Revealing Cryptic Changes of Cyanobacterial Community Structure in Two Eutrophic Lakes Using eDNA Sequencing

Twelve mature (aged 5–16 years) horses and ponies of mixed breed and type were fed restricted (1.25% BM Dry matter) quantities of one of two fiber based diets formulated to be iso-caloric. Diet 1 comprised of 0.8% body mass (BM) of chaff based complete feed plus 0.45% BM low energy grass hay (the same hay used for both diets). Diet 2 comprised 0.1% BM of a nutrient balancer plus 1.15% BM grass hay. Fecal samples were collected at week 10 and week 16. DNA was extracted and the V1-V2 regions of 16SrDNA were 454-pyrosequenced to investigate the bacterial microbiome of the horse. The two most abundant phyla found in both diets and sampling periods were the Firmicutes and Bacteroidetes. There was a clear reduction in Bacteroidetes with a concordant increase in Firmicutes over time. There was a limited degree of stability within the bacterial community of the hindgut of horses, with 65% of bacteria retained, over a 6 week period whilst on a uniform diet. The presence of a core community defined by being present in all samples (each animal/diet combination) included in the study and being present at 0.1% relative abundance (or greater) was identified. In total 65 operational taxonomic units (OTUs) were identified that fit the definition of core making up 21–28% of the total sequences recovered. As with total population the most abundant phyla were the Bacteroidetes followed by the Firmicutes, however there was no obvious shift in phyla due to period. Indeed, when the relative abundance of OTUs was examined across diets and periods there was no significant effect of diet or period alone or in combination on the relative abundance of the core OTUs.

Sustainable development and environmental restoration in Lake Erhai, China

Biodiversity and dynamics of cyanobacterial communities during blooms in temperate lake (Harsha Lake, Ohio, USA)

The use of molecular methods to investigate the community structure and diversity of microalgae has largely replaced the previous morphological methods that were routinely carried out by microscopy. Different DNA polymerases can lead to bias in PCR amplification and affect the downstream community and diversity analysis. In this study, two clone libraries were constructed with two different DNA polymerases, Q5 high-fidelity DNA polymerase and exTaq polymerase, to compare the differences in their capability to accurately reflect the cyanobacterial community structure and diversity in a subtropical deep freshwater reservoir, Dongzhen reservoir. The results indicated that the two cyanobacterial clone libraries constructed by using Q5 high-fidelity DNA polymerase and exTaq DNA polymerase did not show significant differences, although a slightly higher community diversity was revealed by using Q5 high-fidelity DNA polymerase. It is noteworthy that so far Q5 high-fidelity DNA polymerase was the first time to be employed in the genetic analysis of cyanobacterial community. And it is for the first time that the cyanobacterial community structure in Dongzhen reservoir was analyzed using molecular methods. Phylogenetic analysis revealed that most of the clones belonged to Cyanophyta and chloroplast, among which Cyanobium sp. Suigetsu-CR5 made up the largest fraction of cyanobacteria in winter.

417 Background: In pancreaticobiliary (PB) cancers, there is a paucity of data on predictive and pathophysiologic role of the biliary microbiome. We analyzed bile collected from patients with benign and malignant PB diseases to identify microbiomic signatures associated with malignancy. Methods: We collected bile samples from consenting patients during routine endoscopic retrograde cholangiopancreatography at the Cleveland Clinic, approved by the Institutional Review Board. DNA was extracted from bile specimens using PowerViral RNA/DNA Isolation kit. Bacterial 16S rRNA gene amplification and library construction were performed according to the 16S Metagenomic Sequencing Library Preparation guide from Illumina. Post-sequencing analysis was done using QIIME (Quantitative Insights Into Microbial Ecology), Bioconductor phyloseq, microbiomeSeq and mixMC packages. Results: Of 46 enrolled patients, 32 had PB cancers including pancreatic (N = 25), cholangiocarcinoma (N = 6), and gallbladder (N = 1). The rest (N = 14) had benign PB diseases including acute and chronic pancreatitis, and gallstones. Using multivariate approach in mixMC to classify Operational Taxonomic Units (OTUs), we found a predominance of genera Dicekeya (p = 0.0002), [ Eubacterium] hallii group (p = 0.0007) , Bacteroides (p = 0.00099) , Faecalibacterium (p = 0.007) , Facklamia (p = 0.013) , Peptococcus (p = 0.013) , Bergeyella (p = 0.0024) , Lachnospira (p = 0.026) , and Lactobacillus (p = 0.025) in bile samples from PB cancers as compared to benign PB diseases. Furthermore, bile samples from patients with pancreatic cancer showed an increased abundance of genera Enterobacter, Parabacteroides, Atopobium, Alloprevotella, Prevotella 7, Acinetobacter, Bergeyella, Clostridium sensu stricto, Lactobacillus, and Rothia; and a decreased abundance of genera Tannerella, Peptococcus, Colinsella, Capnocytophaga, Achromobacter, Ruminococcus 2, Bacteroides, Alistipes, Barnesiella,, Lachnoclostridium, Lautropia, Akkermansia, and Christensenellaceae R-7 group as compared to bile samples from patients with cholangiocarcinoma. Conclusions: Distinct microbiome signatures are associated with benign and malignant PB diseases. There is a difference in the relative abundance of OTUs in bile samples between patients with benign PB diseases vs PB cancers, and between pancreatic cancer vs cholangiocarcinoma. Our findings raise the possibility that either these OTUs have a role in carcinogenesis, or that changes in the microenvironment of benign PB diseases differ from PB cancers leading to distinct separation of the OTU clusters. Further studies to explore and validate our findings are needed.

Identification of major environmental factors driving phytoplankton community succession before and after the regime shift of Erhai Lake, China

Constructed wetlands are an efficient and cost-effective method of restoring degraded wetlands, in which the microorganisms present make a significant contribution to the ecosystem. In this study, we comprehensively investigated the patterns of diversity and assembly processes of 7 types of constructed wetlands at the rhizosphere and phyllosphere levels. The results showed that the rhizosphere communities of the constructed wetlands exhibited a more balanced structure than that of paddy fields, and 5 types of constructed wetland demonstrated higher potential diversity than that of paddy fields. However, the opposite trend was observed for the phyllosphere communities. Analysis of mean nearest taxon difference indicated that both deterministic and stochastic processes affected the establishment of the rhizosphere and phyllosphere communities, and stochastic processes may have had a larger effect. An iCAMP model showed that dispersal limitation was the most important factor (67% relative contribution) in the rhizosphere community, while drift was the most important (47% relative contribution) in the phyllosphere community. Mantel tests suggested that sucrase, average height, top height, total biomass, belowground biomass, maximum water-holding capacity, and capillary porosity were significantly correlated with processes in the rhizosphere community, whereas factors such as the deterministic process, average height, top height, and SOC were significantly correlated with deterministic processes in the phyllosphere community. Our results can assist in the evaluation of artificial restorations, and can provide understanding of the ecological processes of microbial communities, as well as new insights into the manipulation of microorganisms in polluted wetland ecosystems.

Plant pathogens such as rust fungi (Pucciniales) are of global economic and ecological importance. This means there is a critical need to reliably and cost‐effectively detect, identify, and monitor these fungi at large scales. We investigated and analyzed the causes of differences between next‐generation sequencing (NGS) metabarcoding approaches and traditional DNA cloning in the detection and quantification of recognized species of rust fungi from environmental samples. We found significant differences between observed and expected numbers of shared rust fungal operational taxonomic units (OTUs) among different methods. However, there was no significant difference in relative abundance of OTUs that all methods were capable of detecting. Differences among the methods were mainly driven by the method's ability to detect specific OTUs, likely caused by mismatches with the NGS metabarcoding primers to some Puccinia species. Furthermore, detection ability did not seem to be influenced by differences in sequence lengths among methods, the most appropriate bioinformatic pipeline used for each method, or the ability to detect rare species. Our findings are important to future metabarcoding studies, because they highlight the main sources of difference among methods, and rule out several mechanisms that could drive these differences. Furthermore, strong congruity among three fundamentally different and independent methods demonstrates the promising potential of NGS metabarcoding for tracking important taxa such as rust fungi from within larger NGS metabarcoding communities. Our results support the use of NGS metabarcoding for the large‐scale detection and quantification of rust fungi, but not for confirming the absence of species.

Cyanobacteria act as primary producers of carbon and nitrogen in nutrient-poor ecosystems such as mangroves. This important group of microorganisms plays a critical role in sustaining the productivity of mangrove ecosystems, but the structure and function of cyanobacteria assemblages can be perturbed by anthropogenic influences. The aim of this work was to assess the community structure and ecological drivers that influence the cyanobacterial community harboured in two Brazilian mangrove soils, and examine the long-term effects of oil contamination on these keystone species. Community fingerprinting results showed that, although cyanobacterial communities are distinct between the two mangroves, the structure and diversity of the assemblages exhibit similar responses to environmental gradients. In each ecosystem, cyanobacteria occupying near-shore areas were similar in composition, indicating importance of marine influences for structuring the community. Analysis of 16S rRNA sequences revealed the presence of diverse cyanobacterial communities in mangrove sediments, with clear differences among mangrove habitats along a transect from shore to forest. While near-shore sites in both mangroves were mainly occupied by Prochlorococcus and Synechococcus genera, sequences retrieved from other mangrove niches were mainly affiliated with uncultured cyanobacterial 16S rRNA. The most intriguing finding was the large number of potentially novel cyanobacteria 16S rRNA sequences obtained from a previously oil-contaminated site. The abundance of cyanobacterial 16S rRNA sequences observed in sites with a history of oil contamination was significantly lower than in the unimpacted areas. This study emphasized the role of environmental drivers in determining the structure of cyanobacterial communities in mangrove soils, and suggests that anthropogenic impacts may also act as ecological filters that select cyanobacterial taxa. These results are an important contribution to our understanding of the composition and relative abundance of previously poorly described cyanobacterial assemblages in mangrove ecosystems.

Antioxidant systems are vital in life activities of macrophytes. Species with different life forms need to cope with distinct environments by modifying physiological characters, especially antioxidant systems. In order to find differences among life forms and consequence of lake eutrophication, we studied three antioxidant enzymes activity (superoxide dismutase SOD, ascorbate oxidase APX and catalase CAT) and total soluble phenolics (TP) content in leaves of 26 macrophyte species in September 2013 in Lake Erhai, China. We found that antioxidation varied accordingly with life forms. The activities of SOD and APX in emergent macrophytes (EM) and floating-leaved macrophytes (FM) were much lower than those of submerged macrophytes (SM). On the contrary, TP content was much higher in EM and FM species. There was a negative correlation between TP and antioxidant enzyme activities (CAT and APX). The results suggested that EM and FM species rely on phenolics might to adapt to adverse environments (higher herbivores predation pressure and UV radiation intensity), while SM species more rely on antioxidant enzymes possibly due to lower demand for antioxidation and/or lack of light and inorganic C availability for phenolics synthesis. We also found FM species represent highest fitness in term of antioxidant system, which would lead to overgrowth of FM species and littoral zone bogginess during lake eutrophication. Finally, it is necessary to carry out the verification experiment under the control condition in the later stage, especially for the dominant ones in eutrophic lakes, to understand the exact adaptive mechanisms of them.

The relationship between diversity and productivity from a three-dimensional space view in a natural mesotrophic lake

湖泊和水库是我国的主要饮用水源地，但大都处于富营养化状态，蓝藻水华频发，已成为影响社会稳定、制约区域社会经济可持续发展的重大问题.蓝藻水华的整体应对策略，已经从被动应急转变为主动防御；但要做到主动防御，必须有现代化全方位的蓝藻灾害监测监控手段和体系，及时掌握蓝藻水华及其衍生灾害现状和动向，在未发生或者刚发生时及时觉察.本文针对富营养化湖库富营养化引起的蓝藻水华应急监控问题，按照“整个湖体、重点区域、关键位置”3个监测层次，利用中高分辨率卫星、无人机、岸基/平台视频、自动浮标、人工巡测等技术或手段，围绕“现状掌握、异常报警、原因追溯”的建设目标，提出了天-空-地一体化监控系统建设框架，明确了不同手段的协同方式和业务流程，并正在巢湖进行应用实践.相信本文提出的系统架构不仅在巢湖，在更多的富营养化湖库都具有重要的应用和推广价值.;Lakes and reservoirs are the main sources of drinking water in China, but most of them are in the state of eutrophication with frequent cyanobacterial blooms. This has become major issues affecting social sustainability and restricting the local development. The overall response strategy of cyanobacterial blooms must have modern and comprehensive monitoring methods and systems, so as to timely grasp the status and trends of such blooms and their derived disasters. This article aims at the emergency monitoring of cyanobacterial blooms in eutrophic lakes and reservoirs. According to the three monitoring levels of “the entire lake, key areas, and key locations”, the current satellite, drone, and shore-based /platform video, buoy, manual inspection and other technologies or methods have proposed the construction goals, system framework, and workflow. These are also applied in the Lake Chaohu. It is believed that this framework of monitoring system proposed in this paper has important application and promotion value not only in Lake Chaohu, but also in more eutrophic lakes and reservoirs.

The floodplain ecosystem plays an important role in the mediation or regulation of regional ecological functions, and is the heart of ecosystem functions based on soil microbial communities. In the present study, we investigate the patterns and assembly processes of soil microbial communities along nine profile depths (up to 280 cm in depth) of three stages of floodplain community succession (channel bar, newly deposited river beach and tender floodplain) and farmland in the floodplain of the lower Yellow River. Generally, the bacterial community in agricultural soil had the highest diversity, whereas the archaeal community in the tender floodplain had the highest diversity. Compared to bacteria, the co‐occurrence network of archaea was more complex and stable and had high connectivity. Our data suggest that the resistance of bacteria and archaea to environmental changes in deep soil was higher when compared to that in surface soil. Furthermore, stochastic and deterministic processes acted concurrently to regulate the assembly of the soil microbial community in the floodplain ecosystems. However, the relative contributions of the stochastic process and deterministic process are different between the bacterial community and the archaeal community. The lowest contribution of the stochastic process in the bacterial community was in the subsoil, whereas in the archaeal community, it was the opposite. For ecological selection, bacterial communities in different soil profiles, in the succession stages and in the farmland, were mainly selected through homogeneous selection, whereas archaeal communities were selected through heterogeneous selection. Compared with the three floodplain succession stages, the microbial communities of agricultural lands were more assembled by deterministic processes.Highlights Stochastic and deterministic processes act concurrently to regulate the assembly of soil microbial community in the floodplain ecosystems. The assembly of the bacterial community was mainly affected by the homogeneous selection. The archaeal community was mainly affected by the heterogeneous selection. The microbial communities of agricultural lands were more assembled by deterministic processes.

Multi-proxy approaches to investigate cyanobacteria invasion from a eutrophic lake into the circumjacent groundwater

In the present study, the effects of chemical fertilizer (CF) and organic fertilizer plus chemical fertilizer application (OF–CF) on natural rubber yield, soil properties, and soil bacterial community were systematically investigated in rubber plantations. The rubber dry yield was 26.3% more in the OF treatment group than in the CF treatment group. The contents of total nitrogen (TN), available nitrogen (AN), available phosphorus (AP), and available potassium (AK) as well as soil organic matter (SOM) and pH value were higher following OF–CF treatment. Using Illumina sequencing, a total of 927 operational taxonomic units (OTUs) were obtained following CF treatment, while 955 OTUs were obtained after OF–CF treatment. Relative abundance analysis showed the relative abundances of four phyla (Acidobacteria, Proteobacteria, Actinobacteria, Gemmatimonadetes) were different between the two treatment groups. Correlation analysis revealed Acidobacteria, Bacteroidetes, Thaumarchaeota, Elusimicrobia, Verrucomicrobia were the key taxa that determined the soil properties. Additionally, five OTUs (OTU_506, OTU_391, OTU_189, OTU_278, OTU_1057) were thought to be related to the biodegradation of natural rubber. Taken together, these results improve our understanding of the OF-mediated improvement in soil fertility and contribute to the identification of rubber-degrading bacteria in rubber plantations.