Diagnostic performance, stability, and acceptability of self-collected saliva without additives for SARS-CoV-2 molecular diagnosis.

BACKGROUND: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused the coronavirus disease 2019 (COVID-19), leading to a global pandemic. The molecular diagnosis of this virus is mostly performed by collecting upper respiratory samples, which has many disadvantages, including patient discomfort and the need for trained healthcare professionals. Although saliva has emerged as a more comfortable sample, the use of additives to preserve viral RNA is expensive and, in some cases, difficult for self-collection. METHOD: This study evaluated the diagnostic performance by RT-PCR and stability of self-collected saliva using wide-mouth specimen collection cups without stabilization and/or inactivation buffers for SARS-CoV-2 detection, compared to nasopharyngeal samples and saliva collected with additives. Additionally, the study assessed the acceptability of this sample collection method among participants and healthcare personnel. RESULTS: The study included 1281 volunteers with a 24.6% positive infection rate. Saliva demonstrated comparable diagnostic performance to nasopharyngeal samples, with a sensitivity of 87.6% and specificity of 99.6%, for a total percent agreement of 96.4%. The study also showed that viral RNA in saliva remained stable for at least 72 h at different temperatures. Notably, saliva samples without additives exhibited a lower RdRp Ct compared to samples with additives, suggesting that the absence of stabilization and/or inactivation buffers does not significantly affect its performance. The study highlighted the acceptability of saliva among patients and healthcare personnel due to its noninvasive nature and ease of collection. CONCLUSIONS: This research supports the implementation of self-collected saliva as a comfortable and user-friendly alternative sample for SARS-CoV-2 diagnosis.

Trans-stadial fate of the gut bacterial microbiota in Anopheles albimanus.

Gut microbiota communities in mosquitoes are influenced among others, by developmental stage. There is evidence that the aquatic environment where larvae feed influences the mosquito gut bacterial community composition with only a subgroup of these bacteria been transmitted trans-stadially to adults. This study evaluated the gut bacterial composition of Anopheles albimanus larvae, emerged and circulating mosquitoes, as well as water from the larval habitat, to elucidate transitions in these bacterial communities and determine the final composition in circulating mosquitoes. A 16S rRNA Illumina sequencing allowed to determine that Proteobacteria was the most abundant phylum in larvae (72.4%), emerged mosquitoes (75%), circulating adults (45.4%) and water from the larval habitat (79.1%). A core microbiome analysis evidenced that Enterobacter, Bacillus and Staphylococcus genera were the core bacterial microbiota (OTUs detected in >90%) in the four groups evaluated. PCoA cluster based on Jaccard and Bray Curtis distances showed two main bacterial clusters, one comprising the emerged and circulating adults, and the other the larvae. The results indicated that the gut microbiota of An. albimanus larvae is composed of bacteria acquired from the larval habitat; then, a rearrangement of the bacterial communities occurs in the trans-stadial passage. However, the higher bacterial richness detected in circulating adults suggests bacterial acquisition from the terrestrial environment where the mosquito feeds. Finally, the trans-stadially passage of some bacteria makes of interest their evaluation as candidates for paratransgenic control.

Composition and structure of the culturable gut bacterial communities in Anopheles albimanus from Colombia.

The understanding of factors affecting the gut bacterial communities in malaria vectors is essential for the design of vector control interventions, such as those based on a paratransgenic approach. One of the requirements of this method is the availability of bacteria from the mosquito susceptible to culture. Thus, the aim of this study was to evaluate the composition and structure of the culturable gut bacterial communities in field mosquitoes Anopheles albimanus from Colombia, in addition to generate a bacterial collection to further analyze microbial functional activity. Gut bacteria were isolated from An. albimanus larvae and adult mosquitoes collected in localities of the Atlantic and Pacific Coasts. The bacterial isolates were grouped in 28 morphospecies that corresponded to three phyla, three classes, nine families and 14 genera. The larvae guts from San Antero (Atlantic Coast) and Buenaventura (Pacific Coast) shared the genera Bacillus and Lysinibacillus and in adults, Bacillus and Bacillus cereus Group were registered in both localities. Gut bacterial richness was higher in adults from the Pacific with respect to the Atlantic Coast, while larval richness was similar in samples of both coasts. The Shannon index indicated uniformity in morphospecies abundances in both localities. Finally, the characterization of morphospecies from the gut of Anopheles albimanus mosquitoes from Colombia by culture-dependent methods complemented with 16S rRNA gene sequencing allowed the definition, at a finer resolution, of the composition and structure of these microbial communities. In addition, the obtained bacterial culture collection will allow further evaluation of the microorganisms for their potential as biocontrol agents.

Factors shaping the gut bacterial community assembly in two main Colombian malaria vectors.

BACKGROUND: The understanding of the roles of gut bacteria in the fitness and vectorial capacity of mosquitoes that transmit malaria, is improving; however, the factors shaping the composition and structure of such bacterial communities remain elusive. In this study, a high-throughput 16S rRNA gene sequencing was conducted to understand the effect of developmental stage, feeding status, species, and geography on the composition of the gut bacterial microbiota of two main Colombian malaria vectors, Anopheles nuneztovari and Anopheles darlingi. RESULTS: The results revealed that mosquito developmental stage, followed by geographical location, are more important determinants of the gut bacterial composition than mosquito species or adult feeding status. Further, they showed that mosquito gut is a major filter for environmental bacteria colonization. CONCLUSIONS: The sampling design and analytical approach of this study allowed to untangle the influence of factors that are simultaneously shaping the microbiota composition of two Latin-American malaria vectors, essential aspect for the design of vector biocontrol strategies.