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1.
Mucosal Immunol ; 2024 Jun 24.
Artigo em Inglês | MEDLINE | ID: mdl-38925529

RESUMO

Dietary proteins are taken up by intestinal dendritic cells (DCs), cleaved into peptides, loaded to major histocompatibility complexes, and presented to T cells to generate an immune response. Amino acid (AA)-diets do not have the same effects because AAs cannot bind to major histocompatibility complex to activate T cells. Here, we show that impairment in regulatory T cell generation and loss of tolerance in mice fed a diet lacking whole protein is associated with major transcriptional changes in intestinal DCs including downregulation of genes related to DC maturation, activation and decreased gene expression of immune checkpoint molecules. Moreover, the AA-diet had a profound effect on microbiome composition, including an increase in Akkermansia muciniphilia and Oscillibacter and a decrease in Lactococcus lactis and Bifidobacterium. Although microbiome transfer experiments showed that AA-driven microbiome modulates intestinal DC gene expression, most of the unique transcriptional change in DC was linked to the absence of whole protein in the diet. Our findings highlight the importance of dietary proteins for intestinal DC function and mucosal tolerance.

2.
Fam Cancer ; 23(3): 233-246, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38780705

RESUMO

Pancreatic ductal adenocarcinoma (PDAC) is the fourth leading cause of cancer-related death in the Western world. The number of diagnosed cases and the mortality rate are almost equal as the majority of patients present with advanced disease at diagnosis. Between 4 and 10% of pancreatic cancer cases have an apparent hereditary background, known as hereditary pancreatic cancer (HPC) and familial pancreatic cancer (FPC), when the genetic basis is unknown. Surveillance of high-risk individuals (HRI) from these families by imaging aims to detect PDAC at an early stage to improve prognosis. However, the genetic basis is unknown in the majority of HRIs, with only around 10-13% of families carrying known pathogenic germline mutations. The aim of this study was to assess an individual's genetic cancer risk based on sex and personal and family history of cancer. The Best Linear Unbiased Prediction (BLUP) methodology was used to estimate an individual's predicted risk of developing cancer during their lifetime. The model uses different demographic factors in order to estimate heritability. A reliable estimation of heritability for pancreatic cancer of 0.27 on the liability scale, and 0.07 at the observed data scale as obtained, which is different from zero, indicating a polygenic inheritance pattern of PDAC. BLUP was able to correctly discriminate PDAC cases from healthy individuals and those with other cancer types. Thus, providing an additional tool to assess PDAC risk HRI with an assumed genetic predisposition in the absence of known pathogenic germline mutations.


Assuntos
Carcinoma Ductal Pancreático , Predisposição Genética para Doença , Mutação em Linhagem Germinativa , Neoplasias Pancreáticas , Humanos , Carcinoma Ductal Pancreático/genética , Neoplasias Pancreáticas/genética , Feminino , Masculino , Pessoa de Meia-Idade , Adulto , Idoso , Medição de Risco/métodos , Carcinoma
3.
Fam Cancer ; 23(3): 383-392, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38753287

RESUMO

The Spanish Familial Pancreatic Cancer Registry (PANGENFAM) was established in 2009 and aims to characterize the genotype and phenotype of familial pancreatic cancer (FPC). Furthermore, an early detection screening program for pancreatic ductal adenocarcinoma (PDAC) is provided to healthy high-risk individuals from FPC and hereditary pancreatic cancer families (first-degree relatives). This article describes our experience over the last 10 years in high-risk screening. Hereditary and familial pancreatic cancer families were identified through the oncology and gastroenterology units. High-risk individuals underwent annual screening with endoscopic ultrasound (EUS) and magnetic resonance (MRI) from age 40 or 10 years younger than the youngest affected family member. Results: PANGENFAM has enrolled 290 individuals from 143 families, including 52 PDAC cases and 238 high-risk individuals. All high-risk individuals eligible for screening were offered to enter the surveillance program, with 143 currently participating. Pancreatic abnormalities were detected in 94 individuals (median age 53 years (29-83), with common findings including cystic lesions and inhomogeneous parenchyma. Imaging test concordance was 66%. Surgical intervention was performed in 4 high-risk individuals following highly suspicious lesions detected by imaging. PANGENFAM is a valuable resource for science innovation, such as biobanking, with clinical and imaging data available for analysis. For high-risk families, it may offer a potential for early diagnosis. Collaboration with other national and international registries is needed to increase our understanding of the disease biology and to standardize criteria for inclusion and follow-up, optimizing cost-effectiveness and efficacy.


Assuntos
Carcinoma Ductal Pancreático , Neoplasias Pancreáticas , Sistema de Registros , Humanos , Neoplasias Pancreáticas/genética , Pessoa de Meia-Idade , Espanha , Feminino , Masculino , Adulto , Idoso , Seguimentos , Carcinoma Ductal Pancreático/genética , Carcinoma Ductal Pancreático/diagnóstico por imagem , Carcinoma Ductal Pancreático/diagnóstico , Idoso de 80 Anos ou mais , Predisposição Genética para Doença , Detecção Precoce de Câncer/métodos , Imageamento por Ressonância Magnética , Endossonografia , Fatores de Risco , Carcinoma
4.
Brain Behav Immun ; 117: 242-254, 2024 03.
Artigo em Inglês | MEDLINE | ID: mdl-38281671

RESUMO

Intestinal γδ T cells play an important role in shaping the gut microbiota, which is critical not only for maintaining intestinal homeostasis but also for controlling brain function and behavior. Here, we found that mice deficient for γδ T cells (γδ-/-) developed an abnormal pattern of repetitive/compulsive (R/C) behavior, which was dependent on the gut microbiota. Colonization of WT mice with γδ-/- microbiota induced R/C behavior whereas colonization of γδ-/- mice with WT microbiota abolished the R/C behavior. Moreover, γδ-/- mice had elevated levels of the microbial metabolite 3-phenylpropanoic acid in their cecum, which is a precursor to hippurate (HIP), a metabolite we found to be elevated in the CSF. HIP reaches the striatum and activates dopamine type 1 (D1R)-expressing neurons, leading to R/C behavior. Altogether, these data suggest that intestinal γδ T cells shape the gut microbiota and their metabolites and prevent dysfunctions of the striatum associated with behavior modulation.


Assuntos
Microbioma Gastrointestinal , Hipuratos , Linfócitos T , Animais , Camundongos , Corpo Estriado , Neurônios , Comportamento Compulsivo
5.
J Vis Exp ; (195)2023 05 26.
Artigo em Inglês | MEDLINE | ID: mdl-37306424

RESUMO

Tumor organoids are three-dimensional (3D) ex vivo tumor models that recapitulate the biological key features of the original primary tumor tissues. Patient-derived tumor organoids have been used in translational cancer research and can be applied to assess treatment sensitivity and resistance, cell-cell interactions, and tumor cell interactions with the tumor microenvironment. Tumor organoids are complex culture systems that require advanced cell culture techniques and culture media with specific growth factor cocktails and a biological basement membrane that mimics the extracellular environment. The ability to establish primary tumor cultures highly depends on the tissue of origin, the cellularity, and the clinical features of the tumor, such as the tumor grade. Furthermore, tissue sample collection, material quality and quantity, as well as correct biobanking and storage are crucial elements of this procedure. The technical capabilities of the laboratory are also crucial factors to consider. Here, we report a validated SOP/protocol that is technically and economically feasible for the culture of ex vivo tumor organoids from fresh tissue samples of pancreatic adenocarcinoma origin, either from fresh primary resected patient donor tissue or patient-derived xenografts (PDX). The technique described herein can be performed in laboratories with basic tissue culture and mouse facilities and is tailored for wide application in the translational oncology field.


Assuntos
Adenocarcinoma , Neoplasias Pancreáticas , Humanos , Animais , Camundongos , Bancos de Espécimes Biológicos , Fibroblastos , Organoides , Microambiente Tumoral , Neoplasias Pancreáticas
6.
Microbiome ; 11(1): 32, 2023 02 23.
Artigo em Inglês | MEDLINE | ID: mdl-36814316

RESUMO

BACKGROUND: Gamma-delta (γδ) T cells are a major cell population in the intestinal mucosa and are key mediators of mucosal tolerance and microbiota composition. Little is known about the mechanisms by which intestinal γδ T cells interact with the gut microbiota to maintain tolerance. RESULTS: We found that antibiotic treatment impaired oral tolerance and depleted intestinal γδ T cells, suggesting that the gut microbiota is necessary to maintain γδ T cells. We also found that mice deficient for γδ T cells (γδ-/-) had an altered microbiota composition that led to small intestine (SI) immune dysregulation and impaired tolerance. Accordingly, colonizing WT mice with γδ-/- microbiota resulted in SI immune dysregulation and loss of tolerance whereas colonizing γδ-/- mice with WT microbiota normalized mucosal immune responses and restored mucosal tolerance. Moreover, we found that SI γδ T cells shaped the gut microbiota and regulated intestinal homeostasis by secreting the fecal micro-RNA let-7f. Importantly, oral administration of let-7f to γδ-/- mice rescued mucosal tolerance by promoting the growth of the γδ-/--microbiota-depleted microbe Ruminococcus gnavus. CONCLUSIONS: Taken together, we demonstrate that γδ T cell-selected microbiota is necessary and sufficient to promote mucosal tolerance, is mediated in part by γδ T cell secretion of fecal micro-RNAs, and is mechanistically linked to restoration of mucosal immune responses. Video Abstract.


Assuntos
MicroRNAs , Microbiota , Camundongos , Animais , Linfócitos T , Receptores de Antígenos de Linfócitos T gama-delta/genética , Intestinos , Mucosa Intestinal , Imunidade nas Mucosas
7.
J. coloproctol. (Rio J., Impr.) ; 42(3): 266-272, July-Sept. 2022. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1421981

RESUMO

Abstract Aim This study characterizes Colorectal Cancer (CRC) incidence in the University Hospital Ramon and Cajal, Madrid, and analyzes variations over time. It establishes risk groups, aiming to discover whether diagnosis can be determined in less advanced stages of disease. Method Evolutionary epidemiological study of genetic and environmental factors contributing to the development of CRC in this district that enables the comparison of two cohorts of patients separated by 37 years: G1 (patients of current group) and G2 (patients of historical group). The main risk variables gleaned retrospectively were analyzed and the statistical association between cohorts was determined. Results The mean age of patients increased significantly from 64 to 71 along with the incidence of ascending colon cancer. G1 scored higher than G2 for: the incidence of colon cancer in men, detection of adenomatous polyps (48.1%), percentage of resectability with curative intent (80.4%), and Dukes A stage (34.1%) (p < 0.001). Conclusion Biological aspects of CRC have been compared against its profile three decades earlier. We can confirm the existence of concrete changes in the manifestation and staging at the time of diagnosis or following earlier treatment. (AU)


Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Idoso , Idoso de 80 Anos ou mais , Neoplasias Colorretais/epidemiologia , Fatores de Risco , Pólipos Adenomatosos , Anamnese , Estadiamento de Neoplasias
12.
Nat Commun ; 12(1): 4907, 2021 08 13.
Artigo em Inglês | MEDLINE | ID: mdl-34389726

RESUMO

The intestinal mucosa constitutes an environment of closely regulated immune cells. Dendritic cells (DC) interact with the gut microbiome and antigens and are important in maintaining gut homeostasis. Here, we investigate DC transcriptome, phenotype and function in five anatomical locations of the gut lamina propria (LP) which constitute different antigenic environments. We show that DC from distinct gut LP compartments induce distinct T cell differentiation and cytokine secretion. We also find that PD-L1+ DC in the duodenal LP and XCR1+ DC in the colonic LP comprise distinct tolerogenic DC subsets that are crucial for gut homeostasis. Mice lacking PD-L1+ and XCR1+ DC have a proinflammatory gut milieu associated with an increase in Th1/Th17 cells and a decrease in Treg cells and have exacerbated disease in the models of 5-FU-induced mucositis and DSS-induced colitis. Our findings identify PD-L1+ and XCR1+ DC as region-specific physiologic regulators of intestinal homeostasis.


Assuntos
Antígeno B7-H1/imunologia , Células Dendríticas/imunologia , Homeostase/imunologia , Mucosa Intestinal/imunologia , Receptores de Quimiocinas/imunologia , Animais , Antígeno B7-H1/genética , Antígeno B7-H1/metabolismo , Colite/genética , Colite/imunologia , Colite/metabolismo , Citocinas/imunologia , Citocinas/metabolismo , Células Dendríticas/metabolismo , Fezes/microbiologia , Feminino , Microbioma Gastrointestinal/genética , Microbioma Gastrointestinal/imunologia , Homeostase/genética , Humanos , Mucosa Intestinal/metabolismo , Mucosa Intestinal/microbiologia , Masculino , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Transgênicos , Receptores de Quimiocinas/genética , Receptores de Quimiocinas/metabolismo , Linfócitos T/citologia , Linfócitos T/imunologia , Linfócitos T/metabolismo , Transcriptoma/genética , Transcriptoma/imunologia
14.
Cir. Esp. (Ed. impr.) ; 99(5): 374-378, may. 2021. ilus
Artigo em Espanhol | IBECS | ID: ibc-218147

RESUMO

La fístula pancreática tras la pancreatectomía distal supone una de las complicaciones más frecuentes y con mayor morbilidad asociada en este tipo de cirugía. Hasta el momento ningún método empleado se ha mostrado eficaz en reducir la fístula pancreática de forma contrastada. Presentamos el desarrollo de una nueva técnica quirúrgica que combina la colocación del stent pancreático junto con la colocación de parche autólogo de ligamento redondo sobre el borde pancreático. Tras la realización de la transección pancreática, se introduce de forma anterógrada la prótesis pancreática previo paso de una guía. Se comprueba la colocación de la prótesis y se cierra el conducto de Wirsung. Por último, se realiza una plastia autóloga de ligamento falciforme. A las 6-8 semanas se retira la prótesis vía endoscópica. La técnica ha sido empleada en 2 pacientes sin presentar fístula pancreática postoperatoria. Esta técnica combinada pretende introducir un nuevo elemento en la prevención de la fístula pancreática mediante una técnica reproducible sin una dificultad técnica añadida. (AU)


Postoperative pancreatic fistula in distal pancreatectomy is one of the most important complications in this surgery and it is associated with high morbidity and mortality. Pancreatic fistula after distal pancreatectomy remains an unsolved problem and none preventive procedure has been shown effectively. We present a new technique that combine pancreatic stent placement with round ligament autologous patch over pancreatic edge. A guide is introduced through Wirsung duct prior to stent placement. After stent assessment, Wirsung duct is closed. Finally, falciform ligament autologous patch is placed over pancreatic edge. After 6-8 weeks, the stent is removed by oral endoscopy. This technique introduces a new issue on the pancreatic fistula prevention. (AU)


Assuntos
Humanos , Fístula Pancreática/complicações , Fístula Pancreática/prevenção & controle , Fístula Pancreática/cirurgia , Pancreatectomia , Ligamentos Redondos , Ductos Pancreáticos/cirurgia
15.
Cir Esp (Engl Ed) ; 99(5): 374-378, 2021 May.
Artigo em Inglês, Espanhol | MEDLINE | ID: mdl-33388136

RESUMO

Postoperative pancreatic fistula in distal pancreatectomy is one of the most important complications in this surgery and it is associated with high morbidity and mortality. Pancreatic fistula after distal pancreatectomy remains an unsolved problem and none preventive procedure has been shown effectively. We present a new technique that combine pancreatic stent placement with round ligament autologous patch over pancreatic edge. A guide is introduced through Wirsung duct prior to stent placement. After stent assessment, Wirsung duct is closed. Finally, falciform ligament autologous patch is placed over pancreatic edge. After 6-8 weeks, the stent is removed by oral endoscopy. This technique introduces a new issue on the pancreatic fistula prevention.

16.
Environ Sci Pollut Res Int ; 28(12): 14555-14575, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33211290

RESUMO

In the Basin of Mexico, one of the most important economic regions in the country with over 22 million inhabitants, peri-urban streams have been transformed into sewers, drains, and pipes to avoid flooding or unsanitary conditions; the change affects not only the ecosystem's goods and services but also the aquatic communities that support the natural ecological processes. We aimed to develop a diatom-based diagnosis of the ecological quality of these aquatic ecosystems through the study of epilithic diatom response to regional environmental gradients. Samples of epilithic diatoms and water were collected in 45 sites representing 12 perennial streams, and multivariate analyses were performed on environmental and biological data. The ecological quality gradient to which diatoms responded was related to oxygen saturation, soluble reactive phosphorous, dissolved inorganic nitrogen, and hydromorphological quality. Three species groups were recognized according to their ecological preferences along CCA1 axis, indicators of high and low ecological quality, and tolerant species. By assigning an indicator value to each species group, we calculated the DEQI using the formula adapted from Pantle and Buck, indicating five different ecological quality classes. This index is proposed for complementing the ecological quality evaluation as a biological metric that responds to multiple regional stressors of the structure and function of these peri-urban streams in the Basin of Mexico.


Assuntos
Diatomáceas , Rios , Ecossistema , Monitoramento Ambiental , México
17.
Environ Sci Pollut Res Int ; 28(7): 8698-8708, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33068242

RESUMO

This research aimed at determining reference sites in southern Brazilian lotic systems, using the Water Quality Trophic Index (WQTI), based on epilithic diatom communities. Within the period of 2012 and 2016, 140 epilithic diatom samples from seven sampling sites within the spring area of the Andreas Stream Hydrographic Basin, Vera Cruz County, RS, Brazil, were analyzed quarterly. Similarly, from 2012 and 2014, water samples in these sampling sites were collected for determination of physicochemical and microbiological parameters, such as water temperature, pH, turbidity, dissolved oxygen, biochemical oxygen demand, nitrate, phosphate, total dissolved solids, and fecal coliforms. The results indicated the occurrence of two diatom samples groups with significant differences (p < 0.05). Group 1 (sampling sites P1, P4, and P5) was classified in the oligotrophic category with an average WQTI of 1.3 ± 0.2, while group 2 (sampling sites P2, P3, P6, and P7) was classified in the ß-mesotrophic category with an average value of 2.0 ± 0.4). Achnanthidium minutissimum and Platessa hustedtii the dominant species were classified as oligotrophic taxa. Thus, we proposed the upper limit of the first interval of the WQTI score scale, equal to 1.5, as a reference value to differentiate the limits between the "high" and "good" ecological status to determine "reference sites" for subtropical and temperate Brazilian aquatic systems. The physical, chemical, and microbiological water quality evaluation gives support to this proposal, as far as the results indicated a significant improvement in the water quality, classifying sampling sites P1, P4, and P5 as having good quality appropriate for multiple uses.


Assuntos
Diatomáceas , Brasil , Monitoramento Ambiental , Rios , Qualidade da Água
19.
Water Sci Technol ; 82(6): 1131-1141, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-33055403

RESUMO

The accumulation of environmental pesticides can cause problems in aquatic ecosystems and adverse effects in humans. These compounds can be found in water due to runoff from agricultural, industrial and domestic applications. In Southern Brazil, tobacco cultivation is one of the most important economic agricultural activities. The bifenthrin pesticide, classified as having moderate toxicity (class II), is commonly used as an insecticide in this culture. In this context, the present research aimed to study the performance of microalgae-induced bioremediation processes. Microalgae were isolated from a natural water source in the city of Santa Cruz do Sul, RS, Brazil, which is an artificial reservoir used for public water supply. For this purpose, biodegradation, biosorption, influence of pH, percentage of inoculum and photoperiod were evaluated in batch experiments for 20 cultivation days. After the phycoremediation process, the bifenthrin pesticide (m/z = 181) was quantified by gas chromatography with mass spectrometry (GC-MS). The results indicated that microalgae isolated from the water of the lake were able to contribute to the removal of approximately 99% of bifenthrin through biodegradation and biosorption processes. Photodegradation was identified (>77%) and the best condition for the phycoremediation was 20% inoculum with a photoperiod of 18:6 h.


Assuntos
Microalgas , Piretrinas , Brasil , Ecossistema
20.
C R Biol ; 343(1): 41-52, 2020 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-32720487

RESUMO

Diatom identification is a key step in using these microorganisms as water quality bioindicators. Morphological diagnosis is a difficult task due to the enormous number of species and their microscopic size. This can be overcome using molecular tools to complement the diagnosis. The main goal of this work was to obtain the DNA barcode of Ecuadorian epilithic diatoms with a wide geographical distribution, a well-defined ecological range and characteristics that allow them to be reliable indicator species. Unialgal diatom cultures were obtained from environmental samples of Ecuadorian Andean streams. Morphological characterization of cultures was carried out under SEM microscopy. For molecular characterization, 18SV4 and rbcL barcodes were sequenced from each strain and blasted against a GenBank database. A phylogenetic tree for each barcode was constructed using the ML method including sequences of strains of the studied species from different geographical locations. The results showed the following five species to be suitable as bioindicators and these were isolated. Sellaphora seminulum (strain JA01b, c), Nitzschia fonticola (strain SP02a) and N. palea (strain CA01a) are tolerant to eutrophication; Eolimna minima (strain CH02a) is a mesotrophic water bioindicator, and Achnanthidium minutissimum (strain JA01a) is an oligotrophic water bioindicator. The comparison with the GenBank database of the barcoding regions supported the morphological identification. The barcoding sequences of the strains showed a high percentage of identity with the sequences reported in INSDC databases for the same species. The topology of the phylogenetic trees demonstrates that epilithic diatoms from Ecuador are closely related to those of same species isolated from other geographical regions. This study is a first attempt to establish a morphological and molecular taxonomic reference library for neotropical diatoms. This study demonstrates that it would be feasible to use the existing barcoding data for diatoms to develop molecular tools for the bioassessment of aquatic ecosystems in the Ecuadorian Andean region.


L'identification des diatomées est une étape clé dans l'utilisation de ces microorganismes comme bioindicateurs de la qualité de l'eau. Le diagnostic morphologique est une tâche difficile en raison du nombre considérable d'espèces et de leur dimension microscopique. Il est possible de surmonter cette difficulté en utilisant des techniques moléculaires pour compléter le diagnostic. L'objectif principal de ce travail était d'obtenir le code-barre de l'ADN des diatomées épilithiques équatoriennes ayant une large distribution géographique, une niche écologique bien définie et des caractéristiques leur permettant d'être des espèces indicatrices fiables. Des cultures de diatomées unialgales ont été obtenues à partir d'échantillons environnementaux de cours d'eau des Andes équatoriennes. La caractérisation morphologique des cultures a été réalisée sous microscopie MEB. Pour la caractérisation moléculaire, les codes-barres 18SV4 et rbcL ont été séquencés à partir de chaque souche et comparés à la base de données GenBank. Pour chaque code-barres, un arbre phylogénétique a été construit à partir de la méthode ML comprenant des séquences de souches des espèces étudiées, provenant de différents lieux géographiques. Les résultats ayant montré que les cinq espèces suivantes étaient appropriées comme bioindicateurs, elles ont été isolées. Sellaphora seminulum (souche JA01b, c), Nitzschia fonticola (souche SP02a) et N. palea (souche CA01a) sont tolérantes à l'eutrophisation ; Eolimna minima (souche CH02a) est un bioindicateur d'eau mésotrophe, et Achnanthidium minutissimum (souche JA01a) est un bioindicateur d'eau oligotrophe. La comparaison avec la base de données GenBank des régions de code-barres a supporté leurs identifications morphologiques. Les séquences de code-barres des souches ont montré un pourcentage élevé d'identité génétique avec les séquences signalées dans les bases de données de l'INSDC pour la même espèce. La topologie des arbres phylogénétiques démontre que les diatomées épilithiques de l'Équateur sont étroitement liées à celles des mêmes espèces isolées d'autres régions géographiques. Cette étude est une première tentative d'établir une bibliothèque de référence morphologique et taxonomique moléculaire pour les diatomées néotropicales. Cette étude démontre qu'il serait possible d'utiliser les données de code-barres existantes pour les diatomées afin de développer des instruments moléculaires pour la bioévaluation des écosystèmes aquatiques dans la région andine équatorienne.


Assuntos
Diatomáceas/classificação , Biomarcadores Ambientais , Qualidade da Água , Código de Barras de DNA Taxonômico/métodos , Diatomáceas/genética , Ecossistema , Equador , Eutrofização , Filogenia , Rios
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