RESUMEN
A polyphasic approach was applied to characterize taxonomically a novel endophytic bacterial strain, designated as EP178T, which was previously isolated from Passiflora incarnata leaves and characterized as plant-growth promoter. The strain EP178T forms Gram stain-negative and rod-shaped cells, and circular and yellow-pigmented colonies. Its growth occurs at 10-37 °C, at pH 6.0-8.0, and tolerates up to 7% (w/v) NaCl. The major cellular fatty acids found were summed feature 8 (C18:1 ω7c), summed feature 3 (C16:1 ω6c /C16:1 ω7c), and C16:0, and the predominant ubiquinone was Q-9. The phylogenetic and nucleotide-similarity analysis with 16S rRNA gene sequences showed that strain EP178T belongs to Pseudomonas genus. The genomic-based G + C content was 65.5%. The average nucleotide identity and digital DNA-DNA hybridization values between strains EP178T and the closest type strain, P. oryzihabitans DSM 6835T, were 92.6% and 52.2%, respectively. Various genes associated with plant-growth promoting mechanisms were annotated from genome sequences. Based on the phenotypic, genomic, phylogeny and chemotaxonomic data, strain EP178T represents a new species of the genus Pseudomonas, for which the name Pseudomonas flavocrustae sp. nov. was proposed. The type strain is EP178T (= CBMAI 2609T = ICMP 24844T = MUM 23.01T).
Asunto(s)
Endófitos , Passiflora , Filogenia , Pseudomonas , ARN Ribosómico 16S , Endófitos/genética , Pseudomonas/genética , Pseudomonas/aislamiento & purificación , Passiflora/microbiología , Passiflora/crecimiento & desarrollo , ARN Ribosómico 16S/genética , Composición de Base , Ácidos Grasos/metabolismo , ADN Bacteriano/genética , Hojas de la Planta/microbiología , Hibridación de Ácido NucleicoRESUMEN
The urgent need for new antimicrobials arises from antimicrobial resistance. Actinobacteria, especially Streptomyces genus, are responsible for production of numerous clinical antibiotics and anticancer agents. Genome mining reveals the biosynthetic gene clusters (BGCs) related to secondary metabolites and the genetic potential of a strain to produce natural products. However, this potential may not be expressed under laboratory conditions. In the present study, the Antarctic bacterium was taxonomically affiliated as Streptomyces albidoflavus ANT_B131 (CBMAI 1855). The crude extracts showed antimicrobial activity against both fungi, Gram-positive and Gram-negative bacteria and antiproliferative activity against five human tumor cell lines. Whole-genome sequencing reveals a genome size of 6.96 Mb, and the genome mining identified 24 BGCs, representing 13.3% of the genome. The use of three culture media and three extraction methods reveals the expression and recovery of 20.8% of the BGCs. The natural products identified included compounds, such as surugamide A, surugamide D, desferrioxamine B + Al, desferrioxamine E, and ectoine. This study reveals the potential of S. albidoflavus ANT_B131 as a natural product producer. Yet, the diversity of culture media and extraction methods could enhance the BGCs expression and recovery of natural products, and could be a strategy to intensify the BGC expression of natural products.
Asunto(s)
Antiinfecciosos , Productos Biológicos , Streptomyces , Humanos , Antibacterianos/metabolismo , Bacterias Gramnegativas/genética , Bacterias Grampositivas/genética , Antiinfecciosos/metabolismo , Productos Biológicos/farmacología , Productos Biológicos/metabolismo , Medios de Cultivo/metabolismo , Familia de MultigenesRESUMEN
Microbial biotechnology employes techniques that rely based on the natural interactions that occur in ecosystems. Bacteria, including rhizobacteria, play an important role in plant growth, providing agricultural crops with an alternative that can mitigate the negative effects of abiotic stress, such as those caused by saline environments. In this study, bacterial isolates were obtained from soil and roots of Prosopis limensis Bentham from the department of Lambayeque, Peru. This region has high salinity levels, therefore, the collected samples were used to isolate plant growth-promoting rhizobacteria (PGPR), which were identified through morphological, and physical-biochemical characteristics. These salt tolerant bacteria were screened phosphate solubilization, indole acetic acid, deaminase activity and molecular characterization by 16S rDNA sequencing. Eighteen samples from saline soils of the Prosopis limensis plants in the northern coastal desert of San Jose district, Lambayeque, Peru. The bacterial isolates were screened for salt tolerance ranging from 2 to 10%, a total of 78 isolates were found. Isolates 03, 13 and 31 showed maximum salt tolerance at 10%, in vitro ACC production, phosphate solubilization and IAA production. The three isolates were identified by sequencing the amplified 16S rRNA gene and were found to be Pseudomonas sp. 03 (MW604823), Pseudomonas sp. 13 (MW604824) and Bordetella sp. 31 (MW604826). These microorganisms promoted the germination of radish plants and increased the germination rates for treatments T2, T3 and T4 by 129, 124 and 118% respectively. The beneficial effects of salt tolerant PGPR isolates isolated from saline environments can be new species, used to overcome the detrimental effects of salt stress on plants. The biochemical response and inoculation of the three isolates prove the potential of using these strains as a source of products that can be employed for the development of new compounds proving their potential as biofertilizers for saline environments.
Asunto(s)
Alphaproteobacteria , Prosopis , Raphanus , Prosopis/genética , Ecosistema , ARN Ribosómico 16S/genética , Estrés Salino , Suelo/química , Fosfatos , Alphaproteobacteria/genética , Microbiología del Suelo , Raíces de Plantas/microbiologíaRESUMEN
Sinningia is a genus of plants of Gesneriaceae family with species native to Brazil and is a source of several classes of bioactive secondary metabolites, such as quinones, terpenoids, flavonoids, and phenylethanoid glycosides. However, the diversity of endophytic microorganisms associated with them and the impact of endophytes on the biosynthesis of bioactive substances is unknown. Therefore, we sought to evaluate the microbial diversity, behavior, and frequency of endophytes in leaves blades of S. magnifica, S. schiffneri, and S. speciosa. These plants were collected in different regions and environments of Brazil and were studied comparatively for three consecutive years. The total DNA obtained from the blades of the plant leaves were sequenced by the Illumina MiSeq platform, followed by bioinformatics analysis to assess the microbial diversity of endophytes associated with each plant species and study year. The results of the taxonomic diversity showed a dynamic microbial community, which contained several bacterial phyla among them, Actinomycetota, Bacteroidota, Bacillota, and Pseudomonadota, and for the fungal phyla Ascomycota and Basidiomycota. Comparing the three years of study, the richness of the genera, over time, was decreasing, with signs of recovery towards the third year. The alpha and beta diversity indices confirm a great phylogenetic richness in the endophytic communities of bacteria and fungi associated with the leaf blades of Sinningia. However, these communities are comparatively little conserved, showing population and taxonomic changes of the microorganisms over time, possibly as a measure of adjustment to environmental conditions, evidencing both its fragility and versatility against the effects of environmental change on its endophytic microbial communities.
Asunto(s)
Ascomicetos , Basidiomycota , Filogenia , Bacterias/genética , Hojas de la Planta/microbiología , Endófitos/genética , Hongos/genética , BiodiversidadRESUMEN
Extreme environments Morrope and Bayovar Salt lagoons, several ecosystems and microhabitats remain unexplored, and little is known about the diversity of Actinobacteria. We suggest that the endemic bacteria present in this extreme environment is a source of active molecules with anticancer, antimicrobial, and antiparasitic properties. Using phenotypic and genotypic characterization techniques, including 16S rRNA sequencing, we identified these bacteria as members of the genera Streptomyces, Pseudonocardia, Staphylococcus, Bacillus, and Pseudomonas. Actinobacteria strains were found predominantly. Phylogenetic analysis revealed 13 Actinobacteria clusters of Streptomyces, the main genus. Three Streptomycetes, strains MW562814, MW562805, and MW562807 showed antiproliferative activities against three tumor cell lines: U251 glioma, MCF7 breast, and NCI-H460 lung (non-small cell type); and antibacterial activity against Staphylococcus aureus ATCC 6538, Escherichia coli ATCC 10536, and the multidrug resistant Acinetobacter baumannii AC-972. The antiproliferative activities (measured as total growth inhibition [TGI]) of Streptomyces sp. MW562807 were 0.57 µg/mL, for 0.61 µg/mL, and 0.80 µg/mL for glioma, lung non-small cell type, and breast cancer cell lines, respectively; the methanolic fraction of the crude extract showed a better antiproliferative activity and could inhibit the growth of (U251 (TGI = 38.3 µg/mL), OVCAR-03 (TGI = 62.1 µg/mL), and K562 (TGI = 81.5 µg/mL)) of nine tumor cells types and one nontumor cell type. Extreme enviroments, such as the Morrope and Bayovar Salt saloons are promising sources of new bacteria, whose compounds may be useful for treating various infectious diseases or even some types of cancer.
Asunto(s)
Actinobacteria , Filogenia , Antibacterianos , Pruebas de Sensibilidad Microbiana , ARN Ribosómico 16S , StreptomycesRESUMEN
Bacteria exhibiting beneficial traits like increasing the bioavailability of essential nutrients and modulating hormone levels in plants are known as plant growth promoting (PGP) bacteria. The occurrence of this specific group of bacteria in the endophytic environment may reflect the decisive role they play in a particular condition. This study aimed to determine the taxonomical diversity of the culturable bacterial endophytes, isolated in the vegetative stage of passionflower (Passiflora incarnata), and assess its potential to promote plant growth by phenotypic and genotypic approaches. The sequencing and phylogenetic analysis of the 16S rRNA gene allowed us to classify 58 bacterial endophytes into nine genera. Bacillus (70.7%) was the most dominant genus, followed by Pseudomonas (8.6%) and Pantoea (6.9%). A few isolates belonged to Rhodococcus and Paenibacillus, whereas the genera Lysinibacillus, Microvirga, Xanthomonas, and Leclercia were represented by only one isolate. The strains were tested for nitrogen fixation, phosphate solubilization, indole-acetic-acid synthesis, and siderophore production. Moreover, PGP related genes (nifH, ipdC, asb, and AcPho) were detected by PCR-based screening. Most of the isolates (94.8%) displayed a potential for at least one of the PGP traits tested by biochemical assays or PCR-based screening. Nine strains were selected based on results from both approaches and were evaluated for boosting the Cape gooseberry (Physalis peruviana) germination and growth. All tested isolates improved germination in vitro, and the majority (78%) increased growth parameters in vivo. The results suggested that most of culturable bacteria inhabiting P. incarnata in the vegetative stage could be used as probiotics for agricultural systems. Besides, their occurrence may be associated with specific physiological needs typical of this development stage.
RESUMEN
Plants and endophytic microorganisms have coevolved unique relationships over many generations. Plants show a specific physiological status in each developmental stage, which may determine the occurrence and dominance of specific endophytic populations with a predetermined ecological role. This study aimed to compare and determine the structure and composition of cultivable and uncultivable bacterial endophytic communities in vegetative and reproductive stages (RS) of Passiflora incarnata. To that end, the endophytic communities were assessed by plating and Illumina-based 16S rRNA gene amplicon sequencing. Two hundred and four cultivable bacterial strains were successfully isolated. From the plant's RS, the isolated strains were identified mainly as belonging to the genera Sphingomonas, Curtobacterium, and Methylobacterium, whereas Bacillus was the dominant genus isolated from the vegetative stage (VS). From a total of 133,399 sequences obtained from Illumina-based sequencing, a subset of 25,092 was classified in operational taxonomy units (OTUs). Four hundred and sixteen OTUs were obtained from the VS and 66 from the RS. In the VS, the most abundant families were Pseudoalteromonadaceae and Alicyclobacillaceae, while in the RS, Enterobacteriaceae and Bacillaceae were the most abundant families. The exclusive abundance of specific bacterial populations for each developmental stage suggests that plants may modulate bacterial endophytic community structure in response to different physiological statuses occurring at the different plant developmental stages.
Asunto(s)
Bacterias/clasificación , Endófitos/clasificación , Microbiota , Passiflora/crecimiento & desarrollo , Passiflora/microbiología , Bacterias/genética , Bacterias/crecimiento & desarrollo , Bacterias/aislamiento & purificación , Análisis por Conglomerados , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Endófitos/genética , Endófitos/crecimiento & desarrollo , Endófitos/aislamiento & purificación , Filogenia , Desarrollo de la Planta , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADNRESUMEN
An antibiotic-producing actinobacterium, designated isolate B375T, was isolated from marine sponge Glodia corticostylifera collected from Praia Guaecá, São Paulo, Brazil (23°49S; 45°25W), and its taxonomic position established using data from a polyphasic study. The organism showed a combination of morphological, physiological, biochemical and chemotaxonomic characteristics consistent with its classification in the genus Williamsia. Comparative 16S rRNA gene sequence analysis indicated that the strain B375T was most closely related to Williamsia serinedens DSM 45037T and Williamsia spongiae DSM 46676T and having 99.43% and 98.65% similarities, respectively, but was distinguished from these strains by a low level of DNA-DNA relatedness (53.2-63.2%) and discriminatory phenotypic properties. Chemotaxonomic investigations revealed the presence of cell-wall chemotype IV and N-glycolated muramic acid residues present in the wall cells. The cells contained C16:0 (23.3%), C18:0 10-methyl (23.2%) and C18:1 ω9c (21.6%) as the major cellular fatty acids. The strain B375T inhibited growing of Staphylococcus aureus and Colletotrichum gloeosporioides strains and was considered a producer of antimicrobial compounds. Based on the data obtained, the isolate B375T (= CBMAI 1090T = DSM 46677T) should, therefore, be classified as the type strain of a novel species of the genus Williamsia, for which the name Williamsia aurantiacus sp. nov. is proposed.
Asunto(s)
Actinomycetales/aislamiento & purificación , Actinomycetales/metabolismo , Antibacterianos/metabolismo , Colletotrichum/crecimiento & desarrollo , Poríferos/microbiología , Staphylococcus aureus/crecimiento & desarrollo , Actinomycetales/genética , Animales , Técnicas de Tipificación Bacteriana , Brasil , ADN Bacteriano/genética , Ácidos Grasos/análisis , Ácidos Murámicos/análisis , Hibridación de Ácido Nucleico , Filogenia , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADNRESUMEN
Rapid and reliable identification of bacteria is an important issue in food, medical, forensic, and environmental sciences; however, conventional procedures are time-consuming and often require extensive financial and human resources. Herein, we present a label-free method for bacterial discrimination using surface-enhanced Raman spectroscopy (SERS) and partial least squares discriminant analysis (PLS-DA). Filter paper decorated with gold nanoparticles was fabricated by the dip-coating method and it was utilized as a flexible and highly efficient SERS substrate. Suspensions of bacterial samples from three genera and six species were directly deposited on the filter paper-based SERS substrates before measurements. PLS-DA was successfully employed as a multivariate supervised model to classify and identify bacteria with efficiency, sensitivity, and specificity rates of 100% for all test samples. Variable importance in projection was associated with the presence/absence of some purine metabolites, whereas confidence intervals for each sample in the PLS-DA model were calculated using a resampling bootstrap procedure. Additionally, a potential new species of bacteria was analyzed by the proposed method and the result was in agreement with that obtained via 16S rRNA gene sequence analysis, thereby indicating that the SERS/PLS-DA approach has the potential to be a valuable tool for the discovery of novel bacteria. Graphical abstract This paper describes the discrimination of bacteria at the genus and species levels, after minimal sample preparation, using paper-based SERS substrates and PLS-DA with uncertainty estimation.
Asunto(s)
Bacterias/aislamiento & purificación , Filtración/instrumentación , Papel , Espectrometría Raman/métodos , Incertidumbre , Bacterias/genética , Límite de Detección , Microscopía Electrónica de Rastreo , Filogenia , ARN Ribosómico 16S/genética , Reproducibilidad de los ResultadosRESUMEN
A novel actinobacterium, designated isolate B138T, was isolated from the marine sponge, Amphimedon viridis, which was collected from Praia Guaecá (São Paulo, Brazil), and its taxonomic position was established using data from a polyphasic study. The organism showed a combination of chemotaxonomic and morphological characteristics consistent with its classification in the genus Williamsia and it formed a distinct phyletic line in the Williamsia 16S rRNA gene tree. It was most closely related to Williamsia serinedens DSM 45037T and Williamsia deligens DSM 44902T (99.0â% 16S rRNA gene sequence similarity) and Williamsia maris DSM 44693T (97.5â% 16S rRNA gene sequence similarity), but was distinguished readily from these strains by the low DNA-DNA relatedness values (62.3-64.4â%) and by the discriminatory phenotypic properties. Based on the data obtained, the isolate B138T (=CBMAI 1094T=DSM 46676T) should be classified as the type strain of a novel species of the genus Williamsia, for which the name Williamsia spongiae sp. nov. is proposed.
Asunto(s)
Actinomycetales/clasificación , Filogenia , Poríferos/microbiología , Actinomycetales/genética , Actinomycetales/aislamiento & purificación , Animales , Técnicas de Tipificación Bacteriana , Composición de Base , Brasil , ADN Bacteriano/genética , Hibridación de Ácido Nucleico , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADNRESUMEN
A novel actinobacterium, designated isolate B204(T), was isolated from a marine ascidian Didemnum sp., collected from São Paulo, Brazil, and its taxonomic position established using data from a polyphasic study. The organism showed a combination of chemotaxonomic and morphological characteristics consistent with its classification in the genus Gordonia and formed a distinct phyletic line in the Gordonia 16S rRNA gene tree. It was closely related to Gordonia terrae DSM 43249(T) (99.9 % 16S rRNA gene sequence similarity) and Gordonia lacunae DSM 45085(T) (99.3 % 16S rRNA gene sequence similarity) but was distinguished from these strains by a moderate level of DNA-DNA relatedness (63.0 and 54.7 %) and discriminatory phenotypic properties. Based on the data obtained, the isolate B204(T) (=CBMAI 1069(T) = DSM 46679(T)) should therefore be classified as the type strain of a novel species of the genus Gordonia, for which the name Gordonia didemni sp. nov. is proposed.
Asunto(s)
Actinobacteria/aislamiento & purificación , Agua de Mar/microbiología , Urocordados/microbiología , Actinobacteria/clasificación , Actinobacteria/genética , Animales , Brasil , ADN Bacteriano/genética , ADN Ribosómico/genética , Datos de Secuencia Molecular , Filogenia , ARN Ribosómico 16S/genéticaRESUMEN
Extracts from termite-associated bacteria were evaluated for in vitro antiviral activity against bovine viral diarrhea virus (BVDV). Two bacterial strains were identified as active, with percentages of inhibition (IP) equal to 98%. Both strains were subjected to functional analysis via the addition of virus and extract at different time points in cell culture; the results showed that they were effective as posttreatments. Moreover, we performed MTT colorimetric assays to identify the CC50, IC50, and SI values of these strains, and strain CDPA27 was considered the most promising. In parallel, the isolates were identified as Streptomyces through 16S rRNA gene sequencing analysis. Specifically, CDPA27 was identified as S. chartreusis. The CDPA27 extract was fractionated on a C18-E SPE cartridge, and the fractions were reevaluated. A 100% methanol fraction was identified to contain the compound(s) responsible for antiviral activity, which had an SI of 262.41. GC-MS analysis showed that this activity was likely associated with the compound(s) that had a peak retention time of 5 min. Taken together, the results of the present study provide new information for antiviral research using natural sources, demonstrate the antiviral potential of Streptomyces chartreusis compounds isolated from termite mounds against BVDV, and lay the foundation for further studies on the treatment of HCV infection.
RESUMEN
A novel marine actinomycete, designated B374(T), was isolated from a marine sponge, Glodia corticostylifera, which was collected from São Paulo, Brasil. The taxonomic position of B374(T) was established by using data derived from a polyphasic approach. The organism showed a combination of chemotaxonomic and morphological characteristics consistent with its classification in the genus Marmoricola and it formed a distinct phyletic line in the clade of the genus Marmoricola, based on 16S rRNA gene sequences. Strain B374(T) was most closely related to Marmoricola aequoreus SST-45(T) (98.5% 16S rRNA gene sequence similarity), but was distinguished from this strain and from the other type strains of species of the genus Marmoricola on the basis of a combination of phenotypic properties. The data obtained, therefore, indicates that isolate B374(T) ( = CBMAI 1089(T) = DSM 28169(T)) should be classified as a novel species of the genus Marmoricola, for which the name Marmoricola aquaticus sp. nov. is proposed.
Asunto(s)
Actinomycetales/clasificación , Filogenia , Poríferos/microbiología , Actinomycetales/genética , Actinomycetales/aislamiento & purificación , Animales , Composición de Base , Brasil , ADN Bacteriano/genética , Ácidos Grasos/química , Datos de Secuencia Molecular , Hibridación de Ácido Nucleico , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADNRESUMEN
The taxonomic position of a bacterium isolated from water samples from the Rio Negro, in Amazon, Brazil, was determined by using a polyphasic approach. The organism formed a distinct phyletic line in the Chromobacterium 16S rRNA gene tree and had chemotaxonomic and morphological properties consistent with its classification in this genus. It was found to be closely related to Chromobacterium vaccinii DSM 25150(T) (98.6 % 16S rRNA gene similarity) and shared 98.5 % 16S rRNA gene similarity with Chromobacterium piscinae LGM 3947(T). DNA-DNA relatedness studies showed that isolate CBMAI 310(T) belongs to distinct genomic species. The isolate was readily distinguished from the type strain of these species using a combination of phenotypic and chemotaxonomic properties. Thus, based on genotypic and phenotypic data, it is proposed that isolate CBMAI 310(T) (=DSM 26508(T)) be classified in the genus Chromobacterium as the type strain of a novel species, namely, Chromobacterium amazonense sp. nov.
Asunto(s)
Chromobacterium/clasificación , Chromobacterium/aislamiento & purificación , Microbiología del Agua , Técnicas de Tipificación Bacteriana , Brasil , Chromobacterium/genética , Análisis por Conglomerados , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Ácidos Grasos/análisis , Datos de Secuencia Molecular , Hibridación de Ácido Nucleico , Filogenia , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Homología de Secuencia de Ácido NucleicoRESUMEN
In this work the archaea and eubacteria community of a hypersaline produced water from the Campos Basin that had been transported and discharged to an onshore storage facility was evaluated by 16S recombinant RNA (rRNA) gene sequence analysis. The produced water had a hypersaline salt content of 10 (w/v), had a carbon oxygen demand (COD) of 4,300 mg/l and contains phenol and other aromatic compounds. The high salt and COD content and the presence of toxic phenolic compounds present a problem for conventional discharge to open seawater. In previous studies, we demonstrated that the COD and phenolic content could be largely removed under aerobic conditions, without dilution, by either addition of phenol degrading Haloarchaea or the addition of nutrients alone. In this study our goal was to characterize the microbial community to gain further insight into the persistence of reservoir community members in the produced water and the potential for bioremediation of COD and toxic contaminants. Members of the archaea community were consistent with previously identified communities from mesothermic reservoirs. All identified archaea were located within the phylum Euryarchaeota, with 98 % being identified as methanogens while 2 % could not be affiliated with any known genus. Of the identified archaea, 37 % were identified as members of the strictly carbon-dioxide-reducing genus Methanoplanus and 59 % as members of the acetoclastic genus Methanosaeta. No Haloarchaea were detected, consistent with the need to add these organisms for COD and aromatic removal. Marinobacter and Halomonas dominated the eubacterial community. The presence of these genera is consistent with the ability to stimulate COD and aromatic removal with nutrient addition. In addition, anaerobic members of the phyla Thermotogae, Firmicutes, and unclassified eubacteria were identified and may represent reservoir organisms associated with the conversion hydrocarbons to methane.
Asunto(s)
Archaea/genética , Bacterias/genética , Petróleo/análisis , Filogenia , Salinidad , Aguas Residuales/microbiología , Microbiología del Agua , Biodegradación Ambiental , Análisis de la Demanda Biológica de Oxígeno , Brasil , Datos de Secuencia Molecular , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , TemperaturaRESUMEN
Anopheles darlingi is the principal neotropical malaria vector, responsible for more than a million cases of malaria per year on the American continent. Anopheles darlingi diverged from the African and Asian malaria vectors â¼100 million years ago (mya) and successfully adapted to the New World environment. Here we present an annotated reference A. darlingi genome, sequenced from a wild population of males and females collected in the Brazilian Amazon. A total of 10 481 predicted protein-coding genes were annotated, 72% of which have their closest counterpart in Anopheles gambiae and 21% have highest similarity with other mosquito species. In spite of a long period of divergent evolution, conserved gene synteny was observed between A. darlingi and A. gambiae. More than 10 million single nucleotide polymorphisms and short indels with potential use as genetic markers were identified. Transposable elements correspond to 2.3% of the A. darlingi genome. Genes associated with hematophagy, immunity and insecticide resistance, directly involved in vector-human and vector-parasite interactions, were identified and discussed. This study represents the first effort to sequence the genome of a neotropical malaria vector, and opens a new window through which we can contemplate the evolutionary history of anopheline mosquitoes. It also provides valuable information that may lead to novel strategies to reduce malaria transmission on the South American continent. The A. darlingi genome is accessible at www.labinfo.lncc.br/index.php/anopheles-darlingi.
Asunto(s)
Anopheles/genética , Genoma de los Insectos , Insectos Vectores/genética , Animales , Anopheles/clasificación , Brasil , Cromosomas de Insectos/genética , Elementos Transponibles de ADN , Evolución Molecular , Femenino , Variación Genética , Interacciones Huésped-Parásitos , Proteínas de Insectos/genética , Insectos Vectores/clasificación , Resistencia a los Insecticidas , Insecticidas/farmacología , Malaria/parasitología , Masculino , Anotación de Secuencia Molecular , Filogenia , Sintenía , TranscriptomaRESUMEN
The Hepatitis C virus causes chronic infections in humans, which can develop to liver cirrhosis and hepatocellular carcinoma. The Bovine viral diarrhea virus is used as a surrogate model for antiviral assays for the HCV. From marine invertebrates and microorganisms isolated from them, extracts were prepared for assessment of their possible antiviral activity. Of the 128 tested, 2 were considered active and 1 was considered promising. The best result was obtained from the extracts produced from the Bacillus sp. isolated from the sponge Petromica citrina. The extracts 555 (500 µg/mL, SI>18) and 584 (150 µg/mL, SI 27) showed a percentage of protection of 98% against BVDV, and the extract 616, 90% of protection. All of them showed activity during the viral adsorption. Thus, various substances are active on these studied organisms and may lead to the development of drugs which ensure an alternative therapy for the treatment of hepatitis C.
Asunto(s)
Antivirales/farmacología , Bacillus/química , Virus de la Diarrea Viral Bovina Tipo 1/efectos de los fármacos , Poríferos/microbiología , Acoplamiento Viral/efectos de los fármacos , Animales , Antivirales/aislamiento & purificación , Bacillus/clasificación , Bacillus/aislamiento & purificación , Bovinos , Línea Celular , ADN Bacteriano/química , ADN Bacteriano/genética , Virus de la Diarrea Viral Bovina Tipo 1/fisiología , Pruebas de Sensibilidad Microbiana , Datos de Secuencia Molecular , Análisis de Secuencia de ADNRESUMEN
Endosymbiont-bearing trypanosomatids have been considered excellent models for the study of cell evolution because the host protozoan co-evolves with an intracellular bacterium in a mutualistic relationship. Such protozoa inhabit a single invertebrate host during their entire life cycle and exhibit special characteristics that group them in a particular phylogenetic cluster of the Trypanosomatidae family, thus classified as monoxenics. In an effort to better understand such symbiotic association, we used DNA pyrosequencing and a reference-guided assembly to generate reads that predicted 16,960 and 12,162 open reading frames (ORFs) in two symbiont-bearing trypanosomatids, Angomonas deanei (previously named as Crithidia deanei) and Strigomonas culicis (first known as Blastocrithidia culicis), respectively. Identification of each ORF was based primarily on TriTrypDB using tblastn, and each ORF was confirmed by employing getorf from EMBOSS and Newbler 2.6 when necessary. The monoxenic organisms revealed conserved housekeeping functions when compared to other trypanosomatids, especially compared with Leishmania major. However, major differences were found in ORFs corresponding to the cytoskeleton, the kinetoplast, and the paraflagellar structure. The monoxenic organisms also contain a large number of genes for cytosolic calpain-like and surface gp63 metalloproteases and a reduced number of compartmentalized cysteine proteases in comparison to other TriTryp organisms, reflecting adaptations to the presence of the symbiont. The assembled bacterial endosymbiont sequences exhibit a high A+T content with a total of 787 and 769 ORFs for the Angomonas deanei and Strigomonas culicis endosymbionts, respectively, and indicate that these organisms hold a common ancestor related to the Alcaligenaceae family. Importantly, both symbionts contain enzymes that complement essential host cell biosynthetic pathways, such as those for amino acid, lipid and purine/pyrimidine metabolism. These findings increase our understanding of the intricate symbiotic relationship between the bacterium and the trypanosomatid host and provide clues to better understand eukaryotic cell evolution.
Asunto(s)
Genes Protozoarios , Filogenia , Proteínas Protozoarias/genética , Simbiosis/genética , Trypanosomatina/genética , Bacterias/metabolismo , Composición de Base , Secuencia de Bases , Evolución Biológica , Leishmania major/genética , Redes y Vías Metabólicas , Anotación de Secuencia Molecular , Datos de Secuencia Molecular , Sistemas de Lectura Abierta , Proteínas Protozoarias/metabolismo , Alineación de Secuencia , Análisis de Secuencia de ADN , Trypanosomatina/clasificación , Trypanosomatina/metabolismo , Trypanosomatina/microbiologíaRESUMEN
Little is known about the microbial diversity associated with marine macroorganisms, despite the vital role microorganisms may play in marine ecosystems. The aim of the present study was to investigate the diversity of bacteria and fungi isolated from eight marine invertebrate and one algae samples. Data derived from ARDRA and sequencing analyses allowed the identification of marine-derived microorganisms isolated from those samples. Microbial strains identified up to the genus level revealed 144 distinct ribotypes out of 256 fungal strains and 158 distinct ribotypes out of 181 bacterial strains. Filamentous fungi were distributed among 24 different genera belonging to Ascomycota, Zygomycota and Basidiomycota, some of which had never been reported in the literature as marine invertebrate-inhabiting fungi (Pestalotiopsis, Xylaria, Botrysphaeria and Cunnninghamella). Bacterial isolates were affiliated to 41 different genera, being Bacillus, Ruegeria, Micrococcus, Pseudovibrio and Staphylococcus the most abundant ones. Results revealed an unexpected high microbial diversity associated to the macroorganisms which have been collected and suggested the selection of certain microbial taxonomic groups according to the host. The combined data gathered from this investigation contribute to broaden the knowledge of microbial diversity associated to marine macroorganisms, including as a promising source for the discovery of new natural products.
Asunto(s)
Bacterias/aislamiento & purificación , Biodiversidad , Hongos/aislamiento & purificación , Poríferos/microbiología , Sargassum/microbiología , Urocordados/microbiología , Animales , Bacterias/clasificación , Bacterias/genética , Brasil , Hongos/clasificación , Hongos/genética , Datos de Secuencia Molecular , FilogeniaRESUMEN
Terminal Restriction Fragment Length Polymorphism (T-RFLP) is a culture-independent fingerprinting method for microbial community analysis. Profiles generated by an automated electrophoresis system can be analysed quantitatively using either peak height or peak area data. Statistical testing demontrated that peak height data showed to be more reproducible than peak area data.
Terminal Restriction Fragment Length Polymorphism (T-RFLP) é um método molecular, independente de cultivo, para análise de comunidades microbianas. Perfis gerados por um sistema automatizado de eletroforese podem ser analisados quantitativamente usando dados de altura ou área dos picos. Os dados de altura mostraram-se mais reprodutíveis do que os de área.