Dr. Ivan Nastasijevic
Institute of Meat Hygiene and Technology – INMES

DIBMES (Development and integration of microfluidic biosensors as a tool for meat safety monitoring in farm-to-slaughterhouse continuum) is a project funded through the Proof of Concept program of the Innovation Fund of the Republic of Serbia, led by the Institute of Meat Hygiene and Technology – INMES with the participation of researchers from BioSense Institute.

The main goal of this project is to develop Point-Of-Care, user-friendly and low-cost sensors for health monitoring in the pig farming industry.

The meat supply chain continuum needs to be managed for sufficient levels of consumer protection. Healthy animals are an essential precondition for a safe food supply, since zoonotic diseases are a major threat to public health, as evidenced by the ongoing COVID-19 viral pandemic. Concerning livestock, early information about the prevalence of major health hazards of bacterial origin is of utmost importance for effective control on the farm. Early detection of Salmonella, Campylobacter, STEC, Listeria monocytogenes, etc. in fecal samples, as well as monitoring blood levels of metabolites relevant for animal welfare (hormones) and animal health (acute phase proteins), can provide high-level control in the pig farming industry. Integration of biosensors for pathogens and metabolites in the pig farm-to-slaughterhouse continuum is a practical and cost-efficient tool for early detection of signs that can jeopardize meat safety (Figure 1). Point-Of-Care DIBMES solutions are an advantage in comparison to the commonly used methods ELISA and RT-PCR, as well as other sensors available on the market which do not provide possibilities of early detection and require expensive equipment, trained personnel and significant time for sample transfer and analyses.

Point-Of-Care refers to the systems and tools that can be applied everywhere and anytime without transferring the sample to the laboratory. Therefore, this approach could facilitate monitoring of disease-related parameters in pigs and prevent the further spread of bacterial pathogens, including food (meat) borne agents. Also, the inclusion of the system can improve meat inspection and meat safety controls and with its implementation DIBMES system will be the primary tool for monitoring food safety parameters and will contribute to the modernization of veterinary inspection and meat safety assurance system.

DIBMES project will be conducted in two phases:

• phase 1 – with the emphasis on market analysis and evaluation of the most compatible systems for pathogens and metabolites and
• phase 2 – with the focus on the development of specific biosensors.

Also, this research will reduce the use of chemicals and the possibility of contamination by using microfluidic platforms and nanomaterials such as nanoparticles and dendrimers. Dendrimers have been known for their use in industry and biomedicine. Their successful application lies in their nano-scaled size and structure, as they resemble micelles with specific cavities. Another important property that makes them applicable is biocompatibility based on their functional groups. These properties enable the binding of more analyte or bioreceptor molecules, leading to signal enhancement.

IPANEMA and DIBMES

Point-Of-Care detection and monitoring of foodborne pathogens is being developed within the IPANEMA project, as well.

DIBMES project is led by the Principal Researcher – Dr. Ivan Nastasijevic (INMES). The research team is composed of both, INMES team (Dr. Ivan Nastasijevic, Dr. Radmila Mitrovic, Dr. Sasa Jankovic) – having expertise in detection & enumeration of zoonotic foodborne pathogens, metabolites, risk-based food (meat) safety management system, meat inspection and public health policy development, and several researchers from IPANEMA BioSense team (Dr. Ivana Gadjanski Stanic, Dr. Vasa Radonic, Ph.D. students Mila Djisalov and Teodora Knežić) – having expertise in design and fabrication of microfluidic sensing devices and molecular biology.

A multidisciplinary team aims to optimize different detection methods that can be integrated into microfluidic platforms and to develop innovative bottom-up approaches for the fabrication of multiplex (pathogen, animal welfare & animal health indicator), low-cost devices for Point-Of-Care Testing (POCT), on a pig farm and in a slaughterhouse. Also, in both projects electrochemical and impedimetric methods will be investigated for POCT use.

When it comes to differences in these approaches, the main one is related to the method and target of detection. In the IPANEMA project, the focus is on Nucleic Acid Testing (NAT), as opposed to the whole-cell pathogen detection, further enhanced with dendrimers, which is at the core of the DIBMES project, together with the use of specific monoclonal antibodies for detection of metabolites relevant for animal welfare and animal health. One of the main goals of IPANEMA is the development of user-friendly tools that will be based on rapid and low-cost isothermal nucleic acid amplification methods, facilitating efficient Point-Of-Care detection of pathogen nucleic acids (DNA and/or RNA) in the agri-food sector. These tools, as part of NAT, will be used to detect particular nucleic acid sequences from pathogens, if they are present in the analyzed samples. The main goal of the DIBMES project is to provide multiplex and low-cost POCT biosensor able to provide quantitative detection, with sufficient sensitivity, for the selected pathogen of major importance for pig meat safety (Salmonella) in the production environment (on farm, in a slaughterhouse) in the native sample matrix, as well as animal welfare indicators (selected hormones) and animal health status (selected acute-phase proteins).