Search Results (37)

Nontuberculous mycobacteria (NTM) are environmental organisms that can cause opportunistic infections in humans and animals. Mycobacterium abscessus (Mab) is a rapidly growing Mycobacterium known for its resistance to multiple antibiotics and ability to cause respiratory, skin, and mucosal infections. Understanding the distribution and prevalence of NTM, particularly Mab, in cattle farms and slaughterhouses is crucial for developing effective prevention and control measures. We collected environmental swabs from various surfaces (e.g., feed troughs, sinks, walls, floors, feces, and padding) in cattle farms and slaughterhouses across multiple provinces. High-throughput sequencing technology was utilized to analyze the 16S rDNA V3–V4 region of bacterial DNA extracted from the samples, and qPCR methods were employed to detect and quantify Mycobacterium abscessus in the collected samples. Bioinformatics analysis was performed to identify and classify the NTM species present in the samples. This study compared the abundance and diversity of NTM in different environments and assessed the potential zoonotic risk. A total of 1648 environmental swabs were collected from cattle farms and slaughterhouses in 12 provinces of China in 2023, of which 12 samples tested positive for Mab qPCR detection, yielding a detection rate of 0.73% (12/1648). Among them, the detection rate of environmental samples from cattle farms and slaughterhouses was 0.42% (3/720) and 0.87% (9/928), respectively. This study provides valuable information on the epidemiology of NTM in cattle farms and slaughterhouses, contributing to developing effective strategies for preventing and controlling NTM infections. It also enhances our understanding of the zoonotic potential of Mycobacterium abscessus and other NTM species. Full article

Background/Objectives: Massively parallel sequencing technologies have advanced chronic lymphocytic leukemia (CLL) diagnostics and precision oncology. Illumina platforms, while offering robust performance, require substantial infrastructure investment and a large number of samples for cost-efficiency. Conversely, third-generation long-read nanopore sequencing from Oxford Nanopore Technologies (ONT) can significantly reduce sequencing costs, making it a valuable tool in resource-limited settings. However, nanopore sequencing faces challenges with lower accuracy and throughput than Illumina platforms, necessitating additional computational strategies. In this paper, we demonstrate that integrating publicly available short-read data with in-house generated ONT data, along with the application of machine learning approaches, enables the characterization of the CLL transcriptome landscape, the identification of clinically relevant molecular subtypes, and the assignment of these subtypes to nanopore-sequenced samples. Methods: Public Illumina RNA sequencing data for 608 CLL samples were obtained from the CLL-Map Portal. CLL transcriptome analysis, gene module identification, and transcriptomic subtype classification were performed using the oposSOM R package for high-dimensional data visualization with self-organizing maps. Eight CLL patients were recruited from the Hematology Center After Prof. R. Yeolyan (Yerevan, Armenia). Sequencing libraries were prepared from blood total RNA using the PCR-cDNA sequencing-barcoding kit (SQK-PCB109) following the manufacturer’s protocol and sequenced on an R9.4.1 flow cell for 24–48 h. Raw reads were converted to TPM values. These data were projected into the SOMs space using the supervised SOMs portrayal (supSOM) approach to predict the SOMs portrait of new samples using support vector machine regression. Results: The CLL transcriptomic landscape reveals disruptions in gene modules (spots) associated with T cell cytotoxicity, B and T cell activation, inflammation, cell cycle, DNA repair, proliferation, and splicing. A specific gene module contained genes associated with poor prognosis in CLL. Accordingly, CLL samples were classified into T-cell cytotoxic, immune, proliferative, splicing, and three mixed types: proliferative–immune, proliferative–splicing, and proliferative–immune–splicing. These transcriptomic subtypes were associated with survival orthogonal to gender and mutation status. Using supervised machine learning approaches, transcriptomic subtypes were assigned to patient samples sequenced with nanopore sequencing. Conclusions: This study demonstrates that the CLL transcriptome landscape can be parsed into functional modules, revealing distinct molecular subtypes based on proliferative and immune activity, with important implications for prognosis and treatment that are orthogonal to other molecular classifications. Additionally, the integration of nanopore sequencing with public datasets and machine learning offers a cost-effective approach to molecular subtyping and prognostic prediction, facilitating more accessible and personalized CLL care. Full article

Continuous cropping obstacles pose significant constraints and urgent challenges in the production of Tussilago farfara L. This experiment investigated the effects of consecutive cropping on T. farfara over periods of 1, 2, and 3 years. It assessed the yield and quality of T. farfara flower buds, in addition to the physicochemical properties of the rhizosphere soil. The microbial community in the rhizosphere was analyzed through 16S rDNA and ITS sequencing using Illumina Novaseq high-throughput sequencing technology, while also examining the correlations among these factors. The results reveal that as the duration of continuous cropping increases, the yield of T. farfara flower buds, along with the contents of extract, tussilagone, and total flavonoids, steadily decreased; soil pH, organic matter, available phosphorus, available potassium, alkaline nitrogen, and the activities of sucrose, catalase, and alkaline phosphatase markedly decreased. As the duration of consecutive cropping increases, the quantity and diversity of bacteria in the rhizosphere soil initially increase and then decrease, while the number of fungal species increases by 22.5%. Meanwhile, continuous cropping of T. farfara contributes to a gradual reduction in the relative abundance of beneficial genera such as Ralstonia, Nitrospira, and Trichoderma in the rhizosphere soil, while harmful genera such as Mortierella, Fusarium, and Tricharina accumulate significantly. Correlation analysis shows that changes in microbial communities notably influence the growth of T. farfara and soil quality. This study elucidates the impacts of continuous cropping on the yield and quality of T. farfara flower buds, soil physicochemical properties, and the microbial communities in the rhizosphere, providing a scientific basis for further research on continuous cropping barriers and the selection of beneficial microbial genera for the growth of T. farfara. Full article

Integrative conservation research on animal behavior and nutritional health can contribute to the ex situ conservation of endangered species. Stereotypic behavior, a repetitive behavior without practical function, is associated with animal welfare in its manner and frequency for captive animals. Exploring the potential relationship between stereotypic behavior and internal factors, such as intestinal flora, could improve ex situ conservation, especially for endangered species. In this study, we analyzed the typical behavior characteristics of the endangered sun bears (Helarctos malayanus) under captive conditions based on the behavior sampling method. The seasonal and annual changes in the intestinal flora of H. malayanus in captivity were studied by 16S rDNA high-throughput sequencing technology based on non-invasive fecal sample collection. This study provides the first evidence of a potential association between the gut microbiota and stereotypic behavior characteristics of captive H. malayanus. The results can significantly improve our understanding of the stereotypical behavior of H. malayanus in captivity and contribute to the captive breeding and conservation efforts of this endangered species. Full article

The aim of this study was to investigate the protective effects and potential mechanisms of Tetrahydrocurcumin (THC) on methionine–choline-deficient diet (MCD)-induced MASH in C57BL/6 mice by using multi-omics techniques. The C57BL/6 mice were fed with the MCD for 8 weeks to establish a MASH model, while THC (100 mg·kg⁻¹·d⁻¹) and obeticholic acid (6.5 mg·kg⁻¹·d⁻¹) were administered via gavage to the THC group and the positive control group, respectively. The biochemical indexes of the serum and liver were detected using kits. Liver tissue sections were taken to observe the pathomorphological changes. Serum lipid and bile acid contents were measured via LC-MS, and the changes in ileal intestinal flora were detected by 16S rDNA high-throughput sequencing technology. The results revealed that THC significantly attenuated oxidative stress and lipid accumulation in NCTC-1469 cells and relieved hepatic injury and oxidative stress, reduced hepatic TG content, and improved hepatic steatosis in mice. THC alleviated 34 lipid abnormalities caused by the MCD; increased the abundance and diversity of intestinal flora, the ratio of Firmicutes to Bacteroidota, and the abundance of the probiotic (Verrucomicrobiota, Christensenellaceae, Akkermansiaceae, Lachnospiraceae, Desulfovibrionaceae); and reduced the abundance of obesity-associated pathogenic flora such as Firmicutes. Bile acid analysis showed that THC administration reduced the levels of serum toxic bile acid 7-KDCA and CA. In addition, RT-qPCR studies showed that THC down-regulated the transcript levels of the hepatic lipogenesis-related genes Srebp1c, Acc1, Scd1, and Fas, and up-regulated the transcript levels of the hepatic bile acid secretion-related genes Mrp2 and Bsep. The above results suggest that THC may alleviate MCD-induced MASH by downregulating liver Srebp1c, Acc1, Scd1, and Fas levels to inhibit lipid synthesis, upregulating Mrp2 and Bsep levels to regulate serum toxic BA levels, up-regulating the abundance of intestinal probiotic flora, and down-regulating the abundance of intestinal harmful bacterial flora. The multi-omics findings from the above study identified potential new mechanisms by which THC alleviates MASH, providing new reference targets for the development of anti-MASH drugs. These results also offer a basis for screening clinical diagnostic biomarkers for MASH and provide new directions for personalized diagnosis and treatment. Full article

Background: Soil microbes play a vital role in the ecosystem as they are able to carry out a number of vital tasks. Additionally, metagenomic studies offer valuable insights into the composition and functional potential of soil microbial communities. Furthermore, analyzing the obtained data can improve agricultural restoration practices and aid in developing more effective environmental management strategies. Methodology: In November 2023, sandy soil samples were collected from ten sites of different geographical areas surrounding natural lakes and artificial water points in the Tubaiq conservation area of King Salman Bin Abdulaziz Royal Natural Reserve (KSRNR), Saudi Arabia. In addition, genomic DNA was extracted from the collected soil samples, and 16S rRNA sequencing was conducted using high-throughput Illumina technology. Several computational analysis tools were used for gene prediction and taxonomic classification of the microbial groups. Results: In this study, sandy soil samples from the surroundings of natural and artificial water resources of two distinct natures were used. Based on 16S rRNA sequencing, a total of 24,563 OTUs were detected. The metagenomic information was then categorized into 446 orders, 1036 families, 4102 genera, 213 classes, and 181 phyla. Moreover, the phylum Pseudomonadota was the most dominant microbial community across all samples, representing an average relative abundance of 34%. In addition, Actinomycetes was the most abundant class (26%). The analysis of clustered proteins assigned to COG categories provides a detailed understanding of the functional capabilities and adaptation of microbial communities in soil samples. Amino acid metabolism and transport were the most abundant categories in the soil environment. Conclusions: Metagenome analysis of sandy soils surrounding natural lakes and artificial water points in the Tubaiq conservation area of KSRNR (Saudi Arabia) has unveils rich microbial activity, highlighting the complex interactions and ecological roles of microbial communities in these environments. Full article

Automated nucleic acid extractors are useful instruments for the high-throughput processing of bio-samples and are expected to improve research throughput in addition to decreased inter-sample variability inherent to manual processing. We evaluated three commercial nucleic acid extractors Bioer GenePure Pro (Bioer Technology, Hangzhou, China), Maxwell RSC 16 (Promega Corporation, Madison, WI, USA), and KingFisher Apex (ThermoFisher Scientific, Waltham, MA, USA) based on their DNA yield, DNA purity, and 16S rRNA gene amplicon results using both human fecal samples and a mock community (ZymoBIOMICS Microbial Community Standard (Zymo Research Corp., Irvine, CA, USA)). Bead-beating provided incremental yield to effectively lyse and extract DNA from stool samples compared to lysis buffer alone. Differential abundance analysis and comparison of prevalent bacterial species revealed a greater representation of Gram-positive bacteria in samples subjected to mechanical lysis, regardless of sample type. All three commercial extractors had differences in terms of yield, inter-sample variability, and subsequent sequencing readouts, which we subsequently share in the paper and believe are significant considerations for all researchers undertaking human fecal microbiota research. Full article

Inonotus hispidus (Bull.) P. Karst., is a medicinal fungus, which parasitizes broad-leaved tree such as Morus alba L., Fraxinus mandshurica Rupr., and Ulmus macrocarpa Hance. To elucidate the internal relationship between I. hispidus and its hosts, this study analyzed endophytic bacteria and fungi in the roots of M. alba and F. mandshurica growing I. hispidus using the 16S rDNA and ITS high-throughput sequencing technologies; and conducted widely targeted metabolomics research using UPLC-MS/MS. The results showed that Cyanobacteria and unidentified chloroplasts had the highest relative abundance at the phylum and genus levels, respectively. For endophytic fungi, Ascomycota was dominant at the phylum level, while Pleosporales gen Incertae sedis and Oncopodiella were the dominant genera in the roots of M. alba and F. mandshurica, respectively. Widely targeted metabolomics identified 562 differential metabolites and 46 metabolic pathways. Correlation analysis revealed that Xanthobacteraceae, Pseudorhodoplanes, and Bauldia were potential regulators of phenolic acids and phenylpropanoids biosynthesis. Additionally, the genus Oncopodiella was primarily associated with the enrichment of lipids, amino acids, sugars, phenolic acids, and other compounds. This result provides significant insights into the size of the fruiting body, resource development, and active ingredients of I. hispidus from different tree sources. Full article

The growth environment significantly influences the intestinal microbiota of aquatic organisms. We investigated the composition and functional differences in the intestinal microbiota of red claw crayfish (Cherax quadricarinatus) in rice fields (RB) and ponds (PB) by 16S rDNA high-throughput sequencing technology. The results indicate that the Shannon, Simpson, Sobs, Chao1, and ACE indices of PB are all higher than those of RB, demonstrating greater diversity and richness of intestinal microbiota. The dominant phyla in the intestinal microbiota of the Cherax quadricarinatus were Proteobacteria, Tenericutes, and Firmicutes. Tenericutes and Proteobacteria were significantly more abundant in the RB than in the PB, while Planctomycetes and Firmicutes were significantly more abundant in the PB than in the RB. The results of network correlation analysis indicate that Proteobacteria and Firmicutes exhibit strong connectivity with other microbial groups in the gut microbiota of Cherax quadricarinatus, showing significant centrality. They play an important role in the interactions within the gut microbiota community. The dominant bacterial genera in the Cherax quadricarinatus’s gut were Citrobacter, Candidatus_Bacilloplasma, and Clostridium_sensu_stricto_1. The abundance of the genus Clostridium was significantly higher in the PB than in the RB, whereas the abundance of Candidatus_Hepatoplasma and Vibrio was significantly lower in the PB than in the RB. The prediction function of KEGG enrichment showed that the abundance of Amino acid metabolism, Biosynthesis of Other Secondary Metabolites, Transport and Catabolism, Cancers, and Nervous System, Substance Dependence were significantly higher in the PB, while the infectious diseases pathway was enriched in the RB. In summary, our results revealed significant differences in the composition and diversity of intestinal microbiota in the Cherax quadricarinatus between rice paddy and pond farming environments. The intestinal microbiota of the Cherax quadricarinatus grown in pond environments exhibit higher diversity and stability, manifested by an increase in beneficial bacteria abundance and a decrease in opportunistic pathogens. These findings significantly improve understanding of the complex relationship among Cherax quadricarinatus, intestinal microbiota, and the environment. Full article

Sea urchins play an important role in marine ecosystems. Owing to limitations in previous research methods, there has been insufficient understanding of the food sources and ecological functional value of purple sea urchins, leading to considerable controversy regarding their functional positioning. We focused on Daya Bay as the research area, utilizing stable isotope technology and high-throughput sequencing of 16S rDNA and 18S rDNA to analyze sea urchins and their potential food sources in stone and algae areas. The results showed that the δ¹³C range of purple sea urchins in the stone area is −11.42~−8.17‰, and the δ¹⁵N range is 9.15~10.31‰. However, in the algal area, the δ¹³C range is −13.97~−12.44‰, and the δ¹⁵N range is 8.75~10.14‰. There was a significant difference in δ¹³C between the two areas (p < 0.05), but there was no significant difference in δ¹⁵N (p > 0.05). The main food source for purple sea urchins in both areas is sediment. The sequencing results of 18S rDNA revealed that, in the algal area, the highest proportion in the sea urchin gut was Molluska (57.37%). In the stone area, the highest proportion was Arthropoda (76.71%). The sequencing results of 16S rDNA revealed that, in the algal area, Bacteroidetes was the dominant group in the sea urchin gut (28.87%), whereas, in the stone area, Proteobacteria was the dominant group (37.83%). Diversity detection revealed a significant difference in the number of gut microbes and eukaryotes between the stone and algal areas (p < 0.05). The results revealed that the main food source of purple sea urchins in both areas is sediment, but the organic nutritional value is greater in the algal area, and the richness of microbiota and eukaryotes in the gut of purple sea urchins in the stone area is greater. These results indicated that purple sea urchins are likely omnivores and that the area where they occur impacts their growth and development. The results of this study provide a theoretical basis for the restoration of wild purple sea urchin resources and the selection of areas for restocking and release. Full article

Intercropping, a well-established agroecological technique designed to bolster ecological stability, has been shown to have a significant impact on soil health. However, the specific effects of tea/Trachelospermum jasminoides intercropping on the physicochemical properties and functional microbial community structure in practical cultivation have not been thoroughly investigated. In this study, we utilized high-throughput sequencing technology on the 16S/ITS rDNA genes to assess the impact of tea intercropping with T. jasminoides on the composition, diversity, and potential functions of the soil microbial community in tea gardens. The results indicated that the tea/T. jasminoides intercropping system significantly increased pH levels, soil organic matter, available nitrogen, phosphorus, potassium, and enzyme activity, ultimately augmenting soil nutrient levels. Furthermore, an in-depth analysis of the bacterial co-occurrence network and topological structure portrayed a more intricate and interconnected soil bacterial community in tea gardens. Remarkably, the abundance of beneficial genera, including Burkholderia, Mesorhizobium, Penicillium, and Trichoderma, underwent a substantial increase, whereas the relative abundance of pathogenic fungi such as Aspergillus, Fusarium, and Curvularia experienced a marked decline. Functional predictions also indicated a notable enhancement in the abundance of microorganisms associated with nitrogen and carbon cycling processes. In summary, the intercropping of tea and T. jasminoides holds the potential to enrich soil nutrient content, reshape the microbial community structure, bolster the abundance of functional microorganisms, and mitigate the prevalence of pathogenic fungi. Consequently, this intercropping system offers a promising solution for sustainable tea garden management, overcoming the limitations of traditional cultivation methods and providing valuable insights for sustainable agriculture practices. Full article

As the largest lake in South China, Wanlv Lake is also an important drinking water source for Guangdong and Hong Kong and is responsible for water supplying of more than 40 million people in the Guangdong-Hong Kong-Macao Greater Bay Area. The study of fish diversity in Wanlv Lake can help us to effectively understand the changes and states of its aquatic ecosystem and provide scientific basis for natural ecological protection, biodiversity, sustainable use and scientific management of fishery resources. However, research on the lake’s fish is scarce. The high throughput environmental DNA metabarcoding technology (eDNA technology) of the mitochondrial 12S rRNA gene sequence was herein used to conduct a preliminary exploration of Wanlv Lake fish diversity. A total of 10 sampling sites were set up to monitor fish diversity and analyze the composition and richness of the species. The results revealed a total of 83 genera in 42 families and 17 orders of fish in the lake, with the highest proportion found in Cypriniformes (38 species), accounting for 45.24% of the total. Five alien species and eight rare and endangered species were also detected. The proportion of invasive species was 5.95%, and the combined proportion of rare and endangered species was 9.52%. The results indicated that Wanlv Lake serves as an important drinking water source, and overall, the condition of fish is satisfactory. However, proactive measures should be implemented to control the overpopulation of invasive species, as their unchecked proliferation may lead to a decline in species abundance, particularly among endangered species. This was the first overall assessment report on fish of the Wanlv Lake via 12S rRNA; the results herein lay a foundation for water quality assessment of Wanlv Lake as a drinking water source. Full article

Molecular typing techniques are utilized to determine genetic similarities between bacterial isolates. However, the use of environmental DNA profiling to assess epidemiologic links between patients and their environment has not been fully explored. This work reports the development and validation of two high-throughput short sequence typing (HiSST) schemes targeting the opportunistic pathogens Pseudomonas aeruginosa and Stenotrophomonas maltophilia, along with a modified SM2I selective medium for the specific isolation of S. maltophilia. These HiSST schemes are based on four discriminative loci for each species and demonstrate high discriminating power, comparable to pairwise whole-genome comparisons. Each scheme includes species-specific PCR primers for precise differentiation from closely related taxa, without the need for upstream culture-dependent methods. For example, the primers targeting the bvgS locus make it possible to distinguish P. aeruginosa from the very closely related Pseudomonas paraeruginosa sp. nov. The selected loci included in the schemes are adapted to massive parallel amplicon sequencing technology. An R-based script implemented in the DADA2 pipeline was assembled to facilitate HiSST analyses for efficient and accurate genotyping of P. aeruginosa and S. maltophilia. We demonstrate the performance of both schemes through in silico validations, assessments against reference culture collections, and a case study involving environmental samples. Full article

The crop rhizosphere is the main site of soil microbial activities. Understanding the structure and diversity of microbial communities in the crop rhizosphere will help us reveal interactions between rhizosphere microorganisms and plant growth. In this study, the rhizosphere soil was collected from 35 cultivated barley varieties at the mature stage. To investigate the structure and diversity of bacterial communities in the rhizosphere of different barley varieties, the 16S rDNA gene of microorganisms from the soil was sequenced using Illumina MiSeq next-generation high-throughput sequencing technology. The results showed that 13, 25, 49, and 59 bacterial flora with relative abundance >1% were detected from 35 barley rhizosphere samples at the phylum, class, order, and family levels, respectively. The abundance of bacteria among varieties differed relatively little, but the abundance of the same bacteria in rhizospheres of different varieties was different. In addition, both the cluster analysis and principal component analysis (PCA) divided the 35 samples into three clusters at the phylum level. Groups III and IV showed significantly higher abundance than group II in Proteobacteria, while group II exhibited significantly higher abundance of Chloroflexi than groups III and IV. This finding provides a realistic basis for further using the relationship between barley rhizosphere microorganisms and barley growth to improve the resistance and quality of barley. Full article

The quality changes, dynamic changes in microbial composition, and diversity changes in large yellow croaker (Larimichthys crocea) during 4 °C refrigeration were studied using 16S rDNA high-throughput sequencing technology, and the total viable count (TVC), total volatile basic nitrogen (TVB-N), and thiobarbituric acid-reactive substances (TBARS) were determined. The results revealed a consistent increase in TVC, TVB-N, and TBARS levels over time. On the 9th day, TVC reached 7.43 lg/(CFU/g), while on the 15th day, TVB-N exceeded the upper limit for acceptable quality, reaching 42.56 mg/100 g. Based on the 16S rDNA sequencing results, we categorized the storage period into three phases: early storage (0th and 3rd days), middle storage (6th day), and late storage (9th, 12th, and 15th days). As the storage time increased, both the species richness and diversity exhibited a declining trend. The dominant genus identified among the spoilage bacteria in refrigerated large yellow croaker was Pseudomonas, accounting for a high relative abundance of 82.33%. A comparison was carried out of the spoilage-causing ability of three strains of Pseudomonas screened and isolated from the fish at the end of storage, and they were ranked as follows, from strongest to weakest: P. fluorescen, P. lundensis, and P. psychrophila. This study will provide a theoretical basis for extending the shelf life of large yellow croaker. Full article