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Keywords = T-RFLP fingerprinting

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18 pages, 1999 KiB  
Article
Methodological Aspects of Multiplex Terminal Restriction Fragment Length Polymorphism-Technique to Describe the Genetic Diversity of Soil Bacteria, Archaea and Fungi
by Agata Gryta and Magdalena Frąc
Sensors 2020, 20(11), 3292; https://doi.org/10.3390/s20113292 - 9 Jun 2020
Cited by 14 | Viewed by 4489
Abstract
The molecular fingerprinting methods used to evaluate soil microbial diversity could also be used as effective biosensors for the purposes of monitoring ecological soil status. The biodiversity of microorganisms is a relevant index of soil activity and there is a necessity to develop [...] Read more.
The molecular fingerprinting methods used to evaluate soil microbial diversity could also be used as effective biosensors for the purposes of monitoring ecological soil status. The biodiversity of microorganisms is a relevant index of soil activity and there is a necessity to develop tools to generate reliable results for an emerging approach in the field of environmental control using microbial diversity biosensors. This work reports a method under development for determining soil microbial diversity using high efficiency Multiplex PCR-Terminal Restriction Fragment Length Polymorphism (M-T-RFLP) for the simultaneous detection of bacteria, archaea and fungi. Three different primer sets were used in the reaction and the analytical conditions were optimized. Optimal analytical conditions were achieved using 0.5 µM of primer for bacteria and 1 µM for archaea and fungi, 4 ng of soil DNA template, and HaeIII restriction enzyme. Comparative tests using the proposed analytical approach and a single analysis of each microorganism group were carried out to indicate that both genetic profiles were similar. The Jaccard similarity coefficient between single and multiplexing approach ranged from 0.773 to 0.850 for bacteria and fungi, and 0.208 to 0.905 for archaea. In conclusion, the multiplexing and pooling approaches significantly reduced the costs and time required to perform the analyses, while maintaining a proper effectiveness. Full article
(This article belongs to the Section Biosensors)
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10 pages, 911 KiB  
Article
Effect of Long-Term Cropping Systems on the Diversity of the Soil Bacterial Communities
by Zoltán Mayer, Zita Sasvári, Viktor Szentpéteri, Beatrix Pethőné Rétháti, Balázs Vajna and Katalin Posta
Agronomy 2019, 9(12), 878; https://doi.org/10.3390/agronomy9120878 - 11 Dec 2019
Cited by 26 | Viewed by 4314
Abstract
Soil microbial communities are involved in the maintenance of productivity and health of agricultural systems; therefore an adequate understanding of soil biodiversity plays a key role in ensuring sustainable use of soil. In the present study, we evaluated the influence of different cropping [...] Read more.
Soil microbial communities are involved in the maintenance of productivity and health of agricultural systems; therefore an adequate understanding of soil biodiversity plays a key role in ensuring sustainable use of soil. In the present study, we evaluated the influence of different cropping systems on the biodiversity of the soil bacterial communities, based on a 54-year field experiment established in Martonvásár, Hungary. Terminal restriction fragment length polymorphism (T-RFLP) fingerprinting technique was used to assess soil bacterial diversity and community structure in maize monoculture and three different crop rotations (maize–alfalfa, maize–wheat and the maize–barley–peas–wheat Norfolk type). No differences in richness and diversity were detected between maize monoculture and crop rotations except for the most intense rotation system (Norfolk-type). Although the principal component analysis did not reveal a clear separation between maize monoculture and the other rotation systems, the pairwise tests of analysis of similarity (ANOSIM) revealed that there are significant differences in the composition of bacterial communities between the maize monoculture and maize–alfalfa rotation as well as between wheat–maize and Norfolk-type rotation. Full article
(This article belongs to the Section Innovative Cropping Systems)
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16 pages, 1353 KiB  
Article
Effects of Dietary Yogurt on the Healthy Human Gastrointestinal (GI) Microbiome
by Daniel J. Lisko, G. Patricia Johnston and Carl G. Johnston
Microorganisms 2017, 5(1), 6; https://doi.org/10.3390/microorganisms5010006 - 15 Feb 2017
Cited by 60 | Viewed by 19120
Abstract
The gastrointestinal (GI) tract performs key functions that regulate the relationship between the host and the microbiota. Research has shown numerous benefits of probiotic intake in the modulation of immune responses and human metabolic processes. However, unfavorable attention has been paid to temporal [...] Read more.
The gastrointestinal (GI) tract performs key functions that regulate the relationship between the host and the microbiota. Research has shown numerous benefits of probiotic intake in the modulation of immune responses and human metabolic processes. However, unfavorable attention has been paid to temporal changes of the microbial composition and diversity of the GI tract. This study aimed to investigate the effects of yogurt consumption on the GI microbiome bacteria community composition, structure and diversity during and after a short-term period (42 days). We used a multi-approach combining classical fingerprinting techniques (T-RFLPs), Sanger analyses and Illumina MiSeq 16S rRNA gene amplicon sequencing to elucidate bacterial communities and Lactobacilli and Bifidobacteria populations within healthy adults that consume high doses of yogurt daily. Results indicated that overall GI microbial community and diversity was method-dependent, yet we found individual specific changes in bacterial composition and structure in healthy subjects that consumed high doses of yogurt throughout the study. Full article
(This article belongs to the Special Issue Microbes and Food)
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28 pages, 1040 KiB  
Article
Improved Anaerobic Fermentation of Wheat Straw by Alkaline Pre-Treatment and Addition of Alkali-Tolerant Microorganisms
by Heike Sträuber, Franziska Bühligen, Sabine Kleinsteuber, Marcell Nikolausz and Katharina Porsch
Bioengineering 2015, 2(2), 66-93; https://doi.org/10.3390/bioengineering2020066 - 15 Apr 2015
Cited by 35 | Viewed by 10032
Abstract
The potential of two alkali-tolerant, lignocellulolytic environmental enrichment cultures to improve the anaerobic fermentation of Ca(OH)2-pre-treated wheat straw was studied. The biomethane potential of pre-treated straw was 36% higher than that of untreated straw. The bioaugmentation of pre-treated straw with the [...] Read more.
The potential of two alkali-tolerant, lignocellulolytic environmental enrichment cultures to improve the anaerobic fermentation of Ca(OH)2-pre-treated wheat straw was studied. The biomethane potential of pre-treated straw was 36% higher than that of untreated straw. The bioaugmentation of pre-treated straw with the enrichment cultures did not enhance the methane yield, but accelerated the methane production during the first week. In acidogenic leach-bed fermenters, a 61% higher volatile fatty acid (VFA) production and a 112% higher gas production, mainly CO2, were observed when pre-treated instead of untreated straw was used. With one of the two enrichment cultures as the inoculum, instead of the standard inoculum, the VFA production increased by an additional 36% and the gas production by an additional 110%, again mainly CO2. Analysis of the microbial communities in the leach-bed processes revealed similar bacterial compositions in the fermenters with pre-treated straw, which developed independently of the used inoculum. It was suggested that the positive metabolic effects with the enrichment cultures observed in both systems were due to initial activities of the alkali-tolerant microorganisms tackling the alkaline conditions better than the standard inocula, whereas the latter dominated in the long term. Full article
(This article belongs to the Special Issue Microbial Ecology of Anaerobic Digestion)
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15 pages, 872 KiB  
Article
Seasonal Changes in Microbial Community Structure in Freshwater Stream Sediment in a North Carolina River Basin
by John P. Bucci, Anthony J. Szempruch, Jane M. Caldwell, Joseph C. Ellis and Jay F. Levine
Diversity 2014, 6(1), 18-32; https://doi.org/10.3390/d6010018 - 3 Jan 2014
Cited by 33 | Viewed by 10097
Abstract
This study examined seasonal differences in microbial community structure in the sediment of three streams in North Carolina’s Neuse River Basin. Microbes that reside in sediment are at the base of the food chain and have a profound influence on the health of [...] Read more.
This study examined seasonal differences in microbial community structure in the sediment of three streams in North Carolina’s Neuse River Basin. Microbes that reside in sediment are at the base of the food chain and have a profound influence on the health of freshwater stream environments. Terminal-Restriction Fragment Length Polymorphism (T-RFLP), molecular fingerprint analysis of 16S rRNA genes was used to examine the diversity of bacterial species in stream sediment. Sediment was sampled in both wet and dry seasons from an agricultural (Bear), mixed urban (Crabtree) and forested (Marks) Creek, and the microbiota examined. Gamma, Alpha and Beta proteobacteria were prevalent species of microbial taxa represented among all sites. Actinobacteria was the next most prevalent species observed, with greater occurrence in dry compared to the wet season. Discernable clustering was observed of Marks and Bear Creek samples collected during the wetter period (September–April), which corresponded with a period of higher precipitation and cooler surface water temperatures. Although not statistically significant, microbial community structure appeared different between season (ANOSIM, R = 0.60; p < 0.10). Principal components analysis confirmed this pattern and showed that the bacterial groups were separated by wet and dry seasonal periods. These results suggest seasonal differences among the microbial community structure in sediment of freshwater streams and that these communities may respond to changes in precipitation during wetter periods. Full article
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