Magnesium ions (Mg²⁺) play an important role in animal health, with their concentration in the bloodstream serving as a key indicator for hypomagnesemia diagnosis. In this study, a flexible hydrophobic paper-based microfluidic field-effect biosensor was developed for point-of-care Mg²⁺ detection, which integrated flexible hydrophobic paper, semiconducting single-walled carbon nanotubes (SWNTs) and a Mg²⁺-specific RNA-cleaving DNAzyme(RCD)-based DNA nanostructure. Flexible hydrophobic paper was synthesized by using cellulose paper and octadecyltrichlorosilane, improving mechanical strength and decreasing biological interference. To achieve high sensitivity, the Mg²⁺-specific RCD was functionalized with SWNTs, and then repeatedly self-assembled two different Y-shaped DNAs to construct a DNA nanostructure based on a similar DNA origami technique. This proposed biosensor exhibited a linear detection range from 1 μM to 1000 μM, with a detection limit of 0.57 μM, demonstrating its great stability, selectivity, and anti-interference performance. This innovative design offers promising potential for Mg²⁺ monitoring in real applications.

Monitoring potassium ion (K⁺) concentration is essential in veterinary medicine, particularly for preventing hypokalemia in dairy cows, which can severely impact their health and productivity. While traditional laboratory methods like atomic absorption spectrometry are accurate, they are also time-consuming and require complex sample preparation. Ion-selective electrodes (ISEs) provide an alternative that is faster and more suitable for field measurements, but their performance is often compromised under variable temperature conditions, leading to inaccuracies. To address this, we developed a novel screen-printed ion-selective electrode (SPE) with hydrophobic Ti₃C₂ Mxene and gold nanoparticles (AuNPs), integrated with a temperature sensor. This design improves stability and accuracy across fluctuating temperatures by preventing water layer formation and enhancing conductivity. The sensor was validated across temperatures from 5 °C to 45 °C, achieving a linear detection range of 10⁻⁵ to 10⁻¹ M and a response time of approximately 15 s. It also demonstrated excellent repeatability, selectivity, and stability, making it a robust tool for K⁺ monitoring in complex environments. This advancement could lead to broader applications in other temperature-sensitive analytical fields.

In this paper, screen-printed ion-selective electrodes combined with single-walled carbon nanotubes (SWCNTs) and gold nanoparticles (AuNPs) were used to rapidly and accurately measure serum Ca²⁺ concentration. Due to the susceptibility of cows to hypocalcemia after delivery, this disease can affect the health of cows and reduce milk production. Therefore, the development of an economical and swift detection method holds paramount importance for facilitating early diagnosis and subsequent treatment. In this study, by combining the high electrical conductivity and large surface area of SWCNTs with the strong catalytic activity of AuNPs, a SWCNTs/AuNPs composite with high sensitivity and good stability was prepared, achieving efficient selective recognition and signal conversion of Ca²⁺. The experimental results indicate that the screen-printed electrode modified with SWCNTs/AuNPs exhibited excellent performance in the determination of Ca²⁺ concentration. Its linear response range is 10^−5.5–10⁻¹ M, covering the normal and pathological concentration range of Ca²⁺ in cow blood, and the detection limit is far below the clinical detection requirements. In addition, the electrode also has good anti-interference ability and fast response time (about 15 s), showing good performance in the range of 5–45 °C. In practical applications, the combination of the electrode and portable detection equipment can realize the field rapid determination of cow blood Ca²⁺ concentration. This method is easy to operate, cost-effective, and easy to promote, providing strong technical support for the health management of dairy farms.

Precise quantitative monitoring of 17β-estradiol (E2) is important for reproductive management in precision livestock farming. However, E2 determination in complex biological matrices remains challenging because of matrix-derived background and signal variability. Here, we developed a nanoconfinement-assisted solid-state ratiometric fluorescent aptasensor integrating target-induced strand displacement (TISD), magnetic separation, and anodic aluminum oxide (AAO) nanochannel confinement. The sensing probe consisted of streptavidin-coated magnetic nanoparticles (MNPs) carrying a FAM-labeled cDNA internal reference and a Texas Red-labeled E2 aptamer reporter. E2 binding promoted dissociation of the Texas Red-labeled aptamer from the magnetic probe. Magnetic separation and washing reduced soluble matrix-derived interference, while subsequent deposition of the sensing complexes onto an AAO membrane mitigated coffee-ring-associated nonuniformity and produced a more spatially uniform dual-color fluorescence distribution for ratiometric analysis. Under matrix-matched calibration conditions, linear ranges of 5.0–50.0 pM were obtained in tap water and sow saliva, 5.0–40.0 pM in whole milk, and 5.0–15.0 pM in post-estrus sow urine. The LOD determined in tap water was 3.62 pM. The different calibration slopes obtained among the four matrices indicated that residual matrix-dependent effects remained and that matrix-specific calibration was required for quantitative analysis. Matrix-matched spike recoveries ranged from 86.92% to 119.54% across the investigated matrices. The aptasensor exhibited the strongest response toward 17β-E2 among the tested compounds; however, cross-reactivities of 77.3% for E3 and 47.3% for 17α-E2 indicated preferential rather than exclusive recognition. Molecular docking suggested a putative binding pose but did not experimentally establish the molecular recognition mechanism. Overall, the platform demonstrated laboratory-scale analytical feasibility in pretreated tap water, sow saliva, whole milk, and post-estrus sow urine. Further development of sample preparation, magnetic handling, membrane loading, probe selectivity, and portable fluorescence readout will be required before in situ or on-site application.