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1.
Curr Microbiol ; 80(9): 313, 2023 Aug 05.
Article in English | MEDLINE | ID: mdl-37542533

ABSTRACT

Enterococcus faecalis has emerged as an important opportunistic pathogen due to its increasing resistance to antimicrobials, mainly to vancomycin, which leads substantial cases of therapeutic failures. Wastewater treatment plants (WWTP), in turn, are considered hotpots in the spread of antimicrobial resistance according to One Health perspective. In this study, we present the first report of a vancomycin-resistant E. faecalis strain recovered from treated effluent in Brazil. For this purpose, the whole-genome sequencing (WGS) was carried out aiming to elucidate its molecular mechanisms of antimicrobial resistance and its phylogenetic relationships amongst strains from other sources and countries. According to Multilocus Sequence Typing (MLST) analysis this strain belongs to ST21. The WGS pointed the presence of vanA operon, multiple antibiotic resistance and virulence genes, and a significant pathogenic potential for humans. The phylogenomic analysis of E. faecalis 6805 was performed with ST21 representatives from the PubMLST database, including the E. faecalis IE81 strain from clinical sample in Brazil, which had its genome sequenced in this study. Our results demonstrated a strain showing resistance to vancomycin in treated effluent. To the best of our knowledge, this is an unprecedented report of vanA-carrying E. faecalis ST21. Furthermore, it is the first description of a vanA-harboring strain of this species from environmental sample in Brazil. Our data highlight the role of WWTP in the spread of AMR, since these environments are favorable for the selection of multidrug-resistant pathogens and the treated effluents, carrying antibiotic resistance genes, are directed to receiving water bodies.


Subject(s)
Enterococcus faecium , Gram-Positive Bacterial Infections , Humans , Enterococcus faecalis/genetics , Vancomycin , Multilocus Sequence Typing , Brazil , Phylogeny , Microbial Sensitivity Tests , Anti-Bacterial Agents/pharmacology , Vancomycin Resistance/genetics
2.
J Water Health ; 21(5): 653-662, 2023 May.
Article in English | MEDLINE | ID: mdl-37254912

ABSTRACT

Wastewater-based epidemiology can be a complementary approach for monitoring SARS-CoV-2 prevalence, diversity, and geographic distribution. It is a complementary approach regarding its prevalence and diversity, and geographic distribution. The study aimed to evaluate the genetic diversity of SARS-CoV-2 in two wastewater treatment plants (WWTPs) in Rio de Janeiro, Brazil. Samples were collected over a period of January to December 2021 and were concentrated with PEG8000 and the presence of SARS-CoV-2 was detected using E and N1 genes. Partial sequencing of the SARS-CoV-2 genomes resulted in the identification of variants of concern and variants of interest throughout the collection period. It was possible to identify the Mu, Delta, Gamma and Omicron variants in WWTP1; on the contrary, no variants were observed in WWTP2. To the best of our knowledge, we detected the variant Mu (B.1.621) containing characteristic mutations (S:E484K, S:N501Y) from WWTP, for the first time, in Brazil. Another Mu variant detected from clinical surveillance was announced one month after our finding. The detection of SARS-CoV-2 in wastewater can serve as a tool to monitor the prevalence and epidemiology in each community, helping to understand the spread of the virus among the population.


Subject(s)
COVID-19 , Wastewater , Humans , Brazil/epidemiology , Metagenomics , SARS-CoV-2/genetics , COVID-19/epidemiology
3.
J. bras. patol. med. lab ; 52(5): 284-292, Sept.-Oct. 2016. tab, graf
Article in English | LILACS | ID: biblio-829081

ABSTRACT

ABSTRACT Introduction: The increasing incidence of multi-resistant microorganisms has been considered a public health problem. One of the routines included in hospital practice is the screening of colonized and/or infected patients. Objective: The aim of this study was to evaluate the genetic variability and clonal relationships of extended-spectrum beta-lactamase (ESBL)-producing K. pneumoniae, from surveillance cultures, at an intensive care unit, in Rio de Janeiro, Brazil. Material and methods: Seventy K. pneumoniae isolates were obtained from rectal swabs (March 2013 to March 2014). Antimicrobial susceptibility was assessed by VITEK 2 System. Resistant genes blaSHV, blaTEM, blaOXA-1, blaKPC, blaOXA-48, blaCTX-M-15, blaVIM, blaIMP and blaNDM were investigated by polymerase chain reaction (PCR); genetic diversity, by Enterobacterial Repetitive Intergenic Consensus-PCR (ERIC-PCR). Results: Strains showed high resistance rates to cefepime (94%), ceftazidime (96%), ertapenem (61%), imipenem (54%) meropenem (43%) and ciprofloxacin (69%). The most prevalent genes were blaSHV (69%), blaTEM (63%), blaOXA-1 (60%), blaKPC (57%), blaCTX-M-15 (47%), blaOXA-48 (16%). Genes blaVIM, blaIMP and blaNDM were not detected. Twenty nine profiles of resistance genes were observed, with 23% carrying at least five genes. A great genetic diversity (68 ERIC profiles) was also observed among the strains. Conclusion: Although no clonal relationship was observed within the isolates, this study revealed alarming data on the antimicrobial resistance deficiently monitored for preventive purposes in Brazil. Our data allow us to conclude that the inclusion of surveillance cultures in health facilities is a recommended strategy aiming particularly at preventing the spread of resistance genes in the hospital environment and, consequently, reducing morbidity and mortality.


RESUMO Introdução: O aumento da incidência de microrganismos multirresistentes é considerado um dos principais problemas de saúde pública. Uma das rotinas incluídas na prática hospitalar é a busca de pacientes colonizados e/ou infectados. Objetivo: Avaliar a variabilidade genética e as relações clonais de K. pneumoniae produtoras de betalactamases de espectro estendido (ESBL) em culturas de vigilância de unidade de terapia intensiva (UTI) no Rio de Janeiro, Brasil. Materiais e métodos: Setenta isolados obtidos a partir de swab retal, (março/2013 a março/2014). O perfil de suscetibilidade a antibióticos foi avaliado pelo sistema VITEK 2. Foram pesquisados os genes de resistência: blaSHV, blaTEM, blaOXA-1, blaKPC, blaOXA-48, blaCTX-M-15, blaVIM, blaIMP e blaNDM pela reação em cadeia da polimerase (PCR). A diversidade genética foi avaliada por Enterobacterial Repetitive Intergenic Consensus-PCR (ERIC-PCR). Resultados: Foram detectados altos percentuais de resistência a cefepime (94%), ceftazidima (96%), ertapenem (61%), imipenem (54%) meropenem (43%) e ciprofloxacino (69%). Os genes prevalentes foram: blaSHV (69%), blaTEM (63%), blaOXA-1 (60%), blaKPC (57%), blaCTX-M-15 (47%), blaOXA-48 (16%). Os genes blaVIM, blaIMP e blaNDM não foram detectados. Foram observados 29 perfis em relação aos genes de resistência, com 23% apresentando pelo menos cinco genes. Uma grande diversidade genética (68 perfis) foi observada entre as cepas. Conclusão: Embora não tenha sido observada relação clonal entre os isolados, este estudo revelou dados alarmantes quanto à resistência microbiana em monitoramento preventivo, abordagem ainda pouco adotada no Brasil. Nossos dados permitem concluir que a inclusão de culturas de vigilância nas unidades de saúde é uma estratégia recomendada, visando principalmente à prevenção da disseminação dos genes de resistência no ambiente hospitalar e, consequentemente, redução da morbimortalidade.

4.
Front Microbiol ; 6: 860, 2015.
Article in English | MEDLINE | ID: mdl-26347735

ABSTRACT

The enzymatic hydrolysis of cellulose by cellulases is one of the major limiting steps in the conversion of lignocellulosic biomass to yield bioethanol. To overcome this hindrance, significant efforts are underway to identify novel cellulases. The snail Achatina fulica is a gastropod with high cellulolytic activity, mainly due to the abundance of glycoside hydrolases produced by both the animal and its resident microbiota. In this study, we partially assessed the cellulolytic aerobic bacterial diversity inside the gastrointestinal tract of A. fulica by culture-dependent methods and evaluated the hydrolytic repertoire of the isolates. Forty bacterial isolates were recovered from distinct segments of the snail gut and identified to the genus level by 16S rRNA gene sequence analysis. Additional phenotypic characterization was performed using biochemical tests provided by the Vitek2 identification system. The overall enzymatic repertoire of the isolated strains was investigated by enzymatic plate assays, containing the following substrates: powdered sugarcane bagasse, carboxymethylcellulose (CMC), p-nitrophenyl-ß-D-glucopyranoside (pNPG), p-nitrophenyl-ß-D-cellobioside (pNPC), 4-methylumbelliferyl-ß-D-glucopyranoside (MUG), 4-methylumbelliferyl-ß-D-cellobioside (MUC), and 4-methylumbelliferyl-ß-D-xylopyranoside (MUX). Our results indicate that the snail A. fulica is an attractive source of cultivable bacteria that showed to be valuable resources for the production of different types of biomass-degrading enzymes.

5.
J Biotechnol ; 212: 69-70, 2015 Oct 20.
Article in English | MEDLINE | ID: mdl-26299206

ABSTRACT

Haloferax gibbonsii strain ARA6 is a haloarchaea isolated from saline saltern samples from Vermelha lake, located in Araruama region, Rio de Janeiro, Brazil. Its genome displays 66,2% G+C content and is composed by one circular chromosome of 2,945,391 bp and four circular plasmids comprising 993,063 bp. This genomic information shows H. gibbonsii's potential for biotechnological applications and can also contribute to assign evolutionary traits in the genus Haloferax.


Subject(s)
Genome, Bacterial , Haloferax/genetics , Base Sequence , Brazil , DNA, Bacterial/genetics , Haloferax/isolation & purification , Haloferax/metabolism , Molecular Sequence Data , Peptides/genetics , Polyhydroxyalkanoates/metabolism
6.
J. bras. patol. med. lab ; 51(3): 143-152, May-Jun/2015. tab
Article in English | LILACS | ID: lil-753111

ABSTRACT

ABSTRACT Introduction: Cross-contamination by Staphylococcus aureus among patients, professionals and medical supplies in health facilities is a constant concern, leading many researchers to study the prevalence of this pathogen in asymptomatic carriers. Objective: We investigated the colonization and the antimicrobial susceptibility profile of Staphylococcus spp. on surfaces of medical articles and in professionals from two basic health units in the city of Rio de Janeiro. Materials and methods: Seventy-nine samples resulted in 49 isolates which underwent phenotypic and molecular characterization by polymerase chain reaction (PCR) of coa, mecA and femA genes. Results: According to the phenotypes, the isolates were identified as S. aureus (n = 35, 71.42%) and coagulase-negative Staphylococcus (CoNS) (n = 14, 28.57%). Among these 14 isolates, 42.85% were methicillin-resistant coagulase negative Staphylococcus (MRCoNS). Among the 35 S. aureus, 31.42% were methicillin resistant (MRSA), and 2.8% were vancomycin resistant, characterized as VRSA. Sixty-eight percent were susceptible to methicillin (MSSA). Genes coa, femA and mecA were amplified from 75.51%, 71.42% and 30.61% of the isolates, respectively. After amplification of the mecA gene, 20.41% were characterized as MRSA, and 10.20% as MRCoNS. The vancomycin-resistant strain was characterized as VRSA after detection of the vanB gene. Conclusion: Our results show a higher frequency of MSSA and MRCoNS among S. aureus and CoNS respectively, colonizing devices and health professionals. However, the already described transfer of the staphylococcal cassette chromosome mec (SSCmec) from MRCoNS to MSSA may alter these results, increasing the frequency of MRSA strains. .


RESUMO Introdução: A contaminação cruzada por Staphylococcus aureus entre pacientes, profissionais e materiais de uso médico em unidades de saúde é uma preocupação constante, o que leva pesquisadores a estudar a prevalência desse patógeno em portadores assintomáticos. Objetivos: Investigamos a colonização e o perfil de suscetibilidade aos antimicrobianos de Staphylococcus spp. em superfícies de artigos médicos e em profissionais de duas unidades básicas de saúde no município do Rio de Janeiro. Materiais e métodos: Foram coletadas 79 amostras que resultaram em 49 isolados, submetidos à caracterização fenotípica e molecular por meio da reação em cadeia da polimerase (PCR) dos genes coa, femA e mecA. Resultados: De acordo com os fenótipos apresentados, os isolados foram identificados como S. aureus (n = 35; 71,42%) e Staphylococcus coagulase negativa (CoNS) (n = 14; 28,57%). Destes 14 isolados, 42,85% foram Staphylococcus coagulase negativa resistentes a meticilina (MRCoNS). Dos 35 S. aureus, 31,42% foram resistentes a meticilina (MRSA). Uma cepa foi resistente a vancomicina e identificada como S. aureus resistente a vancomicina (VRSA) após a detecção do gene vanB. Sessenta e oito por cento foram suscetíveis a meticilina (MSSA). Os genes coa, femA e mecA foram amplificados em 75,51%; 71,42% e 30,61% dos isolados, respectivamente. Após amplificação do gene mecA, 20,41% foram classificados como MRSA e 10,20% como MRCoNS. Conclusão: Nossos resultados mostraram frequência maior de MSSA e MRCoNS entre S. aureus e CoNS, respectivamente, colonizando equipamentos e profissionais de saúde. No entanto, a já descrita transferência do cassete cromossômico estafilocócico mec (SSCmec) de MRCoNS para MSSA poderia alterar esses resultados, aumentando a frequência de cepas MRSA. .

7.
Microb Ecol ; 68(3): 441-52, 2014 Oct.
Article in English | MEDLINE | ID: mdl-24821495

ABSTRACT

Bacterial resistance to antibiotics has become a public health issue. Over the years, pathogenic organisms with resistance traits have been studied due to the threat they pose to human well-being. However, several studies raised awareness to the often disregarded importance of environmental bacteria as sources of resistance mechanisms. In this work, we analyze the diversity of antibiotic-resistant bacteria occurring in aquatic environments of the state of Rio de Janeiro, Brazil, that are subjected to distinct degrees of anthropogenic impacts. We access the diversity of aquatic bacteria capable of growing in increasing ampicillin concentrations through 16S rRNA gene libraries. This analysis is complemented by the characterization of antibiotic resistance profiles of isolates obtained from urban aquatic environments. We detect communities capable of tolerating antibiotic concentrations up to 600 times higher than the clinical levels. Among the resistant organisms are included potentially pathogenic species, some of them classified as multiresistant. Our results extend the knowledge of the diversity of antibiotic resistance among environmental microorganisms and provide evidence that the diversity of drug-resistant bacteria in aquatic habitats can be influenced by pollution.


Subject(s)
Ampicillin Resistance , Bacteria/drug effects , Water Microbiology , Ampicillin , Bacteria/classification , Bacteria/genetics , Bathing Beaches , Bays , Brazil , Cities , DNA, Bacterial/genetics , Gene Library , RNA, Ribosomal, 16S/genetics , Rivers/microbiology , Seawater/microbiology
8.
PLoS One ; 8(9): e76321, 2013.
Article in English | MEDLINE | ID: mdl-24086729

ABSTRACT

The Archaea domain is ubiquitously distributed and extremely diverse, however, environmental factors that shape archaeal community structure are not well known. Aquatic environments, including the water column and sediments harbor many new uncultured archaeal species from which metabolic and ecological roles remain elusive. Some environments are especially neglected in terms of archaeal diversity, as is the case of pristine tropical areas. Here we investigate the archaeal composition in marine and freshwater systems from Ilha Grande, a South Atlantic tropical environment. All sampled habitats showed high archaeal diversity. No OTUs were shared between freshwater, marine and mangrove sediment samples, yet these environments are interconnected and geographically close, indicating environment-specific community structuring. Group II Euryarchaeota was the main clade in marine samples, while the new putative phylum Thaumarchaeota and LDS/RCV Euryarchaeota dominated freshwaters. Group III Euryarchaeota, a rare clade, was also retrieved in reasonable abundance in marine samples. The archaeal community from mangrove sediments was composed mainly by members of mesophilic Crenarchaeota and by a distinct clade forming a sister-group to Crenarchaeota and Thaumarchaeota. Our results show strong environment-specific community structuring in tropical aquatic Archaea, as previously seen for Bacteria.


Subject(s)
Adaptation, Biological/physiology , Archaea/physiology , Biota/physiology , Environment , Geologic Sediments/microbiology , Phylogeny , Water Microbiology , Base Sequence , Brazil , DNA Primers/genetics , Gene Library , Likelihood Functions , Models, Genetic , Molecular Sequence Data , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Species Specificity , Tropical Climate
9.
Microb Ecol ; 65(1): 197-204, 2013 Jan.
Article in English | MEDLINE | ID: mdl-22945233

ABSTRACT

Termites inhabit tropical and subtropical areas where they contribute to structure and composition of soils by efficiently degrading biomass with aid of resident gut microbiota. In this study, culture-independent molecular analysis was performed based on bacterial and archaeal 16S rRNA clone libraries to describe the gut microbial communities within Cornitermes cumulans, a South American litter-feeding termite. Our data reveal extensive bacterial diversity, mainly composed of organisms from the phyla Spirochaetes, Bacteroidetes, Firmicutes, Actinobacteria, and Fibrobacteres. In contrast, a low diversity of archaeal 16S rRNA sequences was found, comprising mainly members of the Crenarchaeota phylum. The diversity of archaeal methanogens was further analyzed by sequencing clones from a library for the mcrA gene, which encodes the enzyme methyl coenzyme reductase, responsible for catalyzing the last step in methane production, methane being an important greenhouse gas. The mcrA sequences were diverse and divided phylogenetically into three clades related to uncultured environmental archaea and methanogens found in different termite species. C. cumulans is a litter-feeding, mound-building termite considered a keystone species in natural ecosystems and also a pest in agriculture. Here, we describe the archaeal and bacterial communities within this termite, revealing for the first time its intriguing microbiota.


Subject(s)
Archaea/classification , Bacteria/classification , Gastrointestinal Tract/microbiology , Isoptera/microbiology , Metagenome , Animals , Archaea/genetics , Archaea/isolation & purification , Bacteria/genetics , Bacteria/isolation & purification , DNA, Archaeal/genetics , DNA, Bacterial/genetics , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA
10.
Microb Ecol ; 65(1): 205-13, 2013 Jan.
Article in English | MEDLINE | ID: mdl-22864853

ABSTRACT

The association of metazoan, protist, and microbial communities with Scleractinian corals forms the basis of the coral holobiont. Coral bleaching events have been occurring around the world, introducing changes in the delicate balance of the holobiont symbiotic interactions. In this study, Archaea, bacteria, and eukaryotic phototrophic plastids of bleached colonies of the Brazilian coral Siderastrea stellata were analyzed for the first time, using 16S rRNA gene libraries. Prokaryotic communities were slightly more diverse in healthy than in bleached corals. However, the eukaryotic phototrophic plastids community was more diverse in bleached corals. Archaea phylogenetic analyses revealed a high percentage of Crenarchaeota sequences, mainly related to Nitrosopumilus maritimus and Cenarchaeum symbiosum. Dramatic changes in bacterial community composition were observed in this bleaching episode. The dominant bacterial group was Alphaproteobacteria followed by Gammaproteobacteria in bleached and Betaproteobacteria in healthy samples. Plastid operational taxonomic units (OTUs) from both coral samples were mainly related to red algae chloroplasts (Florideophycea), but we also observed some OTUs related to green algae chloroplasts (Chlorophyta). There seems to be a strong relationship between the Bacillariophyta phylum and our bleached coral samples as clones related to members of the diatom genera Amphora and Nitzschia were detected. The present study reveals information from a poorly investigated coral species and improves the knowledge of coral microbial community shifts that could occur during bleaching episodes.


Subject(s)
Anthozoa/microbiology , Archaea/classification , Bacteria/classification , Chlorophyta/genetics , Rhodophyta/classification , Animals , Archaea/genetics , Bacteria/genetics , Brazil , Chlorophyta/classification , DNA Barcoding, Taxonomic , DNA, Algal/genetics , DNA, Archaeal/genetics , DNA, Bacterial/genetics , Diatoms/classification , Diatoms/genetics , Ecosystem , Gene Library , Phylogeny , Plastids/genetics , RNA, Ribosomal, 16S/genetics , Rhodophyta/genetics , Symbiosis
11.
PLoS One ; 7(11): e51175, 2012.
Article in English | MEDLINE | ID: mdl-23226484

ABSTRACT

Despite a great number of published studies addressing estuarine, freshwater and marine bacterial diversity, few have examined urban coastal lagoons in tropical habitats. There is an increasing interest in monitoring opportunistic pathogens as well as indigenous microbial community members in these water bodies by current molecular and microbiological approaches. In this work, bacterial isolates were obtained through selective plate dilution methods to evaluate antibiotic resistances. In addition, 16S rRNA gene libraries were prepared from environmental waters and mixed cultures grown in BHI medium inoculated with Jacarepaguá lagoon waters. Denaturing gradient gel electrophoresis (DGGE) analyses showed distinct community profiles between environmental communities from each studied site and their cultured counterparts. A total of 497 bacterial sequences were analyzed by MOTHUR, yielding 245 operational taxonomic units (OTUs) grouped at 97% similarity. CCA diagrams showcased how several environmental variables affect the distribution of 18 bacterial orders throughout the three distinct habitats. UniFrac metrics and Venn diagrams revealed that bacterial communities retrieved through each experimental approach were significantly different and that only one OTU, closely related to Vibrio cholerae, was shared between them. Potentially pathogenic bacteria were isolated from most sampled environments, fifty percent of which showed antibiotic resistance.


Subject(s)
Bacteria/genetics , Biodiversity , Cities , Plankton/genetics , Seawater/microbiology , Tropical Climate , Water Pollution/analysis , Anti-Bacterial Agents/pharmacology , Bacteria/classification , Bacteria/drug effects , Brazil , Denaturing Gradient Gel Electrophoresis , Environment , Gene Library , Geography , Microbial Sensitivity Tests , Molecular Sequence Data , Phylogeny , Plankton/classification , Plankton/drug effects , RNA, Ribosomal, 16S/genetics , Water/chemistry , Water Microbiology
12.
Int J Environ Health Res ; 22(6): 560-70, 2012.
Article in English | MEDLINE | ID: mdl-22540159

ABSTRACT

The aim of this study was to evaluate the antimicrobial resistance patterns of Pseudomonas aeruginosa isolates from hospital wastewater treatment system (HWTS) and clinical specimens in a hospital of Rio de Janeiro city, Brazil. Forty-three isolates obtained from four of the five steps of the HWTS (n = 27) and clinical samples (n = 16) from patients were analyzed regarding their susceptibility profiles to 12 antibiotics. Clinical isolates exhibited higher resistance profiles to antibiotics than wastewater isolates. However, out of 27 isolates from sewage, 62.9% showed decreased susceptibility to aztreonam while 50% of clinical isolates were resistant to this antibiotic. Isolates were not detected at the chlorination stage but they were obtained from the following stage of the treatment revealing the capacity of regrowth after chlorinated sewage effluent. To our knowledge, this is the first study reporting decreased aztreonam susceptibility among P. aeruginosa isolates from a hospital wastewater treatment system. Further investigations are being conducted by our laboratory including a larger sampling program in order to obtain more data.


Subject(s)
Anti-Bacterial Agents/pharmacology , Aztreonam/pharmacology , Drug Resistance, Bacterial , Hospitals , Pseudomonas aeruginosa/drug effects , Wastewater/microbiology , Brazil , Humans , Microbial Sensitivity Tests , Pseudomonas Infections/drug therapy , Pseudomonas Infections/microbiology , Pseudomonas aeruginosa/isolation & purification , Waste Disposal, Fluid , Water Pollutants/isolation & purification
13.
PLoS One ; 7(3): e33440, 2012.
Article in English | MEDLINE | ID: mdl-22438932

ABSTRACT

The invasive land snail Achatina fulica is one of the most damaging agricultural pests worldwide representing a potentially serious threat to natural ecosystems and human health. This species is known to carry parasites and harbors a dense and metabolically active microbial community; however, little is known about its diversity and composition. Here, we assessed for the first time the complexity of bacterial communities occurring in the digestive tracts of field-collected snails (FC) by using culture-independent molecular analysis. Crop and intestinal bacteria in FC were then compared to those from groups of snails that were reared in the laboratory (RL) on a sugarcane-based diet. Most of the sequences recovered were novel and related to those reported for herbivorous gut. Changes in the relative abundance of Bacteroidetes and Firmicutes were observed when the snails were fed a high-sugar diet, suggesting that the snail gut microbiota can influence the energy balance equation. Furthermore, this study represents a first step in gaining a better understanding of land snail gut microbiota and shows that this is a complex holobiont system containing diverse, abundant and active microbial communities.


Subject(s)
Metagenome , Snails/microbiology , Animals , Bacteria/classification , Bacteria/genetics , Bacteria/isolation & purification , Base Sequence , DNA Primers/genetics , DNA, Bacterial/genetics , Diet , Digestive System/microbiology , Ecosystem , Humans , Phylogeny , Saccharum , Snails/pathogenicity
14.
Mol Biol Rep ; 39(4): 3401-7, 2012 Apr.
Article in English | MEDLINE | ID: mdl-21717060

ABSTRACT

A culture-independent molecular phylogenetic analysis was carried out to study for the first time the diversity of bacterial ammonia monooxygenase subunit A (amoA) and nitrogenase reductase subunit H (nifH) genes from Urca inlet at Guanabara Bay in Rio de Janeiro, Brazil. Most bacterial amoA and nifH sequences exhibited identities of less than 95% to those in the GenBank database revealing that novel ammonia-oxidizing bacteria and nitrogen-fixing microorganisms may exist in this tropical marine environment. The observation of a large number of clones related to uncultured bacteria also indicates the necessity to describe these microorganisms and to develop new cultivation methodologies.


Subject(s)
Bacteria/genetics , Bays/microbiology , Ecosystem , Genetic Variation , Nitrogen Cycle/genetics , Seawater/microbiology , Tropical Climate , Bacteria/enzymology , Brazil , Genes, Bacterial/genetics , Likelihood Functions , Oxidoreductases/genetics , Phylogeny
16.
PLoS One ; 5(12): e15774, 2010 Dec 30.
Article in English | MEDLINE | ID: mdl-21209889

ABSTRACT

BACKGROUND: Archaea are ubiquitous symbionts of marine sponges but their ecological roles and the influence of environmental factors on these associations are still poorly understood. METHODOLOGY/PRINCIPAL FINDINGS: We compared the diversity and composition of archaea associated with seawater and with the sponges Hymeniacidon heliophila, Paraleucilla magna and Petromica citrina in two distinct environments: Guanabara Bay, a highly impacted estuary in Rio de Janeiro, Brazil, and the nearby Cagarras Archipelago. For this we used metagenomic analyses of 16S rRNA and ammonia monooxygenase (amoA) gene libraries. Hymeniacidon heliophila was more abundant inside the bay, while P. magna was more abundant outside and P. citrina was only recorded at the Cagarras Archipelago. Principal Component Analysis plots (PCA) generated using pairwise unweighted UniFrac distances showed that the archaeal community structure of inner bay seawater and sponges was different from that of coastal Cagarras Archipelago. Rarefaction analyses showed that inner bay archaeaoplankton were more diverse than those from the Cagarras Archipelago. Only members of Crenarchaeota were found in sponge libraries, while in seawater both Crenarchaeota and Euryarchaeota were observed. Although most amoA archaeal genes detected in this study seem to be novel, some clones were affiliated to known ammonia oxidizers such as Nitrosopumilus maritimus and Cenarchaeum symbiosum. CONCLUSION/SIGNIFICANCE: The composition and diversity of archaeal communities associated with pollution-tolerant sponge species can change in a range of few kilometers, probably influenced by eutrophication. The presence of archaeal amoA genes in Porifera suggests that Archaea are involved in the nitrogen cycle within the sponge holobiont, possibly increasing its resistance to anthropogenic impacts. The higher diversity of Crenarchaeota in the polluted area suggests that some marine sponges are able to change the composition of their associated archaeal communities, thereby improving their fitness in impacted environments.


Subject(s)
Archaea/metabolism , Porifera/metabolism , Ammonia/chemistry , Animals , Biodiversity , Brazil , Ecology , Environment , Models, Genetic , Molecular Sequence Data , Phylogeny , Plankton , Principal Component Analysis , RNA, Ribosomal, 16S/metabolism , Seawater
17.
Microb Ecol ; 59(3): 523-32, 2010 Apr.
Article in English | MEDLINE | ID: mdl-20013262

ABSTRACT

Reef-building corals may be seen as holobiont organisms, presenting diverse associated microbial communities. Best known is the symbiotic relationship with zooxanthellae, but Archaea, Bacteria, fungi, viruses, and algal plastids are also abundant. Until now, there is little information concerning microbial communities associated with Brazilian corals. The present study aims to describe the diversity of Archaea, Bacteria, and eukaryotic algal plastid communities associated with two sympatric species, Siderastrea stellata and Mussismilia hispida, from Southeastern Brazil, using 16S rRNA gene libraries. Since corals present a high number of other associated invertebrates, coral barcoding (COI) was performed to confirm the exclusive occurrence of coral DNA in our samples. Our analysis yielded 354 distinct microbial OTUs, represented mainly by novel phylotypes. Richness (Chao1 and ACE) and diversity (H') estimations of the microbial communities associated with both species were high and comparable to other studies. Rarefaction analyses showed that microbial diversity of S. stellata is higher than that of M. hispida. Libshuff comparative analyses showed that the highest microbial community similarity between the two coral species occurred in the bacterial libraries, while archaeal and plastidial communities were significantly different. Crenarchaeota dominated archaeal communities, while Proteobacteria was the most abundant bacterial phylum, dominated by alpha-Proteobacteria. Plastids were also represented by novel phylotypes and did not match with any 16S rRNA sequences of Cyanobacteria and zooxanthellae from GenBank. Our data improves the pool of available information on Brazilian coral microbes and shows corals as sources of diverse prokaryotic and picoeukaryotic communities.


Subject(s)
Anthozoa/microbiology , Archaea/classification , Bacteria/classification , Ecosystem , Eukaryota/classification , Plastids/genetics , Animals , Archaea/genetics , Archaea/isolation & purification , Atlantic Ocean , Bacteria/genetics , Bacteria/isolation & purification , Brazil , DNA, Algal/genetics , DNA, Archaeal/genetics , DNA, Bacterial/genetics , Eukaryota/genetics , Eukaryota/isolation & purification , Gene Library , Phylogeny , Plastids/microbiology , RNA, Ribosomal, 16S/genetics , Seawater/microbiology , Sequence Analysis, DNA , Symbiosis , Water Microbiology
18.
Environ Microbiol ; 10(1): 189-99, 2008 Jan.
Article in English | MEDLINE | ID: mdl-17892478

ABSTRACT

This study is the first to apply a comparative analysis of environmental chemistry, microbiological parameters and bacterioplankton 16S rRNA clone libraries from different areas of a 50 km transect along a trophic gradient in the tropical Guanabara Bay ecosystem. Higher bacterial diversity was found in the coastal area, whereas lower richness was observed in the more polluted inner bay water. The significance of differences between clone libraries was examined with LIBSHUFF statistics. Paired reciprocal comparisons indicated that each of the libraries differs significantly from the others, and this is in agreement with direct interpretation of the phylogenetic tree. Furthermore, correspondence analyses showed that some taxa are related to specific abiotic, trophic and microbiological parameters in Guanabara Bay estuarine system.


Subject(s)
Bacteria/growth & development , Biodiversity , Ecosystem , Plankton/growth & development , Seawater/microbiology , Bacteria/genetics , Base Sequence , Brazil , Fresh Water/microbiology , Genetic Variation/genetics , Geography , Molecular Sequence Data , Phylogeny , Plankton/genetics , RNA, Ribosomal, 16S/genetics , Rivers/microbiology , Salinity , Tropical Climate , Water Microbiology
19.
Microb Ecol ; 54(3): 460-8, 2007 Oct.
Article in English | MEDLINE | ID: mdl-17593423

ABSTRACT

Guanabara Bay is an eutrophic estuarine system located in a humid tropical region surrounded by the second largest metropolitan area of Brazil. This study explores the contrasting environmental chemistry and microbiological parameters that influence the archaeaplankton diversity in a pollution gradient in Guanabara Bay ecosystem. The environments sampled ranged from completely anoxic waters in a polluted inner channel to the adjacent, relatively pristine, coastal Atlantic Ocean. Partial archaeal 16S rDNA sequences in water samples were retrieved by polymerase chain reaction (PCR) and analyzed using denaturing gradient gel electrophoresis (DGGE), cloning, and sequencing. Sequences were subjected to phylogenetic and diversity analyses. Community structure of the free-living archaeal assemblages was different from that of the particle-attached archaea according to DGGE. Gene libraries revealed that phylotype identification was consistent with environmental setting. Archaeal phylotypes found in polluted anoxic waters and in more pristine waters were closely related to organisms that have previously been found in these environments. However, inner bay archaea were related to organisms found in oil, industrial wastes, and sewage, implying that water pollution controls archaea communities in this system. The detection of a substantial number of uncultured phylotypes suggests that Guanabara Bay harbors a pool of novel archaeaplankton taxa.


Subject(s)
Archaea/genetics , Ecosystem , Seawater/microbiology , Archaea/classification , Archaea/growth & development , Brazil , DNA, Archaeal/chemistry , DNA, Archaeal/genetics , Geography , Molecular Sequence Data , Phylogeny , Polymerase Chain Reaction , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA
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