16S rDNA High-throughput Sequencing Research Articles

Hydrocarbons removal and microbial community succession in petroleum-contaminated soil under hydrogen peroxide treatment.

Chemical oxidation as a pretreatment step coupled with bioremediation for petroleum-contaminated soil may pose serious impacts on indigenous microorganisms and the available nutrients. Petroleum-contaminated soil were treated by hydrogen peroxide (H2O2) at initial concentrations of 105mM (HH), 21mM (HL), and 105mM in three equal amounts (HT) without adding any external catalyst. The contents of total petroleum hydrocarbons (TPH) and dissolved nutrients (total organic compounds, nitrogen, and phosphate), and the indigenous bacteria community succession (analyzed by high-throughput sequencing of 16S rDNA) were investigated over 50days. Compared to the control treatment without H2O2 addition, H2O2 treatments for the petroleum-contaminated soil significantly promoted the TPH removal especially in the first 4days and impacted the contents of dissolved nutrients. Both of chemical oxidation and nutrients contributed to microbial community structure changes in alpha diversity. Although the soil microbial community structure had undergone significant changes after different chemical oxidation pretreatments, Firmicutes, Proteobacteria, Gemmatimonadetes, and Actinobacteria were the main bacterial phyla. Compared with adding H2O2 at one time, H2O2 added in stepwise was beneficial to indigenous bacterial diversity recovery and TPH removal. H2O2 oxidation treatments showed a great influence on the microbial community structures in the start-up stage, while recovery time rather than the oxidation treatments presented greater effects on the composition of the microbial community structure with the incubation time extended. Therefore, adding H2O2 as pretreatment for petroleum-contaminated soil showed little effect on the structure of soil indigenous microbial community from a long-term scale, and was conducive to the continuous removal of TPH by indigenous microorganisms.

Antibiotic-induced alternations in gut microflora are associated with the suppression of immune-related pathways in grass carp (Ctenopharyngodon idellus).

Gut microbiota play a vital role in fish health homeostasis. Antibiotics are known to alter microbial community composition and diversity; however, the substantial effects of antibiotics upon the gut microbiome with respect to immune-related pathways in healthy fish remain unclear. Accordingly, here we explored the impact of two antibiotics on the intestinal health, immune response, microbiome dynamics, and transcriptome profiles of grass carp. A two-week feeding trial was carried out in which the basal diet was complemented with enrofloxacin (10 mg/kg) or florfenicol (10 mg/kg). The results showed that: (1) Enrofloxacin and florfenicol both induced intestinal oxidative stress and reduced the digestive enzyme activity of grass carp. (2) High-throughput sequencing of 16S rDNA revealed that enrofloxacin but not the florfenicol treatment influenced gut microbiota diversity in grass carp by shifting α/β-diversity with more abundant pathogens detected. (3) Transcriptome profiling demonstrated that florfenicol down-regulated the immune-related pathways of grass carp, and the network analysis revealed that IgA was negatively correlated with certain pathogens, such as Shewanella and Aeromonas. (4) Antibiotic-induced alternations of gut core microbes were revealed via immune-related transcripts, as were lower mRNA expression levels of mucosal-related genes. (5) Apoptosis and histopathological changes were detected in the enrofloxacin- and florfenicol-treated groups compared with the control group. Overall, administering antibiotics will promote oxidative stress, cause intestinal flora dysbiosis, inhibit the mucosal immune system, and induce apoptosis in grass carp.

Microbiome analysis reveals microecological advantages of emerging ditchless rice-crayfish co-culture mode

Ditchless rice-crayfish co-culture is an emerging model of rice-crayfish farming that circumvents the potential hazards of digging ditches in traditional rice-crayfish farming. However, due to the complex interactions among crayfish, ambient microbiota, and environmental variables, it is necessary to assess the differences in bacterial structure between ditchless and traditional rice-crayfish culture. In this study, the crayfish culture area in the Sichuan basin was selected as the study area, and the bacterial communities of two rice-crayfish culture systems were compared by high-throughput sequencing of 16S rDNA. The results showed that the ditchless system had lower water depth, higher dissolved oxygen, lower total ammonia nitrogen and lower morbidity. There are intuitive differences in the composition of environmental bacterial communities due to environmental changes, even if they are similar in composition at the phylum level. Microbiota in sediments from ditchless systems appear to produce less ammonia nitrogen. The abundance of the pathogens colonizing the intestine of ditchless crayfish was lower than ditched one, and the composition was similar to water. Ditch-farmed crayfish appear to be more susceptible to environmental microbes and have a more fragile intestinal structure. Water depth and dissolved oxygen are the main environmental factors that determine the distribution of microbiota. This study is the first to investigate the bacterial ecology of a ditchless rice- crayfish farming system. The results show that the ditchless rice-crayfish culture model has a more superior bacterial system than the traditional rice-crayfish culture.

A Correlation Between Pericarpium Citri Reticulatae Volatile Components and the Change of the Coexisting Microbial Population Structure Caused by Environmental Factors During Aging.

Pericarpium Citri Reticulatae (PCR) is a natural citrus by-product with beneficial health and nutritive properties that is used widely in food and is an ingredient in traditional Chinese medicine. PCR improves gradually with aging. However, the present research has not yet revealed the reasons for this. Some data prove the important role of microorganisms in the quality of tobacco and fermented tea with the time of the aging of these foods. Our studies further proved that the coexisting Aspergillus niger plays an important role in the change of flavonoids and volatile oil in PCR during this process. Therefore, we put forward that longer storage is better for PCR and is highly correlated with the change of the coexisting microbial population structure caused by environmental factors. Samples of PCR aged in Beijing, Sichuan, Guangdong, and Yunnan were collected at different time points. Using GC/MS and high throughput 16S rDNA and ITS sequencing techniques, massive changes in volatile profile and microbial communities were observed during aging. Spearman correlation analysis indicated that Exobasidium, Xeromyces, Pseudocercospora, Russula, Aspergillus, Herbaspirillum, Sphingomonas, and Streptococcus, which are the dominant microbial genera in Sichuan and Guangdong showed strong connections with volatile components of chemical markers. It was preliminarily verified that the changes of volatile components for PCR are highly correlated with the change of the coexisting microbial population structure caused by environmental factors, providing a new idea for the research on the aging mechanism of PCR and key influencing factors of aging quality.

Using 16S rDNA Sequencing Technology to Preliminarily Analyze Intestinal Flora in Children with Mycoplasma pneumoniae Pneumonia

ObjectiveWe investigated changes in the intestinal flora of children with Mycoplasma pneumoniae pneumonia (MPP). MethodsBetween September 2019 and November 2019, stool samples from 14 children with MPP from The Fourth Hospital of Baotou city, Inner Mongolia Autonomous Region, were collected and divided into general treatment (AF) and probiotic (AFY) groups, according to the treatment of “combined Bifidobacterium, Lactobacillus, Enterococcus, and Bacillus cereus tablets live”. High-throughput 16S rDNA sequencing was used to identify intestinal flora. ResultsIntestinal flora abundance and diversity in children with MPP were decreased. Both Shannon and Simpson indices were lower in the AF group when compared with healthy controls (P < 0.05). When compared with healthy controls, the proportion of Enterorhabdus was lower in the AF group, while the proportion of Lachnoclostridium was higher (P < 0.05). The proportion of Bifidobacteria and Akkermansia was lower in the AFY group but Enterococcus, Lachnoclostridium, Roseburia, and Erysipelatoclostridium proportions were higher. The proportion of Escherichia coli–Shigella in the AFY group after treatment was decreased (P < 0.05). ConclusionsThe intestinal flora of children with MPP is disturbed, manifested as decreased abundance and diversity, and decreased Bifidobacteria. Our probiotic mixture partly improved intestinal flora disorders.

Impact of organic soil amendments in antibiotic levels, antibiotic resistance gene loads, and microbiome composition in corn fields and crops

The potential spreading of antibiotic resistance genes (ARG) into agricultural fields and crops represent a fundamental limitation on the use of organic fertilization in food production systems. We present here a study of the effect of spreading four types of organic soil amendments (raw pig slurry, liquid and solid fractions, and a digested derivative) on demonstrative plots in two consecutive productive cycles of corn harvest (Zea mays), using a mineral fertilizer as a control, following the application of organic amendments at 32–62 T per ha (150 kg total N/ha) and allowing 5–8 months between fertilization and harvest. A combination of qPCR and high-throughput 16S rDNA sequencing methods showed a small, but significant impact of the fertilizers in both ARG loads and microbiomes in soil samples, particularly after the second harvesting cycle. The slurry solid fraction showed the largest impact on both ARG loads and microbiome variation, whereas its digestion derivatives showed a much smaller impact. Soil samples with the highest ARG loads also presented increased levels of tetracyclines, indicating a potential dual hazard by ARG and antibiotic residues linked to some organic amendments. Unlike soils, no accumulation of ARG or antibiotics was observed in corn leaves (used as fodder) or grains, and no grain sample reached detection limits for neither parameter. These results support the use of organic soil amendments in corn crops, while proposing the reduction of the loads of ARGs and antibiotics from the fertilizers to greatly reduce their potential risk.

Subchronic Toxicity of Microcystin-LR on Young Frogs (Xenopus laevis) and Their Gut Microbiota.

Although toxic effects of microcystins (MCs) in mammals and fish have been extensively studied, the effects of MCs on the immune system and gut microbiota of amphibians have not received sufficient attention. As MCs cause general damage to the vertebrate liver and immune system and trigger an inflammatory response, and the gut microbiota is closely related to host metabolism and immunity, we speculated that MCs can cause changes in the immune system and gut microbiota of amphibians. To verify this, we examined the intestinal and liver injury of Xenopus laevis exposed to different microcystin-leucine-arginine (MC-LR) concentrations and the effects on the gut microbiota through high-throughput sequencing of 16S rDNA of the gut microbiota combined with histopathological analysis, enzyme activity determination, and qRT-PCR. Our results showed that MC-LR caused focal infiltration of inflammatory cells and increased the number of T cells and local congestion and vacuolization in X. laevis liver, but reduced the number, density, height, and regularity of villi. These liver and intestinal injuries became more obvious with an increase in MC-LR concentration. MC-LR significantly decreased the activities of malondialdehyde and alkaline phosphatase and the expression of TGF-β in the liver. Moreover, MC-LR significantly altered the gut microbiota of X. laevis. The relative abundance of Firmicutes and Bacteroidetes in high-concentration MC-LR groups was significantly reduced compared to that in low-concentration MC-LR groups, whereas Fusobacteria was significantly enriched. The metabolic gene composition of the gut microbiota in low-concentration MC-LR (≤5 μg/L) groups was significantly different from that in high-concentration MC-LR (≥20 μg/L) groups. These results deepen our understanding of the toxicity of MCs to aquatic organisms and assessment of the ecological risk of MCs in amphibians.

The impact of dromedary camel milk on mice gut microbiota

The gut microbiota plays an important role in the health and disease resistance of the host. Host health depends on the microbiota of the gastrointestinal tract, and imbalance in its composition may leads to certain diseases. This study analyzed the influence that dromedary camel milk has on the gut microbiota of mice. This study characterized the bacterial populations of untreated [Control (C) and camel milk-treated Raw (R), Pasteurized (P), and Fermented (F)] C57BL/6 J mice feces using high-throughput 16S rDNA sequencing on the Illumina MiSeq platform. In total, 286,606 tags were generated, with an average of 71,651 tags being generated per group, and these tags were clustered to the operational taxonomic units (OTUs) at 97% sequence similarity, resulting 1090 OTUs. Significant weight gain was observed among all of the groups, and the total cholesterol level declined in F group followed by in group P compared to group C. The F and P groups demonstrated a correlation between the beneficial microbiota structures that corresponded with lower cholesterol levels than those observed in the other groups. The major dominant bacteria correspond to the phyla Bacteroidetes and Firmicutes. The dromedary camel milk propagated the beneficial bacteria (Allobaculum and Akkermansia) and reduced harmful bacteria such as Proteobacteria, Erysipelotrichaceae, and Desulfovibrionaceae. This study provides a comparative analysis of the gut microbiota of mice based on camel milk, which may be helpful in understanding host health and diverse gut microbial conditions.

Profiling of the Conjunctival Bacterial Microbiota Reveals the Feasibility of Utilizing a Microbiome-Based Machine Learning Model to Differentially Diagnose Microbial Keratitis and the Core Components of the Conjunctival Bacterial Interaction Network.

Both healthy and diseased human ocular surfaces possess their own microbiota. If allowed, opportunistic pathogens within the ocular microbiota may cause microbial keratitis (MK). However, the nonpathogenic component of the ocular microbiota has been proven to undermine the performance of culture, the gold standard of the etiological diagnosis for MK. As the conjunctival bacterial microbiota generates unique alterations with various oculopathies, this study aimed to evaluate the feasibility of distinguishing MK using machine learning based on the characteristics of the conjunctival bacterial microbiome associated with various types of MK. This study also aimed to reveal which bacterial genera constitute the core of the interaction network of the conjunctival bacterial microbiome. Conjunctival swabs collected from the diseased eyes of MK patients and the randomly chosen normal eyes of healthy volunteers were subjected for high-throughput 16S rDNA sequencing. The relative content of each bacterial genus and the composition of bacterial gene functions in every sample were used to establish identification models with the random forest algorithm. Tenfold cross validation was adopted. Accuracy was 96.25% using the bacterial microbiota structure and 93.75% using the bacterial gene functional composition. Therefore, machine learning with the conjunctival bacterial microbiome characteristics might be used for differentiation of MKs as a noninvasive supplementary approach. In addition, this study found that Actinobacteria, Lactobacillus, Clostridium, Helicobacter, and Sphingomonas constitute the core of the interaction network of the conjunctival bacterial microbiome.

Observation of the cervical microbiome in the progression of cervical intraepithelial neoplasia

ObjectiveCervical microbial community in the cervical intraepithelial neoplasia and cervical cancer patients was analysed to study its composition, diversity and signalling pathways by high-throughput 16S rDNA sequencing,and the candidate genes associated with occurrence and progression of cervical intraepithelial neoplasia were screened out and the model was established to predict the evolution of cervical intraepithelial neoplasia malignant transformation from the cervical microbial genes aspect.MethodsCervical tissues of normal, cervical intraepithelial neoplasia and cervical cancer patients without receiving any treatment were collected. The correlation between candidate genes and cervical intraepithelial neoplasia progression was initially determined by analyzing the microbial flora. Real-time fluorescence quantitative PCR was used to detect the expression of candidate genes in different cervical tissues, ROC curve and logistic regression was used to analyse and predict the risk factors related to the occurrence and progression of cervical intraepithelial neoplasia. Finally, the early warning model of cervical intraepithelial neoplasia occurrence and progression is established.ResultsCervical tissues from normal, cervical intraepithelial neoplasia and cervical cancer patients were collected for microbial community high-throughput 16S rDNA sequencing. The analysis revealed five different pathways related to cervical intraepithelial neoplasia. 10 candidate genes were selected by further bioinformatics analysis and preliminary screening. Real time PCR, ROC curve and Logistic regression analysis showed that human papillomavirus infection, TCT severity, ABCG2, TDG, PCNA were independent risk factors for cervical intraepithelial neoplasia. We used these indicators to establish a random forest model. Seven models were built through different combinations. The model 4 (ABCG2 + PCNA + TDG) was the best early warning model for the occurrence and progression of CIN.ConclusionsA total of 5 differential pathways and 10 candidate genes related to occurrence and progression of cervical intraepithelial neoplasia were found in cervical microbial community. This study firstly identified the genes from cervical microbial community that play an important role in the occurrence and progression of cervical intraepithelial neoplasia. At the same time, the early warning model including ABCG2 + PCNA+TDG genes provided a new idea and target for clinical prediction and blocking the evolution of cervical intraepithelial neoplasia malignant transformation from the aspect of cervical microbiological related genes.

Impairment of the gut health in Danio rerio exposed to triclocarban

Triclocarban (TCC) is the principal component in personal and health care products because it is a highly effective, broad-spectrum, and safe antibacterial agent. TCC has recently been discovered in aquatic creatures and has been shown to constitute a health danger to aquatic animals. Although several studies have looked into the toxicological effects of TCC on a variety of aquatic animals from algae to fish, the possible gut-toxicity molecular pathway in zebrafish has never been thoroughly explored. We investigated the gut-toxic effects of TCC on zebrafish by exposing them to different TCC concentrations (100 and 1000 μg/L) for 21 days. We discovered for the first time that the MAPK and TLR signaling pathways related to gut diseases were significantly altered, and inflammation (up-regulation of TNF-α, IL-6, and IL-1β) caused by TCC was confirmed to be largely mediated by the aryl hydrocarbon receptor (AHR) and its related cytokines. This was found using the results of qPCR, a transcriptome analysis, and molecular docking (AHR, AHRR, CYP1A1 and CYP1B1). Furthermore, high-throughput 16S rDNA sequencing demonstrated that TCC exposure reduced the bacterial diversity and changed the gut microbial composition, with the primary phyla Fusobacteria and Proteobacteria, as well as the genera Cetobacterium and Rhodobacteraceae, being the most affected. TCC exposure also caused damage to the gut tissue, including an increase in the number of goblet cells and a reduction in the height of the columnar epithelium and the thickness of the muscular layer, as shown by hematoxylin and eosin staining. Our findings will aid in understanding of the mechanism TCC-induced aquatic toxicity in aquatic species.

Characterization of Conjunctival Sac Microbiome from Patients with Allergic Conjunctivitis.

Conjunctival sac microbiome alterations have been reported to be closely associated with many ocular diseases. However, the characteristic of conjunctival sac microbiome in allergic conjunctivitis (AC) was scarcely described. In this study, we aimed to identify the differences of the conjunctival sac microbiome composition in AC patients compared with normal controls (NCs) using high-throughput 16S rDNA sequencing metagenomic analysis. The conjunctival sac microbiome samples from 28 AC patients and 39 NC patients were collected. The V3-V4 region of 16S rRNA gene high-throughput sequencing was performed on the illumina MiSeq platform. Alpha diversity, beta diversity and the relative abundance at the phylum and genus levels were analyzed using QIIME. Alpha diversity demonstrated by Chao1, Observed_species and PD_whole_tree indexes did not show significant difference between the AC and NC groups, while the Shannon index was higher in the AC group. Beta diversity showed divergent microbiome composition in different groups (p < 0.005). The top five abundant phyla were Firmicutes, Proteobacteria, Actinobacteriota, Bacteroidota and Cyanobacteria in both groups. The top five abundant genera were Bacillus, Staphylococcus, Corynebacterium, Acinetobacter and Ralstonia in the AC group and Acinetobacter, Staphylococcus, Bacillus, Clostridium_sensu_stricto_1, Corynebacterium and Geobacillus in the NC group. The Firmicutes/Bacteroidetes (F/B) ratio at the phylum level was similar between groups (p = 0.144). The Bacillus/Acinetobacter (B/A) ratio at the genus level was higher in the AC group (p = 0.021). The dysbiosis detected in this study might provide further evidence to investigate the mechanism and treatment methods for allergic conjunctivitis.

Methane production and microbial community acclimation of five manure inocula during psychrophilic anaerobic digestion of swine manure

For small-scale farms, the development of rustic and cheap psychrophilic anaerobic digestion systems appears as an opportunity to treat manure, mitigate gaseous emissions and promote decentralized renewable energy production. However, the development of such processes is limited by our understanding of their start-up. In this research, we tested the ability of one mesophilic digestate and four different manure to be used as inoculum for the start-up of psychrophilic anaerobic digestion of swine slurry at 13 °C. The most efficient inoculum appeared to be a swine manure that had been stored for 2 months in a pit. After 9 months of acclimation, the corresponding reactor produced a maximum methane yield of 42L of CH4/kg Volatile Solidesubstrate/day and a CH4 volume of 125L of CH4/kg-Chemical Oxygen Demandadded. The maximum methane production expressed at 13 °C was between 55% and 68% of that obtained at 37 °C. Monitoring of the microbial community dynamics by high throughput 16S rDNA sequencing showed the smooth adaptation of manure microbial species, underlining the transient dominance of the acetogen syntroph candidatus Cloacimonas during acclimation and the enrichment in the Methanosaeta and Methanosarcina methanogens for an efficient methane production.

Seasonal succession of bacterial communities in cultured Caulerpa lentillifera detected by high-throughput sequencing.

An increasing number of microorganisms are being identified as pathogens for diseases in macroalgae, but the species composition of bacteria related to Caulerpa lentillifera, fresh edible green macroalgae worldwide, remains largely unclear. The bacterial communities associated with C. lentillifera were investigated by high-throughput 16S rDNA sequencing, and the bacterial diversities in washed and control groups were compared in this study. A total of 4,388 operational taxonomic units were obtained from all the samples, and the predominant prokaryotic phyla were Proteobacteria, Bacteroidetes, Planctomycetes, Cyanobacteria, Actinobacteria, Verrucomicrobia, Chloroflexi, and Acidobacteria in C. lentillifera. The bacterial diversity changed with seasons and showed an increasing trend of diversity with the rising temperature in C. lentillifera. There were slight reductions in the abundance and diversity of bacteria after washing with tap water for 2 h, indicating that only parts of the bacterial groups could be washed out, and hidden dangers in C. lentillifera still exist. Although the reduction in the abundance of some bacteria revealed a positive significance of washing C. lentillifera with tap water on food safety, more effective cleaning methods still need to be explored.

Study of Rhizosphere Microbial Community Structures of Asian Wild and Cultivated Rice Showed That Cultivated Rice Had Decreased and Enriched Some Functional Microorganisms in the Process of Domestication

Asian cultivated rice (Oryza sativa L.), domesticated from Asian wild rice, is a staple food crop for populations around the world. Asian cultivated rice has undergone physiological changes in the process of its evolution from Asian wild rice, and the closely related rhizosphere microorganisms may have changed in the process of plant domestication. However, the rhizosphere microorganisms of different Asian wild rice species and their related indica and japonica cultivated rice have not yet been illustrated clearly. This study aimed to illustrate the microbial community structures in the rhizosphere of Asian wild rice (common wild rice, nivara wild rice, medicinal wild rice, and spotted wild rice) and Asian cultivated rice (indica and japonica accessions) through the high-throughput sequencing of 16S rDNA, ITS amplifiers and metagenomic data. The results showed that there were significant differences between wild and cultivated rice in their rhizosphere microbial community structures. In view of the indica and japonica rice, the bacterial and fungal community structures of indica rice with the nivara wild rice and medicinal wild rice were more similar than the japonica rice species. The indica and japonica rice had the lowest proportion of Actinobacteria than the wild rice species, and indica rice has the highest relative abundance of Nitrospira. As for the microbial functions, methane metabolism and pyruvate metabolism were found to be the common pathway enriched in the rhizosphere of common and nivara wild rice in comparison with the indica and japonica rice; in addition, though it was found that the relative abundances of the pathogenic fungi in the rhizosphere soil of indica and japonica rice were significantly lower than that of the wild rice, the relative abundances of Magnaporthales and Ustilaginales were significantly higher in indica and japonica rice than that of the wild rice. This study is expected to provide a theoretical basis for the development and utilization of rhizosphere microbial resources for wild and cultivated rice.

Regional discrepancy of microbial community structure in activated sludge system from Chinese WWTPs based on high-throughput 16S rDNA sequencing

Overall understanding of microbial community structure in activated sludge (AS) system at regional level is of great significance for operation regulation of wastewater treatment plants (WWTPs). In this study, 110 AS samples from 21 cities in different Chinese regions were analyzed based on high-throughput 16S ribosomal DNA (rDNA) sequencing to explore effects of different regions on microbial community structure. Results showed that the regions with different characteristics (south and north, coastal and inland, high GDP and low GDP) had great impact on AS bacterial community in China. Core bacterial communities (101 OTUs) in south China were more abundant than those in north China (49 OTUs), and many core species in south China were associated with nutrient removal. Coastal WWTPs possessed unique bacterial communities due to the influence of marine bacteria. Phyla Chloroflexi and Acidobacteria were observed to be main biomarkers in coastal WWTPs. Compared with low GDP regions, more diverse microbial community and effective wastewater treatment were discovered in high GDP regions, and environmental factor analysis suggested that they were mainly correlated with high capacity and influent TP in the WWTPs. β nearest taxon index (βNTI) analysis showed that microbial community assembly in the analyzed AS samples was dominated by deterministic factors (70.67%) and influent quality was observed to be main factor.

Reduced production of the major allergens Bla g 1 and Bla g 2 in Blattella germanica after antibiotic treatment.

Allergens present in the feces or frass of cockroaches can cause allergic sensitization in humans. The use of fecal and frass extracts for immunotherapy has been previously investigated but has not yet been fully standardized. Here, we treated cockroaches with ampicillin to produce extracts with reduced amounts of total bacteria. We performed targeted high-throughput sequencing of 16S rDNA to compare the microbiomes of ampicillin-treated and untreated (control) cockroaches. RNA-seq was performed to identify differentially expressed genes (DEGs) in ampicillin-treated cockroaches. Analysis of the microbiome revealed that alpha diversity was lower in the ampicillin-treated group than in the control group. Beta diversity analysis indicated that ampicillin treatment altered bacterial composition in the microbiome of cockroaches. Quantitative polymerase chain reaction revealed that almost all bacteria were removed from ampicillin-treated cockroaches. RNA-seq analysis revealed 1,236 DEGs in ampicillin-treated cockroaches (compared to untreated cockroaches). Unlike bacterial composition, the DEGs varied between the two groups. Among major allergens, the expression of Bla g 2 decreased significantly in ampicillin-treated cockroaches (compared to untreated group). In this study, the reduced level of allergens observed in cockroaches may be related to lower amounts of total bacteria caused by treatment with antibiotics. It is possible to make a protein extract with few bacteria for use in immunotherapy.

Auricularia auricula-judae (Bull.) polysaccharides improve type 2 diabetes in HFD/STZ-induced mice by regulating the AKT/AMPK signaling pathways and the gut microbiota.

Auricularia auricula-judae is an edible fungus with high nutritional value due to abundant polysaccharides, and is acknowledged as traditional food and medicine in Asia. Polysaccharides from A. auricula (AAPs) are typically fungal polysaccharides and have a wide range of biological activities. It has been shown the potential of AAPs to improve diabetes as an effective adjuvant, but the underlying mechanism remains unclear. In this study, we explored the effects and potential mechanism of AAPs on type 2 diabetes (T2D) using a high-fat diet and streptozotocin (STZ) induced C57BL/6J mice. The results indicated that 50 and 100mg/kg AAPs significantly decreased inflammation, liver injury, and insulin resistance. In addition, AAPs improved glycolipid metabolism disorders by activating the AKT and adenosine 5`monophosphate-activated protein kinase (AMPK) signaling pathways in T2D mice. Furthermore, we investigated the association between changes of gut microbiota and AAPs effects using high-throughput sequencing of 16S rDNA for fecal samples. In our study, AAPs elevated gut microbiota diversity and optimized microbial composition and function in T2D mice, characterized by increased Lactobacillus and Bacteroides abundance and decreased Clostridium and Allobaculum abundance. Particularly, AAPs intervention mainly affected the amino acid metabolism and glycolipid metabolism pathways. Overall, this study confirms that AAPs can improve type 2 diabetes by regulating the AKT and AMPK pathways and modulating intestinal microbiota. PRACTICAL APPLICATION: The article systematically verified the positive effects of AAPs on insulin resistance, glycolipid metabolism disorder, inflammation, and liver injury, key factors closely related to T2D. Furthermore, our study firstly determined the specific underlying mechanism that AAPs ameliorates T2D through regulating AKT/AMPK pathways and modifying the gut microbiota. These results could offer a full explanation and a potential option for the adjuvant therapy of diabetes with AAPs.

Soil bacterial community as impacted by addition of rice straw and biochar

The application of straw and biochar can effectively improve soil quality, but whether such application impacts paddy soil bacterial community development remains to be clarified. Herein, the impacts of three different field amendment strategies were assessed including control (CK) treatment, rice straw (RS) application (9000 kg ha−1), and biochar (BC) application (3150 kg ha−1). Soil samples were collected at five different stages of rice growth, and the bacterial communities therein were characterized via high-throughput 16S rDNA sequencing. The results of these analyses revealed that soil bacterial communities were dominated by three microbial groups (Chloroflexi, Proteobacteria and Acidobacteria). Compared with the CK samples, Chloroflexi, Actinobacteria, Nitrospirae and Gemmatimonadetes levels were dominated phyla in the RS treatment, and Acidobacteria, Actinobacteria, Nitrospirae and Patescibacteria were dominated phyla in the BC treatment. Compared with the RS samples, Chloroflexi, Acidobacteria, Actinobacteria, and Verrucomicrobia levels were increased, however, Proteobacteria, Gemmatimonadetes, Nitrospirae, and Firmicute levels were decreased in the BC samples. Rhizosphere soil bacterial diversity rose significantly following RS and BC amendment, and principal component analyses confirmed that there were significant differences in soil bacterial community composition among treatment groups when comparing all stages of rice growth other than the ripening stage. Relative to the CK treatment, Gemmatimonadaceae, Sphingomonadaceae, Thiovulaceae, Burkholderiaceae, and Clostridiaceae-1 families were dominant following the RS application, while Thiovulaceae and uncultured-bacterium-o-C0119 were dominant following the BC application. These findings suggest that RS and BC application can improve microbial diversity and richness in paddy rice soil in Northeast China.

Salt tolerance of halotolerant bacteria from coastal soils and sediments near saltern field of Hainan Island, China.

Understanding the salt tolerance of microbial communities may help to elucidate the effects of salt concentration and other environmental factors on soil biodiversity. Here, high-throughput sequencing of 16S rDNA and ITS was combined to investigate the distribution and salt tolerance of microbial communities in coastal soils and sediments near the Yinggehai saltern field of Hainan Island, China. The microbial communities in the soils and sediments of the land zone (YGHLS), the intertidal zone (YGHIS), and the inshore zone (YGHWS) were compared. PCoA of weighted and unweighted UniFrac distance revealed obvious differences in soil microbial community among different samples. ANOSIM analysis could clearly separate the three samples from each other. Three halotolerant bacteria, including Halomonas, Halobacillus and Wallemia, were found in the samples, which accounted for 0.0335 ± 0.0586%, 0.0241 ± 0.0304%, and 0.0308 ± 0.0445% of the total microbial community, respectively. The relative abundance of Trk system potassium uptake protein, Kdp operon response regulator, and Na+/H+ antiporter in the samples exceeded 0.09%, 0.06%, and 0.02%, respectively, indicating that the Trk system plays a major role in the salt tolerance of halotolerant bacteria in Yinggehai coastal soils and sediments.