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FFL-JA-Forskningsmidlene for jordbruk og matindustri

Development of a metabolomic- and immunological platform as a precision phenotyping tool for selection of pigs with improved resilience

Alternative title: Utvikling av metabolomikk- og immunologisk analyseplattform for avl på økt motstandsdyktighet og dyrevelferd hos gris

Awarded: NOK 3.8 mill.

Project Number:

281942

Project Period:

2018 - 2022

Location:

Partner countries:

For the international market for pig genetics we have focus on disease resistance and robustness as a response to challenging environments. On a global scale, the Norwegian production conditions are very good, which indirectly might cause limited international growth potential for Norsvin in robustness and disease resistance traits. This is because there are few situations in Norwegian pig production where these traits can be measured as the production conditions and animal health status are good. Hence is it difficult to measure traits that are related to stress tolerance to be included in the breeding program. Due to this, the project shall develop new sampling routines that are related to the existing phenotype recording at the boar station and stressful situations that the animals undergo. Then, new biomarkers related to stress response will be identified from blood samples and analysed. The project shall also investigate the potential of performing in vitro (laboratory) disease challenge tests on cells from healthy young animals. The aim of the project is to identify new methods to evaluate the animals' disease and stress responses, and to investigate genetic variation between animals for future implementation in Norsvins breeding program. Blood samples from the pigs were collected at two time points, one at introduction to test facilities and one at CT scan. Metabolome analysis of the blood from the pigs are analysed at SINTEF in Trondheim, with the aim of finding metabolic markers for stress tolerance, disease resistance and robustness. The metabolome analysis performed is a combination of targeted and non-targeted analysis. Targeted analysis includes analysis of amino acids and neurotransmitters / tryptophan metabolites. Non-targeted analysis was performed in part using LC-MS but mainly with ultra-high-resolution mass spectrometry. Ultra-high-resolution mass spectrometry provides a type of fingerprint of the metabolome with thousands of metabolite signals where the good mass accuracy and high resolution in spectra provide opportunities to identify the metabolites by means of matching against databases (such as human metabolome database (HMBD)). Multivariate data analysis is used to compare and group these complex spectra and to link metabolome analysis with other information that may indicate robustness and resistance to stress and disease. Results for tryptophan metabolism indicate that kynurenine is a potentially interesting marker. Meta-analysis for the ultra-high resolution spectra has started but not finalized. One of the reasons for this is that we are now, somewhat delayed due to the Covid shutdown during much of this reporting period, in the process of collecting significantly more blood samples for metabolic analyzes from the first sample point, ie at the entry of the test. This activity is now ongoing and blood samples are collected every week for shipment to Sintef, Trondheim. Our goal is to get metabolic data from 1000 pigs, with associated breeding data from the animal itself and close relatives. This will give us a very good data set for a high-resolution data analysis. One of the reasons for this is that we are now, somewhat delayed due to the Covid shutdown for much of this reporting period, in the process of collecting significantly more blood samples for metabolic analyzes from the first test time, ie at the robbery test station. This activity is now ongoing and blood samples are collected every week for shipment to Sintef, Trondheim. Our goal is to get metabolic data from 1000 pigs, with associated breeding data from the animal itself and relatives. This will give us a very good data set for a high-resolution data analysis. In addition to this, some of the boars, which were not selected as AI boars, were slaughtered routinely and macrophages from the lungs were in this context collected after slaughter from these individuals. The cells were stress tested with the viral disease PRRSV at Wageningen University in the Netherlands, but the survival of the cells during transport turned out to be too poor, as previously reported. After repeated attempts to gain access to an experimental population in the Netherlands, with Norwegian genetics, we have unfortunately given up this part of the project. If this part of the project is to be carried out, there must be a viral disease that can be studied in a lab in Norway.

The project is intended to lead to new phenotypes and models to create more resilient pigs, better adapted to the international market. Resilience is the ability of the animals to recover quickly from a wide range of disruptive challenges back to the original level of normal health, fertility, and production. According to this definition it is clear that in practice, measuring and then selecting for resilience in livestock is particularly challenging. This is even more challenging in the high health, and fortunate, environment present in Norway, where natural exposure to challenging conditions is limited. The underlying idea in this project is to fill main gaps that must be addressed to be able to breed properly for improved resilience. Norsvin has high ambitions for expansion on international markets. Sustainability and high animal welfare are fundamental elements of Norsvins breeding strategy that have helped to ensure that the ambitious international effort is well on the way to success. However, due to more incidences of diseases worldwide it is essential to develop breeding programs for selection of resilience in order to keep competiveness internationally and improve animal welfare in challenged environments. The main objective of this project is therefore to enhance the ability of pigs to resist and minimize the impact of disease by developing methods to select for disease and stress resilience in a high-health nucleus breeding populations. In this project our ambition is to address these issues by using novel state of the art metabolomic tools combined with immunologic methods and bioinformatics to examine blood plasma for potential novel and known metabolic and immunologic biomarkers, and their role related to resilience. Additionally, instead of direct disease challenge on pigs, the macrophages from blood and lung will be in-vitro challenged with the most important pathogens to obtain genetic variants useful for selection.

Funding scheme:

FFL-JA-Forskningsmidlene for jordbruk og matindustri