PN-III-P4-PCE-2021-0992

PN-III-P4-PCE-2021-0992

Project PCE40

Project Title: Evaluation of the efficiency of agro-industrial by-products with complex matrices of bioactive compounds to replace ZnO; from in vitro to in vivo.

Acronym: Zinc-botanicals

Funding Unit: Executive Unit for Financing Higher Education, Research, Development and Innovation (UEFISCDI)

Project Code: PN-III-P4-PCE-2021-0992

Project Summary:

The importance of zinc oxide in diarrhea control has increased after the ban on the use of antibiotics in animal feed (2006). However, due to the negative impact that zinc can have on the environment, the EU has proposed to withdraw ZnO intervention premixes by 2023. Alternative solutions are needed. In this context, the objective of the project is to investigate the potential of compounds with bactericidal and anti-inflammatory activity derived from oleaginous by-products (mustard) to replace zinc oxide by reducing intestinal inflammation, diarrhea and oxidative stress in piglets after weaning. The project includes three stages and 21 experimental activities. Phase 1 will focus on obtaining and analytical characterization of the mass of compounds with recognized bioactive properties of mustard by-products little investigated in pigs. Phase 2 will include in vitro studies on cell cultures and ex-vivo (explant), in which the efficacy of mustard by-products to replace ZnO will be investigated in a cellular micro-system. Phase 3 will include in vivo studies. The experiments will be performed on piglets after weaning in which the activity of bioactive compounds from mustard compared to ZnO will be investigated. The results will contribute to: i) the development of a new nutritional formula effective in reducing the weaning crisis; (ii) exploitation of mustard by-products as a new natural ZnO replacement agent (iii) understanding the mechanisms of action of bioactive compounds.

Project objective:

The objective of this project is to develop a feed product with antimicrobial potential, including mustard seed meal, a by-product of the oil industry, and to test its ability to replace zinc oxide used to combat microbial infections in farm animals, which will be banned in the EU from 2023. The ability of bioactive compounds from mustard meal to reduce transient intestinal inflammation and oxidative stress in piglets during the weaning period will be investigated.

RESEARCH TEAM

Ionelia Țăranu, project coordinator

Daniela Eliza Marin, project team member

Gina Cecilia Pistol, project team member

Valeria Cristina Bulgaru (Procudin), project team member

Iulian Alexandru Grosu, project team member

Ana-Maria Perțea, project team member

Andrei-Cristian Anghel, project team member

The PCE40 project approached and implementing the principles of circular bioeconomy a very important topic of the European Commission programmes through the recycling of waste.

The mains objectives:

• obtaining a new feed product including mustard seed meal enriched in bioactive compounds with an antibacterial effect, efficient in reducing transient intestinal inflammation caused by the post-weaning crisis in piglets,
• obtaining a new feed product as a viable alternative to zinc oxide (ZnO), banned from 2023 in EU.

Using an experimental contamination model with E.coli-LPS, the project investigated the antibacterial potential of bioactive compounds from mustard by-product (mustard seed meal) as an alternative nutritional solution to replace zinc oxide used in a high-concentration in feed for piglets after weaning as intestinal protector, banned from 2023.

The results of in vitro and ex vivo studies demonstrated that the LPS endotoxin is not only a distinct promoter of inflammation, but also has a pronounced effect on the intestinal epithelium and its barrier functions, such as cell renewal, integrity and permeability.

-E.coli-LPS decreased cell proliferation, increased apoptosis and cell arrest in the G0/G1 phase of the cell cycle.

-LPS altered epithelial permeability by decreasing transepithelial electrical resistance (TEER) and junctional protein expression and by increasing lactate dehydrogenase (LDH) activity, a marker of cell membrane damage.

-LPS exacerbated the gene expression of Toll-like receptors, markers of the innate immune response, the first molecules to respond to bacterial infections.

-The mechanism by which LPS induces all these negative effects at the cellular level involved signalling molecules such as protein kinases PKC (A and B), p38 MAPK and nuclear transcription factor-kB (NF-kB) that were overexpressed.

- LPS treatment induced an increase in IgA and IgM immunoglobulin concentrations in the plasma of piglets inoculated with LPS, indicating an activation of the immune response to counteract the effects of bacterial endotoxin. Feeding with mustard meal or ZnO rebalanced immunoglobulin concentrations toward to control.

-As in vitro, LPS induced an increase in NO and free radical synthesis in the plasma of piglets exposed to bacterial endotoxin, which was counteracted by feeding with mustard meal and zinc oxide.

In vivo results showed that mustard meal has the potential to reduce inflammation and oxidative stress induced by LPS in piglets after weaning. These results suggest that mustard by-product (seed meal) is a promising source in counteracting the harmful effect of bacterial toxins.

Indicators of achieved results

 

ISI articles:

1. Ionelia Taranu, Valeria Cristina Bulgaru, Gina Cecilia Pistol, Mihai Alexandru Gras, Ana-Maria Ciupitu, Iulian Alexandru Grosu, Daniela Eliza Marin.

Extract derived from mustard meal by-product an adequate alternative to zinc oxide in alleviating the negative effect induced by E. coli-LPS on epithelial intestinal barrier. Article accepted in International Journal of Molecular Sciences 2024, Manuscript ID: ijms-3352315.

2. Iulian Alexandru Grosu, Gina Cecilia Pistol, Daniela Eliza Marin, Ionelia Taranu. The Potential of Mustard Seed Meal to Alleviate LPS-Induced Microbial Shifts in Weaned Piglets. Published in Agriculture, 2024, Manuscript ID: agriculture-3337008.
3. Andrei Cristian Anghel, Ionelia Taranu, Alina Ontanu. Optimizing microwave-assisted extraction of polyphenols from mustard seed meal. Pharmacia 71(6), 1174-1180, https://doi.org/10.31925/farmacia.2023.6.7.
4. Andrei Cristian Anghel, Ionelia Țăranu, Alina Orțan, Narcisa Elena Băbeanu. Antimicrobial activity of mustard seed meal polyphenols. Article submitted in Scientific papers, series D, Animal Science, 2024.

BDI Articles:

Andrei Cristian Anghel, Ionelia Taranu. Antimicrobial potential of polyphenols obtained from agro-industrial by-products. Published in Archiva Zootechnica-26(2), 96-115, 2023 DOI: 10.2478/azibna-2023-0017.

Conferences:

1. Gina Cecilia Pistol, Valeria Cristina Bulgaru, Iulian Alexandru Grosu and Ionelia Țăranu. The Modulation of Intestinal Inflammation by a Mustard By-Product in E. Coli-Lypopolysaccharide – Challenged Weaned Piglets. Oral presentation. Life Sciences – Today for Tomorrow International Congress, Iasi, Romania, October 24-25, 2024.
2. Andrei Cristian Anghel, Ionelia TARANU, Alina ORTAN Optimizing microwave-assisted extraction of polyphenols from mustard seed meal. Oral presentation. 17th International symposium of animal biology and nutrition, Balotesti, Romania, September 29, 2023.
3. Ionelia Taranu, Valeria Cristina Bulgaru, Andrei Cristian Anghel, Gina Cecilia Pistol, Alexandra Oancea. Investigating the potential of mustard seed as an alternative source of bioactive compounds in feed for piglets. Poster. Regional Conference of European Federation of Animal Science, EAAP, Nitra, Slovakia, April 26-28, 2024
4. Ionelia Taranu, Gina Cecilia Pistol, Valeria Cristina Bulgaru, Daniela Eliza Marin. Assessing the capacity of mustard by-product to replace medicinal zinc oxide in co-culture cells. Poster. Multidisciplinary Conference on Sustainable Development Section: Animal Resources Bioengineering, Timisoara, Romania, May 25-26, 2023
5. Daniela Eliza Marin, Ionelia Taranu, Valeria Cristina, Andrei Cristian Anghel, Ana Cismileanu. Study concerning the potential use of vegetable waste during the weaning stress in order to improve the gut health of weaned piglets. Poster. 14th European Nutrition Conference FENS 2023 Belgrade, Serbia, November 14-17, 2023
6. Ionelia TARANU, Gina PISTOL, Cristina BULGARU, Iulian GROSU, Ana Maria PERTEA, Daniela MARIN. Ex vivo study on pig jejunum explants exposed to E. coli-LPS and hydrogen peroxide to evaluate the alleviating effect of a mustard by-product extract used as an alternative to medicinal zinc oxide. Poster. 2nd EAAP Regional Meeting, Nicosia, Cyprus, 24th - 26th April 2024.
7. Iulian Alexandru Grosu, Gina Pistol, Daniela Marin, Ionelia Taranu. The effects of a diet containing mustard seeds meal on the microbiota in post-weaning piglets. Poster. Multidisciplinary Conference on Sustainable Development, Timisoara, Romania, 30-31 May 2024
8. Ionelia Taranu, Gina Cecilia Pistol, Daniela Marin, Cristina Bulgaru, Ana-Maria Pertea. Nutritional solutions adapted to the circular bioeconomy to counteract inflammation in piglets after weaning by using mustard seed as a replacer of zinc oxide. Poster. 12th International Conference Agriculture & Food - Burgas, Bulgaria, 12-15 August 2024
9. Ionelia Țăranu, Gina Cecilia Pistol, Cristina Procudin, Iulian Alexandru Grosu, Ana-Maria Ciupitu and Daniela Marin. Polyphenolic extract derived from mustard meal by-product an adequate alternative to zinc oxide in alleviating the negative effect induced by coli LPS on epithelial intestinal barrier. Poster. Life Sciences – Today for Tomorrow International Congress, Iasi, Romania, October 24-25, 2024
10. Mihail Alexandru Gras, Gina Cecilia Pistol, Iulian Alexandru Grosu and Ionelia Țăranu. Mustard by-product prevented the e coli lipopolysaccharide - induced oxidative stress in piglets after weaning. Poster. Life Sciences – Today for Tomorrow International Congress, Iasi, Romania, October 24-25, 2024.

Dissemination Articles

 

1. Ionelia Taranu. Composition of bioactive compounds of mustard meal. Lumea satului

Authors: Ionelia Taranu

2. Ionelia Taranu. Composition of bioactive compounds of mustard meal; use of mustard meal in piglets after weaning. Radio show: Antena Satelor.