OTU Richness Research Articles

Distribution and Environmental Preference of Potential Mercury Methylators in Paddy Soils across China.

The neurotoxin methylmercury (MeHg) is produced mainly from the transformation of inorganic Hg by microorganisms carrying the hgcAB gene pair. Paddy soils are known to harbor diverse microbial communities exhibiting varying abilities in methylating inorganic Hg, but their distribution and environmental drivers remain unknown at a large spatial scale. Using hgcA gene amplicon sequencing, this study examined Hg-methylating communities from major rice-producing paddy soils across a transect of ∼3600 km and an altitude of ∼1300 m in China. Results showed that hgcA+ OTU richness was higher in tropical and subtropical paddy soils compared to temperate zones. Geobacteraceae, Smithellaceae, and Methanoregulaceae were identified as the dominant hgcA+ families associated with MeHg production, collectively accounting for up to 77% of total hgcA+ sequences. Hierarchical partitioning analyses revealed that pH was the main driver of hgcA genes from Geobacteraceae (14.8%) and Methanoregulaceae (16.3%), while altitude accounted for 21.4% of hgcA genes from Smithellaceae. Based on these environmental preferences, a machine-learning algorithm was used to predict the spatial distribution of these dominant hgcA+ families, thereby providing novel insights into important microbial determinants for improved prediction of MeHg production in paddy soils across China.

Composition and functional diversity of soil and water microbial communities in the rice-crab symbiosis system.

Rice-crab co-culture is an environmentally friendly agricultural and aquaculture technology with high economic and ecological value. In order to clarify the structure and function of soil and water microbial communities in the rice-crab symbiosis system, the standard rice-crab field with a ring groove was used as the research object. High-throughput sequencing was performed with rice field water samples to analyze the species and abundance differences of soil bacteria and fungi. The results showed that the OTU richness and community diversity in soil were significantly higher than those in water, while there were significant differences in soil microbial diversity and OTU richness in water sediments. The dominant species at the bacterial phylum level were Amoebacteria, Cyanobacteria, Actinomycetes, Synechococcus and Greenbacteria, and at the genus level the dominant species were norank_f_norank_o_Chloroplast, unclassified_f_Rhodobacteraceae, LD29, Cyanobium_PCC-6307, and norank_f_MWH-UniP1_aquatic_group. The dominant species at the fungal phylum level are unclassified_k_Fungi, Ascomycota, Rozellomycota, Phaeomycota and Stenotrophomonas, and at the genus level the dominant species are unclassified_k_Fungi, unclassified_p_Rozellomycota, Metschnikowia, Cladosporium, unclassified_p_Chytridiomycota. The dominant phylum may rely on mechanisms such as organic matter catabolism, secretion of secondary metabolites and phototrophic autotrophy, as predicted by functional gene analysis. The main functional genes are related to metabolic functions, including secondary product metabolism, energy metabolism, and amino acid metabolism.

Navigating diversity: primer impact on arthropod recovery in molecular environmental monitoring in tropical Amazon

IntroductionThe urgent need for effective environmental monitoring amid the escalating biodiversity crisis has prompted the adoption of molecular techniques like DNA metabarcoding. Through sequencing of taxonomically informative mitochondrial markers in bulk arthropod samples, metabarcoding allows assessment of arthropod diversity, which is crucial for ecosystem health evaluations, especially in threatened regions like the Amazon. However, challenges such as primer biases and reference database limitations persist. MethodsHere, we assess the performance of four metabarcoding primer sets, two COI markers (ZBJ-ArtF1c/ZBJ-ArtR2c [Zeale]: 157bp, targeting arthropods, and mlCOIintF/jgHCO2198 [Leray]: 313bp, targeting metazoans) and two 16S markers (Ins16S_1shortF/Ins16S_1shortR [Ins16S]: 150bp, targeting insects, and Coleop_16Sc/Coleop_16Sd [EPP]: 105bp, targeting arthropods, mainly Coleoptera) in amplifying the taxonomic constituents of bulk arthropod samples, collected across different natural and anthropogenic habitats from the Brazilian Amazon biome. To evaluate primer performance, we used the indicators (i) amplification efficiency, (ii) primer specificity, i.e., the amount of non-target sequences, (iii) detected OTU richness, (iv) group coverage and (v) taxonomic resolution. Finally, we (vi) estimated the refinement in taxa recovery by additional amplifications. ResultsDespite lower specificity and contrasting results regarding OTU richness, the primer pairs returning larger fragments showed higher taxonomic resolution (Ins16S) and broader taxonomic coverage (Leray) than Zeale and EPP did. Furthermore, results demonstrated the complementarity of the Leray and the Ins16S primer sets from taxon-dependent studies. ConclusionsDespite limitations, combining these primers could enhance biodiversity monitoring in the region. Considering incomplete reference gene banks, primers maximizing OTU richness (EPP and Leray) may be the best choice for taxon-independent surveys. These findings underscore the importance of primer selection and highlight the ongoing efforts to refine DNA metabarcoding for robust environmental assessments.

Contrasting gut bacteriomes unveiled between wild Antheraea assamensis Helfer (Lepidoptera: Saturniidae) and domesticated Bombyx mori L. (Lepidoptera: Bombycidae) silkworms.

Insect gut microbiomes play a fundamental role in various aspects of insect physiology, including digestion, nutrient metabolism, detoxification, immunity, growth and development. The wild Muga silkworm,Antheraea assamensisHelfer holds significant economic importance, as it produces golden silk. In the current investigation, we deciphered its intricate gut bacteriome through high-throughput 16S rRNA amplicon sequencing. Further, to understand bacterial community dynamics among silkworms raised under outdoor environmental conditions, we compared its gut bacteriomes with those of the domesticated mulberry silkworm,Bombyx moriL. Most abundant bacterial phyla identified in the gut ofA. assamensiswere Proteobacteria (78.1%), Bacteroidetes (8.0%) and Firmicutes (6.6%), whereas the most-abundant phyla inB. moriwere Firmicutes (49-86%) and Actinobacteria (10-36%). Further, Gammaproteobacteria (57.1%), Alphaproteobacteria (10.47%) and Betaproteobacteria (8.28%) were the dominant bacterial classes found in the gut ofA. assamensis. The predominant bacterial families inA. assamensisgut wereEnterobacteriaceae(27.7%),Comamonadaceae(9.13%),Pseudomonadaceae(9.08%)Flavobacteriaceae(7.59%)Moraxellaceae(7.38%)Alteromonadaceae(6.8%) andEnterococcaceae(4.46%). InB. mori, the most-abundant bacterial families were Peptostreptococcaceae, Enterococcaceae, Lactobacillaceae and Bifidobacteriaceae, though all showed great variability among the samples. The core gut bacteriome ofA. assamensisconsisted ofPseudomonas, Acinetobacter, Variovorax, Myroides, Alteromonas, Enterobacter, Enterococcus, Sphingomonas, Brevundimonas, Oleispira, Comamonas, Oleibacter Vagococcus, Aminobacter, Marinobacter, Cupriavidus, Aeromonas, andBacillus.Comparative gut bacteriome analysis revealed a more complex gut bacterial diversity in wildA. assamensissilkworms than in domesticatedB. morisilkworms, which contained a relatively simple gut bacteriome as estimated by OTU richness. Predictive functional profiling of the gut bacteriome suggested that gut bacteria inA. assamensiswere associated with a wide range of physiological, nutritional, and metabolic functions, including biodegradation of xenobiotics, lipid, amino acid, carbohydrate metabolism, and biosynthesis of secondary metabolites and amino acids. These results showed great differences in the composition and diversity of gut bacteria between the two silkworm species. Both insect species harbored core bacterial taxa commonly found in insects, but the relative abundance and composition of these taxa varied markedly.

Soil mycobiomes in native European aspen forests and hybrid aspen plantations have a similar fungal richness but different compositions, mainly driven by edaphic and floristic factors.

The cultivation of short-rotation tree species on non-forest land is increasing due to the growing demand for woody biomass for the future bioeconomy and to mitigate climate change impacts. However, forest plantations are often seen as a trade-off between climate benefits and low biodiversity. The diversity and composition of soil fungal biota in plantations of hybrid aspen, one of the most planted tree species for short-rotation forestry in Northern Europe, are poorly studied. The goal of this study was to obtain baseline knowledge about the soil fungal biota and the edaphic, floristic and management factors that drive fungal richness and communities in 18-year-old hybrid aspen plantations on former agricultural soils and compare the fungal biota with those of European aspen stands on native forest land in a 130-year chronosequence. Sites were categorized as hybrid aspen (17-18-year-old plantations) and native aspen stands of three age classes (8-29, 30-55, and 65-131-year-old stands). High-throughput sequencing was applied to soil samples to investigate fungal diversity and assemblages. Native aspen forests showed a higher ectomycorrhizal (EcM) fungal OTU richness than plantations, regardless of forest age. Short-distance type EcM genera dominated in both plantations and forests. The richness of saprotrophic fungi was similar between native forest and plantation sites and was highest in the middle-aged class (30-55-year-old stands) in the native aspen stands. The fungal communities of native forests and plantations were significantly different. Community composition varied more, and the natural forest sites were more diverse than the relatively homogeneous plantations. Soil pH was the best explanatory variable to describe soil fungal communities in hybrid aspen stands. Soil fungal community composition did not show any clear patterns between the age classes of native aspen stands. We conclude that edaphic factors are more important in describing fungal communities in both native aspen forest sites and hybrid aspen plantation sites than forest thinning, age, or former land use for plantations. Although first-generation hybrid aspen plantations and native forests are similar in overall fungal diversity, their taxonomic and functional composition is strikingly different. Therefore, hybrid aspen plantations can be used to reduce felling pressure on native forests; however, our knowledge is still insufficient to conclude that plantations could replace native aspen forests from the soil biodiversity perspective.

Foraging ants affect community composition and diversity of phyllosphere fungi on a myrmecophilous plants, Mallotus japonicus.

Many microorganisms inhabit the aboveground parts of plants (i.e. the phyllosphere), which mainly comprise leaves. Understanding the structure of phyllosphere microbial communities and their drivers is important because they influence host plant fitness and ecosystem functions. Despite the high prevalence of ant-plant associations, few studies have used quantitative community data to investigate the effects of ants on phyllosphere microbial communities. In the present study, we investigated the effects of ants on the phyllosphere fungal communities of Mallotus japonicus using high-throughput sequencing. Mallotus japonicus is a myrmecophilous plants that bears extrafloral nectaries, attracting several ant species, but does not provide specific ant species with nest sites like myrmecophytes do. We experimentally excluded ants with sticky resins from the target plants and collected leaf discs to extract fungal DNA. The ribosomal DNA internal transcribed spacer 1 (ITS1) regions of the phyllosphere fungi were amplified and sequenced to obtain fungal community data. Our results showed that the exclusion of ants changed the phyllosphere fungal community composition; however, the effect of ants on OTU richness was not clear. These results indicate that ants can change the community of phyllosphere fungi, even if the plant is not a myrmecophyte.

Application of environmental DNA metabarcoding to identify fish community characteristics in subtropical river systems

AbstractFish are vital in river ecosystems; however, traditional investigations of fish usually cause ecological damage. Extracting DNA from aquatic environments and identifying DNA sequences offer an alternative, noninvasive approach for detecting fish species. In this study, the effects of environmental DNA (eDNA), coupled with PCR and next‐generation sequencing, and electrofishing for identifying fish community composition and diversity were compared. In three subtropical rivers of southern China, fish specimens and eDNA in water were collected along the longitudinal (upstream–downstream) gradient of the rivers. Both fish population parameters, including species abundance and biomass, and eDNA OTU richness grouped 38 sampling sites into eight spatial zones with significant differences in local fish community composition. Compared with order‐/family‐level grouping, genus‐/species‐level grouping could more accurately reveal the differences between upstream zones I−III, midstream zones IV−V, and downstream zones VI–VIII. From the headwaters to the estuary, two environmental gradients significantly influenced the longitudinal distribution of the fish species, including the first gradient composed of habitat and physical water parameters and the second gradient composed of chemical water parameters. The high regression coefficient of alpha diversity between eDNA and electrofishing methods as well as the accurate identification of dominant, alien, and biomarker species in each spatial zone indicated that eDNA could characterize fish community attributes at a level similar to that of traditional approaches. Overall, our results demonstrated that eDNA metabarcoding can be used as an effective tool for revealing fish composition and diversity, which is important for using the eDNA technique in aquatic field monitoring.

Biogeographic effects shape soil bacterial communities across intertidal zones on island beaches through regulating soil properties

Island coastal zones are often mistakenly perceived as “ecological desert”. Actually, they harbour unique communities of organisms. The biodiversity on islands is primarily influenced by the effects of area and isolation (distance from the mainland), which mainly focused on plants and animals, encompassing studies of entire islands. However, the application of area and isolation effects to soil microorganisms on island beaches across the intertidal zones remains largely unexplored. We hypothesized that island area and isolation shape soil bacterial communities by regulating soil properties on island beaches, due to the fact that local soil properties might be strongly influenced by land-use, which may vary among islands of different sizes and isolations. To test this hypothesis, we conducted a study on 108 plots spanning 4 intertidal zones on 9 representative island beaches within Zhoushan Archipelago, eastern China. We employed one-way ANOVA and Tukey's honestly significant difference (HSD) test to assess the differences in diversity, composition of soil bacterial communities and soil properties among intertidal zones. Redundancy analysis and structural equation modelling (SEM) were used to examine the direct and indirect impacts of beach area and isolation on soil bacterial communities. Our findings revealed that the area and isolation did not significantly influence soil bacterial diversity and the relative abundance of dominant soil bacterial phyla. However, soil nitrogen (soil N), phosphorus (soil P), organic carbon (SOC), available potassium content (soil AK), and electrical conductivity (soil EC) showed significant increases with the area and isolation. As the tidal gradient increased on beaches, soil bacterial OTU richness, Chao 1, and relative abundance of Planctomycetota and Crenarchaeota decreased, while relative abundance of other soil bacterial phyla increased. We found that influences of island area and isolation shape soil bacterial communities on beaches by regulating soil properties, particularly soil moisture, salinity, and nutrients, all of which are also influenced by area and isolation. Island with larger areas and in lower intertidal zones, characterized by higher soil water content (SWC), soil EC, and soil AK, exhibited greater soil bacterial diversity and fewer dominant soil bacterial phyla. Conversely, in the higher intertidal zones with vegetation containing higher soil N and SOC, lower soil bacterial diversity and more dominant soil bacterial phyla were observed. These findings have the potential to enhance our new understanding of how island biogeography in interpreting island biome patterns.

Revealing hidden diversity and community dynamics of land snails through DNA barcoding: implications for conservation and ecological studies

IntroductionLand snails play a crucial role in maintaining ecosystem sustainability within their habitats. Therefore, understanding the characteristics of their communities is vital for ecological studies and the development of effective conservation strategies. In this study, land snail communities inhabiting the Hyrcanian forest were identified and the variations in their community composition along elevational gradients were investigated.MethodsSnail samples were collected from three distinct elevations in three different forest locations within the Hyrcanian area of Iran. This study utilized DNA barcoding to identify land snail species. By employing statistical analyses such as ANOVA and PERMANOVA, significant differences in the features of snail communities across different elevations were examined. Concurrently, soil samples were collected from each site to assess soil physicochemical parameters about snail presence.ResultThrough this comprehensive analysis, a total of 10 OTUs, were further classified into seven families, and nine genera were identified. Five of these genera had never been reported in the study region before. We observed a decline in OTU richness with increasing elevation; however, the maximum abundance of snails was found at higher elevations. CCA demonstrated that Ca, Mg, and moisture saturation predominantly shape snail community composition.DiscussionThe unique climatic conditions and spatial distribution of precipitation from lowlands to highlands, as well as from west to east, make the Hyrcanian forests an ideal case study area for understanding the dynamics of land snail communities. In summary, this study provides new insights into the land snail communities thriving in the Hyrcanian forests. The findings from our research can contribute to the development of effective conservation management strategies for forest ecosystems. By understanding the factors influencing the distribution and composition of land snail communities, we can make informed decisions to protect and preserve land snails and the balance they maintain within their habitats.

Detecting kelp-forest associated metazoan biodiversity with eDNA metabarcoding

Environmental DNA (eDNA) metabarcoding is a promising tool for monitoring marine biodiversity, but remains underutilised in Africa. In this study, we evaluated the ability of aquatic eDNA metabarcoding as a tool for detecting biodiversity associated with a South African kelp forest, an ecosystem that harbours high diversity of species, many of which are endemic, but are also sensitive to changing environmental conditions and anthropogenic pressures. Using fine-scale spatial (1 m and 8 m) and temporal (every four hours for 24 h) sampling of aquatic environmental DNA and targeting two gene regions (mtDNA COI and 12S rRNA), metabarcoding detected 880 OTUs representing 75 families in the broader metazoan community with 44 OTUs representing 24 fish families. We show extensive variability in the eDNA signal across space and time and did not recover significant spatio-temporal structure in OTU richness and community assemblages. Metabarcoding detected a broad range of taxonomic groups, including arthropods, ascidians, cnidarians, echinoderms, ctenophores, molluscs, polychaetes, ichthyofauna and sponges, as well as Placozoa, previously not reported from South Africa. Fewer than 3% of OTUs could be identified to species level using available databases (COI = 19 OTUs, 12S = 11 OTUs). Our study emphasizes that kelp-forest associated biodiversity in South Africa is understudied, but that with careful consideration for sampling design in combination with increased barcoding efforts and the construction of regional databases, eDNA metabarcoding will become a powerful biomonitoring tool of kelp-forest associated biodiversity.

The Impact of pH Modulation on the Rhizosphere Bacteriome of Maize and Bean

In soils, pH stands as the main factor modulating bacterial communities’ composition. However, most studies address its effects in bulk soils in natural systems, with few focusing on its effects in the rhizosphere of plants. Predicting pH effects in the rhizosphere is an important step towards successful microbiome manipulation, aiming to increase crop production. Here, we modulated an acidic soil’s pH to four different ranges (4.7, 5.2, 7.5 and 8.2), while correcting for fertility differences among ranges, thus isolating pH effects from other physicochemical characteristics. Then, two crops with distinct metabolisms (maize and bean) were cultivated in a greenhouse experiment and the effects of pH and cover crops on the rhizosphere bacteriome after 90 days explored through 16S rRNA gene sequencing, aiming to characterize pH effects on the rhizosphere of two different plants across this gradient. Alpha diversity indexes (OTU richness and Shannon index) were statistically different with pH but not crop species, with an interaction among factors. For beta diversity, both pH and crop species were significant modulators of community composition, without an interaction, but pH effects were 2.5 times bigger than those of plant species. Additionally, strong and significant positive correlations were observed between pH and Bacteroidetes and Deltaproteobacteria, while Actinobacteria, Planctomycetes and Acidobacteria were negatively correlated with pH. Regarding OTUs, 27 and 46 were correlated with pH in maize and bean’s rhizosphere, respectively, while 11 were shared between them. Altogether, these results provide valuable information on the isolated effect of pH in the rhizosphere of important crop plants, aiding future microbiome manipulation studies.

Primer pairs, PCR conditions, and peptide nucleic acid clamps affect fungal diversity assessment from plant root tissues

ABSTRACT High-throughput sequencing has become a prominent tool to assess plant-associated microbial diversity. Still, some technical challenges remain in characterising these communities, notably due to plant and fungal DNA co-amplification. Fungal-specific primers, Peptide Nucleic Acid (PNA) clamps, or adjusting PCR conditions are approaches to limit plant DNA contamination. However, a systematic comparison of these factors and their interactions, which could limit plant DNA contamination in the study of plant mycobiota, is still lacking. Here, three primers targeting the ITS2 region were evaluated alone or in combination with PNA clamps both on nettle (Urtica dioica) root DNA and a mock community. PNA clamps did not improve the richness or diversity of the fungal communities but increased the number of fungal reads. Among the tested factors, the most significant was the primer pair. Specifically, the 5.8S-Fun/ITS4-Fun pair exhibited a higher OTU richness but fewer fungal reads. Our study demonstrates that the choice of primers is critical for limiting plant and fungal DNA co-amplification. PNA clamps increase the number of fungal reads when ITS2 is targeted but do not result in higher fungal diversity recovery at high sequencing depth. At lower read depths, PNA clamps might enhance microbial diversity quantification for primer pairs lacking fungal specificity.

Increased precipitation alters the effects of nitrogen deposition on soil bacterial and fungal communities in a temperate forest

Anthropogenic nitrogen (N) deposition and increased precipitation are known to alter soil microbial communities. However, the combined effects of elevated N deposition and increased precipitation on soil microbial community dynamics and co-occurrence networks in temperate forests remain elusive. In this study, we conducted a field manipulation experiment by applying N solution and water to the forest canopy to simulate natural N deposition and increased precipitation in a temperate forest. We collected samples in the litter layer, organic soil layer, and mineral soil layer in 2018–2019 after 6–7 years of N and water treatments, and explored how elevated N deposition and increased precipitation regulate soil microbial diversity, community composition, and co-occurrence networks in different soil layers and at different sampling times. We found that the effects of N deposition and increased precipitation on soil microbial communities varied with soil layers and sampling times. Compared to the ambient environment, single canopy N addition (CN) or single canopy water addition (CW) did not affect bacterial Shannon diversity in the mineral soil layer in 2018, but the combined canopy N and water additions (CNW) decreased it in this layer at this time. CN increased fungal OTU richness in the organic and mineral soil layers in 2018; however, CW and CNW did not have an effect on it in the same layer at the same time. CW and CNW, but not CN, significantly affected bacterial and fungal community compositions in the litter layer in 2018 and in the organic soil layer in 2019. In contrast, CN, but not CW or CNW, significantly affected fungal community composition in the litter layer in 2019. CNW exhibited higher complexities of bacterial and fungal co-occurrence networks than CN and the ambient environment, indicating increased precipitation can strengthen the effect of N deposition on the complexity of bacterial and fungal co-occurrence networks. Our findings suggest that increased precipitation alters the effects of atmospheric N deposition on soil bacterial and fungal communities in this temperate forest, depending on soil layer and sampling time. Moreover, both bacterial and fungal community compositions are sensitive to increased precipitation, but the bacterial community composition is more sensitive to N deposition than the fungal community composition in the organic and mineral soil layers in this forest.

Conjunctival and nasal microflora in patients on topical cyclosporine for dry eye.

Introduction: Dry eye is a common ocular condition causing discomfort and visual disturbances. Anti-inflammatory agents like Cyclosporine A (CsA) are often used in its treatment. However, the impact of CsA on ocular flora remains understudied. This research aimed to evaluate changes in conjunctival and nasal microflora in patients receiving topical cyclosporine for dry eye. Methods: In this cross-sectional study, conjunctival and nasal samples were collected from two groups of dry eye patients. Group 1 consisted of 38 patients using CsA eye drops, while Group 2 included 34 patients using preservative-free artificial tear drops. Bacterial cultures were grown from the samples, and the identified organisms underwent antibiotic susceptibility testing. Additionally, alpha diversity metrics were employed to assess the diversity of bacterial species in the samples. Results: Bacterial growth was observed in 75% of conjunctival samples and 97.22% of nasal samples. Staphylococcus epidermidis was the predominant organism in both groups. Alpha diversity analysis showed no significant differences in Shannon diversity and OTU richness between the groups for most bacterial species. Antibiotic susceptibility tests revealed no substantial variations in resistance patterns between the groups. Conclusion: This study provides valuable insights into the impact of CsA eye drops on conjunctival and nasal flora in dry eye patients. The findings suggest that CsA does not significantly influence the composition, diversity, or antibiotic resistance patterns of ocular flora. Long-term topical cyclosporine treatment for dry eye does not significantly impact conjunctival microflora or lead to antibiotic resistance. These results have important implications for the safe use of CsA in patients undergoing ocular treatments, particularly those at risk of intraocular infections.

Non-significant influence between aerobic and anaerobic sample transport materials on gut (fecal) microbiota in healthy and fat-metabolic disorder Thai adults.

The appropriate sample handling for human fecal microbiota studies is essential to prevent changes in bacterial composition and quantities that could lead to misinterpretation of the data. This study firstly identified the potential effect of aerobic and anaerobic fecal sample collection and transport materials on microbiota and quantitative microbiota in healthy and fat-metabolic disorder Thai adults aged 23-43 years. We employed metagenomics followed by 16S rRNA gene sequencing and 16S rRNA gene qPCR, to analyze taxonomic composition, alpha diversity, beta diversity, bacterial quantification, Pearson's correlation with clinical factors for fat-metabolic disorder, and the microbial community and species potential metabolic functions. Our study successfully obtained microbiota results in percent and quantitative compositions. Each sample exhibited quality sequences with a >99% Good's coverage index, and a relatively plateau rarefaction curve. Alpha diversity indices showed no statistical difference in percent and quantitative microbiota OTU richness and evenness, between aerobic and anaerobic sample transport materials. Obligate and facultative anaerobic species were analyzed and no statistical difference was observed. Supportively, the beta diversity analysis by non-metric multidimensional scale (NMDS) constructed using various beta diversity coefficients showed resembling microbiota community structures between aerobic and anaerobic sample transport groups (P = 0.86). On the other hand, the beta diversity could distinguish microbiota community structures between healthy and fat-metabolic disorder groups (P = 0.02), along with Pearson's correlated clinical parameters (i.e., age, liver stiffness, GGT, BMI, and TC), the significantly associated bacterial species and their microbial metabolic functions. For example, genera such as Ruminococcus and Bifidobacterium in healthy human gut provide functions in metabolisms of cofactors and vitamins, biosynthesis of secondary metabolites against gut pathogens, energy metabolisms, digestive system, and carbohydrate metabolism. These microbial functional characteristics were also predicted as healthy individual biomarkers by LEfSe scores. In conclusion, this study demonstrated that aerobic sample collection and transport (<48 h) did not statistically affect the microbiota and quantitative microbiota analyses in alpha and beta diversity measurements. The study also showed that the short-term aerobic sample collection and transport still allowed fecal microbiota differentiation between healthy and fat-metabolic disorder subjects, similar to anaerobic sample collection and transport. The core microbiota were analyzed, and the findings were consistent. Moreover, the microbiota-related metabolic potentials and bacterial species biomarkers in healthy and fat-metabolic disorder were suggested with statistical bioinformatics (i.e., Bacteroides plebeius).

Active microeukaryotes hold clues of effects of global warming on benthic diversity and connectivity in the coastal sediments

The marine flora and fauna along the China seas include the Indo-West Pacific warm-water fauna and the North Pacific temperate fauna. The latitude of 32°N is recognized as the ecological barrier for benthic fauna in the China Seas. However, the ecological barrier on macrobenthos community gradually weakened because of global warming. Microeukaryotes are major components of marine food webs and can quickly response to environmental changes. We hypothesis that microeukaryotic benthos can stride over the ecological barrier of 32°N from the East China Sea (ECS) to the Yellow Sea (YS) under the global change, but their distribution is still limited by depth gradients between the South China Sea (SCS) and the ESC. Therefore, we investigated the distribution of benthic microeukaryotes in the China Seas using RNA metabarcoding. Higher OTU richness and phylogenetic diversity were detected in the SCS than those in the YS and ECS. The YS and ECS communities clustered together firstly and then clustered with the SCS community. Unexpectedly, communities from the 33°N in the YS clustered with those from the ECS. Similar pattern was also detected for Cercozoa, Diatomea, Amoebozoa and Fungi. These findings indicated that microeukaryotic benthos strode over the ecological barrier of 32°N under the global change. Source-sink analyses indicated more than 50% sources of the 33°N community were attributed to the ECS community, while only 12.5% from the YS community. Depth, temperature and latitude showed significant influence on communities in the China Seas. This study highlights the response of active microeukaryotes to the global warming in the coastal sediments.

Gut microbiota of Suncus murinus, a naturally obesity-resistant animal, improves the ecological diversity of the gut microbiota in high-fat-diet-induced obese mice.

The global population of obese individuals is increasing, affecting human health. High-fat diets are a leading cause of this epidemic, and animal models, such as mice, are often used in related research. Obese individuals have a different gut microbiota composition from non-obese ones, characterized by a sizeable population of certain bacteria associated with fat storage. The gut microbiome plays a significant role in regulating human physiological and metabolic functions. Links between obesity, high-fat diets and gut microbiota have become hot topics of discussion. Recently, research on the modulation of the gut microbiota has focused on fecal microbiota transplantation (FMT), which has been recognized as an effective method of studying the function of gut microbiota. The purpose of this study was to investigate how the gut microbiota of Suncus murinus, a naturally obesity-resistant animal, through FMT, affected the ecology of the gut microbiota of high-fat diet induced obese mice. In this study, Suncus murinus was used as a donor for FMT. High-fat diet induced C57BL/6NCrSIc mice were used as recipients, the body weight changes were measured and changes in their gut flora were analyzed using a 16S rRNA gene analysis. The study found that, after the FMT procedure, the FMT group tended to have a lower body weight than the control group. At the phylum level, the most predominant phyla in all groups were Firmicutes and Proteobacteria, while Deferribacteres was not detected in the FMT or antibiotic administration groups, and Bacteroidetes was not present in the antibiotic administration group. At the genus level, the FMT group had significantly lower OTU richness than the control group but greater diversity than the control group. These results indicate that FMT from Suncus murinus can help reorganize and improve the gut microbiota of mice in a balanced and diverse ecosystem.

Cropping sequence affects the structure and diversity of pathogenic and non-pathogenic soil microbial communities

AimsCurrent understanding of how cropping sequence affects pathogen-suppressive microbiomes in soil is limited. We investigated the effects of several cropping sequences from the 2020–2021 growing seasons, including cereals, pulses, and an oilseed, on microbial communities in rhizosphere and bulk soils in two western Canadian field locations.MethodsThe fungi and bacteria were characterized by Internal Transcribed Spacer (ITS) and 16S rRNA gene sequencing, respectively. The QIIME 2™ bioinformatic pipeline was used to measure the diversity and abundance of microbial species. Additionally, the concentration of the soil mineral chemicals, including macro and micro nutrients, was determined by colorimetric analysis.ResultsAscomycota (62.5%) was the most common fungal phylum, followed by Glomeromycota (11.1%), Mucoromycota (8.9%), and Basidiomycota (6.8%). Pseudomonadota (35.0%), Actinomycetota (21.1%), and Bacillota (10.1%) were the three most common bacterial phyla. Fungal OTU richness and phylogenetic diversity were highest in the cereal-pulse cropping sequencing, and bacterial OTU richness was highest in the pulse-oilseed sequences. Fusarium was the fungal genus most commonly associated with cereal-cereal monoculture and least common in the oilseed-pulse cropping sequences. The fungi (Mortierella, Funneliformis, and Diversispora) and bacteria (Rhizobium, Bradyrhizobium, Flavobacterium, and Candidatus) were higher in the cropping sequences involving pulses. The most prevalent bacteria were Streptomyces in cereal-related sequences and Solirubrobacter and Pseudomonas in oilseed-related sequences. Among soil mineral chemicals, nitrate-nitrogen, copper, calcium, potassium, and chlorine were associated with a number of beneficial fungal and bacterial genera but not with pathogenic fungal genera.ConclusionsThe results highlight the consequences of crop species selection in cropping sequences and the management of agrochemicals in the agricultural production system.

Soil Alveolata diversity in the undisturbed steppe and wheat agrocenoses under different tillage.

Microeukaryotes are vital for maintaining soil quality and ecosystem functioning, however, their communities are less studied than bacterial and fungal ones, especially by high throughput sequencing techniques. Alveolates are important members of soil microbial communities, being consumers and/or prey for other microorganisms. We studied alveolate diversity in soil under the undisturbed steppe (US) and cropped for wheat using two tillage practices (conventional, CT, and no-till, NT) by amplifying the ITS2 marker with ITS3_KYO2/ITS4 primers and sequencing amplicons using Illumina MiSeq. A total of 198 Alveolata OTUs were identified, with 158 OTUs attributed to the Ciliophora phylum, containing five classes: Litostomatea, Spirotrichea and Oligohymenophorea, Nassophorea and Phyllopharyngea. Litostomatea and Phyllopharyngea were more abundant in US as compared with CT and NT. The observed OTU richness was higher in US than in CT and NT. The β-biodiversity of soil ciliates also very distinctly differentiated the US field from CT and NT. In the US, Nassophorea and Spirotrichea correlated positively with sand and negatively with clay, silt and SOM contents. This is the first report about soil ciliates diversity in Siberia as assessed by metabarcoding technique. The revealed clear effect of land use on the relative abundance of some taxa and a lack of tillage effect suggest the importance of the quantity and quality of plant material input for shaping the prey for ciliates. The ITS-metabarcoding technique was used for the first time in the research of ciliates diversity; further studies, embracing diverse aspects of soil ciliates by combining -omics methodology with the traditional one, are needed to get a better insight on the ecological roles of the main ciliate taxa in the complex soil system.

Fungi Inhabiting Stem Wounds of Quercus robur following Bark Stripping by Deer Animals

We investigated fungal communities in oak wounds to determine how fungal species richness and community composition changes depending on the age of wounds. The sampling of wood cores was carried out from 10-, 20-, 30-, 40-, and 50-year-old wounds. The fungal community was analyzed using high-throughput sequencing of ITS2 rDNA. Sequence analysis showed the presence of 534 fungal OTUs, which were 83.4% Ascomycota, 16.3% Basidiomycota, and 0.3% Mucoromycota. The fungal OTU richness changed over time: it increased as compared between 10- and 20-year-old wounds, remained similar in 20- to 40-year-old wounds, and decreased in 50-year-old wounds. The fungal community composition also changed over time with the largest differences detected between 10-year-old and older wounds (p &lt; 0.001). The most common representatives of Basidiomycota were Laetiporus sulphureus (34.7%), Mycena galericulata (17.0%), and Cylindrobasidium evolvens (6.5%), and the most common of Ascomycota were Aposphaeria corallinolutea (13.6%), Sclerostagonospora cycadis (7.6%), and Cadophora malorum (5.8%). In conclusion, oak wounds of different ages were colonized by a high diversity of fungi including oak-associated species. Fungal communities in oak wounds underwent qualitative and quantitative changes over time, which led to the gradual shift from fungal generalists in young wounds to oak specialists in older wounds.