DEVELOPMENT OF A NEW ANIMAL FEED SUPPLEMENT FOR IMPROVING ANIMALS PERFORMANCE

Acronim: ZINCOPPYEAST

PROJECT NUMBER : NR. 94 ⁄ 2017

Project Coordonator : AVICOLA LUMINA SA

PROJECT MANAGER :Drd. Ing. VASILE BUNDUC, director AVICOLA LUMINA SA

Financed by  UEFISCDI – Program  PN III: Cooperare Europeană și Internațională

Subprogram name: Subprogram 3.5. Other European and international initiatives and programs  EUREKA Tradiţional, EUREKA-Cluster Project , Eurostars

Project value: Link to the page UEFISCDI: www.uefiscdi.gov.ro

Budget RO 333.333 EURO

SELF FUNDING RO: 212.360 EURO

Total : 545.693,00 EURO

Start date of the contract: 03/11/2017

Date of finalization of the contract: 03/11/2020

Project end date (including the reporting period for economic effects: 03/11/2023

Consortium component:

Project Coordonator: AVICOLA LUMINA SA . Reprezentant Legal  Drd.ing.Vasile Bunduc . Director Proiect  Drd.ing.Vasile Bunduc            

Partner 1 : PHARMACORP INNOVATION. Reprezentant Legal  Dr.ing.Diana BARBULESCU . Responsabil de proiect P1 Dr.ing.Diana BARBULESCU. 

Partner2 : AGSIRA SRL . Reprezentant Legal  Ing.NEGRILA  Radian  Nicolae . Responsabil de proiect P 2 Ing.NEGRILA  Radian
Nicolae.

Partner3 :  INSTITUTUL NATIONAL DE CERCETARE-DEZVOLTARE PENTRU BIOLOGIE SI NUTRITIE ANIMALA –
IBNA BALOTESTI . Reprezentant Legal  Prof. Dr. Horia GROSU. Responsabil de proiect P3 Dr. Daniela MARIN .

Partner 4: Szent István University, Dept. Appl. Chem., Faculty of Food Science (HUNGARY)–P4 . Represented by  János Tőzsér, Dr. Rector and  Ferenc Magyar Chancellor. Jókainé Dr. Zsuzsanna Szatura. 

Partner 5 : ADEXGO Kft. Represented by Tamás Tóth Dr. (Managing Director). Dr. Hajnalka Hingyi .                                                            

Contact : Ing. VASILE BUNDUC – tel:0040 241 251 050, email: vasile.bunduc@avicolalumina.ro

Abstract:

The main scientific and technical goal of this project is to obtain innovative products: an animal feed supplement for new feed recipes, mainly based on a blend between spent brewer yeast from the brewers industries with the wet yeast biomass enriched with microelements (organic zinc), polyphenols and vitamins and also a blend between spent brewing yeast with inorganic zinc. The comparative test will be made in vivo on the laying hens and the antioxidant activity will be evaluated.

Through these activities the ZINCOPPYEAST consortium will contribute to a better characterization of the new products and will provide the scientific knowledge concerning the effect of the new product on the general health of the laying hens and their capacity to improve the response to stress. The valorisation of spent brewing yeast will be very important for the environment as it was considered a waste. Also, in this project, we will determine the homogeneity, shelf life and ingredient stability of the zinc, vitamin and yeast-containing supplement in different laying hen premixes, concentrates and compound feeds.

During the production of poultry feed several feed technological procedures are carried out which involve the use of high temperature (pelletizing-crumbing, hygienization, expansion). The organoleptic (sensory) characteristics of eggs produced by hens. The innovative bio-ingredients as new feed supplement for new feed receipts will be patented.

Through the fundamental objectives, the project will increase multidisciplinary transnational technological cooperation between technological research centers from countries such as Romania and Hungary.

Project objective:

The main scientific and technical goal of this project is to obtain innovative products: an animal feed supplements for new feed receipts, mainly based on blended between spent brewer’s yeast from the brewers industries with the wet yeast biomass enriched with microelements (organic zinc),polyphenols, and vitamins and also blended between spent brewing yeast with inorganic zinc.The comparative test will be made in vivo on the laying hens and the antioxidant activity will be evaluated.

The main objectives of the project are:

– To obtain a new feed bio-ingredient, an yeast biomass enriched in organic zinc, polyphenols and valorization of spent brewing yeast;

This new product based on blended between spent brewer’s yeast from the brewers industries with the wet yeast biomass enriched in zinc organic, vitamin, polyphenols will drive to an innovative feed supplement for laying hens.

This will be comparative tested with other new feed supplement based on

dry spent brewing yeast with inorganic zinc in the new recipe for laying hens.

-To identify and characterize the in vivo effects of the new feed supplement on laying hen’s health.

The project will investigate the effect of the new combinations, new mixtures between spent brewing yeast and  wine or with yeast consortium (wine and beer yeast) wet biomass enriched with Zn and polyphenols, in order to produce an innovative feed receipt, a new feed supplement which will be tested on the laying hens with further commercialization on the market.

Estinated results:

By using the  mentioned technologies in project the consortium will develop two categories of products:

– bioingredient/bioingredients, which are considered innovative for the feed supplement industry

– a new feed recipes for the laying hen’s feed.

 

Work plan stages

WP I. Development and new design of bio ingredients and proposed  new feed receipes.Nutrition program.  Dissemination of information. Design and proiection  of new bioingrediente new feed recipes and nutrition program design.- Partial

WP II. Development and new design of bio ingredients and proposed  new feed receipes.Nutrition program.  Dissemination of information Design and proiection  of new bioingrediente new feed recipes and nutrition program design. – Completion of activity  and  Development of new bio ingredients based on zinc and polyphenols . Experimental evaluation of them in vivo, to be used as a feed additive for laying hens antioxidant-enriched in zinc and polyphenols.(partial)

WP III SCALE UP  at pilot level. Testing of new feed receipes at pilot level) (partial)

WP  IV  Testing of new feed receipes at pilot level and  scale up  at industrial level

WP V Economic effect

WP VI Economic effect

WP  VIII  Economic effect

Stage 1. Development and design of new bio ingredients, new feed recipes and designing the nutrition program -Partially- Design of the project web page. Dissemination of information. (3.11.2017-31.12.2017).

In stage 1 of the “ZINCOPPYEAST” project, more than 30 colonies of wine and beer yeast were isolated. The majority have been tested for the influence of the carbon source used both for the development of inoculum and for the fermentation medium over the biomass development.

– the influence of the zinc on the biomass development was also tested;

– the technical specification for the beer yeast was made regarding the protein content, and the total number of germs;

– the main objective of this step was to optimize the extraction methods of total polyphenols  from yeast residues and to characterize some yeast residues from the beer industry in terms of content in total polyphenols.

– also, new nutritional solutions that use the yeast in the feed of laying hens have been achieved and a nutrition program was proposed;

– the project’s web page was created.

 

 

 

 

 

 

 

Wine and beer yeast colonies

 

 

 

 

 

 

 

 

 

 

Isolated wine and beer yeast colonies

Activity I.1. PARTIALIsolation of wine and beer yeast strains in order to be used alone or in a consortium for obtaining new bioingredients based on yeast consortium biomass enriched in zinc, that will be used to valorise  through  mixture with the used beer yeast yielding a product based on organic zinc, polyphenols,  and  bioingredients based on yeast biomass rich in zinc –

Activity I.2 Elaboration of nutrition program- PARTIAL

Activity 1.3 Dissemination of information Project Website

STAGE II. Elaboration and design of new bioingredients, new feed recipes and nutrition program design – (FINALIZATION) and Development of new bioingredients based on biomass of zinc / organic zinc yeasts, polyphenols and their experimental evaluation, in vivo, to be used as a supplement with an antioxidant role in the feed laying hens, enriched in zinc and polyphenols. Results dissemination – article / presentation at a conference (Partial)

ACTIVITIES OF STAGE 2

Act 2.1 – Isolation of brewer’s yeast strains to obtain new bioingredients based on zinc and polyphenols enriched yeasts, to be used by blending with the spent beer yeast. COMPLETION

Act 2.2 – Elaboration of nutrition program – Completion

Act 2.3 – Selection of isolated wine and beer yeasts and identification of those selected for obtaining bioingredients based on zinc, polyphenols yeast biomass that uses spent beer yeast and the effect on performances, health status, the oxidative status and polyphenol content at hens. Selection of raw materials for new formulas for obtaining feed additives with the help of innovative bioingredients based on yeasts enriched in zinc and polyphenols. Screening bioproducts. -Setting the parameters that define the effectiveness of new recipes.

Act 2.4 – Optimizing processes and new feed recipes at pilot level and verifying through nutrition experiments the effectiveness of new optimal recipes used to produce eggs enriched with zinc,  polyphenols. Publication

Act 2.5 – Study of the effect of bio-ingredients on performances, health status on oxidative status and polyphenol content in laying hens – partially

The main objective of this step was to isolate and identify yeast strains by molecular sequencing in order to obtain new bio-ingredients based on zinc-enriched yeasts (P1) mixed with the spent brewing yeast from (P2) obtaining the product (ZINCOPPYEAST), tested by the Project Coordinator (CO) in the feed of laying hens and analyzed by partner (P3) in terms of polyphenol content and antioxidant activity.

Analyzes of total zinc, organic zinc and polyphenols were made by partner P4.

By scale-up by optimization, the zinc enriched yeast mixture with the yeast enriched with zinc with spent beer yeast was made obtaining a mg / kg yeast content of  Organic Zn> 65, a total of 85 zinc total mg / kg of yeast.

-ZINCOPPYEAST presents, after the mixture, a total of 250 mg / 100g total polyphenols (referred to gallic acid).

-The total zinc content of the ZINCOPPYEAST feed was 160 mg / kg of yeast Zn Total and> 140 mg / kg  Organic Zn yeast.

-The polyphenol content of the ZINCOPPYEAST feed was 170 mg / 100g

– total polyphenol analysis determined a significant increase of polyphenols in the batch (ZINCOPPYEAST)

– Optimized recipes for the feed of laying hens have been achieved by CO.

– Different premixes and concentrate recipes were made by partner P5.

 

 

 

 

 

 

 

 

Selected  yeasts for ZINCOPPYEAST’biongredient obtainment

WP III DEVELOPMENT OF NEW BIOINGREDIENTS (FINALIZATION) AND VALIDATION AT PILOT LEVEL OF BIOINGREDIENTS AND FODDER RECIPES (PARTIALLY) Dissemination of results. Presentation of the paper at a conference / published article (01.01.2019-31.12.2019)

Tasks for WP.3

Task 3.1Optimization of processes and new feed recipes at pilot level and Verification through carried out nutrition experiments of the efficiency of the new optimal recipes used to obtain eggs enriched in zinc, polyphenols. Publishing – completion of activity

Task 3.2Study of the effect of bioingredients over the performance and health status on oxidative status and polyphenol content in laying hens – completion of activity

Task 3.3.Validation of bioingredients in order to obtain eggs enriched in organic zinc, polyphenols. Verification of the effect of the use of feed on the health of the laying hens and the microbiological control of the eggs.

Task 3.4.Pilot validation of bioingredients

Task 3.5.Study of the effect of the bioingredients on the performances, the health status on the oxidative status and the content of polyphenols in the laying hens. Microbiological control of the eggs. Partially

Task 3.6.Pilot validation of fodder recipes for laying hens at pilot level. Partially

The main objective of this stage was to complete the optimization of the technologies in order to obtain new bioingredients based on yeasts enriched in zinc in combination with the spent brewing yeast (ZINCOPPYEAST) and the optimization of fodder recipes. Another objective was to demonstrate and validate both technologies and feed recipes (partially).

Avicola Lumina has developed feed recipes that have been tested on 5 lots of 7000 layen hens / lot. The results were disseminated both at conferences and as an article.

Partner P 1 together with Partner P2 participated in obtaining the Zincoppyeast product. The yeast product enriched in organic zinc obtained by P1 was mixed with the spent brewing yeast and dried on a large scale on a tumble dryer by partner P2. Yeast samples were delivered to determine the content of polyphenols, total zinc and organic zinc to Hungary’s P4 Partner.

– The effect of technological procedures for premixes and concentrated on the stability of yeast-based products was analyzed by P5, partner from Hungary.

Partner P3 performed biochemical analyzes for blood samples/ each experimental layen hens batch. Various premix recipes were tested by Partner P5.

In conclusion, products based on yeasts enriched in organic zinc and polyphenols were made

by P1 in collaboration with P2, which have been studied in different ratios in new feed recipes by the coordinator (CO).

Following the optimization of the process and the demonstration, a content of 78% of organic zinc in the bioingredient based on yeasts enriched in zinc was revealed.

The highest polyphenol content was 246 mg/100g gallic acid for the bio-ingredients obtained through fermentations with yeast consortia (wine and beer).

The Zincoppyeast mixture was analyzed with regard to the content of Zinc by P4 77mg / kg Zn. This Zincoppyeast was tested by the CO in the recipe with 2%  Zincoppyeast.

Zincopyeast polyphenols concentration was 620 mg / 100g gallic acid.

Zincoppyeast: regarding the Zinc content it was analyzed by P4; 95 mg / kg Zinc was found for this mixture and was tested by the CO in the feed recipe with 3% Zincoppyeast.

 

 

 

 

 

 

 

 

 

Experimental lots of new fodder recipes obtained with different quantity of ZINCOPPYEAST, inorganic zinc, chelated zinc. Lots of eggs obtained after testing

  • A higher accumulation of Zn was observed in eggs from laying hens fed with „ZYNCOPPYEAST”.
  • Feeding hens with diets with spent brewing yeast mixed with Zn-enriched yeast („ZINCOPPYEAST”) can improve the egg quality through an increase of the zinc con tent (Mihály Dernovics, Vasile Bunduc, et al, 2019).
  • Mihály Dernovics, Vasile Bunduc, Daniela E.Marin, Ana Maria Pertea ,Zsuzsa Jókai, Márta Üveges,Simona Ioana Marinescu, Mihaela Begea, László Abrankó, Radian Nicolae Negrila, Hajnalka Hingyi ,Iuliana Diana Bărbulescu, Study of the effect of zinc-enriched spent brewing yeast on the performance characteristics and health status of laying hens, 7th International Symposium on Metallomics, 30 June – 3 July 2019, prezentare poster Warsaw | POLAND, BOOK OF ABSTRACTS, pp 83, ISBN: 978-83-950720-0-0

 

WP IV Testing of new feed receipes at PILOT LEVEL AND SCALE UP AT INDUSTRIAL LEVEL regarding testing of new feed receipes on layen hens

Task 4.1. Tests for obtaining and analyzing bioingredientes and final testing of the feed receipes for the laying hens
Act 4.1 – Study of the effect of bioingredients performance, health status on oxidative status and polyphenol content in laying hens
-Microbiological control of eggs. The polyphenol content will be achieved by partner P5 from Hungary
-Study of the effect of bioingredients on performance, health on oxidative status and polyphenol content in laying hens
-Microbiological control of eggs.
-The polyphenol content will be made by partner P5 from Hungary

Task 4.2. Validation of of the bioingredients and feed receipes for layen hens
Validation of pilot level for feed receipes for layen hens

Task 4.3.Tests for obtaining and analyzing bioingredientes and final testing of the feed receipes for the laying hens
Obtaining bioingredients for testing
Testing of feed networks at industrial level using the new bio-ingredients that contain spent brewing yeast

Task IV.4. Capitalizing on the results. Check the organoleptic and nutritional qualities of eggs by determining chemical nutrients
Setting parameters defining the quality of the new type of eggs (Test and verify organoleptic and nutritional qualities of eggs and keeping them in long time (shelf).
Evaluation of the organoleptic characteristics of eggs with human and instrumental sensory analysis – preliminary phase: feeding of layer hens with the developed products; sensory analyses (Developed supplements (ZINCOPPYEAST) can influence the important organoleptic (sensory) characteristics of eggs produced by hens. In the frame of the study the possible changes in the fragrance and flavour of eggs would be examined by qualified reviewers and with a technology that is considered novel (as known sensory)
-Verification of the organoleptic and nutritional qualities of eggs by chemical determination of nutrients
-Establishing the defining parameters for the quality of the new type of eggs
Food safety studies of bioproducts, microbiological control of eggs.

Activity IV.5.Industrial level validation
Technical and economical evaluation based on yeast bioingredients with zinc and polyphenols
– Production, at industrial level, of zinc-enriched eggs using the patented solution / solutions that include used brewer’s yeast and bio-ingredients enriched in zinc / organic zinc, polyphenols, Mos, used as antioxidant
-Safety of new feed recipes

Task IV.6. Submission of a strain / samples of wine and / or beer strain strains that have been molecularly identified to a Collection of Microorganism Cultures recognized by the IDA (international Depositary Authority)

Activity IV.7. Protecting intellectual property rights for the new bioingredients;
Design documentation and filing the patent application OSIM minimum nutritional solution of the two tested through the project
• The main objective of this stage was to validate the technologies in order to obtain new bio-ingredients based on zinc-enriched yeasts mixed with used brewer’s yeast (ZINCOPPYEAST) and to validate feed recipes.

• Blood samples were taken from laying hens which were fed with zinc feed from zincoppyeast and zinc oxide and were sent to IBNA where enzymological and biochemical analyzes were performed on the blood and microbiological analyzes on the eggs.

• Blood samples were collected by Avicola Lumina and sent to IBNA Balotești immediately after collection. A total of 90 blood samples were received from the following groups: Control, Experimental 1 with ZnO and Experimental 2 with Zincoopyeast

-Serum analyzes regarding the energetic, protein, mineral and enzymatic metabolic profile showed that the blood parameters fall within the normal values for the age of the animals.
-As observed, supplementation of ZINCOPPYEAST yeast feed resulted in a decrease in plasma cholesterol and triglyceride levels. As a result, the decrease in the concentration of cholesterol in the plasma can lead to a decrease in the concentration of cholesterol in the eggs (yolk).
-Glucose concentration at the end of the experiment decreased in the groups tested with Zincoppyeast compared to the control
– No development of Samonella colonies was observed after the cultivation of egg samples from the experimental batches. Zinc from ZINCOPPYEAST and plants (control) is better absorbed than zinc in inorganic form as zinc oxide with yeast and zinc chelate.

• The yeast product enriched in organic zinc obtained by Pharmacorp Innovation was mixed with used brewer’s yeast and was dried on a large scale on a roller dryer by partner Agsira and ZINCOPPYEAST was obtained
• ZINCOPPYEAST EXPERIMENTS – Development of new bioingredients based on zinc biomass enriched in zinc / organic zinc, polyphenols and their experimental evaluation, in vivo to be used as an antioxidant supplement in feed chickens enriched in zinc, polyphenols were made in experimental trials by Avicola Lumina
• Yeast samples were delivered to determine the content of polyphenols, total zinc and organic zinc to the Partner Szent István University (acting entity: Faculty of Food Science, Department of Applied Chemistry) from Hungary.
• Other samples of Zincoppyeast were delivered for testing to partner Adexgo Ipari
• Analytical and microbiological specifications were performed for the Zincoppyeast samples.
Experimental recipes were developed and validated.

The results were disseminated both at conferences and in the form of an article:

1. BEGEA, Iuliana Diana BĂRBULESCU*, Mihaela Violeta GHICA, Zsuzsa JÓKAI, Simona-Ioana (MARINESCU) MĂRCULESCU, Márta ÜVEGES, Alexandru Ionuţ CÎRÎC, Vasile BUNDUC, Hajnalka HINGYI, Sanziana Ioana PRUNDIANU, Radian Nicolae NEGRILĂ, POST-FERMENTATION BREWING YEAST AS A FEED SOLUTION FOR LAYING HENS, Scientific Papers. Series D. Animal Science. Vol. LXIII, No. 1, 2020
ISSN 2285-5750; ISSN CD-ROM 2285-5769; ISSN Online 2393-2260; ISSN-L 2285-5750 –

2. UTILIZAREA UNOR FORME DIFERITE DE ZINC LA OBȚINEREA DE INGREDIENTE FURAJERE CU APLICATII ÎN NUTRITIA GAINILOR OUĂTOARE –REVIEW,Bărbulescu I.D., Begea M.,Jókai Zs, Cîrîc A.I., Mărculescu (Marinescu) S.I,Üveges M., Bunduc V., Hajnalka H), Matei P.M.,Prundianu S.I –articol in extenso acceptat la la INTERNATIONAL SYMPOSIUM, 10thEdition ISB-INMA TECH INTERNATIONAL SYMPOSIUM, 10thEdition Jubilee Edition -30 octombrie 2020

3.UTILIZAREA UNOR FORME DIFERITE DE ZINC LA OBȚINEREA DE INGREDIENTE FURAJERE CU APLICATII ÎN NUTRITIA GAINILOR OUĂTOARE –REVIEW,Bărbulescu I.D., Begea M.,Jókai Zs, Cîrîc A.I., Mărculescu (Marinescu) S.I,Üveges M., Bunduc V., Hajnalka H), Matei P.M.,Prundianu S.I
Prezentare orala la sectiunea
SECTION 3 -FOOD INDUSTRY(ORAL PRESENTATION) la INTERNATIONAL SYMPOSIUM, 10thEdition ISB-INMA TECH INTERNATIONAL SYMPOSIUM, 10thEdition Jubilee Edition -30 octombrie 2020

– Submission of a strain of wine yeast strain that has been molecularly identified in a Collection of Microorganism Cultures recognized by IDA, prior to the filing of patent applications and Accession number 43P was obtained

• The patent application A00684 “Product based on yeast biomass enriched in organic zinc and process of obtaining” was filed with OSIM on 30.10.2020

• The patent application was submitted to OSIM: “New nutritional solutions by obtaining feed recipes made with different forms of organic and inorganic zinc, with use in laying hen feed” with no.A00690 / 2.11.2020