Academic literature on the topic 'BIOLOG™ EcoPlate'

Pliek u is an Acehnese traditional condiment made from fermented coconut (Cocos nucifera) endosperm. The traditional pliek u fermentation process typically involves a diverse bacterial community. This research aimed to discover the physiological profile of the bacterial community diversity in pliek u fermentation. BIOLOGTM EcoPlates was used to obtain the physiological functions of the bacterial community during the pliek u fermentation process. The bacteria were then isolated from EcoPlate substrate to determine the predominant microorganism. Results from the analysis showed that the value of the Average Well Colour Development (AWCD) increased during the EcoPlates incubation period. The AWCD values in sample IV were higher than the AWCD values in samples I, II, and III. PCA analysis showed that the use of EcoPlate substrate by the bacterial community at the beginning of the fermentation was correlated with the substrate groups of carbohydrate and polymer, and with the substrate groups of carbohydrate and the amino acid at the end of the fermentation. The phylogenetic analysis showed that EC1 had a close relation with Pseudomonas azotoformans strain NBRC, while EC3 had a close relation with Psedomonas lundensis strain ATCC 49968. In conclusion, there were changes in the use of EcoPlate substrate and the activities of the bacterial community during the pliek u fermentation process.

Carbon utilization of bacterial communities is a key factor of the biomineralization process in limestone-rich curst areas. An efficient carbon catabolism of the microbial community is associated with the availability of carbon sources in such an ecological niche. As cave environments promote oligotrophic (carbon source stress) situations, the present study investigated the variations of different carbon substrate utilization patterns of soil and rock microbial communities between outside and inside cave environments in limestone-rich crust topography by Biolog EcoPlate™ assay and categorized their taxonomical structure and predicted functional metabolic pathways based on 16S rRNA amplicon sequencing. Community level physiological profiling (CLPP) analysis by Biolog EcoPlate™ assay revealed that microbes from outside of the cave were metabolically active and had higher carbon source utilization rate than the microbial community inside the cave. 16S rRNA amplicon sequence analysis demonstrated, among eight predominant bacterial phylum Planctomycetes, Proteobacteria, Cyanobacteria, and Nitrospirae were predominantly associated with outside-cave samples, whereas Acidobacteria, Actinobacteria, Chloroflexi, and Gemmatimonadetes were associated with inside-cave samples. Functional prediction showed bacterial communities both inside and outside of the cave were functionally involved in the metabolism of carbohydrates, amino acids, lipids, xenobiotic compounds, energy metabolism, and environmental information processing. However, the amino acid and carbohydrate metabolic pathways were predominantly linked to the outside-cave samples, while xenobiotic compounds, lipids, other amino acids, and energy metabolism were associated with inside-cave samples. Overall, a positive correlation was observed between Biolog EcoPlate™ assay carbon utilization and the abundance of functional metabolic pathways in this study.

The aim of this research was to compare the effects of organic (ORG, no mineral fertilisers and pesticides) and conventional (CON, with mineral fertilisation and plant protection chemicals) crop production systems on some soil properties: soil organic matter (SOM) and particulate organic matter (POM) contents, dehydrogenase activity and metabolic microbial diversity (Biolog EcoPlate™). Samples of Haplic Luvisol soil were collected from 23-year-old experimental fields under winter wheat from the depth layers of 0–5, 5–10, 15–20 and 30–35cm. Soil under ORG showed higher contents of SOM, POM and POM expressed as a percentage of SOM by 16.0%, 25.6% and 23.9% respectively, in particular down to 5cm depth, compared with the CON system. Also, dehydrogenase activity was 47.5% higher under the ORG than the CON system. The Biolog EcoPlate revealed higher metabolic microbial diversity in soil under ORG crop production system compared with CON. This result was supported by the average well colour development and Shannon’s diversity index values. Significant correlations between soil quality parameters and winter wheat yield confirmed their effectiveness as indicators of soil disturbance. The results showed that the ORG system much better maintained SOM and soil fertility compared with CON.

The application of Biolog EcoPlate™ for community-level physiological profiling of soils is well documented; however, the functional diversity of aquatic bacterial communities has been hardly studied. The objective of this study was to investigate the applicability of the Biolog EcoPlate™ technique and evaluate comparatively the applied endpoints, for the characterisation of the effects of metal oxide nanoparticles (MONPs) on freshwater microbial communities. Microcosm experiments were run to assess the effect of nano ZnO and nano TiO2 in freshwater at 0.8–100 mg/L concentration range. The average well colour development, substrate average well colour development, substrate richness, Shannon index and evenness, Simpson index, McIntosh index and Gini coefficient were determined to quantify the metabolic capabilities and functional diversity. Comprehensive analysis of the experimental data demonstrated that short-term exposure to TiO2 and ZnO NPs affected the metabolic activity at different extent and through different mechanisms of action. TiO2 NPs displayed lower impact on the metabolic profile showing up to 30% inhibition. However, the inhibitory effect of ZnO NPs reached 99% with clearly concentration-dependent responses. This study demonstrated that the McIntosh and Gini coefficients were well applicable and sensitive diversity indices. The parallel use of general metabolic capabilities and functional diversity indices may improve the output information of the ecological studies on microbial communities.

The quality of a soil environment affects the microbial community that inhabits it. We decided to examine whether soils formed from river sediments, located in an area of high biodiversity of organisms, are fertile and microbiologically diverse. Fluvisols are considered to be one of the most fertile soils. In this research, bacterial and metabolic diversity, as well as physico–chemical parameters, in three Fluvisols from the Vistula River Gorge of Lesser Poland was investigated. The analysis of physico–chemical and biological parameters demonstrated statistically significant differences between the three Fluvisols examined. While determining the metabolic potential of soil microbiomes with the use of the EcoPlate™ Biolog® technique, we also noted variation between the Fluvisols; however, they were arranged in a significantly different manner from other properties. The next generation sequencing method enabled us to determine the microorganisms common to three Fluvisols, and we identified bacteria specific to individual soils. These results corresponded with the data obtained through EcoPlate™, indicating that the structural diversity and metabolic potential of the microbiome does not always depend on soil quality parameters. Meanwhile, the increased structural diversity of the microbiome was found to improve the metabolic potential of soil microorganisms.

Bacterial communities in rhizosphere and root nodules have significant contributions to the growth and productivity of the soybean (Glycine max (L.) Merr.). In this report, we analyzed the physiological properties and dynamics of bacterial community structure in rhizosphere and root nodules at different growth stages using BioLog EcoPlate and high-throughput sequencing technology, respectively. The BioLog assay found that the metabolic capability of rhizosphere is in increasing trend in the growth of soybeans as compared to the bulk soil. As a result of the Illumina sequencing analysis, the microbial community structure of rhizosphere and root nodules was found to be influenced by the variety and growth stage of the soybean. At the phylum level, Actinobacteria were the most abundant in rhizosphere at all growth stages, followed by Alphaproteobacteria and Acidobacteria, and the phylum Bacteroidetes showed the greatest change. But, in the root nodules Alphaproteobacteria were dominant. The results of the OTU analysis exhibited the dominance of Bradyrhizobium during the entire stage of growth, but the ratio of non-rhizobial bacteria showed an increasing trend as the soybean growth progressed. These findings revealed that bacterial community in the rhizosphere and root nodules changed according to both the variety and growth stages of soybean in the field.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
O interesse no estudo de comunidades microbianas de ambientes pouco avaliados, impactados e/ou extremos tem aumentado significativamente. Dentre os ambientes estudados, encontram-se as águas residuais de áreas de mineração. Com o grande número de cavas e lagoas de rejeitos gerados através da atividade mineradora e o pouco estudo deste tema no Brasil, foi desenvolvido este trabalho, onde avaliamos a taxonomia e capacidade funcional da comunidade microbiana. As amostras de água foram coletadas de uma cava de mineração e uma lagoa de rejeitos de uma região anteriormente utilizada para extração de minério de ferro no Centro de Biodiversidade (CeBio) da empresa VALE/SA, em Sabará/MG. Análises do perfil de atividade metabólica indicaram um excelente desempenho quanto à degradação de diferentes compostos. Além disso, o isolamento com métodos tradicionais de cultivo mostrou que algumas bactérias apresentam capacidade em produzir diferentes enzimas de interesse biotecnológico. O DNA foi extraído e sequenciado através de técnicas metagenômicas, o sequenciamento do gene de 16S rRNA foi realizado na plataforma Ion PGM™. Esta análise, além de revelar a taxonomia da comunidade presente na água da cava e lagoa, identificou o perfil funcional indicando enriquecimento de subsistemas importantes para ambas as amostras. Adicionalmente para a água da cava de mineração, foi realizado o sequenciamento do DNA total na plataforma Ion Proton™, que através de análises com o banco de dados SEED revelou genes associados à manutenção celular e ao ciclo do nitrogênio, sugerindo que a comunidade microbiana está bem adaptada ao processo de recuperação da área. Além disso, as ORfs preditas do metagenoma foram utilizadas para uma análise no banco de dados BacMet, que revelou uma abundância de genes relacionados a resistência a metais. Assim, nossos dados expandem os conhecimentos sobre a estrutura e capacidade funcional das comunidades microbianas de ambientes aquáticos impactados, fornecendo uma melhor compreensão do papel dos micro-organismos em processos de remediação natural.
The interest in the study of microbial communities of low evaluated, impacted or extreme environments has increased significantly. Among the studied environments, there is the wastewater from mining areas. Due to the large number of reject pits and lagoons generated by the mining activity and the lack of study of this theme in Brazil, this study was developed to evaluate the taxonomy and functional capacity of the microbial community. The water samples were collected from a mining pit and a reject lagoon from a region formerly used for iron extraction at the Biodiversity Center (CeBio) from VALE/SA, in Sabará/MG. Analyzes of the metabolic activity profile indicated an excellent performance regarding the degradation of different compounds. Moreover, the isolation with traditional methods of culture showed that some bacteria have the capacity to produce different enzymes of biotechnological interest. The DNA was extracted and sequenced using metagenomic techniques, the 16S rRNA gene sequencing was performed on the Ion PGM™ platform. This analysis, in addition to revealing the taxonomy of the community present in the water of the pit and the lagoon, identified the functional profile indicating enrichment of important subsystems for the studied samples. In addition to the water from the mining pit, the total DNA sequencing was carried out on the ION Proton™ platform, which, through analysis with the SEED database, revealed genes associated with cell maintenance and the nitrogen cycle, suggesting that the microbial community is well adapted to the recovery process of the area. Also, the predicted ORfs from metagenomic were used for an analysis in the BacMet database that revealed an abundance of genes related to resistance to metals. Thus, our data expand the knowledge about the structure and functional capacity of microbial communities in impacted aquatic environments, providing a better understanding of the role of microorganisms in natural remediation processes.
FAPESP: 2014/07592-3

The need for new antibiotics has been highlighted recently with the increasing pace of emergence of drug resistant pathogens (MRSA, XDR-TB, etc.). Modification of existing antibiotics with the additions of side chains or other chemical groups and genomics based drug targeting have been the preferred method of drug development at the corporate level in recent years. These approaches have yielded few viable antibiotics and natural products are once again becoming an area of interest for drug discovery. We examined the antimicrobial â Red Soilsâ of the Hashemite Kingdom of Jordan that have historically been used to prevent infection and cure rashes by the native peoples. Antimicrobial producing bacteria were present in these soils and found to be the reason for their antibiotic activity. After isolation, these bacteria were found to excrete their antimicrobials into the liquid culture media which we could then attempt to isolate for further study. Adsorbent resins were employed to capture the antimicrobial compounds and then elute them in a more concentrated solution. As part of a drug discovery program, we sought a way to quickly characterize other soils for potential antibiotic producing bacteria. The community level physiologic profile was examined to determine if this approach would allow for a rapid categorizing of soils based on their probability of containing antimicrobial producing microorganisms. This method proved to have a high level of variability that could not be overcome even after mixing using a commercial blender. The role of these antimicrobial producing bacteria within their natural microbial community has largely been confined to microbe-plant interactions. The role of antimicrobial-producing microorganisms in driving the diversity of their community has not been a focus of considerable study. The potential of an antimicrobial-producing bacterium to act as a driver of diversity was examined using an artificial microbial community based in a sand microcosm. The changes in the microbial assemblage indicate that antimicrobial-producing bacteria may act in an allelopathic manner rather than in a predatory role.
Ph. D.

Visible lesions on coral colonies are potential indicators that environmental stressors are influencing a reef. To test this hypothesis, pairs of near-shore reefs on Taiwan were surveyed along an anthropogenically influenced gradient that included locations near the cities of Taipei and Taitung, and more remote reefs off Green Island. Two fringing reefs at Sanya, Hainan Island, a popular Chinese resort area, were also assessed. Field surveys were undertaken to detect, quantify and visually describe the occurrence of lesions at each site. Coral mucus samples were collected from both normal-appearing polyps and lesion-afflicted areas of colonies to assess carbon requirements of associated microbes. Tissue samples were also collected to identify bacterial communities inhabiting healthy tissue for comparison with those associated with lesions; denaturing gradient gel electrophoresis and 16S rRNA sequencing for bacterial identification were utilized in these analyses. In addition, tissue samples were collected in the vicinity of lesions and prepared for histological examination. At sites in Taiwan, lesions were encountered twice as often at the sites near Taipei and Taitung than at Green Island. The fewest (15/72 sightings) lesions were encountered at the reefs near Sanya, primarily because there has been nearly an 80% loss of coral cover at Sanya in recent decades. Overall, tissue loss was the most common lesion recorded (52%), followed by pink discoloration (27%) and color loss (i.e., bleaching, 15%). Porites was the taxon most commonly observed with one or more lesions (45% of sightings). Microbes within mucus from lesioned areas utilized similar carbon sources as microbes from mucus from healthy polyps, but utilized those sources more than twice as often. Examples of carbon sources utilized by microbes in >50% of the lesion samples were D-cellobiose, D-mannitol, N-acetyl-D-glucosamine, alpha-cyclodextrin, and glycogen. Bacterial assemblages on corals were significantly different between Taiwan and China, among sites, and between water samples and coral samples, but not between healthy samples and lesions. Bacterial sequences identified in tissue samples from lesions revealed the presence of well-known disease-related genera, such as Clostridium and Vibrio. Microbes specifically indicating anthropogenic sources, included Bacillus sp. (sewage sludge) and Geobacillus thermolevorans (irritable bowel syndrome). Histological examination of tissue samples, particularly those from lesions characterized as tissue loss, revealed fragmentation and detachment from the mesoglea of gastrodermis and epidermis, as well as brown granular material, and the presence of ciliates and small crustaceans. Corals are susceptible to a variety of diseases. For reefs in the western Atlantic and Caribbean, occurrences of lesions and characterization of coral diseases have been relatively well documented. In contrast, many areas in the vast Indo-Pacific, including the reefs of Taiwan and China, have received much less attention. This study of lesions and associated microbiomes on nearshore reefs of Taiwan and Hainan Island supports previous research that has revealed higher incidences of coral lesions and disease in reefs near extensive human populations. The results also support the hypothesis that many of the microbes associated with coral lesions are part of the natural coral microbiome and that some microbes can become opportunistic when the host corals are stressed.

The objective of this work was to understand microbial population density and diversity, both spatially and temporally, in wetland mesocosms to gain a better fundamental understanding for use in the optimization and design of constructed wetlands (CWs). A standardized community level physiological profiling (CLPP) data analysis protocol was adapted and utilized for CW mesocosms. A new one-dimensional metric was developed to track community divergence using BIOLOGTM ECO plate data. The method proved easy to use, did not require a background in multivariate statistics, and accurately described community divergence in mesocosm systems. To study mesocosm biofilm-bound bacterial communities an appropriate detachment protocol was required. Various shaking protocols were evaluated for their effectiveness in the detachment of bacteria from mesocosm pea gravel, with a focus on detachment of viable and representative bacterial communities. A protocol based on mechanical shaking with buffer and enzymes was identified as an optimal approach and used further in this study. The bacterial communities associated with the interstitial water, pea gravel media, and rhizospheric regions from both planted and unplanted CW mesocosms were profiled using the CLPP method and compared. Vertical community stratification was observed for all mesocosm systems. Rhizospheric communities were found to be significantly more active than their gravel-associated counterparts, suggesting that although rhizospheric bacteria were less abundant in the mesocosms they may play a more significant role in the removal and fate of water born contaminants. The start-up dynamics of CW mesocosms was investigated using the CLPP and standard CW characterization methods over an eight month period. All mesocosms showed a steep increase in interstitial community divergence until day 75-100, at which point a steady-state was reached. The interstitial communities were also characterized in terms of similarity based on experimental design treatments (planted/unplanted and origin of seeding inoculum). Four stages were identified during the start-up consisting of an initial stage where mesocosm communities were differentiated based on origin of the inoculum, a period where adjustments and shifts occurred in all mesocosm, a time where all mesocosm communities were quite similar, and a final state where community differentiations were made based plant presence in the mesocosms.

Carrion decomposition is an essential ecosystem function as it is an important component of nutrient cycling. Carrion decomposition has primarily been attributed to insect consumption, with little attention given to microbial communities or their potential interactions with insects. The first objective was to use passive insect-trapping methods to assess primary colonizer communities on swine carcasses between two treatments: 1) carrion with access to insects and 2) carrion excluded from insect access for five days using exclusion cages. Despite similarities between succession patterns within each treatment, carcasses initially exposed to insects had significantly fewer insect taxa. Therefore, collections of adult insect communities associated with carrion are promising as an indication of whether or not there has been a delay in insect colonization of a resource. There has yet to be a study documenting bacterial communities during carrion decomposition. The second objective was to describe bacterial community succession and composition during decomposition in the presence and absence of naturally occurring insects. Total genomic DNA was used to identify bacterial community composition via a modified bacterial tagged encoded FLX amplicon pyrosequencing. I obtained 378,904 sequences and documented distinct bacterial community successional trajectories associated with insect access and exclusion carcasses. By the fifth day of decomposition, Proteus was the dominant (72%) bacterial genus on exclusion carcasses while Psychrobacillus (58%) and Ignatzschineria (18%) were dominant bacterial genera on insect carcasses. These data are the first to document bacterial community composition and succession on carrion. My final objective was to assess microbial community function in response to carrion insect colonization using metabolic profiling. I characterized microbial community metabolic function in the presence and absence of the primary necrophagous insects. I documented significant microbial community metabolic profile changes during active decomposition of carcasses. Mean carcass microbial community metabolic function with insect access continuously decreased over decomposition during both field seasons. Thus demonstrating microbial metabolic activity may have discriminatory power to differentiate early and late stages of decomposition. Overall, my data contributes to an understudied area of microbial research important to organic matter decomposition, forensic entomology, and microbial-insect ecological interactions.