Modifying soil bacterial communities in saline mudflats with organic acids and substrates.

Aims: The high salinity of soil, nutrient scarcity, and poor aggregate structure limit the exploitation and utilization of coastal mudflat resources and the sustainable development of saline soil agriculture. In this paper, the effects of applying exogenous organic acids combined with biological substrate on the composition and diversity of soil bacterial community were studied in moderately saline mudflats in Jiangsu Province. Methods: A combination of three exogenous organic acids (humic acid, fulvic acid, and citric acid) and four biological substrates (cottonseed hull, cow manure, grass charcoal, and pine needle) was set up set up on a coastal saline mudflat planted with a salt-tolerant forage grass, sweet sorghum. A total of 120 kg ha-1 of organic acids and 5,000 kg ha-1 of substrates were used, plus two treatments, CK without application of organic acids and substrates and CK0 in bare ground, for a total of 14 treatments. Results: No significant difference was found in the alpha diversity of soil bacterial community among all treatments (p ≥ 0.05), with the fulvic acid composite pine needle (FPN) treatment showing the largest increase in each index. The beta diversity differed significantly (p < 0.05) among all treatments, and the difference between citric acid-grass charcoal (CGC) and CK treatments was greater than that of other treatments. All treatments were effective in increasing the number of bacterial ASVs and affecting the structural composition of the community. Citric acid-cow manure (CCM), FPN, and CGC treatments were found to be beneficial for increasing the relative abundance of Proteobacteria, Chloroflexi, and Actinobacteria, respectively. By contrast, all treatments triggered a decrease in the relative abundance of Acidobacteria. Conclusion: Among the 12 different combinations of exogenous organic acid composite biomass substrates applied to the coastal beach, the CGC treatment was more conducive to increasing the relative abundance of the salt-tolerant bacteria Proteobacteria, Chloroflexi and Actinobacteria, and improving the community structure of soil bacteria. The FPN treatment was more conducive to increase the species diversity of the soil bacterial community and adjust the species composition of the bacterial community.

Intestinal microbiota and gene expression alterations in leopard coral grouper (Plectropomus leopardus) under enteritis.

Enteritis poses a significant threat to fish farming, characterized by symptoms of intestinal and hepatic inflammation, physiological dysfunction, and dysbiosis. Focused on the leopard coral grouper (Plectropomus leopardus) with an enteritis outbreak on a South China Sea farm, our prior scrutiny did not find any abnormalities in feeding or conventional water quality factors, nor were any specific pathogen infections related to enteritis identified. This study further elucidates their intestinal flora alterations, host responses, and their interactions to uncover the underlying pathogenetic mechanisms and facilitate effective prevention and management strategies. Enteritis-affected fish exhibited substantial differences in intestinal flora compared to control fish (P = 0.001). Notably, norank_f_Alcaligenaceae, which has a negative impact on fish health, predominated in enteritis-affected fish (91.76 %), while the probiotic genus Lactococcus dominated in controls (93.90 %). Additionally, certain genera with pathogenesis potentials like Achromobacter, Sphingomonas, and Streptococcus were more abundant in diseased fish, whereas Enterococcus and Clostridium_sensu_stricto with probiotic potentials were enriched in control fish. At the transcriptomic level, strong inflammatory responses, accompanied by impaired metabolic functions, tissue damage, and iron death signaling activation were observed in the intestines and liver during enteritis. Furthermore, correlation analysis highlighted that potential pathogen groups were positively associated with inflammation and tissue damage genes while presenting negatively correlated with metabolic function-related genes. In conclusion, dysbiosis in the intestinal microbiome, particularly an aberrantly high abundance of Alcaligenaceae with pathogenic potential may be the main trigger for this enteritis outbreak. Alcaligenaceae alongside Achromobacter, Sphingomonas, and Streptococcus emerged as biomarkers for enteritis, whereas some species of Lactococcus, Clostridium_sensu_stricto, and Enterococcus showed promise as probiotics to alleviate enteritis symptoms. These findings enhance our understanding of enteritis pathogenesis, highlight intestinal microbiota shifts in leopard coral grouper, and propose biomarkers for monitoring, probiotic selection, and enteritis management.

Tea green leafhopper infestations affect tea plant growth by altering the synthesis of brassinolide.

Tea green leafhoppers are insects widely distributed in major tea-growing areas. At present, less attention has been paid to the study on effect of tea green leafhopper infestation on tea growth phenotype. In this study, tea green leafhoppers were used to treat tea branches in laboratory and co-treated with brassinolide (BL), the highest bioactivity of brassinosteroids (BRs), in tea garden. The results showed that the expression of genes related to BRs synthesis was inhibited and BL content was reduced in tea shoots after infestation by tea green leafhoppers. In addition, area of each leaf position, length and diameter of internodes, and the biomass of the tender shoots of tea plant were decreased after infestation by tea green leafhoppers. The number of trichomes, leaf thickness, palisade tissue thickness and cuticle thickness of tea shoots were increased after tea green leafhoppers infestation. BL spraying could partially recover the phenotypic changes of tea branches caused by tea green leafhoppers infestation. Further studies showed that tea green leafhoppers infestation may regulate the expression of CsDWF4 (a key gene for BL synthesis) through transcription factors CsFP1 and CsTCP1a, which finally affect the BL content. Moreover, BL was applied to inhibit the tea green leafhoppers infestation on tea shoots. In conclusion, our study revealed the effect of plant hormone BL-mediated tea green leafhoppers infestation on the growth phenotype of tea plants.

Learning data distribution of three-dimensional ocean sound speed fields via diffusion models.

The probability distribution of three-dimensional sound speed fields (3D SSFs) in an ocean region encapsulates vital information about their variations, serving as valuable data-driven priors for SSF inversion tasks. However, learning such a distribution is challenging due to the high dimensionality and complexity of 3D SSFs. To tackle this challenge, we propose employing the diffusion model, a cutting-edge deep generative model that has showcased remarkable performance in diverse domains, including image and audio processing. Nonetheless, applying this approach to 3D ocean SSFs encounters two primary hurdles. First, the lack of publicly available well-crafted 3D SSF datasets impedes training and evaluation. Second, 3D SSF data consist of multiple 2D layers with varying variances, which can lead to uneven denoising during the reverse process. To surmount these obstacles, we introduce a novel 3D SSF dataset called 3DSSF, specifically designed for training and evaluating deep generative models. In addition, we devise a high-capacity neural architecture for the diffusion model to effectively handle variations in 3D sound speeds. Furthermore, we employ state-of-the-art continuous-time-based optimization method and predictor-corrector scheme for high-performance training and sampling. Notably, this paper presents the first evaluation of the diffusion model's effectiveness in generating 3D SSF data. Numerical experiments validate the proposed method's strong ability to learn the underlying data distribution of 3D SSFs, and highlight its effectiveness in assisting SSF inversion tasks and subsequently characterizing the transmission loss of underwater acoustics.

Zero-shot reconstruction of ocean sound speed field tensors: A deep plug-and-play approach.

Reconstructing a three-dimensional ocean sound speed field (SSF) from limited and noisy measurements presents an ill-posed and challenging inverse problem. Existing methods used a number of pre-specified priors (e.g., low-rank tensor and tensor neural network structures) to address this issue. However, the SSFs are often too complex to be accurately described by these pre-defined priors. While utilizing neural network-based priors trained on historical SSF data may be a viable workaround, acquiring SSF data remains a nontrivial task. This work starts with a key observation: Although natural images and SSFs admit fairly different characteristics, their denoising processes appear to share similar traits-as both remove random components from more structured signals. This observation allows us to incorporate deep denoisers trained using extensive natural images to realize zero-shot SSF reconstruction, without any extra training or network modifications. To implement this idea, an alternating direction method of multipliers (ADMM) algorithm using such a deep denoiser is proposed, which is reminiscent of the plug-and-play scheme from medical imaging. Our plug-and-play framework is tailored for SSF recovery such that the learned denoiser can be simultaneously used with other handcrafted SSF priors. Extensive numerical studies show that the new framework largely outperforms state-of-the-art baselines, especially under widely recognized challenging scenarios, e.g., when the SSF samples are taken as tensor fibers. The code is available at https://github.com/OceanSTARLab/DeepPnP.

Whole-genome sequencing of Sphingobium baderi SC-1 and identification of a crucial 3-phenoxybenzoic acid-degrading gene.

As an efficient degradation strain, Sphingobium baderi SC-1 can breakdown 3-phenoxybenzoic acid (3-PBA) with high proficiency. To investigate the internal factors that regulate this process, we conducted whole-genome sequencing and successfully identified the pivotal 3-PBA-degrading gene sca (1,230 bp). After sca was expressed in engineered bacteria, a remarkable degradation efficiency was observed, as 20 mg/L 3-PBA was almost completely decomposed within 24 h. The phenol was formed as one of the degradation products. Notably, in addition to their ability to degrade 3-PBA, the resting cells proficiently degraded 4'-HO-3-PBA and 3'-HO-4-PBA. In conclusion, we successfully identified and validated sca as the pivotal enzyme responsible for the efficient degradation of 3-PBA from Sphingomonas baderi, providing a crucial theoretical foundation for further explorations on the degradation potential of SC-1.

Indirect Estimation of Heavy Metal Contamination in Rice Soil Using Spectral Techniques.

The rapid growth of industrialization and urbanization in China has led to an increase in soil heavy metal pollution, which poses a serious threat to ecosystem safety and human health. The advancement of spectral technology offers a way to rapidly and non-destructively monitor soil heavy metal content. In order to explore the potential of rice leaf spectra to indirectly estimate soil heavy metal content. We collected farmland soil samples and measured rice leaf spectra in Xushe Town, Yixing City, Jiangsu Province, China. In the laboratory, the heavy metals Cd and As were determined. In order to establish an estimation model between the pre-processed spectra and the soil heavy metals Cd and As content, a genetic algorithm (GA) was used to optimise the partial least squares regression (PLSR). The model's accuracy was evaluated and the best estimation model was obtained. The results showed that spectral pre-processing techniques can extract hidden information from the spectra. The first-order derivative of absorbance was more effective in extracting spectral sensitive information from rice leaf spectra. The GA-PLSR model selects only about 10% of the bands and has better accuracy in spectral modeling than the PLSR model. The spectral reflectance of rice leaves has the capacity to estimate Cd content in the soil (relative percent difference [RPD] = 2.09) and a good capacity to estimate As content in the soil (RPD = 2.97). Therefore, the content of the heavy metals Cd and As in the soil can be estimated indirectly from the spectral data of rice leaves. This study provides a reference for future remote sensing monitoring of soil heavy metal pollution in farmland that is quantitative, dynamic, and non-destructive over a large area.

Occurrence of ochratoxin A in Sichuan bacon from different geographical regions and characterization and biocontrol of ochratoxigenic Aspergillus westerdijkiae strain 21G2-1A.

Sichuan bacon represents the most prevalent dry-cured meat product across Southwest China, but it is vulnerable to fungal spoilage. In the present study, a total of 47 Sichuan bacons were obtained from different regions of the Sichuan Province and analyzed for the presence of ochratoxin A (OTA), yielding a positive rate of 23.4 % (11/47). All the observed OTA concentrations exceeded the maximum admissible dose in meat products (1 µg/kg) established by some EU countries, with the highest OTA concentration being 250.75 µg/kg, which raises a food safety concern and reveals the need for a standardized scientific processing protocol. Then, an OTA-producing fungus named 21G2-1A was isolated from positive samples and found to be Aspergillus westerdijkiae. Further characterization suggested a positive correlation between fungal growth and OTA production. The optimal temperature for the former was 25 °C, while it was 20 °C for the latter. Although the A. westerdijkiae strain 21G2-1A demonstrated greater mycelium growth in the presence of NaCl, OTA production was significantly dismissed when the salinity was greater than 5 %. Four lactic acid bacteria (LAB) were screened out as antagonists against the ochratoxigenic fungus. In vitro evaluation of the antagonists revealed that live cells inhibited fungal growth, and adsorption also contributed to OTA removal at different levels. This study sheds some light on OTA control in Sichuan bacon through a biological approach.

Lactiplantibacillus plantarum RS20D Alleviates Male Reproductive Toxicity Induced by Pubertal Exposure to Di-n-butyl Phthalate and Mono-n-butyl Phthalate.

Phthalate acid esters (PAEs) and their metabolites, such as di-n-butyl phthalate (DBP) and mono-n-butyl phthalate (MBP), are known to cause male reproductive damage. Lactiplantibacillus plantarum RS20D has demonstrated the ability to remove both DBP and MBP in vitro, suggesting its potential as a detoxifying agent against these compounds. This study aimed to investigate the protective effects of RS20D on DBP or MBP-induced male reproductive toxicity in adolescent rats. Oral administration of RS20D significantly mitigated the histological damage to the testes caused by MBP or DBP, restored sperm concentration, morphological abnormalities, and the proliferation index in MBP-exposed rats, and partially reversed spermatogenic damage in DBP-exposed rats. Furthermore, RS20D restored serum levels of estradiol (E2) and testosterone, and superoxide dismutase (SOD) activity in DBP-exposed rats, significantly increased testosterone levels in MBP-exposed rats, and restored copper (Cu) concentrations in the testes after exposure to DBP or MBP. Additionally, RS20D effectively modulated the intestinal microbiota in DBP-exposed rats and partially ameliorated dysbiosis induced by MBP, which may be associated with the alleviation of reproductive toxic effects induced by DBP or MBP. In conclusion, this study demonstrates that RS20D administration can alleviate male reproductive toxicity and gut dysbacteriosis induced by DBP or MBP exposure, providing a dietary strategy for the bioremediation of PAEs and their metabolites.

Identification, synthesis, and field evaluation of components of the female-produced sex pheromone of Helopeltis cinchonae (Hemiptera: Miridae), an emerging pest of tea.

BACKGROUND: Helopeltis cinchonae (Hemiptera: Miridae) is a major pest of tea plantations in Asia. Conventional control of pests with pesticides is unsustainable. Therefore, safe and eco-friendly alternatives, such as pheromones, are required to manage the pest. RESULTS: In gas chromatography-electroantennographic detection (GC-EAD) analysis of whole-body extracts of virgin female H. cinchonae, two compounds elicited electroantennogram (EAG) responses from male antennae. These were identified as hexyl (R)-3-acetoxybutyrate and (R)-1-acetoxy-5-butyroxyhexane using gas chromatography-mass spectrometry (GC-MS) analysis compared to synthetic compounds. This is the first study to report 1-acetoxy-5-butyroxyhexane as an insect pheromone component. The synthetic compounds elicited dose-dependent EAG responses from the antennae of male H. cinchonae. In two field trapping experiments, the individual compounds were highly attractive to male H. cinchonae when dispensed from polyethylene vials. However, higher catches were obtained with blends of the two compounds in a 1:10 ratio. The blend of racemic compounds was as attractive as the blend of (R)-enantiomers. CONCLUSIONS: We reported that 1-acetoxy-5-butyroxyhexane and hexyl 3-acetoxybutyrate are components of the female-produced sex pheromone of H. cinchonae, but further work is required on the blend and loading of pheromone and on trap design to provide an optimized system for monitoring and control of this pest. The results may also facilitate the identification of the pheromones of other Helopeltis species, which are major pests in many crops. © 2024 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Enhancing the acid stability of the recombinant GH11 xylanase xynA through N-terminal substitution to facilitate its application in apple juice clarification.

The utilization of xylanase in juice clarification is contingent upon its stability within acidic environments. We generated a mutant xynA-1 by substituting the N-terminal segment of the recombinant xylanase xynA to investigate the correlation between the N-terminal region of xylanase and its acid stability. The enzymatic activity of xynA-1 was found to be superior under acidic conditions (pH 5.0). It exhibited enhanced acid stability, surpassing the residual enzyme activity values of xynA at pH 4.0 (53.07 %), pH 4.5 (69.8 %), and pH 5.0 (82.4 %), with values of 60.16 %, 77.74 %, and 87.3 %, respectively. Additionally, the catalytic efficiency of xynA was concurrently improved. Through molecular dynamics simulation, we observed that N-terminal shortening induced a reduction in motility across most regions of the protein structure while enhancing its stability, particularly Lys131-Phe146 and Leu176-Gly206. Furthermore, the application of treated xynA-1 in the process of apple juice clarification led to a significant increase in clarity within a short duration of 20 min at 35 °C while ensuring the quality of the apple juice. This study not only enhances the understanding of the N-terminal region of xylanase but also establishes a theoretical basis for augmenting xylanase resources employed in fruit juice clarification.

T-YOLO: a lightweight and efficient detection model for nutrient buds in complex tea-plantation environments.

BACKGROUND: Quick and accurate detection of nutrient buds is essential for yield prediction and field management in tea plantations. However, the complexity of tea plantation environments and the similarity in color between nutrient buds and older leaves make the location of tea nutrient buds challenging. RESULTS: This research presents a lightweight and efficient detection model, T-YOLO, for the accurate detection of tea nutrient buds in unstructured environments. First, a lightweight module, C2fG2, and an efficient feature extraction module, DBS, are introduced into the backbone and neck of the YOLOv5 baseline model. Second, the head network of the model is pruned to achieve further lightweighting. Finally, the dynamic detection head is integrated to mitigate the feature loss caused by lightweighting. The experimental data show that T-YOLO achieves a mean average precision (mAP) of 84.1%, the total number of parameters for model training (Params) is 11.26 million (M), and the number of floating-point operations (FLOPs) is 17.2 Giga (G). Compared with the baseline YOLOv5 model, T-YOLO reduces Params by 47% and lowers FLOPs by 65%. T-YOLO also outperforms the existing optimal detection YOLOv8 model by 7.5% in terms of mAP. CONCLUSION: The T-YOLO model proposed in this study performs well in detecting small tea nutrient buds. It provides a decision-making basis for tea farmers to manage smart tea gardens. The T-YOLO model outperforms mainstream detection models on the public dataset, Global Wheat Head Detection (GWHD), which offers a reference for the construction of lightweight and efficient detection models for other small target crops. © 2024 Society of Chemical Industry.

Synthesis of 1,4-epoxy-2-aryltetrahydro-1-benzazepines via rhodium(III)-catalyzed C-H allylation/intramolecular 1,3-dipolar cycloaddition.

Herein, we report a new synthetic route to 1,4-epoxy-2-aryltetrahydro-1-benzazepine derivatives with high efficiency, namely the Rh(III)-catalyzed C-H allylation of nitrones with allyl precursors, followed by subsequent intramolecular 1,3-dipolar cycloaddition, to deliver the title compounds. This reaction is regio- and stereo-selective, generating the cis-isomer with a broad substrate scope and good functional group tolerance.

Biosynthetic Pathway and Bioactivity of Vanillin, a Highly Abundant Metabolite Distributed in the Root Cortex of Tea Plants (Camellia sinensis).

Volatiles are important for plant root stress resistance. The diseases in tea root are serious, causing major losses. The volatile composition in tea root and whether it can resist diseases remain unclear. In this study, the volatile composition in different tea tissues was revealed. The vanillin content was higher in the root (mainly in root cortex) than in aerial parts. The antifungal effects of vanillin on pathogenic fungi in tea root were equal to or greater than those of other metabolites. O-methyltransferase (CsOMT), a key enzyme in one of two biosynthetic pathways of vanillin, converted protocatechualdehyde to vanillin in vitro. Furthermore, its characteristics and kinetic parameters were studied. In Arabidopsis thaliana protoplasts, the transiently expressed CsOMT was localized in the cytoplasm and nucleus. These findings have clarified the formation and bioactivities of volatiles in tea roots and provided a theoretical basis for understanding how tea plants resist root diseases.

Microbial characterization of Sichuan Baoning vinegar: lactic acid bacteria, acetic acid bacteria and yeasts.

Sichuan Baoning vinegar, a typical representative of Sichuan bran vinegar, is a famous traditional fermented food made from cereals in China. At present, there are few studies on microbial characterization of culturable microorganisms in solid-state fermentation of Sichuan bran vinegar. To comprehensively understand the diversity of lactic acid bacteria, acetic acid bacteria and yeasts, which play an important role in the fermentation of Sichuan bran vinegar, traditional culture-dependent methods combined with morphological, biochemical, and molecular identification techniques were employed to screen and identify these isolates. A total of 34 lactic acid bacteria isolates, 39 acetic acid bacteria isolates, and 48 yeast isolates were obtained. Lactic acid bacteria were dominated by Enterococcus durans, Leuconostoc citreum, Lactococcus lactis, and Lactiplantibacillus plantarum, respectively. Latilactobacillus sakei was the first discovery in cereal vinegar. Acetic acid bacteria were mainly Acetobacter pomorum and A. pasteurianus. The dominant yeast isolates were Saccharomyces cerevisiae, in addition to four non-Saccharomyces yeasts. DNA fingerprinting revealed that isolates belonging to the same species exhibited intraspecific diversity, and there were differences between phenotypic and genotypic classification results. This study further enriches studies on cereal vinegar and lays a foundation for the development of vinegar starters.

Metabolomics reveals how spinach plants reprogram metabolites to cope with intense stress responses induced by photoaged polystyrene nanoplastics (PSNPs).

While land-based sources have been recognized as significant long-term sinks for micro- and nanoplastics, there is limited knowledge about the uptake, translocation, and phytotoxicity of nanoplastics (NPs) in terrestrial environments, especially aged NPs. In this study, we investigated the impact of aged polystyrene nanoplastics (PSNPs) on the uptake, physiology, and metabolism of spinach. Our findings revealed that both pristine and aged PSNPs can accumulate in the roots and subsequently translocate to the aboveground tissues, thereby influencing numerous key growth indicators in spinach plants. A more pronounced impact was observed in the treatment of aged PSNPs, triggering more significant and extensive changes in metabolite levels. Furthermore, alterations in targeted pathways, specifically aminoacyl-tRNA biosynthesis and phenylpropanoid biosynthesis, were induced by aged PSNPs, while pristine PSNPs influenced pathways related to sulfur metabolism, biosynthesis of unsaturated fatty acids, and tryptophan metabolism. Additionally, tissue-specific responses were observed at the metabolomics level in both roots and leaves. These results highlight the existence of diverse and tissue-specific metabolic responses in spinach plants exposed to pristine and aged PSNPs, providing insights into the mechanisms of defense and detoxification against NP-induced stress.

Effect of Lactiplantibacillus plantarum fermentation on the physicochemical, antioxidant activity and immunomodulatory ability of polysaccharides from Lvjian okra.

Okra polysaccharides exhibits a range of biological activities. To date, its processing using microbial fermentation has not been explored. This study investigated the fermentation of okra juice with various lactic acid bacteria, followed by the extraction and characterization of crude polysaccharides (termed OPS-F), in contrast to their non-fermented counterpart (OPS). Changes in physicochemical properties, antioxidant activity and immunomodulatory ability were noted. The results demonstrated that OPS-F had a 7.42-12.53 % increase in total polysaccharides content compared to OPS. However, high-performance size-exclusion chromatography indicated a reduction in the molecular weight of OPS-F (7.9-9.5 × 105 Da) relative to OPS (1.66 × 106 Da). Compared to OPS, OPS-F had reduced levels of mannose, glucose, glucuronic acid and arabinose, but increased rhamnose, galacturonic acid and galactose, exhibiting enhanced solubility and lower apparent viscosity. Fourier transform infrared spectroscopy and nuclear magnetic resonance analysis showed minimal changes in polysaccharide structure post-fermentation. Moreover, despite a decrease in antioxidant activity post-fermentation, OPS-F exhibited superior immunomodulatory potential. In conclusion, fermenting okra juice with lactic acid bacteria alters the physicochemical properties of crude polysaccharides and enhances their immunomodulatory activity, offering a promising approach for developing new functional food resources.

Insight into a natural novel histidine decarboxylase gene deletion in Enterobacter hormaechei RH3 from traditional Sichuan-style sausage.

Histamine (HIS) is primarily formed from decarboxylated histidine by certain bacteria with histidine decarboxylase (hdc) activity and is the most toxic biogenic amine. Hdc, which is encoded by the hdc gene, serves as a key enzyme that controls HIS production in bacteria. In this paper, we characterized the changes in microbial and biogenic amines content of traditional Sichuan-style sausage before and after storage and demonstrated that Enterobacteriaceae play an important role in the formation of HIS. To screen for Enterobacteriaceae with high levels of HIS production, we isolated strain RH3 which has a HIS production of 2.27 mg/mL from sausages stored at 37°C for 180 days, using selective media and high-performance liquid chromatography. The strain RH3 can produce a high level of HIS after 28 h of fermentation with a significant hysteresis. Analysis of the physicochemical factors revealed that RH3 still retained its ability to partially produce HIS in extreme environments with pH 3.5 and 10.0. In addition, RH3 exhibited excellent salt tolerance (6.0% NaCl and 1.0% NaNO2 ). Subsequently, RH3 was confirmed as Enterobacter hormaechei with hdc gene deletion by PCR, western blot, and whole-genome sequencing analysis. Furthermore, RH3 exhibited pathogenicity rate of 75.60% toward the organism, indicating that it was not a food-grade safe strain, and demonstrated a high level of conservation in intraspecific evolution. The results of this experiment provide a new reference for studying the mechanism of HIS formation in microorganisms. PRACTICAL APPLICATION: This study provides a new direction for investigating the mechanism of histamine (HIS) formation by microorganisms and provides new insights for further controlling HIS levels in meat products. Further research can control the key enzymes that form HIS to control HIS levels in food.

Insights into antibiotic and heavy metal resistance interactions in Escherichia coli isolated from livestock manure and fertilized soil.

Heavy metal and antibiotic-resistant bacteria from livestock feces are ecological and public health problems. However, the distribution and relationships of antibiotic resistance genes (ARGs), heavy metal resistance genes (HMRGs), and virulence factors (VFs) and their transmission mechanisms remain unclear. Therefore, we investigated the resistance of Escherichia coli, the prevalence of its ARGs, HMRGs, and VFs, and their transmission mechanisms in livestock fresh feces (FF), composted feces (CF), and fertilized soil (FS). In total, 99.54% (n = 221) and 91.44% (n = 203) of E. coli were resistant to at least one antibiotic and one heavy metal, respectively. Additionally, 72.52% (n = 161) were multi-drug resistant (MDR), of which Cu-resistant E. coli accounted for 72.67% (117/161). More than 99.34% (88/89) of E. coli carried multidrug ARGs, VFs, and the Cu resistance genes cueO and cusABCRFS. The Cu resistance genes cueO and cusABCRFS were mainly located on chromosomes, and cueO and cusF were positively associated with HMRGs, ARGs, and VFs. The Cu resistance genes pcoABCDRS were located on the plasmid pLKYL-P02 flanked by ARGs in PF18C from FF group and on chromosomes flanked by HMRGs in SAXZ1-1 from FS group. These results improved our understanding of bacterial multidrug and heavy metal resistance in the environment.

A portable all-in-one microfluidic device with real-time colorimetric LAMP for HPV16 and HPV18 DNA point-of-care testing.

Screening for high-risk human papillomavirus (HPV) infection is one of the most important preventative measures for cervical cancer. However, fast, convenient, and low-cost HPV detection remains challenging, especially in resource-limited settings. Here, we report a portable all-in-one device (PAD) for point-of-care testing (POCT) for HPV16 and HPV18 DNA in cervical swabs. The PAD was engineered to integrate modules for extraction-free sample lysis, loop-mediated isothermal amplification (LAMP) with lyophilized reagent beads, and real-time colorimetric signal sensing into a single miniaturized device, considerably shortening the sample-to-result time to 15 min. The precision liquid handling in the completely sealed microfluidic chip is achieved by a uniquely designed pressure-balanced automatic liquid flow mechanism, thereby eliminating the need for manual manipulation of liquids and thus the risk of biohazards. The PAD employs an improved real-time colorimetric LAMP (rcLAMP) assay with a limit of detection (LOD) of 1 copy/µL, enabled by enhanced assay chemistry to maximize the reaction kinetics. To validate this device for clinical application, we tested 206 clinical cervical swab samples and obtained a sensitivity of 92.1% and a specificity of 99.0%. This custom PAD enabled by microfluidic and electronic engineering techniques can be configured for the simultaneous detection of HPV16 and HPV18 or other pathogens in point-of-care applications.