scholarly journals Microbial Ecology of Artisanal Feta and Kefalograviera Cheeses, Part I: Bacterial Community and Its Functional Characteristics with Focus on Lactic Acid Bacteria as Determined by Culture-Dependent Methods and Phenotype Microarrays

2022 ◽  
Vol 10 (1) ◽  
pp. 161
Author(s):  
Markella Tsigkrimani ◽  
Magdalini Bakogianni ◽  
Spiros Paramithiotis ◽  
Loulouda Bosnea ◽  
Eleni Pappa ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Bacterial Community ◽  
Microbial Communities ◽  
Carbon Sources ◽  
Rrna Gene ◽  
Ripening Stage ◽  
Biolog Ecoplates ◽  
Feta Cheese ◽  
Main Carbon Source ◽  
Early Ripening

Artisanal cheesemaking is still performed using practices and conditions derived from tradition. Feta and Kefalograviera cheeses are very popular in Greece and have met worldwide commercial success. However, there is a lack of knowledge regarding their lactic acid microecosystem composition and species dynamics during ripening. Thus, the aim of the present study was to assess the microecosystem as well as the autochthonous lactic acid microbiota during the ripening of artisanal Feta and Kefalograviera cheeses. For that purpose, raw sheep’s milk intended for cheesemaking, as well as Feta and Kefalograviera cheeses during early and late ripening were analyzed, and the lactic acid microbiota was identified using the classical phenotypic approach, clustering with PCR-RAPD and identification with sequencing of the 16S-rRNA gene, as well as with the Biolog GEN III microplates. In addition, the functional properties of the bacterial community were evaluated using the Biolog EcoPlates, which consists of 31 different carbon sources. In general, concordance between the techniques used was achieved. The most frequently isolated species from raw sheep’s milk were Enteroroccus faecium, Lactiplantibacillus plantarum and Pediococcus pentosaceus. The microecosystem of Feta cheese in the early ripening stage was dominated by Lp. plantarum and E. faecium, whereas, in late ripening, the microecosystem was dominated by Weissella paramesenteroides. The microecosystem of Kefalograviera cheese in the early ripening stage was dominated by Levilactobacillus brevis and E. faecium, and in late ripening by W. paramesenteroides and E. faecium. Finally, Carbohydrates was the main carbon source category that metabolized by all microbial communities, but the extent of their utilization was varied. Kefalograviera samples, especially at early ripening, demonstrated higher metabolic activity compared to Feta cheese. However, dominating species within microbial communities of the cheese samples were not significantly different.

scholarly journals Assessing the Microbial Communities in Four Different Daqusby Using PCR-DGGE, PLFA, and Biolog Analyses

2020 ◽  
Vol 69 (1) ◽  
pp. 27-37
Author(s):  
YUXI LING ◽  
WENYING LI ◽  
TONG TONG ◽  
ZUMING LI ◽  
QIAN LI ◽  
...  
Keyword(s):  
Microbial Communities ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Phospholipid Fatty Acid ◽  
Bacterial Species ◽  
Carbon Sources ◽  
Bacterial Biomass ◽  
Biolog Ecoplates ◽  
Different Types ◽  
Gradient Gel Electrophoresis ◽  
Plfa Analysis

Daqu made from raw wheat, barley or pea is used as an inoculum for the fermentation of Chinese Baijiu. In this study, the microbial communities of four different types of Daqus (sauce-flavor Wuling Daqu, sauce and strong-flavor Baisha Daqu, strong-flavor Deshan Daqu, and light-flavor Niulanshan Daqu) were analyzed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE), phospholipid fatty acid (PLFA) analysis, and Biolog EcoPlates analysis (Biolog). Clear differences were seen between the microbial communities of the four Daqus. PCR-DGGE showed differences in the number and brightness of bands between the Daqus, indicating the presence of unique bacterial species in Deshan Daqu, Wuling Daqu, and Niulanshan Daqu. Lactobacillus sanfranciscensis, Bacillus thermoamylovorans, and some unclassified bacteria were unique to Wuling Daqu, Deshan Daqu, and Niulanshan Daqu, respectively. Moreover, some bacterial species were observed in all four Daqus. A total of 26 PLFAs between C12 to C20 were detected from the four Daqus by PLFA analysis. Wuling Daqu had the highest total and fungal biomasses, Baisha Daqu had the highest bacterial biomass, and Niulanshan Daqu had the highest ratio of fungal biomass to bacterial biomass. The Biolog results indicated differences in the carbon source use and mode of the four Daqus, and also demonstrated that each Daqu had varying abilities to utilize different types of carbon sources. The cluster analysis of the three methods showed that the microbial communities of the four Daqus were different. This study also demonstrates the applicability of the three analytical methods in the evaluating of the microbial communities of Daqus.

scholarly journals Causal Relationship between Microbial Ecology Dynamics and Proteolysis during Manufacture and Ripening of Protected Designation of Origin (PDO) Cheese Canestrato Pugliese

2014 ◽  
Vol 80 (14) ◽  
pp. 4085-4094 ◽  
Author(s):  
Ilaria De Pasquale ◽  
Maria Calasso ◽  
Leonardo Mancini ◽  
Danilo Ercolini ◽  
Antonietta La Storia ◽  
...  
Keyword(s):  
Causal Relationship ◽  
Carbon Sources ◽  
Rrna Gene ◽  
Microbial Composition ◽  
Content Type ◽  
Bacterial Phyla ◽  
Biolog Ecoplates ◽  
Protected Designation Of Origin ◽  
Physiological Profiles ◽  
Microbial Groups

ABSTRACTPyrosequencing of the 16S rRNA gene, community-level physiological profiles determined by the use of Biolog EcoPlates, and proteolysis analyses were used to characterize Canestrato Pugliese Protected Designation of Origin (PDO) cheese. The number of presumptive mesophilic lactococci in raw ewes' milk was higher than that of presumptive mesophilic lactobacilli. The numbers of these microbial groups increased during ripening, showing temporal and numerical differences. Urea-PAGE showed limited primary proteolysis, whereas the analysis of the pH 4.6-soluble fraction of the cheese revealed that secondary proteolysis increased mainly from 45 to 75 days of ripening. This agreed with the concentration of free amino acids. Raw ewes' milk was contaminated by several bacterial phyla:Proteobacteria(68%; mainlyPseudomonas),Firmicutes(30%; mainlyCarnobacteriumandLactococcus),Bacteroidetes(0.05%), andActinobacteria(0.02%). Almost the same microbial composition persisted in the curd after molding. From day 1 of ripening onwards, the phylumFirmicutesdominated.Lactococcusdominated throughout ripening, and most of theLactobacillusspecies appeared only at 7 or 15 days. At 90 days,Lactococcus(87.2%),Lactobacillus(4.8%; mainlyLactobacillus plantarumandLactobacillus sakei), andLeuconostoc(3.9%) dominated. The relative utilization of carbon sources by the bacterial community reflected the succession. This study identified strategic phases that characterized the manufacture and ripening of Canestrato Pugliese cheese and established a causal relationship between mesophilic lactobacilli and proteolysis.

scholarly journals Assessment of the bacterial community structure in shallow and deep sediments of the Perdido Fold Belt region in the Gulf of Mexico

PeerJ ◽  
2018 ◽  
Vol 6 ◽  
pp. e5583 ◽  
Author(s):  
Ma. Fernanda Sánchez-Soto Jiménez ◽  
Daniel Cerqueda-García ◽  
Jorge L. Montero-Muñoz ◽  
Ma. Leopoldina Aguirre-Macedo ◽  
José Q. García-Maldonado
Keyword(s):  
Community Structure ◽  
Bacterial Community ◽  
Gulf Of Mexico ◽  
Microbial Communities ◽  
Bacterial Community Structure ◽  
High Throughput Sequencing ◽  
Hydrocarbon Contamination ◽  
Rrna Gene ◽  
Fold Belt ◽  
Abundant Otus

The Mexican region of the Perdido Fold Belt (PFB), in northwestern Gulf of Mexico (GoM), is a geological province with important oil reservoirs that will be subjected to forthcoming oil exploration and extraction activities. To date, little is known about the native microbial communities of this region, and how these change relative to water depth. In this study we assessed the bacterial community structure of surficial sediments by high-throughput sequencing of the 16S rRNA gene at 11 sites in the PFB, along a water column depth gradient from 20 to 3,700 m, including five shallow (20–600 m) and six deep (2,800–3,700 m) samples. The results indicated that OTUs richness and diversity were higher for shallow sites (OTUs = 2,888.2 ± 567.88;H′ = 9.6 ± 0.85) than for deep sites (OTUs = 1,884.7 ± 464.2;H′ = 7.74 ± 1.02). Nonmetric multidimensional scaling (NMDS) ordination revealed that shallow microbial communities grouped separately from deep samples. Additionally, the shallow sites plotted further from each other on the NMDS whereas samples from the deeper sites (abyssal plains) plotted much more closely to each other. These differences were related to depth, redox potential, sulfur concentration, and grain size (lime and clay), based on the environmental variables fitted with the axis of the NMDS ordination. In addition, differential abundance analysis identified 147 OTUs with significant fold changes among the zones (107 from shallow and 40 from deep sites), which constituted 10 to 40% of the total relative abundances of the microbial communities. The most abundant OTUs with significant fold changes in shallow samples corresponded toKordiimonadales, Rhodospirillales,Desulfobacterales(Desulfococcus), Syntrophobacterales and Nitrospirales(GOUTA 19,BD2-6,LCP-6), whilstChromatiales,Oceanospirillales(Amphritea,Alcanivorax),Methylococcales,Flavobacteriales,Alteromonadales(Shewanella,ZD0117) andRhodobacteraleswere the better represented taxa in deep samples. Several of the OTUs detected in both deep and shallow sites have been previously related to hydrocarbons consumption. Thus, this metabolism seems to be well represented in the studied sites, and it could abate future hydrocarbon contamination in this ecosystem. The results presented herein, along with biological and physicochemical data, constitute an available reference for further monitoring of the bacterial communities in this economically important region in the GoM.

scholarly journals Monitoring soil microorganisms with community-level physiological profiles using Biolog EcoPlates™ in Chaohu lakeside wetland, east China

2019 ◽  
Author(s):  
Zhen Teng ◽  
Wei Fan ◽  
Huiling Wang ◽  
Xiaoqing Cao ◽  
Xiaoniu Xu
Keyword(s):  
Microbial Communities ◽  
Carbon Sources ◽  
Growing Season ◽  
Wiener Index ◽  
Utilization Efficiency ◽  
Growth Trend ◽  
Biolog Ecoplates ◽  
Growing Seasons ◽  
Dormant Season ◽  
The Impact

AbstractUnder the circumstance of wetland degradation, we used Biolog EcoPlates™ method to investigate the impact of ecological restoration on the function of topsoil microbial communities by monitoring their metabolic diversity around Chaohu lakeside wetland. Four restoration patterns including reed shoaly land (RL), poplar plantation land (PL), abandoned shoaly grassland (GL) and cultivated flower land (FL) were selected. The result showed a rapid growth trend at the initial stage of incubation, following the fastest change rate at 72 h in both dormant and growing seasons, and the AWCD values of RL pattern was the highest at the detection points of each culture time, while the GL were the lowest. The calculation of diversity indicators also displayed significant lower McIntosh index in dormant season and Shannon-Wiener index in growing season in GL than in the others (P < 0.05). Carbohydrates and carboxylic acids were found to be the dominant substrates used in dormant season, whereas amino acids, polymers and phenolic acids were increasingly utilized by the microbial communities in growing season. We observed soil total potassium as the key factor that significantly affected the utilization efficiency of different carbon sources in both seasons (P < 0.05).

scholarly journals Denaturing gradient gel electrophoresis and multi-SIR profiles of soil microbial communities from a karst doline at Aggtelek National Park, Hungary

2020 ◽  
Author(s):  
Márton Mucsi ◽  
Gergely Krett ◽  
Tibor Szili-Kovács ◽  
János Móga ◽  
Andrea K. Borsodi
Keyword(s):  
Microbial Communities ◽  
Bacterial Communities ◽  
Gel Electrophoresis ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Vegetation Type ◽  
Carbon Sources ◽  
Soil Microbial Communities ◽  
Rrna Gene ◽  
Soil Microbial ◽  
Gradient Gel Electrophoresis

Abstract Soils play an important role in the ecosystem of karstic landscapes both as a buffer zone and as a source of acidity to belowground water. Although the microbiota of karstic soils is known to have a great effect on karstification processes, the activity and composition of these communities are largely unknown. This study gives a comparative analysis of soil microbial profiles from different parts of a doline located at Aggtelek, Hungary. The aim was to reveal the relationships between the vegetation type and genetic fingerprints and substrate utilisation (multi-SIR) profiles of the soil microbiota. Soil samples were collected in early and late springs along a transect in a doline covered with different types of vegetation. Genetic fingerprints of bacterial communities were examined by denaturing gradient gel electrophoresis (DGGE) based on the 16S rRNA gene, along with multi-SIR profiles of the microbial communities measured by the MicroResp method using 15 different carbon sources. Genetic fingerprinting indicated that vegetation cover had a strong effect on the composition of soil bacterial communities. Procrustean analysis showed only a weak connection between DGGE and multi-SIR profiles, probably due to the high functional redundancy of the communities. Seasonality had a significant effect on substrate usage, which can be an important factor to consider in future studies.

scholarly journals Analysis of BIOLOG GN Substrate Utilization Patterns by Microbial Communities

1998 ◽  
Vol 64 (4) ◽  
pp. 1220-1225 ◽  
Author(s):  
Kornelia Smalla ◽  
Ute Wachtendorf ◽  
Holger Heuer ◽  
Wen-tso Liu ◽  
Larry Forney
Keyword(s):  
Activated Sludge ◽  
Microbial Communities ◽  
Gel Electrophoresis ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Carbon Sources ◽  
Rrna Gene ◽  
Activated Sludge Reactor ◽  
Gradient Gel Electrophoresis ◽  
Temperature Gradient Gel Electrophoresis ◽  
Biolog Gn

ABSTRACT BIOLOG GN plates are increasingly used to characterize microbial communities by determining the ability of the communities to oxidize various carbon sources. Studies were done to determine whether the BIOLOG GN plate assay accurately reflects the catabolic potential of the inoculum used. To gain insight into which populations of microbial communities contribute to the BIOLOG patterns, denaturing gradient gel electrophoresis and temperature gradient gel electrophoresis (TGGE) were used to assess the diversity of ribotypes in the inocula and individual wells of BIOLOG plates following incubation. These studies were done with microbial communities from the rhizosphere of potatoes and an activated sludge reactor fed with glucose and peptone. TGGE analyses of BIOLOG wells inoculated with cell suspensions from the potato rhizosphere revealed that, compared with the inoculum, there was a decrease in the number of 16S rRNA gene fragments obtained from various wells, as well as a concomitant loss of populations that had been numerically dominant in the inoculum. The dominant fragments in TGGE gels could be assigned to the γ subclass of the classProteobacteria, suggesting that fast-growing bacteria adapted to high substrate concentrations were numerically dominant in the wells and may have been primarily responsible for the patterns of substrate use that were observed. Similarly, the community structure changed in wells inoculated with cells from activated sludge; one or more populations were enriched, but all dominant populations of the inoculum could be detected in at least one well. This study showed that carbon source utilization profiles obtained with BIOLOG GN plates do not necessarily reflect the functional potential of the numerically dominant members of the microbial community used as the inoculum.

Seasonal effects of river flow on microbial community coalescence and diversity in a riverine network

2020 ◽  
Vol 96 (8) ◽  
Author(s):  
Xia Luo ◽  
Xinyi Xiang ◽  
Yuanhao Yang ◽  
Guoyi Huang ◽  
Kaidao Fu ◽  
...  
Keyword(s):  
Community Structure ◽  
Microbial Community ◽  
Bacterial Community ◽  
Microbial Communities ◽  
Flow Rate ◽  
Bacterial Communities ◽  
Bacterial Community Structure ◽  
Rrna Gene ◽  
Strong Impact ◽  
Running Waters

ABSTRACT Terrestrial microbial communities may take advantage of running waters and runoff to enter rivers and mix with aquatic microorganisms. However, the environmental factors governing the interchange of the microbial community within a watercourse and its surrounding environment and the composition of the resulting community are often underestimated. The present study investigated the effect of flow rate on the mixing of water, soil, sediment and biofilm at four sites along the Lancang River and one branch of the river in winter and summer and, in turn, the resultant changes in the microbial community within each habitat. 16S rRNA gene-based Illumina high-throughput sequencing illustrated that bacterial communities were apparently distinct among biofilm, water, soil and sediment. Biofilms had the lowest richness, Shannon diversity and evenness indices compared with other habitats, and those three indices in all habitats increased significantly from winter to summer. SourceTracker analysis showed a significant coalescence between the bacterial communities of sediment, water and biofilm samples at lower flow rates. Additionally, the proportion of Betaproteobacteria in sediment and biofilms increased with a decrease in flow rate, suggesting the flow rate had a strong impact on microbial community composition and exchange among aquatic habitats. These results were further confirmed by a Mantel test and linear regression analysis. Microbial communities in all samples exhibited a significant but very weak distance–decay relationship (r = 0.093, P = 0.024). Turbidity played a much more important role on water bacterial community structure in summer (i.e. rainy season) (BIOENV, r = 0.92). Together, these results suggest that dispersal is an important factor affecting bacterial community structure in this system.

scholarly journals Pollution Gradients Altered the Bacterial Community Composition and Stochastic Process of Rural Polluted Ponds

2020 ◽  
Vol 8 (2) ◽  
pp. 311
Author(s):  
Xin Tai ◽  
Rui Li ◽  
Bao Zhang ◽  
Hao Yu ◽  
Xiao Kong ◽  
...  
Keyword(s):  
Bacterial Community ◽  
Microbial Communities ◽  
Community Composition ◽  
Community Assembly ◽  
High Throughput Sequencing ◽  
Bacterial Community Composition ◽  
Rrna Gene ◽  
Sediment Samples ◽  
Pollution Levels ◽  
Water And Sediment

Understanding the effects of pollution on ecological communities and the underlying mechanisms that drive them will helpful for selecting a method to mediate polluted ecosystems. Quantifying the relative importance of deterministic and stochastic processes is a very important issue in ecology. However, little is known about their effects on the succession of microbial communities in different pollution levels rural ponds. Also, the processes that govern bacterial communities in polluted ponds are poorly understood. In this study, the microbial communities in water and sediment from the ponds were investigated by using the 16S rRNA gene high-throughput sequencing technology. Meanwhile, we used null model analyses based on a taxonomic and phylogenetic metrics approach to test the microbial community assembly processes. Pollution levels were found to significantly alter the community composition and diversity of bacteria. In the sediment samples, the bacterial diversity indices decreased with increasing pollutant levels. Between-community analysis revealed that community assembly processes among water and sediment samples stochastic ratio both gradually decreased with the increased pollution levels, indicating a potential deterministic environmental filtering that is elicited by pollution. Our results identified assemblage drivers of bacterial community is important for improving the efficacies of ecological evaluation and remediation for contaminated freshwater systems.

scholarly journals Niche and neutral processes both shape community structure in parallelized, aerobic, single carbon-source enrichments

2017 ◽  
Author(s):  
Theodore M. Flynn ◽  
Jason C. Koval ◽  
Stephanie M. Greenwald ◽  
Sarah M. Owens ◽  
Kenneth M. Kemner ◽  
...  
Keyword(s):  
Carbon Source ◽  
Microbial Communities ◽  
Forest Soil ◽  
Carbon Sources ◽  
Subalpine Forest ◽  
Rrna Gene ◽  
Initial Structure ◽  
Inoculum Source ◽  
Alpha And Beta Diversity ◽  
Carbon Amendment

AbstractHere we seek to test the extent to which laboratory enrichments mimic natural community processes and the degree to which the initial structure of a community determines its response to a press disturbance via the addition of environmentally-relevant carbon compounds. By utilizing aerobic substrate arrays to examine the effect of carbon amendment on microbial communities taken from six distinct environments (soil from a temperate prairie and forest, tropical forest soil, subalpine forest soil, and surface water and soil from a palustrine emergent wetland), we examined how carbon amendment and inoculum source shape the composition of the community in each enrichment. Dilute subsamples from each environment were used to inoculate 96-well microtiter plates containing triplicate wells amended with one of 31 carbon sources from 6 different classes of organic compound (phenols, polymers, carbohydrates, carboxylic acids, amines, amino acids). After incubating each well aerobically in the dark for 72 hours, we analyzed the composition of the microbial communities on the substrate arrays as well as the initial inocula by sequencing 16S rRNA gene amplicons using the Illumina MiSeq platform. Comparisons of alpha and beta diversity in these systems showed that, while the composition of the communities that grow to inhabit the wells in each substrate array diverges sharply from that of the original community in the inoculum, these enrichment communities are still is strongly affected by the inoculum source. We found most enrichments were dominated by one or several OTUs most closely related to aerobes or facultative anaerobes from theProteobacteria(e.g.Pseudomonas,Burkholderia, andRalstonia) orBacteroidetes(e.g.Chryseobacterium). Comparisons within each substrate array based on the class of carbon source further show that the communities inhabiting wells amended with a carbohydrate differ significantly from those enriched with a phenolic compound. Niche selection therefore seems to play a strong role in shaping the communities in the substrate arrays, although some stochasticity is seen whereby several replicate wells within a single substrate array display strongly divergent community compositions. Overall, the use of highly parallel substrate arrays offers a promising path forward to study the response of microbial communities to a changing environment.

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