Dual-Modal Immunochromatographic Test for Sensitive Detection of Zearalenone in Food Samples Based On Biosynthetic Staphylococcus aureus-Mediated Polymer Dot Nanocomposites.

The rapid detection of toxins is of great significance to food security and human health. In this work, a dual-modality immunochromatographic test (DICT) mediated by Staphylococcus aureus (SA)-biosynthesized polymer dots (SABPDs) was constructed for sensitive monitoring of zearalenone (ZEN) in agro products. The SABPDs as potent microorganism nanoscaffolds with excellent solubility, brightness, and stability were ingeniously fabricated employing hydroquinone and SA as precursors in the Schiff base reaction and a self-assembly technique. Thanks to the fact that they not only preserved an intact microsphere for loading Fc regions of monoclonal antibodies (mAbs) and the affinity of their labeled mAbs to antigen but also generated superb colorimetric-fluorescent dual signals, the versatile SABPDs manifested unique possibilities as the new carriers for dual-readout ICT with remarkable enhancement in sensitivity in ZEN screening (limit of detection = 0.036 ng/mL, which was 31-fold lower than that of traditional gold nanoparticle-based ICT). Ultimately, the proposed immunosensor performed well in millet and corn samples with satisfactory recoveries, demonstrating its potential for point-of-care testing. This work offers a bio-friendly strategy for biosynthesizing cell-based PD vehicles with bimodal signals for food safety analysis.

Polydopamine nanospheres-assisted direct PCR for rapid detection of Escherichia coli O157:H7.

Polymerase chain reaction (PCR) is one of the most common methods for rapid monitoring of foodborne pathogens; however, it requires purified nucleic acid as a template. Conventional nucleic acid purification is a time-consuming and laborious process. To overcome this, we developed polydopamine nanospheres (PDA NPs)-assisted direct PCR for detecting Escherichia coli O157:H7 (E. coli O157:H7). PDA NPs significantly enhanced PCR efficiency because of their strong interaction with PCR reagents, including polymerase and primers, thereby enabling regulation of the PCR performance. The optimal concentration and diameter for PDA NPs were 0.10 µg/µL and 504 nm, respectively. The PDA NPs-assisted direct PCR exhibited high sensitivity in E.coli O157:H7 detection. The detection limit of PDA NPs-assisted direct PCR was 6.7 × 104 CFU/mL, which was 10-fold lower than that of direct PCR (6.7 × 105 CFU/mL). Moreover, the sensor demonstrated excellent selectivity against E. coli O157:H7, with a negative reaction to eight other common pathogens. Most importantly, the PDA NPs-assisted direct PCR detected the order of 104-5 CFU/mL E.coli O157:H7 in milk, beef, and watermelon samples. No cultural enrichment was required, with the whole process taking <3 h. Therefore, PDA NPs-assisted direct PCR has tremendous potential in the rapid and sensitive detection of pathogens.

The establishment of clonally derived chicken embryonic fibroblast cell line (CSC) with high transfection efficiency and ability as a feeder cell.

This study established a single cloned chicken embryonic fibroblast (CEF) cell line. It solves the main problem of the instability of a cultured primary cell and its impact on the experiment. In this study, CEF pass through this crisis and formed a continuous cell line after subculture. We isolated single postcrisis CEF by a mouth pipette under a convert microscope then established a single cloned cell line named CSC-1-5 which passaged continuously from 96-well plates to 60 mm culture plates. CSC has a normal chicken diploid karyotype, no tumorigenicity, and a high G2/M phase cell ratio. We found that Fugene could mediate the transfection of CSCs efficiently; it was significantly improved compared with the primary cells. It could also promote the proliferation of chicken embryonic stem cell as a feeder layer.

Cholestyramine alleviates bone and muscle loss in irritable bowel syndrome via regulating bile acid metabolism.

Irritable bowel syndrome (IBS) is a widespread gastrointestinal disorder known for its multifaceted pathogenesis and varied extraintestinal manifestations, yet its implications for bone and muscle health are underexplored. Recent studies suggest a link between IBS and musculoskeletal disorders, but a comprehensive understanding remains elusive, especially concerning the role of bile acids (BAs) in this context. This study aimed to elucidate the potential contribution of IBS to bone and muscle deterioration via alterations in gut microbiota and BA profiles, hypothesizing that cholestyramine could counteract these adverse effects. We employed a mouse model to characterize IBS and analysed its impact on bone and muscle health. Our results revealed that IBS promotes bone and muscle loss, accompanied by microbial dysbiosis and elevated BAs. Administering cholestyramine significantly mitigated these effects, highlighting its therapeutic potential. This research not only confirms the critical role of BAs and gut microbiota in IBS-associated bone and muscle loss but also demonstrates the efficacy of cholestyramine in ameliorating these conditions, thereby contributing significantly to the field's understanding and offering a promising avenue for treatment.

Validation of the KangarooSci® Aerobic Count Plate for the Enumeration of Meosphilic Aerobic Bacteria in Selected Foods and on Stainless Steel Environmental Surfaces: AOAC Performance Tested MethodSM 062301.

BACKGROUND: The KangarooSci® Aerobic Count Plate (ACP) is a sample-ready culture medium system for direct counting of aerobic bacteria colonies after 48-72 h of incubation. OBJECTIVE: The KangarooSci ACP was evaluated for AOAC Performance Tested MethodsSM certification. METHODS: The KangarooSci ACP was evaluated through matrix studies and product consistency/stability study and robustness testing. For the matrix study, nine food products (nonfat dry milk powder, fresh raw bovine milk, pasteurized liquid bovine milk, fresh raw ground beef, frozen uncooked chicken breast, cooked shredded pork, apple juice, ice cream, and fresh strawberries), and one environmental surface (stainless steel) were evaluated following the KangarooSci ACP product instructions and compared to the ISO 4833-1:2013, Microbiology of food and animal feeding stuffs-Horizontal methods for the enumeration of microorganisms-Part 1: Colony count at 30 °C by the pour plate technique reference standard. The product consistency and stability testing evaluated three separate production lots of the KangarooSci ACP. The robustness testing examined three test parameters, volume of sample plated, incubation time, and incubation temperature, using a factorial study design. RESULTS: Results from the matrix study demonstrated equivalent performance between the KangarooSci ACP and the ISO 4833-1:2013 reference standard. The product consistency and stability testing showed that the performance of the assay was equivalent over time up to 12 months and between production lots. Minor changes to the operational test conditions showed no significant impact on performance during the robustness testing. CONCLUSION: The KangarooSci ACP is an effective method for aerobic plate count for all matrixes evaluated. HIGHLIGHTS: The KangarooSci ACP allows for fast, reliable enumeration of aerobic bacteria. Utilizing the alternative method takes up less space in incubators, requires no sample spreader, and requires fewer consumables compared to the reference method.

Surface engineering of carbon selenide nanofilms on carbon cloth: An advanced and ultrasensitive self-supporting binder-free electrode for nitrite sensing.

Large uptakes of nitrite have been proven to be detrimental to human health, therefore, the development of high-performance nitrite sensors is highly emergent. Herein, a carbon selenide nanofilms modified carbon fiber cloth (CSe2 NF/CC) electrode was obtained via in-situ synthesis to detect nitrite. The electrode integrates the collective merits of macroporous CC and pleated carbon selenide nanofilms, possessing a low overpotential of 0.83 V, a high electrochemical active surface area (EASA) of 5.39 cm2, great electrical conductivity, and fast charge transport as well as ion diffusion. The proposed electrode achieved a low limit of detection of 0.04 µmol L-1 (S/N = 3), a high sensitivity of 2048.56 µA mmol L-1 cm-2, excellent selectivity, and long-term stability. Additionally, the CSe2 NF/CC was successfully used for nitrite detection in different food samples such as pickled vegetables and sausage samples.

Metal-polydopamine framework based lateral flow assay for high sensitive detection of tetracycline in food samples.

Gold nanoparticles (AuNPs)-based lateral flow assay (LFA) enables a rapid detection of tetracycline (TET) in food samples but suffers from low sensitivity. Herein, metal-polydopamine framework (MPF), as a label, was employed to load monoclonal antibodies (mAbs) directly as the probe in LFA for highly sensitive sensing of TET. Combining zeolitic imidazolate framework (ZIF-67) and polydopamine (PDA), a stable MPF with large size, well water-dispersible, excellent affinity and optical properties was prepared through a versatile layer-by-layer assembly (LLA) strategy. Under optimized conditions, the biosensor (MPF-LFA) exhibited a great linear relationship in the range of 0.09-6 ng/mL and a detection limit of 0.045 ng/mL for TET detection, which was over 66-fold more sensitive than traditional AuNPs based LFA. Furthermore, the MPF-LFA was successfully applied to the screening of TET in fish, chicken, milk and shrimp samples with satisfied recoveries from 91% to 114%.

Polydopamine coated zirconium metal-organic frameworks-based immunochromatographic assay for highly sensitive detection of deoxynivalenol.

Conventional immunochromatographic assays (ICAs) based on gold nanoparticles (GNPs) suffer from the disadvantage of low sensitivity. In this work, a highly sensitive ICA based on polydopamine coated zirconium metal-organic frameworks labeled antibodies (ZrPA-Ab) as a novel probe was developed for visual determination of deoxynivalenol (DON). The ZrPA was synthesized via an oxidative self-polymerized assembly (OPMA) strategy using porphyrin functionalized zirconium metal-organic frameworks (Zr-MOFs, MOF-525) and polydopamine (PDA). The Abs could directly attach to the ZrPA surface owing to the large specific surface area, excellent water-stability and bio-compatibility of the ZrPA, on this basis, a sensitive, precise and reliable immunoassay method can be developed for rapid and selective detection of DON. Under optimized conditions, a non-linear calibration curve was obtained in the range of 0-50 ng/mL DON concentration with an IC50 of 1.22 ng/mL, and the visual detection limit was 0.18 ng/mL, which was about 8-times more sensitive than that of the conventional GNPs-based ICA. Finally, the proposed ZrPA-ICA was successfully applied for the detection of DON in pig hind legs meat, green bean, maize and millet samples, revealing the feasible and reliable application of this biosensor in different food matrices. Thus, this work broadens the possibilities for the use of MOFs as a novel labeling carrier in immunoassays.