N-acetyl-β-D-glucosaminidase biomarker quantification in milk using Ag-porous Si SERS platform for mastitis severity evaluation

Abstract

Bovine mammary gland is susceptible to numerous bacterial infections resulting in inflammatory disease condition of bovine mastitis (BM) with a staggering economic impact on the dairy industry worldwide. Early BM detection is crucial for infection control within the dairy herd, which can be alleviated by antimicrobial therapy. N-acetyl-β-D-glucosaminidase (NAGase) is a prominent BM inflammatory biomarker secreted onto the blood circulation upon pathogenesis and then released into milk, capable of separating healthy quarters from subclinical and clinical BM cases. Herein, we report a sensitive, cost-effective and handy-to-use BM severity assay based on NAGase inherent content found in whole milk samples. Silver-coated porous Si (Ag-PSi) Fabry–Pérot interferometers were employed for quantifying the lysosomal activity in different milk qualities using surface-enhanced Raman spectroscopy (SERS). The optical response was proportional to the occurring pathogenesis induced by predominant bacteria found in dairy farms at different somatic cell levels. The optimized Ag-PSi SERS substrates were utilized for quantitative analysis of NAGase levels found at clinically relevant BM concentrations while presenting the detection limit of 0.27 μM min−1. The optical performances were associated with the conventional standardized approach in terms of recovery values (85–98%), thus presenting sufficient adaptability to complex matrix analysis as whole milk.