To identify relationships between animal, dietary and management factors and the resulting methane (CH₄) emissions, and to identify potential mitigation strategies for CH₄ production, it is vital to develop reliable and accurate CH₄ measurement techniques. This review outlines various methods for measuring enteric CH₄ emissions from ruminants such as respiration chambers (RC), sulphur hexafluoride (SF₆) tracer, GreenFeed, sniffer method, ventilated hood, facemask, laser CH₄ detector and portable accumulation chamber. The advantages and disadvantages of these techniques are discussed. In general, RC, SF₆ and ventilated hood are capable of 24 h continuous measurements for each individual animal, providing accurate reference methods used for research and inventory purposes. However, they require high labor input, animal training and are time consuming. In contrast, short-term measurement techniques (i.e., GreenFeed, sniffer method, facemask, laser CH₄ detector and portable accumulation chamber) contain additional variations in timing and frequency of measurements obtained relative to the 24 h feeding cycle. However, they are suitable for large-scale measurements under commercial conditions due to their simplicity and high throughput. Successful use of these techniques relies on optimal matching between the objectives of the studies and the mechanism of each method with consideration of animal behavior and welfare. This review can provide useful information in selecting suitable techniques for CH₄ emission measurement in ruminants.

For more than 50% of multiparous cows, it is difficult to adapt to the sudden increase in calcium demand for milk production, which is highly likely to cause hypocalcemia. An electrochemical biosensor is a portable and efficient method to sense Ca²⁺ concentrations, but biomaterial is easily affected by the pH of the analyte solution. Here, an electrochemical biosensor was fabricated using a glassy carbon electrode (GCE) and single-walled carbon nanotube (SWNT), which amplified the impedance signal by changing the structure and length of the DNAzyme. Aiming at the interference of the pH, the electrochemical biosensor (GCE/SWNT/DNAzyme) was coupled with a pH meter to form an electrochemical device. It was used to collect data at different Ca²⁺ concentrations and pH values, and then was processed using different mathematical models, of which GPR showed higher detecting accuracy. After optimizing the detecting parameters, the electrochemical device could determine the Ca²⁺ concentration ranging from 5 μM to 25 mM, with a detection limit of 4.2 μM at pH values ranging from 4.0 to 7.5. Finally, the electrochemical device was used to determine the Ca²⁺ concentrations in different blood and milk samples, which can overcome the influence of the pH.

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.