Biology subjects

Alves, G.

Publications and source records attributed to Alves, G..

13 recordsLinked to original sources

bioRxiv2026

A novel qPCR assay to detect the presence of Anopheles gambiae complex mosquitoes

BackgroundAnopheles mosquitoes vector pathogens responsible for more than 600,000 human deaths annually. Ecological studies of these insects are important to guide effective vector-control campaigns and to understand their broader ecological consequences. Molecular ecology methods, particularly qPCR, provide a valuable tool in such studies. By detecting trace DNA of a taxon of interest within mixed or environmental samples, qPCR can facilitate identification of prey taxa of interest in the diets of consumers. However, no protocol for the detection of An. gambiae complex mosquitoes in dietary samples has been available. MethodsWe introduce a new set of qPCR primers (Agam_CO1_F1 and Agam_CO1_R1) and a probe-based assay for detection of Anopheles gambiae-complex mosquitoes, even with short reads common in dietary and environmental samples. The primers were tested in vitro for their specificity and sensitivity, and in silico using Primer-BLAST to assess potential off-target amplification. ResultsThe qPCR primers amplified An. gambiae DNA even at low starting concentrations (5 copies {micro}l-1). The primers did not amplify any non-target DNA in either the in vitro or in silico tests, but consistently amplified An. gambiae complex DNA. The primers can therefore provide reliable tests for the presence or absence of An. gambiae complex in dietary or eDNA samples. ConclusionsThe new qPCR primers should allow advances in research into mosquito ecology by allowing detection of even trace amounts of An. gambiae DNA in dietary and environmental samples. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=72 SRC="FIGDIR/small/707393v1_ufig1.gif" ALT="Figure 1"> View larger version (11K): org.highwire.dtl.DTLVardef@1fcce3corg.highwire.dtl.DTLVardef@47e5f5org.highwire.dtl.DTLVardef@4a7063org.highwire.dtl.DTLVardef@1188d60_HPS_FORMAT_FIGEXP M_FIG C_FIG

molecular biology↗

bioRxiv2026

Anopheles azevedoi (Ribeiro, 1969) in Angola: New geographical records, molecular characterization, and insecticide susceptibility profile of a commonly misidentified species

BACKGROUNDAngola ranks among the five countries with the highest malaria burden globally. The Ministry of Health in Angola has consistently partnered with international donors to sustain entomological surveillance and vector control strategies in a context of high malaria burden. METHODSVector surveillance was carried out in Luanda, Benguela, Namibe and Cuanza Sul provinces from 2016-2022. Collected adult mosquitoes were tested to assess the presence of Plasmodium parasites and determine blood sources. Larvae collections provided live material to test insecticide susceptibility in local Anopheles populations. Taxonomic determination of mosquitoes was based on external morphology and confirmed with molecular assays. The presence of Anopheles azevedoi was confirmed through morphology and genetic sequences, and errors in the original species determination were detected, discussed and corrected. OBJECTIVESThe study aimed to update the geographical range of Anopheles azevedoi in Angola and monitor the species susceptibility to public health insecticides. FINDINGS and MAIN CONCLUSIONSWe report on populations of Anopheles azevedoi occurring along the western coast of Angola, a highly abundant species with anthropophilic behavior in urban areas. Anopheles azevedoi is widely resistant to pyrethroids and DDT but fully susceptible to chlorfenapyr. We contribute with COI and ITS-2 barcoding sequences for future species identification and explain the reasons for which this species has been for long misidentified in Angola.

zoology↗

bioRxiv2025

Phenotypic and Genotypic Characterization of the Population of Phytophthora infestans in Brazil

Recently, phenotypic variations and differences in virulence have been observed among Brazilian isolates of P. infestans. Analysis of phenotypic variability can be useful to understand the pathogen population and how variants may affect late blight epidemics. Thus, this study aimed to characterize the population of P. infestans in Brazil using mating type, mitochondrial DNA haplotype, microsatellite genotype, and mefenoxam sensitivity. Isolates of the A2 mating type from tomato are reported for the first time. Isolates from potato plants were either of A1 or A2 mating types. The Ia and Ib mitochondrial DNA haplotypes were detected in Brazil. The Ia is associated with isolates from potato and the Ib with isolates obtained from potato and tomato. All tomato isolates were of a new genotype, SA_1_A2, while the EU_37_A2 and EU_2_A1 genotypes were detected among isolates sampled from potato fields. The sensitivity to mefenoxam varied: sensitive, intermediately insensitive, and insensitive isolates are present in potato and tomato crops. The population of P. infestans in Brazil remains structured according to host, but genotypes associated with potato or tomato plants seem to vary periodically.

plant biology↗

bioRxiv2025

Biological Function Assignment Across Taxonomic Levels in Mass-Spectrometry-Based Metaproteomics via a Modified Expectation Maximization Algorithm

A major challenge in mass-spectrometry-based metaproteomics is accurately identifying and quantifying biological functions across the full taxonomic lineage of microorganisms. This issue stems from what we refer to as the "shared confidently identified peptide problem". To address this issue, most metaproteomics tools rely on the lowest common ancestor (LCA) algorithm to assign biological functions, which often leads to incomplete biological function assignments across the full taxonomic lineage of identified microorganisms. To overcome this limitation, we implemented an expectation-maximization (EM) algorithm, along with a biological function database, within MiCId workflow. Using synthetic datasets, our study demonstrates that the enhanced MiCId workflow achieves better control over false discoveries and improved accuracy in microorganism identification and biomass estimation compared to Unipept and MetaGOmics. Additionally, the updated MiCId offers improved accuracy and better control of false discoveries in biological function identification compared to Unipept, along with reliable computation of function abundances across the full taxonomic lineage of identified microorganisms. Reanalyzing human oral and gut microbiome datasets using the enhanced MiCId workflow, we show that the results are consistent with those reported in the original publications, which were analyzed using the Galaxy-P platform with MEGAN5 and the MetaPro-IQ approach with Unipept, respectively.

bioinformatics↗

bioRxiv2024

Detection of bacteria through taste receptors primes the cellular immune response

Animals use their sensory system to detect cues in their external environment, then communicate, process, and integrate these cues through the nervous system in order to elicit a specific response. Taste is an important cue used by animals to explore their external environment and can modulate various aspects of animal behavior and physiology. A major ongoing challenge for animals is to detect and respond to the presence of a variety of microbes in their environment. However, to date, the links between the sensory system and the response to pathogenic threats remain poorly understood. Here we show that Drosophila melanogaster larvae use their taste system to detect bacterial peptidoglycans in their environment and respond by modulating the activity of their cellular immune system. We show that specific PeptidoGlycan Receptor Proteins (PGRPs) act in aversive taste neurons, via a non-canonical Immune Deficiency (Imd) pathway. These PGRPs mediate signaling in taste neurons and control immune cells production in the larval hematopoietic organ, the lymph gland. Taste-mediated sensing of bacteria in larvae primes the immune system, and improves survival after infection in adult flies. These results demonstrate that sensory inputs such as taste play an important role in protecting animals from bacterial infection by providing a powerful adaptive response to potential pathogens. Overall, our findings add to the growing list of examples of crosstalk between the nervous and immune systems and provide novel and important mechanisms for linking them. One Sentence SummaryNajera Mazariegos et al. demonstrate that organisms can use taste to monitor their environment for potential immune challenges and activate their immune system if they detect bacteria.

immunology↗

bioRxiv2023

Mitochondrial complex I deficiency stratifies idiopathic Parkinson's disease

Idiopathic Parkinsons disease (iPD) is believed to have a heterogeneous pathophysiology, but molecular disease subtypes have not been identified. Here, we show that iPD can be stratified according to the severity of neuronal respiratory complex I (CI) deficiency, and identify two emerging disease subtypes with distinct molecular and clinical profiles. The CI deficient (CI-PD) subtype accounts for approximately a fourth of all cases, and is characterized by anatomically widespread neuronal CI deficiency, a distinct cell type-specific gene expression profile, increased load of neuronal mtDNA deletions, and a predilection for non-tremor dominant motor phenotypes. In contrast, the non-CI deficient (nCI-PD) subtype exhibits no evidence of mitochondrial impairment outside the dopaminergic substantia nigra and has a predilection for a tremor dominant phenotype. These findings constitute a step towards resolving the biological heterogeneity of iPD with implications for both mechanistic understanding and treatment strategies.

neuroscience↗

bioRxiv2022

The Brazilian population of Fusarium oxysporum f. sp. cubense is not structured by VCG or by geographic origin

Fusarium wilt, caused by the soil-borne fungus Fusarium oxysporum f. sp. cubense (Foc), is considered one of the most destructive diseases of bananas. Paradoxically, knowledge of the genetics of the pathogen population in the Americas is very limited. In this study, a collection of 178 monosporic isolates from several banana producing regions, located in different climatic zones along a South to North transect in Brazil, was formed to assess the genetic structure of the population of Foc. The isolates underwent pathogenicity tests, PCR diagnosis for the detection of Tropical race 4 and screening to SIX homologs. The VCG of 119 isolates was determined by pairing against 17 testers. A group of 158 isolates was selected for microsatellite genotyping. There was moderate diversity of Foc in Brazil. Eight VCGs were identified: 0120, 0122, 0124, 0125, 0128, 01215, 01220, and 01222, of which 78% of isolates belong to a single VCG, while 22% of isolates belong to complexes of VCGs. The distribution of VCGs is uneven and independent of the banana genotype. VCGs were correlated with homologs of the SIX genes and varied according to geographic regions. Four SSR loci were polymorphic and on average 7.5 alleles were detected per locus. Thirty-five multilocus genotypes (MLGs) were identified. There was no association between VCG and MLGs and no genetic structure of the population of Foc in Brazil was detected.

microbiology↗

bioRxiv2021

Identification of antibiotic resistance proteins via MiCId's augmented workflow. A mass spectrometry-based proteomics approach

Fast and accurate identifications of pathogenic bacteria along with their associated antibiotic resistance proteins are of paramount importance for patient treatments and public health. While mass spectrometry has become an important, technique for diagnostics of infectious disease, there is a need for mass spectrometry workflows offering this capability. To meet this need, we have augmented the previously published Microorganism Classification and Identification (MiCId) workflow for this capability. To evaluate the performance of the newly augmented MiCId workflow, we have used MS/MS datafiles from samples of 10 antibiotic resistance bacterial strains belonging to three different species: Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa. The evaluation results show that MiCIds workflow has a sensitivity value around 85% (with a lower bound at about 72%) and a precision greater than 95% in the identification of antibiotic resistance proteins. Using MS/MS datasets from samples of two bacterial clonal isolates, one being antibiotic-sensitive while the other (obtained from the same patient at different times) being multidrug-resistant, we applied MiCIds workflow to investigate possible mechanisms of antibiotic resistance in these pathogenic bacteria; the results showed that MiCIds conclusions are in agreement with the published study. Furthermore, we show that MiCIds workflow is fast. It pro-vides microorganismal identifications, protein identifications, sample biomass estimates, and antibiotic resistance protein identifications in 6-17 minutes per MS/MS sample using computing resources that are available in most desktop and laptop computers, making it a highly portable workflow. This study demonstrated that MiCIds workflow is fast, portable, and with high sensitivity and high precision, making it a valuable tool for rapid identifications of bacteria as well as detection of their antibiotic resistance proteins. The new version of MiCId (v.07.01.2021) is freely available for download at https://www.ncbi.nlm.nih.gov/CBBresearch/Yu/downloads.html.

systems biology↗

bioRxiv2021

Weevil borers affect the spatio-temporal dynamics of banana Fusarium wilt

Dispersal of propagules of a pathogen has remarkable effects on the development of epidemics. Previous studies suggested that insect pests play a role in the development of Fusarium wilt (FW) epidemics in banana fields. We provided complementary evidence for the involvement of two insect pests of banana, the weevil borer (Cosmopolites sordidus L. - WB) and the false weevil borer (Metamasius hemipterus L. - FWB), in the dispersal of Fusarium oxysporum f. sp. cubense (Foc) using a comparative epidemiology approach under field conditions. Two banana plots located in a field with historical records of FW epidemics were used, one was managed with Beauveria bassiana to reduce the population of weevils, and the other was left without B. bassiana applications. The number of WB and FWB was monitored biweekly and the FW incidence was quantified bimonthly during two years. The population of WB and the incidence (6.7%) of FW in the plot managed with B. bassiana were lower than in the plot left unmanaged (13%). The monomolecular model best fitted the FW disease progress data and, as expected, the average estimated disease progress rate was lower in the plot managed with the entomopathogenic fungus (r = 0.0024) compared to the unmanaged plot (r = 0.0056). Aggregation of FW was higher in the field with WB management. WB affected the spatial and temporal dynamics of FW epidemics under field conditions and brought evidence that managing the insects may reduce FW of bananas intensity.

systems biology↗

bioRxiv2021

Ultra-deep whole genome bisulfite sequencing reveals a single methylation hotspot in human brain mitochondrial DNA

While DNA methylation is established as a major regulator of gene expression in the nucleus, the existence of mitochondrial DNA (mtDNA) methylation remains controversial. Here, we characterised the mtDNA methylation landscape in the prefrontal cortex of neurological healthy individuals (n=26) and patients with Parkinsons disease (n=27), using a combination of whole genome bisulfite sequencing (WGBS) and bisulfite-independent methods. Accurate mtDNA mapping from WGBS data required alignment to an mtDNA reference only, to avoid misalignment to nuclear mitochondrial pseudogenes. Once correctly aligned, WGBS data provided ultra-deep mtDNA coverage (16,723{+/-}7,711), and revealed overall very low levels of cytosine methylation. The highest methylation levels (5.49{+/-}0.97%) were found on CpG position m.545, located in the heavy-strand promoter 1 region. The m.545 methylation was validated using a combination of methylation-sensitive DNA digestion and quantitative PCR analysis. We detected no association between mtDNA methylation profile and Parkinsons disease. Interestingly, m.545 methylation correlated with the levels of mtDNA transcripts, suggesting a putative role in regulating mtDNA gene expression. In addition, we propose a robust framework for methylation analysis of mtDNA from WGBS data, which is less prone to false-positive findings due to misalignment of nuclear mitochondrial pseudogene sequences. Graphical abstract of the analyses and main findingsFresh-frozen brain tissue was obtained from the prefrontal cortex (Brodmann area 9) of 53 individuals, comprising 27 patients with idiopathic PD and 26 healthy controls. Tissue from the same samples was used in three different downstream analyses. WGBS was conducted on all 53 samples and the data were analysed using three different alignment strategies. Alignment against an mtDNA reference only was clearly superior as it gave the highest and most even depth of coverage. WGBS analysis revealed that mtDNA harbours very low levels of cytosine methylation, with the exception of the CpG position m.545 within the HSP1 region (lower right inset). The m.545 methylation was confirmed by bisulfite- and sequencing-independent methods, employing methylation-specific MspJI DNA digestion, followed by quantification with qPCR or fluorescent PCR and capillary electrophoresis. Finally, mtDNA transcript levels were determined by RT-qPCR and correlated to m.545 methylation levels, showing a positive association. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=196 SRC="FIGDIR/small/437685v1_ufig1.gif" ALT="Figure 1"> View larger version (33K): org.highwire.dtl.DTLVardef@1d478e3org.highwire.dtl.DTLVardef@4cbf9forg.highwire.dtl.DTLVardef@b863faorg.highwire.dtl.DTLVardef@1abe6c5_HPS_FORMAT_FIGEXP M_FIG C_FIG

genomics↗

bioRxiv2019

Dysregulation of histone acetylation and decoupling from transcription in Parkinson disease

Parkinson disease (PD) is a complex neurodegenerative disorder of largely unknown etiology. While several genetic risk factors have been identified, the involvement of epigenetics in the pathophysiology of PD is mostly unaccounted for. We conducted a histone acetylome-wide association study in PD, using brain tissue from two independent cohorts of cases and controls. Immunoblotting revealed increased acetylation at several histone sites in PD, with the most prominent change observed for H3K27, a marker of active promoters and enhancers. Chromatin immunoprecipitation sequencing (ChIP-seq) further indicated that H3K27 hyperacetylation in the PD brain is a genome-wide phenomenon, with a strong predilection for genes implicated in the disease, including SNCA, PARK7, PRKN and MAPT. Integration of the ChIP-seq with transcriptomic data revealed that the correlation between promoter H3K27 acetylation and gene expression is attenuated in PD patients, suggesting that H3K27 acetylation may be decoupled from transcription in the PD brain. Our findings strongly suggest that dysregulation of histone acetylation plays an important role in the pathophysiology of PD and identify novel epigenetic signatures associated with the disease.

genomics↗

bioRxiv2019

Common gene expression signatures in Parkinson’s disease are driven by changes in cell composition

BackgroundThe etiology of Parkinsons disease (PD) is largely unknown. Genome-wide transcriptomic studies in bulk brain tissue have identified several molecular signatures associated with the disease. While these studies have the potential to shed light into the pathogenesis of PD, they are also limited by two major confounders: RNA post mortem degradation and heterogeneous cell type composition of bulk tissue samples. We performed RNA sequencing following ribosomal RNA depletion in the prefrontal cortex of 49 individuals from two independent case-control cohorts. Using cell-type specific markers, we estimated the cell-type composition for each sample and included this in our analysis models to compensate for the variation in cell-type proportions.\n\nResultsRibosomal RNA depletion results in substantially more even transcript coverage, compared to poly(A) capture, in post mortem tissue. Moreover, we show that cell-type composition is a major confounder of differential gene expression analysis in the PD brain. Correcting for cell-type proportions attenuates numerous transcriptomic signatures that have been previously associated with PD, including vesicle trafficking, synaptic transmission, immune and mitochondrial function. Conversely, pathways related to endoplasmic reticulum, lipid oxidation and unfolded protein response are strengthened and surface as the top differential gene expression signatures in the PD prefrontal cortex.\n\nConclusionsDifferential gene expression signatures in PD bulk brain tissue are significantly confounded by underlying differences in cell-type composition. Modeling cell-type heterogeneity is crucial in order to unveil transcriptomic signatures that represent regulatory changes in the PD brain and are, therefore, more likely to be associated with underlying disease mechanisms.

neuroscience↗

bioRxiv2019

Genome-wide association study of Parkinson's disease progression biomarkers in 12 longitudinal patients' cohorts

BackgroundSeveral reports have identified different patterns of Parkinsons disease progression in individuals carrying missense variants in the GBA or LRRK2 genes. The overall contribution of genetic factors to the severity and progression of Parkinsons disease, however, has not been well studied. ObjectivesTo test the association between genetic variants and the clinical features and progression of Parkinsons disease on a genome-wide scale. MethodsWe accumulated individual data from 12 longitudinal cohorts in a total of 4,093 patients with 25,254 observations over a median of 3.81 years. Genome-wide associations were evaluated for 25 cross-sectional and longitudinal phenotypes. Specific variants of interest, including 90 recently-identified disease risk variants, were also investigated for the associations with these phenotypes. ResultsTwo variants were genome-wide significant. Rs382940(T>A), within the intron of SLC44A1, was associated with reaching Hoehn and Yahr stage 3 or higher faster (HR 2.04 [1.58, 2.62], P-value = 3.46E-8). Rs61863020(G>A), an intergenic variant and eQTL for ADRA2A, was associated with a lower prevalence of insomnia at baseline (OR 0.63 [0,52, 0.75], P-value = 4.74E-8). In the targeted analysis, we found nine associations between known Parkinsons risk variants and more severe motor/cognitive symptoms. Also, we replicated previous reports of GBA coding variants (rs2230288: p.E365K, rs75548401: p.T408M) being associated with greater motor and cognitive decline over time, and APOE E4 tagging variant (rs429358) being associated with greater cognitive deficits in patients. ConclusionsWe identified novel genetic factors associated with heterogeneity of progression in Parkinsons disease. The results provide new insights into the pathogenesis of Parkinsons disease as well as patient stratification for clinical trials.

genetics↗