El papel del laboratorio de microbiología en la práctica clínica / The role of the microbiology laboratory in clinical practice

Los seres humanos albergamos en el cuerpo a cientos de miles de microbios (bacterias, virus, hongos, archaea). Con la mayoría convivimos armónicamente, y obtenemos beneficios mutuos. Ellos participan activamente en nuestros procesos metabólicos e inmunológicos, y nosotros les damos vivienda y nutrientes [1]. Si el equilibrio se altera, como ocurre cuando el sistema inmune se suprime, algunos de estos microorganismos aprovechan la oportunidad y desencadenan infección [2], siendo este el origen de la mayoría de las infecciones, pero no el único, pues los ambientes externos al cuerpo humano también están poblados de microbios, y si bien pocos se consideran patógenos, al entrar en contacto con ellos adquirimos infecciones que llamamos exógenas. De acuerdo con la Organización Mundial de la Salud (OMS), las enfermedades transmisibles están entre las 10 principales causas de muerte en el mundo, y excluyendo la infección por SARS-CoV-2 (COVID-19) en 2021, las infecciones respiratorias agudas fueron la cuarta causa más frecuente de defunción a nivel mundial [3]. El laboratorio de microbiología clínica tiene un papel crucial en el proceso diagnóstico, terapéutico y preventivo de las enfermedades infecciosas causadas por bacterias, hongos, virus y parásitos, pues determinar el agente causal de manera eficaz permite tomar acciones oportunas para definir el manejo individual de las personas infectadas y cortar la cadena de transmisión cuando se requiera [2]

The influence of primer choice on archaeal phylogenetic analyses based on 16S rRNA gene PCR / A influência da escolha de iniciadores na filogenia de Archaea em ensaios de PCR para o gene rRNA 16S

Abstract Polymerase chain reaction (PCR) assays targeting 16S rRNA genes followed by DNA sequencing are still important tools to characterize microbial communities present in environmental samples. However, despite the crescent number of deposited archaeal DNA sequences in databases, until now we do not have a clear picture of the effectiveness and specificity of the universal primers widely used to describe archaeal communities from different natural habitats. Therefore, in this study, we compared the phylogenetic profile obtained when Cerrado lake sediment DNA samples were submitted to 16S rDNA PCR employing three Archaea-specific primer sets commonly used. Our findings reveal that specificity of primers differed depending on the source of the analyzed DNA. Furthermore, archaeal communities revealed by each primer pair varied greatly, indicating that 16S rRNA gene primer choice affects the community profile obtained, with differences in both taxon detection and operational taxonomic unit (OTU) estimates.

Resumo A amplificação de genes que codificam o rRNA 16S por reação em cadeia da polimerase (PCR) e o seu subsequente sequenciamento consistem em uma ferramenta importante na caracterização de comunidades microbianas presentes em amostras ambientais. No entanto, apesar do crescente número de sequências de DNA de Archaea depositadas em bancos de dados, a especificidade e efetividade dos iniciadores de PCR descritos como universais e amplamente utilizados na descrição desse grupo ainda não está clara. Neste estudo foram comparados os perfis filogenéticos de comunidades de arqueias obtidos a partir amostras de DNA de sedimentos lacustres do Cerrado submetidas a ensaios de PCR empregando três pares de iniciadores específicos para Archaea, comumente utilizados neste tipo de estudo. Nossos resultados indicam que as comunidades de arqueias detectadas com cada par de iniciadores apresentaram grande variação filogenética, sugerindo que a escolha de iniciadores dirigidos ao gene de rRNA 16S tem efeito significativo no perfil da comunidade descrita, com diferenças tanto em relação aos táxons detectados, como nas estimativas de unidades taxonômicas operacionais (OTU).

Identification by molecular techniques of halophilic bacteria producing important enzymes from pristine area in Campeche, Mexico / Identificação por técnicas moleculares de bactérias halofílicas que produzem enzimas importantes em área intocada de Campeche, México

Abstract Isla Arena is located in the coordinate 20° 70´ N - 90° 45´ W, from Campeche, Mexico. In these estuaries, the ocean mixes with fresh water, and ecosystems are concentrated where petenes and pink flamingos proliferate. Crustaceans and mollusks abound in the sea. Despite its enormous marine wealth, there are no studies carried out on which halophilic microorganisms are present in these waters. In this work, the diversity and structure of the microbial community was investigated through a metagenomics approach and corroborated for sequencing of 16S rRNA genes. It was found that the phylum Fimicutes predominates with more than 50%, in almost the same proportion of the class Bacilli and with almost 41% of relative abundance of the order Bacillales. The sequencing results showed that one of the samples presented a high percentage of similarity (99.75%) using the Nucleotide BLAST program with a peculiar microorganism: Bacillus subtilis. This microorganism is one of the best characterized bacteria among the gram-positive ones. Our results demonstrate that B. subtilis can be an efficient source of proteases, lipases and cellulases, from halophilic microbial communities located in poorly explored areas.

Resumo Isla Arena está localizada na coordenada 20°70'N - 90°45'W, de Campeche, México. Nesses estuários, o oceano se mistura com a água doce e os ecossistemas se concentram onde proliferam petenos e flamingos rosa. Crustáceos e moluscos abundam no mar. Apesar de sua enorme riqueza marinha, não há estudos realizados sobre a presença de microrganismos halofílicos nessas águas. Neste trabalho, a diversidade e estrutura da comunidade microbiana foram investigadas através de uma abordagem metagenômica e corroboradas para o sequenciamento de genes 16S rRNA. Verificou-se que o filo Fimicutes predomina com mais de 50%, quase na mesma proporção da classe Bacilli e com quase 41% de abundância relativa da ordem Bacillales. Os resultados do sequenciamento mostraram que uma das amostras apresentou alto percentual de similaridade (99,75%) pelo programa Nucleotide BLAST com um microrganismo peculiar: Bacillus subtilis. Nossos resultados demonstram que B. subtilis pode ser uma fonte eficiente de proteases, lipases e celulases, provenientes de comunidades microbianas halofílicas localizadas em áreas pouco exploradas.

Characterization of highly stable extracellular lipase from the extremely halophilic archaeon Halolamina sp

Enzymes of the archaea living in extreme environments are resistant to the challenging conditions. Lipase is among the important enzymes used in the industry and agriculture. In this study, the extracellular lipase from extremely halophilic archaeon Halolamina sp. was characterized for the first time. Optimum temperature for the enzyme activity was determined as 70oC, optimum pH was 7.0, and the optimum salt concentration was 3.6 M. Additionally, more than 70% of the enzyme activity was remained between pH 3.0-10.0 for 48 h as well as incubation of the enzyme at 70oC for 30 min increased its activity for 44%, and no activity loss was observed after incubation at 80oC. Also, presence of the metals increased the enzyme activity up to 88%. The enzyme was highly resistant to the organic solvents acetone, methanol, and DMSO while strong inhibition was caused by n-butanol. Among the detergents, the enzyme kept its activity substantially in the presence of SDS; however, other detergents caused inhibition of the enzyme activity. This characterization study showed that the lipase from the haloarchaeon Halolamina sp. is highly stable at the wide ranges of temperature and pH values as well as in the presence of diverse inhibitors. This enzyme is promising to be used in biotechnological applications.

Genetic diversity of 16S rRNA and mcrA genes from methanogenic archaeas

Methanogenic archaeas are found in aquatic and terrestrial environments and are fundamental in the conversion of organic matter into methane, a gas that has a potential use as renewable source of energy, which is also considered as one of the main agents of the greenhouse effect. The vast majority of microbial genomes can be identified by a conservative molecular marker, the 16S ribosomal gene. However, the mcrA gene have been using in studies of methanogenic archaea diversity as an alternative marker, highly conserved and present only in methanogens. This gene allows the expression of the enzyme Methyl-coenzyme M reductase, the main agent in converting by-products of anaerobic digestion into methane. In this context, we aimed to study the genetic diversity of mcrA and 16S rRNA genes sequences available in databases. The nucleotide sequences were selected from the NCBI. The heterozygosity and molecular diversity indexes were calculated using the Arlequin 3.5 software, with plots generated by package R v3.0. The diversity and heterozygosity indices for both genes may have been influenced by the number and size of the sequences. Descriptive analysis of genetic diversity generated by sequences deposited in databases allowed a detailed study of these molecules. It is known that the organisms in a population are genetically distinct, and that, despite having similarities in their gene composition, the differences are essential for their adaptation to different environments.

The effect of introduction of chicken manure on the biodiversity and performance of an anaerobic digester

Background: Ammonium stress is a prime limiting phenomenon that occurs during methane formation from poultry manure. It is caused by elevated ammonium nitrogen concentrations that result from substrate decomposition. The amounts of methane formed depend on the activity of methanogenic microbes. Results: During the research reported in this paper, the response of a mesophilic consortium inhabiting a biogas reactor to rising load of poultry manure was investigated. The taxonomic composition of bacterial population was mostly typical, however syntrophic bacteria were not detected. This absence resulted in limitation of succession of some methanogenic microorganisms, especially obligate hydrogenotrophs. The methanogenic activity of the consortium was totally dependent on the activity of Methanosaeta. Inhibition of methanoganesis was noticed at ammonium nitrogen concentration of 3.68 g/L, total cessation occurred at 5.45 g/L. Significant amounts of acetic acid in the fermentation pulp accompanied the inhibition. Conclusions: The effectiveness of the consortium was totally dependent on the metabolic activity of the acetoclastic Methanoseata genus and lack of SAOB did not allow hydrogenotrophic methanogens to propagate and lead to cessation of biogas production at an elevated ammonium concentration at which acetoclastic methanogens were inhibited.

Correlación entre los microorganismos oxidantes del amonio y los factores ambientales durante el compostaje de estiércol de ganado / Correlation between ammonia-oxidizing microorganisms and environmental factors during cattle manure composting

Abstract Cattle manure composting was performed in an aerated vessel. Community structure and diversity of ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) were investigated using polymerase chain reaction and denaturing gradient gel electrophoresis (PCR-DGGE) techniques targeting the ammonia monooxygenase alpha subunit (amoA) gene and the correlation between AOB and AOA communities and environmental factors was explored. Thirteen (13) AOB sequences were obtained, which were closely related to Nitrosomonas spp., Nitrosomonas eutropha, and Nitrosospira spp. and uncultured bacteria, among which Nitrosomonas spp. were predominant. Excessively high temperature and high ammonium concentration were not favorable for AOB growth. Five AOA sequences, belonging to Candidatus Nitrososphaera gargensis and to an uncultured archaeon, were obtained. During composting, community diversity of AOB and AOA fluctuated, with AOA showing a higher Shannon-Wiener index. The AOB community changed more dramatically in the mesophilic stage and the early thermophilic stage, whereas the most obvious AOA community succession occurred in the late thermophilic stage, the cooling stage and the maturity stage. Water content, total nitrogen (TN) and ammonium concentration were more relevant to the AOB community structure, while higher correlations were observed between ammonia, nitrate and TN and the AOA community. AOB community diversity was negatively correlated with pH (r = -0.938, p < 0.01) and water content (r = -0.765, p < 0.05), while positively correlated with TN (r = 0.894, p < 0.01). AOA community diversity was negatively correlated with ammonium concentration (r = -0.901, p < 0.01). Ammonium concentration played an important role in the succession of AOB and AOA communities during composting.

Resumen Se llevó a cabo un compostaje de estiércol de ganado en un recipiente aireado. Se investigó la estructura de la comunidad y la diversidad de bacterias oxidantes del amoníaco (AOB) y las arqueas oxidantes del amoníaco (AOA) mediante el uso de las técnicas de reacción en cadena de la polimerasa y la electroforesis en gel con gradiente de desnaturalización (PCR-DGGE) dirigidas al gen de la subunidad alfa de la amonio monooxigenasa (amoA), y se exploró la correlación entre las comunidades AOB, AOA y los factores ambientales. Se obtuvieron 13 secuencias de AOB, las cuales se relacionaron estrechamente con Nitrosomonas spp., Nitrosomonas eutropha y Nitrosospira spp., y bacterias no cultivadas, entre las cuales fueron predominantes las Nitrosomonas spp. La temperatura excesivamente alta y la concentración de amonio elevada no fueron favorables para el crecimiento de las AOB. Se obtuvieron 5 secuencias de AOA, pertenecientes a Candidatus Nitrososphaera gargensis y un Archaeon no cultivado. Durante el compostaje, la diversidad de AOB y AOA fluctuó y las AOA mostraron un índice de Shannon-Wiener más alto. La comunidad de AOB cambió significativamente en la etapa mesofílica y la etapa termofílica temprana, mientras que la sucesión más obvia de la comunidad AOA ocurrió en la etapa termofílica tardía y las etapas de enfriamiento y de maduración. El contenido de agua, el nitrógeno total (TN) y la concentración de amonio fueron más relevantes para la estructura de la comunidad AOB, mientras que se observaron correlaciones mayores entre amoníaco, nitrato y TN, y la comunidad AOA. La diversidad de la comunidad AOB se correlacionó negativamente con el pH (r= -0,938; p < 0,01) y el contenido de agua (r = -0,765; p < 0,05), mientras que se relacionó positivamente con TN (r = 0,894; p < 0,01). La diversidad de la comunidad AOA se correlacionó negativamente con la concentración de amonio (r = -0,901; p < 0,01). La concentración de amonio desempenó un papel importante en la sucesión de las comunidades AOB y AOA durante el compostaje.

Exploring tRNA gene cluster in archaea

BACKGROUND Shared traits between prokaryotes and eukaryotes are helpful in the understanding of the tree of life evolution. In bacteria and eukaryotes, it has been shown a particular organisation of tRNA genes as clusters, but this trait has not been explored in the archaea domain. OBJECTIVE Explore the occurrence of tRNA gene clusters in archaea. METHODS In-silico analyses of complete and draft archaeal genomes based on tRNA gene isotype and synteny, tRNA gene cluster content and mobilome elements. FINDINGS We demonstrated the prevalence of tRNA gene clusters in archaea. tRNA gene clusters, composed of archaeal-type tRNAs, were identified in two Archaea class, Halobacteria and Methanobacteria from Euryarchaeota supergroup. Genomic analyses also revealed evidence of the association between tRNA gene clusters to mobile genetic elements and intra-domain horizontal gene transfer. MAIN CONCLUSIONS tRNA gene cluster occurs in the three domains of life, suggesting a role of this type of tRNA gene organisation in the biology of the living organisms.

Outstanding impact of soil tillage on the abundance of soil hydrolases revealed by a metagenomic approach

ABSTRACT The soil represents the main source of novel biocatalysts and biomolecules of industrial relevance. We searched for hydrolases in silico in four shotgun metagenomes (4,079,223 sequences) obtained in a 13-year field trial carried out in southern Brazil, under the no-tillage (NT), or conventional tillage (CT) managements, with crop succession (CS, soybean/wheat), or crop rotation (CR, soybean/maize/wheat/lupine/oat). We identified 42,631 hydrolases belonging to five classes by comparing with the KEGG database, and 44,928 sequences by comparing with the NCBI-NR database. The abundance followed the order: lipases > laccases > cellulases > proteases > amylases > pectinases. Statistically significant differences were attributed to the tillage system, with the NT showing about five times more hydrolases than the CT system. The outstanding differences can be attributed to the management of crop residues, left on the soil surface in the NT, and mechanically broken and incorporated into the soil in the CT. Differences between the CS and the CR were slighter, 10% higher for the CS, but not statistically different. Most of the sequences belonged to fungi (Verticillium, and Colletotrichum for lipases and laccases, and Aspergillus for proteases), and to the archaea Sulfolobus acidocaldarius for amylases. Our results indicate that agricultural soils under conservative managements may represent a hotspot for bioprospection of hydrolases.

Archaea diversity in vegetation gradients from the Brazilian Cerrado

Abstract We used 16S rRNA sequencing to assess the archaeal communities across a gradient of Cerrado. The archaeal communities differed across the gradient. Crenarcheota was the most abundant phyla, with Nitrosphaerales and NRPJ as the predominant classes. Euryachaeota was also found across the Cerrado gradient, including the classes Metanocellales and Methanomassiliicoccaceae.

Biotechnological applications of archaeal enzymes from extreme environments

To date, many industrial processes are performed using chemical compounds, which are harmful to nature. An alternative to overcome this problem is biocatalysis, which uses whole cells or enzymes to carry out chemical reactions in an environmentally friendly manner. Enzymes can be used as biocatalyst in food and feed, pharmaceutical, textile, detergent and beverage industries, among others. Since industrial processes require harsh reaction conditions to be performed, these enzymes must possess several characteristics that make them suitable for this purpose. Currently the best option is to use enzymes from extremophilic microorganisms, particularly archaea because of their special characteristics, such as stability to elevated temperatures, extremes of pH, organic solvents, and high ionic strength. Extremozymes, are being used in biotechnological industry and improved through modern technologies, such as protein engineering for best performance. Despite the wide distribution of archaea, exist only few reports about these microorganisms isolated from Antarctica and very little is known about thermophilic or hyperthermophilic archaeal enzymes particularly from Antarctica. This review summarizes current knowledge of archaeal enzymes with biotechnological applications, including two extremozymes from Antarctic archaea with potential industrial use, which are being studied in our laboratory. Both enzymes have been discovered through conventional screening and genome sequencing, respectively.

Effects of volatile fatty acids in biohydrogen effluent on biohythane production from palm oil mill effluent under thermophilic condition

Background: Biohydrogen effluent contains a high concentration of volatile fatty acid (VFA) mainly as butyric, acetic, lactic and propionic acids. The presence of various VFAs (mixture VFAs) and their cooperative effects on two-stage biohythane production need to be further studied. The effect of VFA concentrations in biohydrogen effluent of palm oil mill effluent (POME) on methane yield in methane stage of biohythane production was investigated. Results: The methane yield obtained in low VFA loading (0.9 and 1.8 g/L) was 15­20% times greater than that of high VFA loading (3.6 and 4.7 g/L). Butyric acid at high concentrations (8 g/L) has the individual significantly negative effect the methane production process (P b 0.05). Lactic, acetic and butyric acid mixed with propionic acid at a concentration higher than 0.5 g/L has an interaction significantly negative effect on the methanogenesis process (P b 0.05). Inhibition condition had a negative effect on both bacteria and archaea with inhibited on Geobacillus sp., Thermoanaerobacterium thermosaccharolyticum, Methanoculleus thermophilus and Methanothermobacter delfuvii resulting in low methane yield. Conclusion: Preventing the high concentration of butyric acid, and propionic acid in the hydrogenic effluent could enhance methane production in two-stage anaerobic digestion for biohythane production.

Characterization of a hyperthermophilic sulphur-oxidizing biofilm produced by archaea isolated from a hot spring

Background: Sulphur-oxidizing microorganisms are widely used in the biofiltration of total reduced sulphur compounds (odorous and neurotoxic) produced by industries such as the cellulose and petrochemical industries, which include high-temperature process steps. Some hyperthermophilic microorganisms have the capability to oxidize these compounds at high temperatures (N60°C), and archaea of this group, for example, Sulfolobus metallicus, are commonly used in biofiltration technology. Results: In this study, a hyperthermophilic sulphur-oxidizing strain of archaea was isolated from a hot spring (Chillán, Chile) and designated as M1. It was identified as archaea of the genus Sulfolobus (99% homology with S. solfataricus 16S rDNA). Biofilms of this culture grown on polyethylene rings showed an elemental sulphur oxidation rate of 95.15 ± 15.39 mg S l-1 d-1, higher than the rate exhibited by the biofilm of the sulphur-oxidizing archaea S. metallicus (56.8 ± 10.91 mg l-1 d-1). Conclusions: The results suggest that the culture M1 is useful for the biofiltration of total reduced sulphur gases at high temperatures and for other biotechnological applications.

Anaerobic microbiota: spatial-temporal changes in the sediment of a tropical coastal lagoon with ephemeral inlet in the Gulf of Mexico / Microbiota anaerobia: cambios espacio-temporales en el sedimento de una laguna costera tropical con boca efímera en el Golfo de México

Abstract:Sulfate-reducing bacteria (SRB) and methanogenic archaea (MA) share common niches in coastal sediments during the terminal phases of the anaerobic mineralization of organic matter. The purpose of this study was to analyze the spatial - temporal variation of SRB and MA in the sediments of a tropical coastal lagoon with ephemeral inlet (La Mancha, Veracruz, Gulf of Mexico) and its relationship with environmental changes. A total of 24 sediment samples were collected during the dry (April, May), rainy (July, September) and Northern (November, February) seasons in the period 2013-2014. Microbiological analyses included the quantification of the viable SRB and MA with different substrates, as well as mineralization experiments to determine the effect of sulfate on acetate oxidation. The analyzed environmental variables in the sediments included: temperature, pH, Eh, salinity, sulfates, H2S, volatile solids, carbohydrates, and granulometric characteristics. Major changes occurred between the dry and rainy seasons. During the dry season, sulfate-reducing abundance was significantly greater with lactate (8.3x105 - 1.2x107 cells / g) and propionate (1.8x105 - 6.6x106 cells / g) as substrates, while the MA that use methanol were dominant (4.2x105 - 9.1x106 cells / g). In contrast, during the rainy season, hydrogenophylic (2.6x105 - 8.3x106 cells/g) and acetoclastic (5.4x105-6.4x106 cells / g) MA increased significantly and SRB decreased in the analyzed substrates. An apparent competition for acetate was observed, with a greater oxidation in the media with sulfates in the dry season (0.06 mM acetate / g sediment / day), and a greater oxidation in the media without sulfates in the rainy season (0.02 mM acetate / g sediment / day). SRB and MA were present throughout the sediment column, however SRB dominated in the first centimeters of the sediment while MA were abundant in deeper layers. In conclusion, SRB and MA together played a role in the mineralization of organic matter in the sediments of La Mancha lagoon, with sulfate-reduction dominating in the dry season (closed inlet) and methanogenesis during the rainy season (open inlet). Changes in rainfall and river input in this lagoon significantly affect salinity and sulfate content, the main factors that regulate the dynamics of SRB and MA in the sediments. Rev. Biol. Trop. 64 (4): 1759-1770. Epub 2016 December 01.

Resumen:Las bacterias sulfatorreductoras (BSR) y las arqueas metanogénicas (AM) comparten nichos comunes en los sedimentos costeros durante las fases terminales de la mineralización anaerobia de la materia orgánica. El objetivo del presente estudio fue analizar la variación espacio-temporal de las BSR y AM en los sedimentos de una laguna costera tropical con una boca efímera (La Mancha, Veracruz, Golfo de México) y su relación con los cambios ambientales. Un total de 24 muestras de sedimentos fueron recolectadas en las temporadas de secas (abril, mayo), lluvias (julio, septiembre) y nortes (noviembre, febrero) en el periodo 2013 - 2014. Los análisis microbiológicos incluyeron la cuantificación de las BSR y AM viables con diferentes sustratos, así como experimentos de mineralización para determinar el efecto de los sulfatos en la oxidación del acetato. Las variables ambientales analizadas en los sedimentos incluyeron la temperatura, pH, Eh, salinidad, sulfatos, H2S, sólidos volátiles, carbohidratos y las características granulométricas. Los principales cambios se presentaron entre las estaciones de secas y lluvias. En la temporada de secas la abundancia sulfatorreductora fue significativamente superior con lactato (8.3x105 - 1.2x107 células / g) y propionato (1.8x105 - 6.6x106 células / g) como sustratos, mientras que las AM que emplean metanol dominaron (4.2x105 - 9.1x106 células / g). Por el contrario, en la época de lluvias aumentaron significativamente las AM hidrogenofílicas (2.6x105-8.3x106 células / g) y acetoclásticas (5.4x105-6.4x106 células / g), disminuyendo las BSR con los sustratos analizados. Se determinó una competencia aparente por el acetato. Su oxidación fue mayor en los medios con presencia de sulfatos en las temporadas de secas (0.06 mM acetato / g sedimento / día), mientras que aumentó en los medios sin sulfatos en la época de lluvias (0.02 mM acetato / g sedimento / día). Las BSR y las AM estuvieron presentes a lo largo de la columna sedimentaria; sin embargo, las BSR dominaron en los primeros centímetros del sedimento y las AM abundaron a mayor profundidad. En conclusión las BSR y AM participaron de manera conjunta en la mineralización de la materia orgánica en los sedimentos de la laguna de La Mancha, dominando la sulfatorreducción en la temporada de secas (boca cerrada) y la metanogénesis en la época de lluvias (boca abierta). Los cambios en el aporte fluvial y la precipitación en esta laguna afectaron significativamente la salinidad y el contenido de sulfatos, siendo los principales factores que regularon la dinámica de las BSR y AM en los sedimentos.

Habitat conditions drive phylogenetic structure of dominant bacterial phyla of microbialite communities from several locations in Mexico / Las condiciones del hábitat determinan la estructura filogenética de filos (phyla) bacterianos dominantes de comunidades de microbialitos de diferentes regiones en México

Abstract:Community structure and composition are dictated by evolutionary and ecological assembly processes which are manifested in signals of, species diversity, species abundance and species relatedness. Analysis of species coexisting relatedness, has received attention as a tool to identify the processes that influence the composition of a community within a particular habitat. In this study, we tested if microbialite genetic composition is dependent on random events versus biological/abiotical factors. This study was based on a large genetic data set of two hypervariable regions (V5 and V6) from previously generated barcoded 16S rRNA amplicons from nine microbialite communities distributed in Northeastern, Central and Southeastern Mexico collected in May and June of 2009. Genetic data of the most abundant phyla (Proteobacteria, Planctomycetes, Verrucomicrobia, Bacteroidetes, and Cyanobacteria) were investigated in order to state the phylogenetic structure of the complete communities as well as each phylum. For the complete dataset, Webb NTI index showed positive and significant values in the nine communities analysed, where values ranged from 31.5 in Pozas Azules I to 57.2 in Bacalar Pirate Channel; meanwhile, NRI index were positive and significant in six of the nine communities analysed with values ranging from 18.1 in Pozas Azules I to 45.1 in Río Mesquites. On the other hand, when comparing each individual phylum, NTI index were positive and significant in all groups, except in Cyanobacteria for which positive and significant values were only found in three localities; finally, NRI index was significant in only a few of the comparisons performed. The results suggest that habitat filtering is the main process that drives phylogenetic structure in bacterial communities associated to microbialites with the exception of Cyanobacteria where different lineages can contribute to microbialite formation and growth. Rev. Biol. Trop. 64 (3): 10571065. Epub 2016 September 01.

ResumenLa estructura y composición de las comunidades son determinadas por procesos evolutivos y ecológicos que se manifiestan en señales de diversidad, abundancia y la relación de especies. El análisis de la relación de especies que coexisten ha recibido atención como una herramienta para identificar los procesos que influyen en la composición de una comunidad dentro de un hábitat particular. En este estudio, evaluamos si la composición genética de bacterias microbialíticas depende de acontecimientos al azar vs factores biológicos/ abióticos. Este estudio se basa en un conjunto de datos genéticos de dos regiones hipervariables (V5 y V6) de gen 16S rRNA generados previamente de nueve comunidades de microbialitos distribuidos en el Noreste, Centro y Sureste de México, recolectados en mayo y junio 2009. Los datos genéticos de los filos más abundantes (Proteobacteria, Planctomycetes, Verrucomicrobia, Bacteroidetes y Cyanobacteria) fueron analizados para determinar la estructura filogenética de la comunidad y de cada filo por separado. Para el análisis conjunto, el índice NTI de Webb mostró valores positivos y significativos en las nueve comunidades analizadas, en donde los valores oscilaron entre 31.5 en Pozas Azules I y 57.2 en el Canal Pirata en Bacalar; en contraste, los valores del índice NRI fueron positivos y significativos en seis de las nueve comunidades analizadas con valores oscilando desde 18.1 en Pozas Azules I hasta 45.1 en Río Mezquites. Por otro lado, en la comparación de cada filo individual, el índice NTI fue positivo y significativo en todos los grupos excepto en Cyanobacteria, en donde valores positivos y significativos fueron encontrados sólo en tres localidades; finalmente, el índice NRI fue significativo sólo en unas cuantas de las comparaciones realizadas. Los resultados sugieren que el filtrado del hábitat es el proceso principal que determina la estructura filogenética de las comunidades bacterianas asociadas a microbialitos con la excepción de las cianobacterias en donde diferentes linajes pueden contribuir a la formación y crecimiento del microbialito.

The hyporheic zone and its functions: revision and research status in Neotropical regions / A zona hiporréica e as suas funções: revisão e estado da arte da pesquisa na região neotropical

AbstractThe hyporheic zone (HZ), as the connecting ecotone between surface- and groundwater, is functionally part of both fluvial and groundwater ecosystems. Its hydrological, chemical, biological and metabolic features are specific of this zone, not belonging truly neither to surface- nor to groundwater. Exchanges of water, nutrients, and organic matter occur in response to variations in discharge and bed topography and porosity. Dynamic gradients exist at all scales and vary temporally. Across all scales, the functional significance of the HZ relates to its activity and connection with the surface stream. The HZ is a relatively rich environment and almost all invertebrate groups have colonized this habitat. This fauna, so-called hyporheos, is composed of species typical from interstitial environment, and also of benthic epigean and phreatic species. The hyporheic microbiocenose consists in bacteria, archaea, protozoa and fungi. The HZ provides several ecosystem services, playing a pivotal role in mediating exchange processes, including both matter and energy, between surface and subterranean ecosystems, functioning as regulator of water flow, benthic invertebrates refuge and place of storage, source and transformation of organic matter. The hyporheic zone is one of the most threatened aquatic environments, being strongly influenced by human activities, and the least protected by legislation worldwide. Its maintenance and conservation is compelling in order to preserve the ecological interconnectivity among the three spatial dimensions of the aquatic environment. Although several researchers addressed the importance of the hyporheic zone early, and most contemporary stream ecosystem models explicitly include it, very little is known about the HZ of Neotropical regions. From a biological standpoint, hyporheos fauna in Neotropical regions are still largely underestimated. This review focuses on a brief presentation of the hyporheic zone and its functions and significance as an ecotone. We also highlighted the key aspects considering also the current status of research in Neotropical regions.

ResumoA zona hiporréica, como ecótono de ligação entre a superfície e as águas subterrâneas, é parte funcional seja dos ecossistemas fluviais seja das águas subterrâneas. As características hidrológicas, as características químicas, biológicas e metabólicas são específicas desta zona, não pertencendo verdadeiramente nem a superfície nem às águas subterrâneas. Trocas de água, nutrientes e matéria orgânica ocorrem em resposta a variações na descarga, topografia do álveo e porosidade. Gradientes dinâmicos existem em todas as escalas e variam temporalmente. Em todas as escalas, o significado funcional da zona hyporheic relaciona-se com a sua conexão e atividades com a água superficial. O HZ é um ambiente relativamente rico e quase todos os grupos de invertebrados colonizaram este habitat. Esta fauna, chamada hyporheos, é composta por espécies típicas do ambiente intersticial, e também de espécies bentônicas epígeas e freáticas. A microbiocenose consiste em bactérias, arqueobactérias, fungos e protozoários. O HZ fornece vários serviços para o ecossistema, desempenhando um papel fundamental na mediação de processos de troca, incluindo seja a matéria, seja a energia, entre os ecossistemas superfíciais e os subterrâneos, funcionando como regulador do fluxo de água, de refúgio para invertebrados bentônicos e local de armazenagem, fonte e transformação de matéria orgânica. A zona hyporheic é um dos ambientes aquáticos mais ameaçados, sendo fortemente influenciado pelas atividades humanas, e um dos menos protegidos pela legislação em todo o mundo. A sua manutenção e conservação é necessaria para preservar a interconectividade ecológica entre as três dimensões espaciais do ambiente aquático. Apesar de vários pesquisadores aborem a importância da zona hyporheic a tempo, e a maioria dos modelos de ecossistemas atualmente incluí-lo de forma explicita, muito pouco se sabe sobre o HZ das regiões neotropicais. Do ponto de vista biológico, a fauna hiporréica das regiões neotropicais é ainda largamente subestimada. Esta revisão visa apresentar de forma resumida a zona hiporréica, suas funções e importância como ecótono. Também visa destacar os aspectos principais considerando também o estado actual da investigação em regiões neotropicais.

Large scale distribution of bacterial communities in the upper Paraná River floodplain

A bacterial community has a central role in nutrient cycle in aquatic habitats. Therefore, it is important to analyze how this community is distributed throughout different locations. Thirty-six different sites in the upper Paraná River floodplain were surveyed to determine the influence of environmental variable in bacterial community composition. The sites are classified as rivers, channels, and floodplain lakes connected or unconnected to the main river channel. The bacterial community structure was analyzed by fluorescent in situ hybridization (FISH) technique, based on frequency of the main domains Bacteria and Archaea, and subdivisions of the phylum Proteobacteria (Alpha-proteobacteria, Beta-proteobacteria, Gamma-proteobacteria) and the Cytophaga-Flavobacterium cluster. It has been demonstrated that the bacterial community differed in density and frequency of the studied groups. And these differences responded to distinct characteristics of the three main rivers of the floodplain as well as to the classification of the environments found in this floodplain. We conclude that dissimilarities in the bacterial community structure are related to environmental heterogeneity, and the limnological variables that most predicted bacterial communities in the upper Paraná River floodplain was total and ammoniacal nitrogen, orthophosphate and chlorophyll-a.

Diversity and three-dimensional structures of the alpha Mcr of the methanogenic Archaea from the anoxic region of Tucuruí Lake, in Eastern Brazilian Amazonia

Methanogenic archaeans are organisms of considerable ecological and biotechnological interest that produce methane through a restricted metabolic pathway, which culminates in the reaction catalyzed by the Methyl-coenzyme M reductase (Mcr) enzyme, and results in the release of methane. Using a metagenomic approach, the gene of the a subunit of mcr (mcrα) was isolated from sediment sample from an anoxic zone, rich in decomposing organic material, obtained from the Tucuruí hydroelectric dam reservoir in eastern Brazilian Amazonia. The partial nucleotide sequences obtained were 83 to 95 percent similar to those available in databases, indicating a low diversity of archaeans in the reservoir. Two orders were identified -the Methanomicrobiales, and a unique Operational Taxonomic Unit (OTU) forming a clade with the Methanosarcinales according to low bootstrap values. Homology modeling was used to determine the three-dimensional (3D) structures, for this the partial nucleotide sequence of the mcrα were isolated and translated on their partial amino acid sequences. The 3D structures of the archaean mcrα observed in the present study varied little, and presented approximately 70 percent identity in comparison with the mcrα of Methanopyrus klanderi. The results demonstrated that the community of methanogenic archaeans of the anoxic C1 region of the Tucurui reservoir is relatively homogeneous.

Molecular identification of methanogenic archaea from Surti buffaloes (bubalus bubalis), reveals more hydrogenotrophic methanogens phylotypes

Methane emissions from ruminant livestock are considered to be one of the more potent forms of greenhouses gases contributing to global warming. Many strategies to reduce emissions are targeting the methanogens that inhabit the rumen, but such an approach can only be successful if it targets all the major groups of ruminant methanogens. Therefore, a thorough knowledge of the diversity of these microbes in breeds of buffaloes, as well as in response to geographical location and different diets, is required. Therefore, molecular diversity of rumen methanogens in Surti buffaloes was investigated using 16S rRNA gene libraries prepared from pooled rumen contents from three Surti buffaloes. A total of 171 clones were identified revealing 23 different sequences (phylotypes). Of these 23 sequences, twelve sequences (12 OTUs, 83 clones) and 10 sequences (10 OTUs, 83 clones) were similar to methanogens belonging to the orders Methanomicrobiales and Methanobacteriales, and the remaining 1 phylotype (5 clones) were similar to Methanosarcina barkeri. These unique sequences clustered within a distinct and strongly supported phylogenetic group. Further studies and effective strategies can be made to inhibit the growth of Methanomicrobiales and Methanobacteriales phylotypes to reduce the methane emission from rumen and thus help in preventing global warming.

Prevalência de Archaea em indivíduos com doença periodontal e saúde periodontal / Prevalence of Archaea in subjets with periodontal disease and periodontally healthy

Apesar de micro-organismos do domínio Archaea já terem sido detectados em amostras de biofilme subgengival de indivíduos com periodontite, nenhum estudo comparou diretamente a associação de Archaea com as periodontites crônica e agressiva. Assim sendo, o objetivo desse estudo foi avaliar a prevalência do domínio Archaea em indivíduos com periodontite crônica generalizada (PCrG), periodontite agressiva generalizada (PAgG) e indivíduos com saúde periodontal (SP), e correlacionar os achados microbiológicos com o aumento da profundidade de sondagem (PS). Foram selecionados 15 indivíduos com PCrG, 15 com PAgG e 15 com SP. Inicialmente, os indivíduos foram submetidos ao exame clínico-periodontal. Em seguida, amostras de biofilme subgengival foram coletadas de 9 sítios nos indivíduos do grupo SP e de 9 sítios (PS <3mm, 3 PS 4-6mm e 3 PS >7mm) nos indivíduos dos grupos PCrG e PAgG. Após a extração do DNA, as amostras foram submetidas à reação de amplificação do gene 16S rRNA de Archaea. A presença de Archaeafoi detectada em 11/15 indivíduos do grupo PCrG, em 9/15 indivíduos com PAgG e 12/15 indivíduos com SP (p>0,05). Em relação ao número de amostras positivas, foram 21 no grupo SP, 39 no PAgG e 43 no PCrG observados. Não foram observadas diferenças estatísticas entre os diferentes grupos experimentais (p>0,05) e também não houve correlação entre a presença de Archaea e as diferentes categorias de profundidade de sondagem. Em conclusão, sugere-se ausência de associação entre a detecção de Archaea e a periodontite.

Archaea has been detected in subgingival biofilm samples from individuals with periodontitis, however, no studies were found to enable a direct comparison of the association of Archaea with chronic and aggressive periodontitis. Therefore, the aim of the present study was to evaluate the prevalence of the Archaea domine in individuals with generalized chronic periodontitis (GCrP), generalized aggressive periodontitis (GAgP) and periodontal healthy (PH) and correlation these finding with the increase of probing depth (PD). Fifteen individuals with GCrP, fifteen individuals with GAgP and fifteen individuals with PH were selected. The individuals were submitted a clinical periodontal examination. Subgingival biofilm samples were collected from 9 sites in the PH group and 9 sites (3 PD ¡Ü3mm, 3 PD 4-6mm and 3 PD ¡Ý7mm) in the 30 individuals in the GAgP and GCrP groups. After the DNA extraction, the presence of Archaea was analyzed by polymerase chair reaction (PCR). The occurrence of Archaea was detected in 11/15 individuals in the GCrP group, 9/15 individuals with GAgP and 12/15 individuals with SP (p>0.05). Among the individuals of with SP, PAgG and PCrG, 21, 39 and 43 samples were positive for Archaea, respectively. However, there were no statistical differences observed among experimental groups (p>0.05). There was no correlation between the presence of Archaea and the different probing depth categories. In conclusion, these results suggest the absence of association between the detection of Archaeaand periodontitis.