16S rRNA Sequence Analysis Research Articles

Effects of xylo-oligosaccharide supplementation on the production performance, intestinal morphology, cecal short-chain fatty acid levels, and gut microbiota of laying hens

This study investigated the effects of xylo-oligosaccharide supplementation on the production performance, intestinal morphology, cecal short-chain fatty acid levels, and gut microbiota of laying hens. A total of 800 Lohmann pink layers, each 48 wk old, were randomly divided into 5 dietary treatment groups, namely XOS at 0 (CON), 100 (XOS1), 200 (XOS2), 300 (XOS3) and 400 (X0S4) mg/kg. The experimental period was 24 wk. The results revealed that the egg production rate and the number of eggs laid by each layer between 1 to 12 wk increased as the XOS concentration increased (Plinear < 0.05). The sand-shell egg percentage decreased significantly from 1 to 12 wk in the XOS1, XOS2, and XOS3 groups (PANOVA < 0.05). Compared with the CON group, the 4 XOS dosage groups presented significant increases in the villus height and the ratio of villus height to crypt depth in the jejunum (PANOVA < 0.05), whereas a linear decrease in jejunal crypt depth (Plinear < 0.05) was noted. In addition, XOS supplementation significantly increased the concentrations of butyric acid and isovaleric acid in the caeca (PANOVA < 0.05). High-throughput sequencing analysis of bacterial 16S rRNA revealed that dietary XOS supplementation influenced the cecal microbiota. The alpha diversity analysis indicated that the richness of cecal bacteria was greater in the laying hens fed XOS. The modulation of the cecal microbiota composition upon the addition of XOS was characterized by an increased abundance of Firmicutes and Bifidobacteriaceae, and decreased abundance of Bacteroidetes. At the genus level, dietary XOS supplementation resulted in decreases in the abundances of Bacteroides and Rikenellaceae_RC9_gut_group and an increase in the abundance of Lactobacillus. In conclusion, dietary XOS supplementation improved the production performance of laying hens by increasing the production of short-chain fatty acids and improving their intestinal morphology, which was achieved mainly through changes in the composition of the intestinal microbiota. The recommended level of XOS in the diet of laying hens is 200 mg/kg.

Molecular characterization of ‘Candidatus Phytoplasma australasiaticum’ associated with ornamentals, amaranthus, French bean and sunhemp crops and development of rapid LAMP assay for detection

Phytoplasma is one of the most economically important pathogens, causing significant yield losses in many crops worldwide. During a survey in Bengaluru rural and Chikkabalpura areas of Karnataka, India, a total of 33 symptomatic plant samples including ornamentals, amaranthus, french bean, and sunhemp displaying phytoplasma symptoms were collected and confirmed as infected with phytoplasma through PCR targeting the 16S rRNA and secY genes. Pairwise sequence comparison and phylogenetic analysis of 16S rRNA and secY gene sequences confirmed the presence of ‘Candidatus Phytoplasma australasiaticum’ in antirrhinum, chrysanthemum, china aster, dianthus, amaranthus, french bean, and sunhemp. In-silico restriction fragment length polymorphism (RFLP) analysis indicated that phytoplasma strains infecting Antirrhinum, China aster, Dianthus, French bean, and Sunhemp represented distinct variants within the 16SrII phytoplasma group, with a threshold similarity coefficient of less than 0.98. A loop-mediated isothermal amplification (LAMP) assay was developed for detecting phytoplasma strains infecting the four ornamentals and other studied crops, targeting the phytoplasma 16S rRNA gene sequence with detection in 60 min. The comparative testing of LAMP products using various nucleic acid dyes and a colorimetric dye (hydroxy naphthol blue) confirmed the presence of phytoplasmas in symptomatic samples. This LAMP assay offers superior ease of use, cost-effectiveness, and rapid results for detecting phytoplasmas in symptomatic plants.

PSIX-18 Multiomic analysis to identify host and microbiome contributions to digestibility in beef cattle

Abstract This study evaluated beef heifers selected for high (efficient) or low (inefficient) digestible fiber intake (DFI). Initial analysis showed that high DFI animals had reduced methane production versus low-DFI under a high forage diet. Using the same cohort of animals maintained on a further 4 diets of varying forage:concentrate ratios, we employed multi-kingdom amplicon sequencing and metagenome shotgun sequencing of rumen digesta and feces alongside RNA sequencing of rumen epimural samples to evaluate the compositional and functional interplay between different microbial groups, and their relationship with host gene expression in cattle divergent for DFI. Samples were collected from 16 cattle during 2 metabolism trials, comprising 5 diets. Amplicon sequencing analysis was conducted using QIIME2; 16S rRNA and 18S rRNA reads were analyzed using the SILVA database, while LSU (fungal) sequences were analyzed using a custom D1/D2 database. Additional analysis of archaeal 16S rRNA sequences was conducted using the Rumen and Intestinal Methanogens (RIM) database. Metagenome shotgun reads underwent a two-pass classification with Kraken2 using a database of prokaryotic genomes derived from the GTDB taxonomy, with the unclassified output undergoing classification using a custom database containing all NCBI protozoa, fungi, and phage genomes, enriched with selected rumen-specific ciliate and fungal genomes. Downstream analysis of taxonomic data from all microbiome work was conducted in R, and differentially abundant taxa were identified using ANCOM-BC and Aldex2. Functional analysis of metagenome contigs using the CAZY database implemented in dbCAN3 is ongoing. RNA-seq data were analyzed using the ARS-UCD reference genome, with identification of DE genes conducted using DeSeq2. Preliminary results indicate no major effect of DFI ranking on host gene expression, bacterial 16S rRNA, or metagenome compositional profiles. Several bacterial genera were differentially abundant between digestibility groups (P &amp;lt; 0.05), but these were all minor (&amp;lt; 0.01%) members of the microbiome. Fungal and methanogen communities different significantly (P &amp;lt; 0.05) according to DFI group, with efficient (high DFI) containing and more diverse communities under high-grain diet (P &amp;lt; 0.05). The same difference showed a tendency toward significance for the 18S rRNA protozoa data (P &amp;lt; 0.1). These preliminary data indicate that the microbial factors underpinning divergence in efficiency measured by DFI vary according to diet and may be more prominent in the non-bacterial fraction of the microbiome. Ongoing functional analysis of metagenome data as well as integration of multiomic data will provide deeper insight into these relationships and how they contribute to feed digestibility and efficiency in cattle.

Isolation and identification of Acinetobacter lwoffii from the Chinese Giant Salamander (Andrias davidianus)

This study aimed to identify the primary pathogenic bacteria responsible for the mortality of the Chinese giant salamander (Andriasd davidianus). A pathogenic bacterium was isolated from a diseased Chinese giant salamander exhibiting typical symptoms under aseptic conditions and then identified by morphological examinations, biochemical analysis, and the sequence analysis of bacterial 16S rRNA. Artificial infection testing was then conducted to determine the pathogenicity of the isolated bacteria strain. Drug susceptibility tests were conducted using the agar diffusion method. The isolated pathogenic bacteria, named DN-2, was successfully identified as Acinetobacter lwoffii. The artificial infection showed that the typical symptoms of the disease could be replicated as the ones originally occurred, and this bacterium exhibited high pathogenicity to the Chinese giant salamander. In the Chinese giant salamander, the median lethal dosage (LD50) of A. lwoffii DN-2 for A. davidianus was determined to be 4.63×104 CFU/g. Drug sensitivity testing showed that these bacteria were highly sensitive to erythromycin, gentamicin, neomycin, streptomycin, midecamycin, ciprofloxacin, piperacillin, florfenicol, doxycycline, carbenicillin, and sulfanilamide. In summary, A. lwoffii was identified as the primary pathogen responsible for the demise of A. davidianus. Our study first presents how these bacteria harm Chinese giant salamanders.

RFLP-kenzy: a new bioinformatics tool for in silico detection of key restriction enzyme in RFLP technique

BackgroundToday, several bioinformatics tools are available for analyzing restriction fragment length data. RFLP-kenzy is a new bioinformatic tool for identifying restriction key enzyme that cut at least 1 sequence and a maximum of n-1 sequence.ResultsThis bioinformatic tool helps researchers to select appropriate enzymes that yield different RFLP patterns, especially from overly identical sequences with single nucleotide mutation or other small variations. By using RFLP-kenzy, multiple DNA sequences could be analyzed simultaneously and the key enzymes list is provided. The present paper also demonstrates the ability of RFLP-kenzy to identify the key enzymes through the analysis of 16S rRNA sequences and the complete genome of various genera of microorganisms.ConclusionFrom the results, several key enzymes were provided indicating the importance of this new tool in the selection of appropriate restriction enzymes.

Utilization of 16Sr RNA and secA genes for molecular discernment of 'Candidatus Phytoplasma australasiaticum' strain associated with linseed germplasm in India.

The growing prevalence of phytoplasma associated symptoms on linseed or flax (Linum usitatissimum L.) germplasm at Indian Council of Agricultural Research- National Bureau of Plant Genetic Resources (ICAR-NBPGR) fields was noticed during the 2019-22 growing seasons. The characteristic phytoplasma symptoms of phyllody, stem fasciation, stunting, along with floral and capsule malformations were observed in 41 linseed accessions grown at experimental fields of ICAR-NBPGR, Delhi. During 3years, the presence of phytoplasma in symptomatic linseed accessions was confirmed by nested-PCR assays utilizing 16S rRNA and secA gene-specific primers. The 16S rRNA and secA gene sequences of linseed phytoplasma strains from the representative symptomatic 41 linseed accessions exhibited 100% sequence identity among themselves and 99.93% and 99.82% sequence homology with reference strain, 'Candidatus Phytoplasma australasiaticum' (GenBank Accession: Y10097). Phylogenetic analysis of 16S rRNA and secA gene sequences clustered the linseed isolates with the peanut witches' broom group belonging to 'Ca. P. australasiaticum' strains. The virtual RFLP analysis of 16S rRNA F2nR2 fragment (~1.2kb) of linseed phytoplasma strains further classified it into 16Sr group II, subgroup D. Our results suggested confirmation of the association of 'Ca. P. australasiaticum' strain (16SrII-D) in the linseed germplasm accessions from North India, which is the first report from India. The phytoplasma infection also reduced the growth and yield parameters of two linseed accessions (IC0498748 and EC0718851).

Characterization of Antibiotic Actinomycin D Isolated from Streptomyces parvulus Collected from Marine Sponge of Barrang Lompo Island, Makassar, Indonesia.

Microorganisms associated with sea sponges have proven to be good natural product resources that are biologically active and pharmaceutically important. This research aimed to identify actinomycetes related to a sponge from Bar-ranglompo Island Makassar and the antibacterial compounds. Identification of actinomycetes was based on molecular characterization of sequence gen16S rRNA. The antibacterial compound was separated using vacuum liquid chromatog-raphy and preparative Thin Layer Chromatography (TLC). The structure determination was done based on spectroscopy 1H-NMR, 13C-NMR, 2D NMR, and mass spectra. Molecular characterization showed that actinomycetes strain BLP 20 had the closest relationship with Streptomyces parvulus and Uncultured Streptomyces sp. with a similarity value of 83%. The results obtained from the characterization of antibacterial compounds based on spectroscopic data indicate that these compounds lead to Actinomycin D. Characterization and identification of Strain 20 / BLP by molecular phylogenetic analysis of 16S rRNA sequences revealed the closest relationship with Uncultured Streptomy-ces sp and S. parvulus with a similarity value of 83 %, which indicated a new species. The structure of the active compound isolated from actinomycetes strain 20 / BLP leads to Actino-mycin D.

Distinct changes in tomato-associated multi-kingdom microbiomes during Meloidogyne incognita parasitism

BackgroundThe interplay between root-knot nematode (RKN) parasitism and the complex web of host-associated microbiota has been recognized as pivotal for effective management of the pest. However, studies assessing this relationship have focussed on the bacterial and fungal communities, neglecting the unicellular eukaryotic members. Here, we employed amplicon sequencing analysis of the bacterial 16S rRNA, fungal ITS and eukaryotic 18S rRNA genes, and comprehensively examined how the microbiome composition, diversity and networking developed with time in the rhizospheres and roots of RKN-inoculated and non-inoculated tomato plants.ResultsAs expected, infection with the RKN Meloidogyne incognita decreased plant growth. At individual timepoints, we found distinct bacterial, fungal and eukaryote community structures in the RKN-inoculated and non-inoculated rhizospheres and roots, and RKN inoculation affected several taxa in the root-associated microbiome differentially. Correlation analysis revealed several bacterial and fungal and few protist taxa that correlated negatively or positively with M. incognita. Moreover, network analysis using bacterial, fungal and eukaryotic data revealed more dynamic networks with higher robustness to disturbances in the RKN-inoculated than in the non-inoculated rhizospheres/roots. Hub taxa displayed a noticeable successional pattern that coincided with different phases of M. incognita parasitism. We found that fungal hubs had strong negative correlations with bacteria and eukaryotes, while positive correlations characterized hub members within individual kingdoms.ConclusionOur results reveal dynamic tomato-associated microbiomes that develop along different trajectories in plants suffering M. incognita infestation and non-infested plants. Overall, the results identify stronger associations between RKN and bacterial and fungal taxa than between eukaryotic taxa and RKN, suggesting that fungal and bacterial communities could play a larger role in the regulation of RKN. The study identifies several putative RKN-antagonistic bacterial and fungal taxa and confirms the antagonistic potential previously identified in other taxa.

Effectiveness of vaginal probiotics Lactobacillus crispatus chen-01 in women with high-risk HPV infection: a prospective controlled pilot study.

Female genital tract infection with high-risk human papilloma virus (HR-HPV) has the risk of developing into cervical cancer, and there is still a lack of effective therapeutic strategies. Probiotic intervention is considered as a potential intervention for HR-HPV, while exploration into living probiotic preparations for specific diseases remains limited and insufficient. This prospective controlled pilot study was conducted to observe the effect of intravaginal transplantation of a vaginal isolated natural probiotic strain, Lactobacillus crispatus chen-01, on the clearance of high-risk HPV infection. 100 women with high-risk HPV infection were enrolled and randomly divided into placebo group and probiotic treatment group, which received intravaginal transplantation of L. crispatus chen-01. Cervical exfoliated cells were collected 6 months later for detecting DNA load, typing of HPV, and cytological analysis. Our results showed that vaginal transplantation with L. crispatus chen-01 significantly reduced viral load of HPV, ameliorated HPV clearance rate, and improved vaginal inflammation state without causing obvious adverse reactions. Analysis of 16S rRNA sequencing revealed that L. crispatus chen-01 could effectively reconstitute the vaginal microbiota in women with high-risk HPV, which might be one of the underlying mechanisms of the beneficial effect of L. crispatus chen-01 transplantation. Our results suggested that vaginal transplantation of L. crispatus chen-01 might be a promising treatment for patients with high-risk HPV infection.

Bacillus Paralicheniformis: A potential candidate for natural antibacterial agents from marine macro-algae associated bacteria.

Antibiotic resistance poses a major global health threat, necessitating new approaches to treat bacterial infections. We isolated bacteria from the brown macroalgae Sargassum swartzii collected along the Indian coast. Of 279 isolates, 47.6% showed antibacterial activity against Staphylococcus aureus and Vibrio parahaemolyticus when screened. Ten potent isolates underwent analysis of biosynthetic gene clusters and 16S rRNA sequencing. Isolate MP-99 carried polyketide synthase (pks) and non-ribosomal peptide synthetase (nrps) genes and exhibited antibacterial effects.

Biocontrol potentiality of &lt;i&gt;Burkholderia vietnamiensis&lt;/i&gt; nrv12 against the rice blast fungus &lt;i&gt;magnaporthe oryzae&lt;/i&gt;

Rice blast disease, caused by the pathogenic fungus Magnaporthe oryzae, is a widespread infection leading to serious crop loss worldwide. In order to achieve sustainable agriculture, root-associated bacteria have been applied to manage fungal diseases and promote growth. The present study aimed to evaluate in vitro the growth-promoting ability and in vivo biocontrol activity against M. oryzae of rice rhizosphere bacterium. Out of sixty-eight isolates recovered from the rhizosphere of blast-infected rice plants, isolate NRV12 exhibited the highest antifungal activity against M. oryzae SH, with an inhibition percentage of 72.7±3.44%. By analysis of 16S rRNA sequence associated with morphology, physiological and biochemical tests, the strain was identified as Burkholderia vietnamiensis. In addition, NRV12 produced hydrolytic enzymes (amylase, cellulase, protease), indole acetic acid (IAA), exhibited nitrogen-fixing potential and the ability to solubilize phosphate and zinc. Innoculation with NRV12 significantly promoted in vivo rice seedling growth to 23.3% as compared to the non-bacteria-treated seedlings. Importantly, infected rice seedlings treated with NRV12 led to a 40% disease reduction in rice blast. These findings suggest that NRV12 is a valuable and promising isolate with biocontrol potential against rice blast caused by M. oryzae.

Gut microbiota dysbiosis in hyperuricaemia promotes renal injury through the activation of NLRP3 inflammasome

BackgroundThe prevalence of hyperuricaemia (HUA), a metabolic disorder characterized by elevated levels of uric acid, is on the rise and is frequently associated with renal injury. Gut microbiota and gut-derived uremic toxins are critical mediators in the gut-kidney axis that can cause damage to kidney function. Gut dysbiosis has been implicated in various kidney diseases. However, the role and underlying mechanism of the gut microbiota in HUA-induced renal injury remain unknown.ResultsA HUA rat model was first established by knocking out the uricase (UOX). HUA rats exhibited apparent renal dysfunction, renal tubular injury, fibrosis, NLRP3 inflammasome activation, and impaired intestinal barrier functions. Analysis of 16S rRNA sequencing and functional prediction data revealed an abnormal gut microbiota profile and activation of pathways associated with uremic toxin production. A metabolomic analysis showed evident accumulation of gut-derived uremic toxins in the kidneys of HUA rats. Furthermore, faecal microbiota transplantation (FMT) was performed to confirm the effects of HUA-induced gut dysbiosis on renal injury. Mice recolonized with HUA microbiota exhibited severe renal injury and impaired intestinal barrier functions following renal ischemia/reperfusion (I/R) surgery. Notably, in NLRP3-knockout (NLRP3−/−) I/R mice, the deleterious effects of the HUA microbiota on renal injury and the intestinal barrier were eliminated.ConclusionOur results demonstrate that HUA-induced gut dysbiosis contributes to the development of renal injury, possibly by promoting the production of gut-derived uremic toxins and subsequently activating the NLRP3 inflammasome. Our data suggest a potential therapeutic strategy for the treatment of renal diseases by targeting the gut microbiota and the NLRP3 inflammasome.-txnFZYkVzJbh2g8RkPjrkVideo Graphical

Combined analysis of metabolomics and 16S rRNA sequencing for ankylosing spondylitis patients before and after secukinumab therapy.

Alterations in gut microbiota have been implicated in the pathogenesis of ankylosing spondylitis (AS), but the underlying mechanisms remain elusive. This study aims to investigate changes in gut microbiota and metabolites in individuals with AS before and after treatment with secukinumab, to identify the biological characteristics specific to AS patients and investigate the potential biomarkers, for optimizing therapeutic strategies more effectively. Fecal microbiome data were collected from 30 AS patients before and after secukinumab therapy and compared with data from 40 healthy controls (HC). Additionally, we analyzed the metabolic profile of both groups from plasma. Findings indicated that the treatment-induced changes in the composition of several crucial bacterial groups, including Megamonas, Prevotella_9, Faecalibacterium, Roseburia, Bacteroides, and Agathobacter. Post-treatment, these groups exhibited a distribution more akin to that of the healthy populations compared with their pretreatment status. We identified three gut microbial taxa, namely Prevotellaceae_bacterium_Marseille_P2831, Prevotella_buccae, and Elusimicrobiota, as potential biomarkers for diagnosing individuals at a higher risk of developing AS and assessing disease outcomes. Plasma metabolomics analysis revealed 479 distinct metabolites and highlighted three disrupted metabolic pathways. Integration of microbiome and metabolomics datasets demonstrated a significant degree of correlation, underscoring the impact of the microbiome on metabolic activity. Secukinumab can restore the balance of the gut microbiome and metabolites in AS patients, rendering them more similar to those found in the healthy population. The analysis of microbiome and metabolomics data have unveiled some candidate biomarkers capable of evaluating treatment efficacy.

Faecalibacterium taiwanense sp. nov., isolated from human faeces.

Two Gram-stain-negative, straight rods, non-motile, asporogenous, catalase-negative and obligately anaerobic butyrate-producing strains, HLW78T and CYL33, were isolated from faecal samples of two healthy Taiwanese adults. Phylogenetic analyses of 16S rRNA and DNA mismatch repair protein MutL (mutL) gene sequences revealed that these two novel strains belonged to the genus Faecalibacterium. On the basis of 16S rRNA and mutL gene sequence similarities, the type strains Faecalibacterium butyricigenerans AF52-21T(98.3-98.1 % and 79.0-79.5 % similarity), Faecalibacterium duncaniae A2-165T(97.8-97.9 % and 70.9-80.1 %), Faecalibacterium hattorii APC922/41-1T(97.1-97.3 % and 80.3-80.5 %), Faecalibacterium longum CM04-06T(97.8-98.0% and 78.3 %) and Faecalibacterium prausnitzii ATCC 27768T(97.3-97.4 % and 82.7-82.9 %) were the closest neighbours to the novel strains HLW78T and CYL33. Strains HLW78T and CYL33 had 99.4 % both the 16S rRNA and mutL gene sequence similarities, 97.9 % average nucleotide identity (ANI), 96.3 % average amino acid identity (AAI), and 80.5 % digital DNA-DNA hybridization (dDDH) values, indicating that these two strains are members of the same species. Phylogenomic tree analysis indicated that strains HLW78T and CYL33 formed an independent robust cluster together with F. prausnitzii ATCC 27768T. The ANI, AAI and dDDH values between strain HLW78T and its closest neighbours were below the species delineation thresholds of 77.6-85.1 %, 71.4-85.2 % and 28.3-30.9 %, respectively. The two novel strains could be differentiated from the type strains of their closest Faecalibacterium species based on their cellular fatty acid compositions, which contained C18 : 1 ω7c and lacked C15 : 0 and C17 : 1 ω6c, respectively. Phenotypic, chemotaxonomic and genotypic test results demonstrated that the two novel strains HLW78T and CYL33 represented a single, novel species within the genus Faecalibacterium, for which the name Faecalibacterium taiwanense sp. nov. is proposed. The type strain is HLW78T (=BCRC 81397T=NBRC 116372T).

Petroleum ether extract of Schisandra sphenanthera prevents hyperglycemia and insulin resistance in association with modulation of sweet taste receptors and gut microbiota in T2DM rats

Ethnopharmacological relevanceSchisandra sphenanthera (Schisandra sphenanthera Rehd. et Wils.) is the dried mature fruit of Schisandra sphenanthera, a plant in the Magnoliaceae family. It was used in the treatment of diabetes mellitus in the Jade Fluid Decoction and the Xiaoke pills, which were recorded in ancient books. However, its mechanism of action in the treatment of type 2 diabetes mellitus (T2DM) was unclear and needs further study. Aim of the studyThis research aimed to investigate the chemical composition and lignan content of Schisandra sphenanthera petroleum ether parts (SPEP) and to evaluate the effects of SPEP on sweet taste receptors (STRs) and intestinal flora in rats on a high-fat diet (HFD). Additionally, the relationships between SPEP and hyperglycemia and insulin resistance were examined. Materials and methodsGC-MS was used to determine the chemical composition of SPEP, and HPLC was used to determine the lignin content. A combination of the HFD and the administration of streptozotocin (STZ) was employed to generate a rat model of T2DM. Petroleum ether extracts from Schisandra sphenanthera were used as the focus of the research to evaluate the effects of these extracts on the glucolipid metabolism of T2DM rats, as well as the underlying mechanisms. ResultsAnalysis of the GC-MS spectrum of SESP revealed a total of 58 compounds. HPLC analysis revealed that SPEP had the highest concentration of Schisandrin A and the lowest concentration of Schisandrol A. The drug administration intervention resulted in a significant decrease in body weight and pancreatic weight of diabetic rats compared to the Normal group. When compared to the Model group, the body weight of rats in the drug administration group and the Metformin group had a more moderate decrease, while the pancreatic weight and pancreatic-to-body ratio increased. The Model group shown significant increases in FBG, OGTT, GHb, TC, TG, LDL-C, ALT, AST, MDA, FINS, and NEFA, as well as significant decreases in HDL-C and SOD, when compared to the Normal group (P < 0.05). The administration of each group was found to be significantly effective in decreasing FBG, OGTT, GHb, TC, TG, LDL-C, ALT, AST, MDA, FINS, NEFA, while increasing HDL-C and SOD when compared to the Model group. The application of SPEP had a positive impact on hepatocyte swelling, hepatocyte degeneration, and necrosis, as well as the morphological structure of pancreatic islet cells. Furthermore, the protein expression levels of T1R2, TRPM5 and GLP-1 in the small intestine of the Model group were reduced. After a period of six weeks, the protein expression levels began to align more closely with those of the Normal group of rats. Analysis of 16S rRNA sequencing revealed that the intestinal microbiota of diabetic rats was significantly disrupted, with a decrease in the abundance of the Firmicutes phylum and an increase in the abundance of the Bacteroidetes phylum. Furthermore, the composition of the dominant genus was distinct from that of the control group. After the drug intervention, the microbiota of diabetic rats was significantly altered, exhibiting a higher abundance and diversity, as well as a significant enrichment of the community. The SPEP treatment resulted in a significant increase in acetic acid, propionic acid, and butyric acid. ConclusionsThe findings of this research indicated that SPEP could be effective in treating T2DM through the regulation of STRs, the adjustment of disturbed metabolite levels, and the alteration of intestinal flora.

Berberine alleviates Alzheimer's disease by regulating the gut microenvironment, restoring the gut barrier and brain-gut axis balance

BackgroundAlzheimer's disease (AD) is the most common neurodegenerative disease. Intestinal flora and its metabolism play a significant role in ameliorating central nervous system disorders, including AD, through bidirectional interactions between the gut-brain axis. A naturally occurring alkaloid compound called berberine (BBR) has neuroprotective properties and prevents Aβ-induced microglial activation. Additionally, BBR can suppress the synthesis of Aβ and decrease BACE1 expression. However, it is still unclear if BBR therapy can alleviate AD by changing the gut flora. PurposeIn this study, we examined whether a partial alleviation of AD could be achieved with BBR treatment and the molecular mechanisms involved. MethodsWe did this by analyzing alterations in Aβ plaques, neurons, and related neuroinflammation-related markers in the brain and the transcriptome of the mouse brain. The relationship between the intestinal flora of 5xFAD model mice and BBR treatment was investigated using high-throughput sequencing analysis of 16S rRNA from mouse feces. ResultsThe findings demonstrated that treatment with BBR cleared Aβ plaques, alleviated neuroinflammation, and ameliorated spatial memory dysfunction in AD. BBR significantly alleviated intestinal inflammation, decreased intestinal permeability, and could improve intestinal microbiota composition in 5xFAD mice.

Enhancing biomass and lipid accumulation by a novel microalga for unsterilized piggery biogas slurry remediation.

The cost and efficiency of an algal-BS treatment system are determined by the specific microalgal species and BS pretreatment method. This study examines the growth of a novel algae Chlorella sp. YSD-2 and the removal of nutrients from the BS using different pretreatment methods, including dilution ratio and sterilization. The highest biomass production (1.84 g L-1) was achieved in the 1:2 unsterilized biogas slurry, which was 2.03 times higher than that in the sterilized group, as well as higher lipid productivity (17.29 mg L-1 d-1). Nevertheless, the sterilized biogas slurry at a 1:1 dilution ratio exhibited the most notable nutrient-removal efficiency, with COD at 71.97%, TP at 91.32%, and TN at 88.80%. Additionally, the analysis of 16S rRNA sequencing revealed a significant alteration in the indigenous bacterial composition of the biogas slurry by microalgal treatment, with Proteobacteria and Cyanobacteria emerging as the predominant phyla, and unidentified_Cyanobacteria as the primary genus. These findings suggest that Chlorella sp. YSD-2 exhibits favorable tolerance and nutrient-removal capabilities in unsterilized, high-strength biogas slurry, along with high productivity of biomass and lipids. Consequently, these results offer a theoretical foundation for the development of an efficient and economically viable treatment method for algal-BS.

First report of Pseudomonas marginalis causing tuber soft rot of potato in Iran

In 2016, potato tuber soft rot symptoms were observed in major potato production fields in Jiroft county (southern Kerman province, Iran). Bacteria, which appeared as white to creamy colonies, were isolated from diseased tissues and shown to be pathogenic on potato tubers after inoculation and re-isolation of the isolates. Based on the assessment of colony morphology, biochemical characteristics, and analysis of 16S rRNA and gyrB gene sequences, the pathogen was identified as Pseudomonas marginalis. To our knowledge, this is the first report of P. marginalis causing potato tuber soft rot disease in Iran.

Domestic dogs in indigenous Amazonian communities: Key players in Leptospira cycling and transmission?

Leptospirosis is the world's most common zoonotic disease. Mitigation and control rely on pathogen identification and understanding the roles of potential reservoirs in cycling and transmission. Underreporting and misdiagnosis obscure the magnitude of the problem and confound efforts to understand key epidemiological components. Difficulties in culturing hamper the use of serological diagnostics and delay the development of DNA detection methods. As a result, especially in complex ecosystems, we know very little about the importance of different mammalian host species in cycling and transmission to humans. We sampled dogs from five indigenous Kichwa communities living in the Yasuní National Park in the Ecuadorian Amazon basin. Blood and urine samples from domestic dogs were collected to assess the exposure of these animals to Leptospira and to identify the circulating species. Microscopic Agglutination Tests with a panel of 22 different serovars showed anti-leptospira antibodies in 36 sampled dogs (75%), and 7 serogroups were detected. Two DNA-based detection assays revealed pathogenic Leptospira DNA in 18 of 19 dog urine samples (94.7%). Amplicon sequencing and phylogenetic analysis of 16S rRNA and SecY genes from 15 urine samples revealed genetic diversity within two of three different Leptospira species: noguchii (n = 7), santarosai (n = 7), and interrogans (n = 1). The high prevalence of antibodies and Leptospira DNA provides strong evidence for high rates of past and current infections. Such high prevalence has not been previously reported for dogs. These dogs live in the peridomestic environment in close contact with humans, yet they are free-ranging animals that interact with wildlife. This complex web of interactions may explain the diverse types of pathogenic Leptospira observed in this study. Our results suggest that domestic dogs are likely to play an important role in the cycling and transmission of Leptospira. Future studies in areas with complex ecoepidemiology will enable better parsing of the significance of genotypic, environmental, and host characteristics.

Searching for bacterial plastitrophs in modified Winogradsky columns

IntroductionPlastic pollution has surged due to increased human consumption and disposal of plastic products. Microbial communities capable of utilizing plastic as a carbon source may play a crucial role in degrading and consuming environmental plastic. In this study, we investigated the potential of a modified Winogradsky column (WC) to enrich Connecticut landfill soil for plastic-degrading bacteria and genes.MethodsBy filling WCs with landfill soil and inorganic Bushnell Haas medium, and incorporating polyethylene (PE) strips at different soil layers, we aimed to identify bacterial taxa capable of degrading PE. We employed high-throughput 16S rRNA sequencing to identify the microbes cultivated on the plastic strips and the intervening landfill soil. We used PICRUSt2 to estimate the functional attributes of each community from 16S rRNA sequences.Results and discussionAfter 12 months of incubation, distinct colors were observed along the WC layers, indicating successful cultivation. Sequencing revealed significant differences in bacterial communities between the plastic strips and the intervening landfill-soil habitats, including increased abundance of the phyla Verrucomicrobiota and Pseudomonadota (néé Proteobacteria) on the strips. Based on inferred genomic content, the most highly abundant proteins in PE strip communities tended to be associated with plastic degradation pathways. Phylogenetic analysis of 16S rRNA sequences showed novel unclassified phyla and genera enriched on the plastic strips. Our findings suggest PE-supplemented Winogradsky columns can enrich for plastic-degrading microbes, offering insights into bioremediation strategies.