Mastitis Pathogen Identification Methods in Raw Milk
Mastitis infections in dairy herds can escalate from subclinical to clinical states within 24-48 hours, with somatic cell counts rising from baseline levels of 100,000 cells/mL to over 1 million cells/mL. Traditional culture-based identification methods require 24-72 hours for results, creating a critical gap between detection and targeted treatment that impacts both animal welfare and dairy economics.
The challenge lies in developing rapid, sensitive detection methods that can identify specific pathogens in raw milk samples while maintaining accuracy across varying somatic cell counts and milk composition profiles.
This page brings together solutions from recent research—including luminescence-based gram-positive detection systems, multiplex LAMP primer sets for simultaneous pathogen identification, integrated milking line monitoring systems, and novel immunoassay platforms. These and other approaches focus on providing actionable results within the critical early intervention window while maintaining compatibility with on-farm testing environments.
1. Substrate-Based Multiplex Immunoassay with Antigen-Specific Capture Elements and Colorimetric Detection System
THE UNIVERSITY OF MELBOURNE, 2024
Multiplex immunoassay for detecting Mycoplasma bovis infection in cattle using a substrate with multiple capture elements. Each capture element is specific to Mycoplasma bovis antigens, including proteins, lipoproteins, glycoproteins, and peptides, and can bind to target antibodies. The substrate also includes fiduciary markers, control elements, and a colorimetric detection system to monitor assay performance. The assay provides a comprehensive view of Mycoplasma bovis antigens and antibodies in milk or serum samples, enabling accurate detection of the pathogen in clinical samples.
2. Method for Luminescence-Based Detection of Gram Positive Bacteria in Bovine Milk Using Enzymatic Lysis and ATP Quantification
TRANSFORMATIVE TECHNOLOGIES, 2022
A rapid, in-stall method for detecting gram positive mastitis pathogens in bovine milk using luminescence. The method involves: 1) filtering raw milk to remove large particles, 2) rupturing gram positive bacteria in the milk using enzymes like lysostaphin and phage endolysins, 3) eliminating endogenous milk ATP to prevent interference, 4) adding luciferin-luciferase reagent to measure bacterial ATP, and 5) detecting luminescence to quantify gram positive bacteria. The method provides a "cow side" rapid diagnostic for mastitis within 10 minutes using simple reagents and devices that could significantly reduce antibiotic use and costs by enabling targeted treatment of gram positive mastitis.
3. Diagnostic Kit with Somatic Cell Count and Pathogen-Specific Fluorescence Analysis for Mastitis Detection
ARBILIM BIYOTEKNOLOJI SAN VE DIS TIC A S, 2019
A rapid diagnostic kit for mastitis that enables quick identification of pathogenic bacteria causing the disease, while preventing unnecessary antibiotic use. The diagnostic system combines a somatic cell count (SCC) measurement with a unique pathogen-specific fluorescence pattern analysis to provide rapid and accurate identification of mastitis-causing pathogens. The system uses a proprietary fluorescence-based detection method that is more sensitive than traditional somatic cell count methods, allowing for faster and more reliable identification of pathogenic bacteria compared to traditional methods.
4. Integrated Milking Line Somatic Cell Count Monitoring System with Continuous Cell Analysis
HI IMPACTS LTD, 2019
Real-time somatic cell count monitoring system for dairy farms during milking, enabling early detection of mastitis through continuous cell analysis. The system integrates into the milking process, employing a portable, automated somatic cell counter that continuously measures cell counts at various levels along the milking line. This real-time monitoring enables early intervention before clinical mastitis develops, significantly reducing treatment costs and improving herd health. The system operates in parallel with the milking process, with the cell counter positioned between the milk source and the collective tank, allowing continuous testing throughout the milking cycle.
5. Nucleotide Primer Set for Multiplex LAMP Detection of Eight Mastitis Pathogens
WISCONSIN ALUMNI RESEARCH FOUNDATION, 2015
A nucleotide primer set for simultaneous detection and identification of eight mastitis pathogens through loop-mediated isothermal DNA amplification (LAMP). The primer set enables simultaneous detection of E. coli, Staphylococcus aureus, Klebsiella pneumonia, Streptococcus uberis, Streptococcus dysgalactiae, Streptococcus agalactiae, Mycoplasma bovis, and coagulase-negative Staphylococci (CNS) tuf including subtypes S. epidermidis, S. chromogenes, and S. simulans.
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