Changes in Metabolic Regulation and the Microbiota Composition after Supplementation with Different Fatty Acids in db/db Mice

Introduction. The effects of fatty acids on health vary and depend on the type, amount, and route of consumption. EPA and DHA have a defined role in health, unlike coconut oil. Objective. The aim was to investigate the changes in metabolic regulation and the composition of the culture-dependent microbiota after supplementation with different fatty acids in db/db mice. Material and Methods. We were using 32 8-week-old db/db mice, supplemented for eight weeks with EPA/DHA derived from microalgae as well as coconut oil. The lipid, hormonal profiles, and composition of the culture-dependent microbiota and the phylogenetic analysis based on the 16S rRNA gene sequencing were determined for identification of the intestinal microbiota. Results. Enriched diet with EPA/DHA reduced TNF-α, C-peptide, insulin resistance, resistin, and the plasma atherogenic index, but increased TC, LDL-c, VLDL-c, and TG without changes in HDL-c. Coconut oil raised the HDL-c, GIP, and TNF-α, with TG, insulin resistance, adiponectin, and C-peptide reduced. Conclusion. The most abundant microbial populations were Firmicutes and the least Proteobacteria. EPA/DHA derived from microalgae contributes to improving the systemic inflammatory status, but depressed the diversity of the small intestine microbiota. Coconut oil only decreased the C-peptide, raising TNF-α, with an unfavorable hormonal and lipid profile.

Cultivable Microbial Diversity in Speleothems Using MALDI-TOF Spectrometry and DNA Sequencing from Krem Soitan, Krem Lawbah, Krem Mawpun, Khasi Hills, Meghalaya, India

This study reports on the culturable microbial communities in caves from the Indian sub-continent. A high bacterial diversity and a greater bacterial taxonomic diversity is reported using MALDI-TOF spectrometry and 16S rRNA gene sequencing. This approach helped to detect a number bacterial strains from the Indian caves. The microbial diversity in the Indian caves is inadequately characterized. The study aims to expand the current understanding of bacterial diversity in the speleothems from Krem Soitan, Krem Lawbah, Krem Mawpun in Khasi Hills, Meghalaya, India. High microbial enumerations were observed on dilute nutrient agar (5.3 × 103 to 8.8 × 105) followed by M9 minimal medium (4 × 104 to 1.7 × 105) and R2A medium (1.0 × 104 to 5.7 × 105). A total of 826 bacterial isolates were selected and preserved for the study. 295 bacterial isolates were identified using MALDI-TOF spectrometry and the isolates which showed no reliable peaks were further identified by 16S rRNA gene sequencing. 91% of the total bacterial diversity was dominated by Proteobacteria and Actinobacteria. The other important phyla detected include the Firmicutes (7.45%), Deinococcus-Thermus (0.33%) and Bacteroidetes (0.67%). At the genus level, Pseudomonas (55%) and Arthrobacter (23%) were ubiquitous followed by Acinetobacter, Bacillus, Brevundimonas, Deinococcus, Flavobacterium, Paenibacillus, Pseudarthrobacter. Multivariate statistical analysis indicated that the bacterial genera formed separate clusters depending on the geochemical constituents in the spring waters suitable for their growth and metabolism. A culture-dependent approach was employed for elucidating the community structure colonizing the speleothems and wall deposits in the caves using MALDI-TOF and 16S rRNA gene sequencing. To the best of our knowledge, there are no previous geomicrobiological investigations in these caves and this study is a pioneering culture dependent study of the microbial community with many cultured isolates.

Culture-Dependent and Metagenomic Profiling of Eukaryal Diversity in Petroleum Hydrocarbon-Polluted Soil from B-Dere, Gokana, Rivers State, Nigeria

B-Dere is one of the communities in Ogoniland and a major oil producing area in Rivers State where oil exploration and production activities commenced 50 years ago and is now characterized by oil fields and installations that have remained dormant for several decades. Past spills, lack of maintenance, oil trapping and damage to oil infrastructures have been common sight in this region and the environment has been without remediation over the years. B-Dere community has suffered surface water pollution throughout its creeks with massive hydrocarbons for years without remediation. The aim of this study was to determine the Culture- dependent and Metagenomic studies of fungal diversity in petroleum polluted soils in B-Dere community in Gokana LGA of Rivers State, Nigeria. This is to profile fungal communities through next-generation techniques by shotgun sequencing of total DNA isolates directly from the oil polluted environment. Soil samples were collected aseptically with hand auger at a depths of 0-15 and 15-30 cm and made up to a composite sample and transported to the laboratory for analysis using standard microbiological methods for culture- dependent analysis while the Metagenomic studies was carried out at the Microbial Insights, Incorporated; United State of America. In this study next-generation sequencing techniques by shotgun sequencing of total DNA methods were used for identification of fungal isolates from the crude oil polluted soils in B-Dere. Deoxyribonucleic acid (DNA) extraction from crude oil polluted soil samples was performed using ZymoBiomics DNA extraction kits (Zymo Research, Inc; USA). DNA sequencing was performed by the next generation sequencing technique to determine the nucleotide sequence of all eukaryal community structure present in the polluted soil sample using ITS region. Results of the culture-dependent technique showed that only two fungal genera namely Penicillium sp and Aspergillus sp were isolated and identified while the soil was mainly dominated by the genera Penicillium (73.33%), followed by the Rhodotorula (6.54%), Dactylellina(5.09%), Kalmanozyma(2.56%), Fereydounia(1.89%), Xerochrysium(1.36%), Arthrobotrys (1.14%) and Diutina (0.77%) by the metagenomic analysis. However the three major groups were classified as Ascomycota, Basidiomycota and Mucoromycota with Ascomycota having the highest taxonomic reads of 86.76%. However, a total of 60 eukaryal species were identified, in the metagenomic study. In conclusion, these fungal strains can be used in bioremediation process and oil pollution reduction in soil ecosystems because of their high activity in aliphatic hydrocarbon degradation and cell surface hydrophobicity. The next-generation techniques by shotgun sequencing assays appear to be suitable alternatives for rapid identification of the above mentioned fungal isolates.

Comparison of culture-dependent and culture-independent techniques in the detection of lactic acid bacteria biodiversity and dynamics throughout the ripening process: The case of Turkish artisanal Tulum cheese produced in the Anamur region

Our objective was to analyze the diversity of the microbiota over 180 d of ripening of eight batches of artisanal goatskin Tulum cheeses by culture-dependent and culture-independent (PCR-DGGE) methods. V3 region of the bacterial 16S rRNA gene was amplified with the PCR after direct DNA isolation from the cheese samples. Nine different species and five genera were determined by culturing, while 11 species were identified in the PCR-DGGE technique. This diversity revealed the uniqueness of artisanal cheese varieties. The dominant genera in all the cheese samples were composed of Enterococcus species. The culture-dependent method revealed five genera (Enterococcus,Bacillus,Lactococcus,Lactobacillus, Sphingomonas) while three genera (Enterococcus, Streptococcus, Lactococcus) were detected in the culture-independent method. It was concluded that combining the two methods is important for characterizing the whole microbiota of the Tulum cheese varieties produced in the Anamur region.

Contents of lobetyolin, syringin, and atractylolide III in Codonopsis pilosula are related to dynamic changes of endophytes under drought stress

Background Codonopsis pilosula, an important medicinal plant, can accumulate certain metabolites under moderate drought stress. Endophytes are involved in the metabolite accumulations within medicinal plants. It is still unknown that the endophytes of C. pilosula are associated with the accumulations of metabolites. This study aims to investigate the promoting effect of endophytes on the accumulations of active substances in C. pilosula under drought stress. Methods High–performance liquid chromatography and high–throughput sequencing technology were performed to investigate changes in the contents of secondary metabolite and endophyte abundances of C. pilosula under drought stress, respectively. Spearman’s correlation analysis was further conducted to identify the endophytic biomarkers related to accumulations of pharmacodynamic compounds. Culture-dependent experiments were performed to confirm the functions of endophytes in metabolite accumulations. Results The distribution of pharmacological components and diversity and composition of endophytes showed tissue specificity within C. pilosula. The contents of lobetyolin, syringin, and atractylolide III in C. pilosula under drought stress were increased by 8.47%‒86.47%, 28.78%‒230.98%, and 32.17%‒177.86%, respectively, in comparison with those in untreated groups. The Chao 1 and Shannon indices in different parts of drought–stressed C. pilosula increased compared with those in untreated parts. The composition of endophytic communities in drought treatment parts of C. pilosula was different from that in control parts. A total of 226 microbial taxa were identified as potential biomarkers, of which the abundances of 42 taxa were significantly and positively correlated to the pharmacodynamic contents. Culture-dependent experiments confirmed that the contents of lobetyolin and atractylolide III were increased by the application of Epicoccum thailandicum, Filobasidium magnum, and Paraphoma rhaphiolepidis at the rates of 11.12%‒46.02%, and that the content of syringin was increased by Pseudomonas nitroreducens at the rates of 118.61%‒119.36%. Conclusions Certain endophytes participated in the accumulations of bioactive metabolites, which provided a scientific evidence for the development and application of microorganisms to improve the quality of traditional Chinese medicine.

Biological potential of Ascophyllum nodosum extract on rhizobial diversity in nodules of mothbean Vigna aconitifolia Jacq. via Amplified Ribosomal DNA Restriction Analysis

Aim: The aim of this study was to use Ascophyllum nodosum for potentially increasing the growth and rhizobial diversity in nodulating rhizobia in Vigna aconitifolia. Methodology: Different concentrations of Ascophyllum nodosum extracts (0.01%, 0.02%, 0.05%, 0.10% and 0.50%) were applied via foliar spray and on roots of Vigna aconitifolia. Growth characteristics and Amplified Ribosomal DNA Restriction Analysis were conducted to detect the morphological and molecular changes in rhizobial diversity. The restriction profiles thus obtained were used to study the rhizobial communities via Cluster analysis and Dendrogram using NTSYS-PC program and UPGMA constructed. Results: Roots treated with 0.05% Ascophyllum nodosum extract showed best growth of plants. This concentration not only proved best for the aggregation of nodules but also for obtaining enormous rhizobial diversity. Interpretation: Ascophyllum nodosum is a modern, cheap, non-toxic natural biofertilizer and Amplified Ribosomal DNA Restriction Analysis represents a favorable alternative to culture dependent method for assessing rhizobial diversity in nodulating bacteria.

Health Seeking Behaviors among Displaced Populations/Refugees

The United States resettles refugees every year. Their population in south-central Kentucky (KY) is diverse and fitting to research into culture-dependent healthcare utilization patterns. A mixed study was conducted with one hundred and ten semi-structured questionnaires and three individual interviews. Significant differences were observed between the use of healthcare services and refugees’ nationality (F (5, 98) = 4.29, p < 0.001), acculturation (t = −2.03, p < 0.04), and interpreters (t = 1.92, p < 0.05). Beliefs affect use through the level of cultural competency of the healthcare provider. These findings contribute to the health policy debates surrounding this culturally diverse population.