Fungal diversity in shade-coffee plantations in Soconusco, Mexico

Background As forested natural habitats disappear in the world, traditional, shade-coffee plantations offer an opportunity to conserve biodiversity and ecosystem services. Traditional coffee plantations maintain a diversity of tree species that provide shade for coffee bushes and, at the same time, are important repositories for plants and animals that inhabited the original cloud forest. However, there is still little information about shade-coffee plantation’s fungal diversity despite their relevance for ecosystem functioning as decomposers, symbionts and pathogens. Specifically, it is unknown if and what mycorrhizae-forming fungi can be found on the branches and trunks of coffee bushes and trees, which hold a diversity of epiphytes. Here, we evaluate fungal communities on specific plant microsites on both coffee bushes and shade trees. We investigate the ecological roles played by this diversity, with a special focus on mycorrhizae-forming fungi that may enable the establishment and development of epiphytic plants. Methods We collected 48 bark samples from coffee bushes and shade trees (coffee; tree), from four plant microsites (upper and lower trunks, branches and twigs), in two shade-coffee plantations in the Soconusco region in southern Mexico, at different altitudes. We obtained ITS amplicon sequences that served to estimate alpha and beta diversity, to assign taxonomy and to infer the potential ecological role played by the detected taxa. Results The bark of shade trees and coffee bushes supported high fungal diversity (3,783 amplicon sequence variants). There were no strong associations between community species richness and collection site, plant type or microsite. However, we detected differences in beta diversity between collection sites. All trophic modes defined by FUNGuild database were represented in both plant types. However, when looking into guilds that involve mycorrhizae formation, the CLAM test suggests that coffee bushes are more likely to host taxa that may function as mycorrhizae. Discussion We detected high fungal diversity in shade-coffee plantations in Soconusco, Chiapas, possibly remnants of the original cloud forest ecosystem. Several mycorrhiza forming fungi guilds occur on the bark of coffee bushes and shade trees in this agroecosystem, with the potential of supporting epiphyte establishment and development. Thus, traditional coffee cultivation could be part of an integrated strategy for restoration and conservation of epiphytic populations. This is particularly relevant for conservation of threatened species of Orchidaceae that are highly dependent on mycorrhizae formation

Tween 80 Improves the Acid-Fast Bacilli Quantification in the Magnetic Nanoparticle-Based Colorimetric Biosensing Assay (NCBA)

Despite its reduced sensitivity, sputum smear microscopy (SSM) remains the main diagnostic test for detecting tuberculosis in many parts of the world. A new diagnostic technique, the magnetic nanoparticle-based colorimetric biosensing assay (NCBA) was optimized by evaluating different concentrations of glycan-functionalized magnetic nanoparticles (GMNP) and Tween 80 to improve the acid-fast bacilli (AFB) count. Comparative analysis was performed on 225 sputum smears: 30 with SSM, 107 with NCBA at different GMNP concentrations, and 88 with NCBA-Tween 80 at various concentrations and incubation times. AFB quantification was performed by adding the total number of AFB in all fields per smear and classified according to standard guidelines (scanty, 1+, 2+ and 3+). Smears by NCBA with low GMNP concentrations (≤1.5 mg/mL) showed higher AFB quantification compared to SSM. Cell enrichment of sputum samples by combining NCBA-GMNP, incubated with Tween 80 (5%) for three minutes, improved capture efficiency and increased AFB detection up to 445% over SSM. NCBA with Tween 80 offers the opportunity to improve TB diagnostics, mainly in paucibacillary cases. As this method provides biosafety with a simple and inexpensive methodology that obtains results in a short time, it might be considered as a point-of-care TB diagnostic method in regions where resources are limited

Time to Switch to Second-line Antiretroviral Therapy in Children With Human Immunodeficiency Virus in Europe and Thailand.

Background: Data on durability of first-line antiretroviral therapy (ART) in children with human immunodeficiency virus (HIV) are limited. We assessed time to switch to second-line therapy in 16 European countries and Thailand. Methods: Children aged <18 years initiating combination ART (≥2 nucleoside reverse transcriptase inhibitors [NRTIs] plus nonnucleoside reverse transcriptase inhibitor [NNRTI] or boosted protease inhibitor [PI]) were included. Switch to second-line was defined as (i) change across drug class (PI to NNRTI or vice versa) or within PI class plus change of ≥1 NRTI; (ii) change from single to dual PI; or (iii) addition of a new drug class. Cumulative incidence of switch was calculated with death and loss to follow-up as competing risks. Results: Of 3668 children included, median age at ART initiation was 6.1 (interquartile range (IQR), 1.7-10.5) years. Initial regimens were 32% PI based, 34% nevirapine (NVP) based, and 33% efavirenz based. Median duration of follow-up was 5.4 (IQR, 2.9-8.3) years. Cumulative incidence of switch at 5 years was 21% (95% confidence interval, 20%-23%), with significant regional variations. Median time to switch was 30 (IQR, 16-58) months; two-thirds of switches were related to treatment failure. In multivariable analysis, older age, severe immunosuppression and higher viral load (VL) at ART start, and NVP-based initial regimens were associated with increased risk of switch. Conclusions: One in 5 children switched to a second-line regimen by 5 years of ART, with two-thirds failure related. Advanced HIV, older age, and NVP-based regimens were associated with increased risk of switch

Microbios ¿amigos o enemigos?

Cuando escuchamos o leemos la palabra “microbio”, casi siempre la asociamos con enfermedades, epidemias, mugre o algo que hay que evitar. Entonces, parecerá increíble saber que muchas personas dedicadas a la actividad científica coinciden en que la vida en el planeta, tal como la conocemos hoy, no hubiera sido posible de no ser por los microbios, y que incluso si lográramos eliminarlos de nuestro propio organismo con desinfectantes, antibióticos y demás productos, simplemente dejaríamos de existir

Microbios ¿amigos o enemigos?

Cuando escuchamos o leemos la palabra “microbio”, casi siempre la asociamos con enfermedades, epidemias, mugre o algo que hay que evitar. Entonces, parecerá increíble saber que muchas personas dedicadas a la actividad científica coinciden en que la vida en el planeta, tal como la conocemos hoy, no hubiera sido posible de no ser por los microbios, y que incluso si lográramos eliminarlos de nuestro propio organismo con desinfectantes, antibióticos y demás productos, simplemente dejaríamos de existir

Unveiling the Microbial Ecology behind Mezcal: A Spirit Drink with a Growing Global Demand

The advent of omics has expanded our knowledge of microbial ecology behind Mezcal, a fermented spirit made from the juices of cooked Agave plants (Agave spp., Asparagaceae). Mezcal has been produced in Mexico for over 200 years, however, has been in high demand since its discovery by international markets in the last decade. Mezcal is appreciated for its diverse and complex sensory profile, which is tied to the geographic and environmental diversity of the different Mezcal-producing regions. This regional typicity is brought about by spontaneous fermentation consortia that act in loosely controlled artisanal fermentation processes. Previous works have mainly concentrated on microorganisms involved in the biosynthesis of alcohol and other volatile compounds, or from a different perspective, on culturable microorganisms (mainly yeasts) influencing the taste profile. Attention has been aimed at the richness of microbial populations in point events or under laboratory conditions, which leaves much of the biological richness out of account. Omics techniques have become powerful tools for characterizing the composition of autochthonous fermentation microbiota, regional or endemic features, and ecological processes that determine the dynamics of Mezcal fermentation. The analyses of genetic material, proteins, and metabolites allow disentangling the biological complexity of Mezcal production. This review presents the reader with an up-to-date overview of publications that discuss microbial communities in Mezcal fermentation, metabolic pathways regulated by microbial interactions, and the application of omics to characterize the spontaneous fermenting microbiota conformation and dynamics considering the subjacent ecological processes

Metagenomic analysis of plant viruses in tropical fresh and wastewater

Abstract Due to considerable losses in the agricultural sector, phytopathogenic viruses are of significant economic importance. Several phytopathogenic viruses have been detected in water bodies. These can be transmitted to their hosts through water contact. However, relatively few studies concentrate on the presence and impact of plant‐infecting viruses in water environments. In the present study, we used a metagenomic approach to determine viral phytopathogen diversity in wastewater, freshwater, and sludge from the southernmost tropical region in Mexico. We detected 15 different phytopathogenic viruses that belonged to the family Virgaviridae, genus Tobamovirus. Pepper mild mottle virus, tomato brown rugose fruit virus, and tropical soda apple mosaic virus had the highest number of reads. Phylogenetic analysis of the coat protein (CP) amino acid sequence revealed a close relationship between the solanaceous plant‐infecting viruses and the reference sequences found in GenBank. On the other hand, the CP amino acid sequence of Fabaceae and Cucurbitaceae‐infecting viruses did not show such a close relationship when compared with GenBank reference sequences. These results suggested that detected viral sequences could originate from new strains or species of local viruses likely from the consumption of locally cultivated produce, whereas those infecting solanaceous plants might come from consumption of processed or imported food to Mexico. The prevalence of a high diversity of plant viruses in wastewater and rivers and their potential infectivity raise concerns about the use of contaminated freshwater and wastewater in agriculture in Mexico

Complex regulation of Hsf1-Skn7 activities by the catalytic subunits of PKA in Saccharomyces cerevisiae: experimental and computational evidences

Background. The cAMP-dependent protein kinase regulatory network (PKA-RN) regulates metabolism, memory, learning, development, and response to stress. Previous models of this network considered the catalytic subunits (CS) as a single entity, overlooking their functional individualities. Furthermore, PKA-RN dynamics are often measured through cAMP levels in nutrient-depleted cells shortly after being fed with glucose, dismissing downstream physiological processes. Results Here we show that temperature stress, along with deletion of PKA-RN genes, significantly affected HSE-dependent gene expression and the dynamics of the PKA-RN in cells growing in exponential phase. Our genetic analysis revealed complex regulatory interactions between the CS that influenced the inhibition of Hsf1/Skn7 transcription factors. Accordingly, we found new roles in growth control and stress response for Hsf1/Skn7 when PKA activity was low (cdc25Δ cells). Experimental results were used to propose an interaction scheme for the PKA-RN and to build an extension of a classic synchronous discrete modeling framework. Our computational model reproduced the experimental data and predicted complex interactions between the CS and the existence of a repressor of Hsf1/Skn7 that is activated by the CS. Additional genetic analysis identified Ssa1 and Ssa2 chaperones as such repressors. Further modeling of the new data foresaw a third repressor of Hsf1/Skn7, active only in theabsence of Tpk2. By averaging the network state over all its attractors, a good quantitative agreement between computational and experimental results was obtained, as the averages reflected more accurately the population measurements. Conclusions The assumption of PKA being one molecular entity has hindered the study of a wide range of behaviors. Additionally, the dynamics of HSE-dependent gene expression cannot be simulated accurately by considering the activity of single PKA-RN components (i.e., cAMP, individual CS, Bcy1, etc.). We show that the differential roles of the CS are essential to understand the dynamics of the PKA-RN and its targets. Our systems level approach, which combined experimental results with theoretical modeling, unveils the relevance of the interaction scheme for the CS and offers quantitative predictions for several scenarios (WT vs. mutants in PKA-RN genes and growth at optimal temperature vs. heat shock).Anesthesiology, Pharmacology and Therapeutics, Department ofMedicine, Faculty ofNon UBCReviewedFacult