composition considering lactation stages (colostrum, transition, and mature HM) and free glutamate concentrations in Ecuadorian mothers. Methods: We recruited 20 mothers that gave birth to normal full-term babies and donated colostrum, transition, and mature milk. Samples were assessed by 16S rRNA gene sequencing by Oxford Nanopore Technologies (ONT). Free glutamate concentrations were measured by proton nuclear magnetic resonance (NMR) spectroscopy. Results: For each lactation stage and in order of frequency, the majority of ASVs were assigned to Staphylococcus, Streptococcus, (Firmicutes); Escherichia, Acinetobacter, (Proteobacteria); Corynebacterium, Lactobacillus, Cutibacterium, (Actinobacteriota); Chryseobacterium, and Flavobacterium (Bacteroidota). Alfa diversity was similar in HM samples and tended to be higher in milk intended for male infants. We observed significant differences in qualitative β-diversity metrics between samples with low and high glutamate concentrations. Functional predictions of HM microbiota demonstrated the presence of polyamine biosynthesis II super pathway in samples with high glutamate concentrations. Conclusions: The core bacterial components of the HM microbiota in Ecuadorian women were similar to those reported from different parts of the world, with variations at the genus level. Free glutamate dynamics in HM need to be studied considering maternal production and bacterial metabolism to better understand HM composition for optimal infant nutrition.

Ecuador, a tropical country with frequent dengue outbreaks, including a surge from 16,017 cases in 2022 to 61,329 in 2024, was the focus of this study. The study was conducted in Borbon, a semi-urban rural town in the Esmeraldas province. Genomic analysis, alongside entomo-virological surveillance, provides valuable insights into DENV-2 genotypes. Five pools of female Aedes aegypti mosquitoes from Borbon tested positive for DENV serotype 2 through RT-qPCR. One positive pool (CT = 16.13) was sequenced using Illumina MiSeq, and genotyping was conducted via the Dengue Typing Tool and Maximum Likelihood phylogenetic tree. The genotype assigned was III Southern Asian-American. Comparison with other genomes revealed genetic similarity to a human dengue genome sequenced in 2021, also from Esmeraldas, clustering with genomes reported across the Americas, particularly from Colombia and Venezuela. This study enhances our understanding of dengue virus epidemiology in rural areas, emphasizing the critical role of clinical case surveillance and vector monitoring in guiding evidence-based interventions.

Mosquitoes are major vectors of human and animal diseases, making their accurate identification essential for vector surveillance and control. However, morphological identification has often been challenging, requiring taxonomic expertise and well-preserved specimens. Molecular markers, particularly DNA barcoding, offer an effective alternative for identifying both adult and immature stages. Ecuador is one of the most biodiverse countries in the world, a diversity that is also evident in its Culicidae fauna. This study provides a comprehensive revision of Ecuadorian mosquitoes, updating the national checklist and emphasizing species of public health importance. For species identification, an integrative approach was used combining morphology and DNA barcoding (COI and ITS2 regions). We list 266 species in 22 genera, of which 17 species are new national records, and 33 species are validated through molecular analysis. The updated checklist highlights Ecuador’s Culicidae diversity across its biogeographic regions, which represent 7% of the world’s mosquito diversity. These findings provide a critical foundation for future entomological research and vector control in the country.