MOET Si Bibliografie Vaci

Procedia Economics and Finance 8 ( 2014 ) 696 703

2212-5671 2014 The Authors. Published by Elsevier B.V.Selection and peer-review under responsibility of the Organizing Committee of ESPERA 2013doi: 10.1016/S2212-5671(14)00147-6

ScienceDirect

1st International Conference 'Economic Scientific Research - Theoretical, Empirical and Practical Approaches', ESPERA 2013

The bio-economic impact of the MOET program for the development of vulnerable genetic resources through eco-innovative

reproduction biotechnologies

George Florea Toba*, Marcel Theodor Paraschivescua, Alexandru Bogdana, Mariana Sandua

aRomanian Academy, National Institute for Economic Research "Costin C. Kiritescu", Centre of Studies and Research in Agricultural and Forestry Biodiversity, Bucharest, Romania

Abstract

MOET Program (Multiple Ovulation Embryo Transfer) has more important bioeconomy significance by biodiversity profile metabolic and hormonal of the donor and recipient female of transferred embryos and allowed access to animal genetic biotechnologies, with low costs and minimal risk of introducing diseases towards imports with live animals. The authors present synthesis postdoctoral research in program POSTDRU ID 63258, included in the Research Plan of the National Institute of Economic Research Costin C Kiriescu on 2013 with original contributions to the application of systems MOET (with open circuit, closed circuit and Nucleus herd) which allowed some elite farms (ex.: Pantelimon, Otopeni - Ilfov County, Ardud Satu Mare County and Matca Galati County) to obtain genetic progress accelerated, evaluation by different methods biostatistics trough BLUP (Best Linear Unbiased Prediction).MOET Program was apply in Romania by imported embryos of SUA from Dr. Edwin Robertson of Harrogate Genetic International achieved a pregnancy rate after E.T. between 42-47%. Following the application of MOET program in this research made banks to preserve genetic resources with major risk of breed Pinzgauer and breed Grey Steppe in Country Haeg -Retezat, Dancu- Iasi and CSCBAS-Bucharest by risk management of the reduction extinction of these animal breeds. In this paper the authors develop the idea of the establishment of gene banks based on their own experience and especially mobile containers laboratories have been described in a scientific paper presented at the annual meeting of D.A.G.E.N.E. (Danubian Countries Alliance for Conservation of Genes in Animal Species) of Timisoara this year

* Corresponding author. Tel.: +40723.504.339..

E-mail address: tobageorgeflorea@ yahoo.com

Available online at www.sciencedirect.com

Open access under CC BY-NC-ND license.

697 George Florea Tob et al. / Procedia Economics and Finance 8 ( 2014 ) 696 703

2014 The Authors. Published by Elsevier B.V. Selection and peer-review under responsibility of the Organizing Committee of ESPERA 2013.

Keywords: MOET program, bioeconomic significances, farm;

Introduction

The MOET program (Multiple Ovulation Embrio Transfer) has bioeconomic significances by the biodiversity of

the metabolic and hormonal profile of embryon donor and recipient female and allowed the access to animal genetic biotechnologies, with low costs and a minimum risk of introducing diseases towards imports with live animals.

The authors present synthesis postdoctoral research in program POSTDRU ID 63258, included in the Research Plan of the National Institute of Economic Research Costin C Kiriescu on 2013 with original contributions to the application of systems MOET (with open circuit, closed circuit and Nucleus herd) which allowed some elite farms (ex.: Pantelimon, Otopeni - Ilfov County, Ardud Satu Mare County and Matca Galati County) to obtain genetic progress accelerated, evaluation by different methods biostatistics through BLUP (Best Linear Unbiased Prediction).

To highlight the fact that the use of BLUP for bull related female allows the obtaining of differences between complete brothers of a family only if their daughters also are included among such relatives, as the collateral relatives are the same (M.Th. Paraschivescu, 2010)

The use of MOET as potential instrument for the improvement of milch cows in a nucleus heads was imagined for the first time, in detail, in 1983, by Nicholas and Smith, quoted by Ruane J. and Thomson R (Ruane, J, 1990). They describe two types of MOET charts: the juvenile charte, with donor selection before the first insemination and the adult chart with the donor selection according to their pwn performances regarding milk production in control days. The aspects aiming at the bio-economic aspect of the MOET program for the development of the zootechnical genetic resources vulnerable through eco-innovative reproduction biotechnologies, are approached by the Manager of the Postdoctoral School, Prof.univ.dr. Alexandru T. Bogdan, c.m. of the Romanian Academy, in several scientific works (Bogdan A.T. 2009, 2011).

In this paper, the authors develop the idea of the establishment of a gene bank based on their own experience and, in particular, on the experience of the mobile container laboratories, which were described in a scientific work presented at the annual meeting of D.A.G.E.N.E. in Timioara, this year, and published in The 12 th International Symposium Prospects for the 3rd Millennium Agriculture al U.S.A.M.V.-Cluj Napoca ( Toba G. F . 2013).

In our country an estimation work of the probable effects of the application of MOET for the improvement of a milk producing cattle population was published in 1989 by Prof.dr.ing. Paraschivescu M. and collaborators (Paraschivescu, M, 1989). In the paper, it is estimated the possible genetic progress through I.A. or through E.T., as compared to that achievable through natural freshening. In the same year, 1989, Prof.dr.ing. Paraschivescu M. and collaborators imagine and publish a technological project, hypothetical, for a MOET farm, in fact a nucleus of the population where, by reproduction in closed circuit it is achieved the selection of donor adults resulted from E.T. young beef used successively as receivers, as heifers and as primiparae. The chart of the incubation farm for taurine, imagined by Prof.dr.ing .M.Paraschivescu and collaborators, does not isolate the nucleus from a genetic point of view from the population to which it belongs but isolates it from a sanitary- veterinary point of view as the reproduction is assured through E.T. and without the access of other animals in the farms heads. The name of incubation expresses the fact that the purpose of this farm is to provide recipients for the embryo-transfer. The character of veterinary isolation of the reproduction results from the fact that the recipients are the young beef coming from embryo-transfer. As the young beef cannot maintain the womb heads it is also necessary for the primiparae to be used as recipients, as the work is with non-sex embryopns and a female cannot assure its refreshening but from two gestations which it must transit.

This restriction, without which the farm cannot isolate itself from a veterinary point of view, results in the fact that all the young beef coming from embryo-transfer are evident in their milk production at the first lactation, which

2014 The Authors. Published by Elsevier B.V.Selection and peer-review under responsibility of the Organizing Committee of ESPERA 2013

Open access under CC BY-NC-ND license.


means that we can select among them embryopn donors with the desired selection intensity. Thus, such a herd becomes a MOET farm with closed circuit reproduction.

The first advantage of this type of farm is that of a severe veterinary prophylaxis. The only contamination source of the farm remains in a first stage the embryon and subsequently the seminal materual inseminating the donors. The second advantage is that, from the improvement point of view, the herd remains within the populations gearing, enjoying the effects of the high selection intensity allowed by a numerous population

The farm has, as main product, male testing calves, necessary for the artificial insemination organizations. Secondary to this purpose there is also a second merchandise, namely cows on second lactation, with valuable origin and well known production performances, of which some may become bull mothers. The maintenance of bull mothers in the nucleus farm is not excluded.

The dimensioning of such a farm is more rigid than that presented above and it is adaptable to the fodder resources owned by an interested entrepreneur.

In order to set up a MOET herd, of the type Nucleus Herd or with closed circuit, we can start with any type of recipients which hope to have a good gynecological situation, generally the young beef, but we must procure first rank genetic value embryons. From the descendency obtained, the best female verified by its own production performances and corporal performances the donor females are selected, stimulated for super-ovulation, inseminated with seminal material from the best available bulls and washed for the sampling of embryons, which are then frozen. These operations are repeated to have multiple ovulations. Thus, the incubation farm proposed by Prof.dr.ing. M.Paraschivescu becomes a closed circuit MOET farm where only young beef and primiparae from its own breeder are used as recipients, not to allow the penetration of foreign animals in the farm heads. Some progress regarding the MOET performances could be obtained by sexing the spermatozoa from the seminal material inseminating the donors or by sexing the embryons before being frozen (Paraschivescu, M.; Tibr, Dana,1989 ). The E.T. young beef resulted are retained in the units own breeder, subject to corporal type reliability and production control and, depending on the case, will be used as donors, being inseminated nominalized with seminal material from tested bulls or, others, with testing bulls for obtaining collateral relatives. If the young beef entries allow it, certain cows will be delivered outside the farm. An interesting development of MOET, privately considered, applicable within the Nucleus farms or closed circuit farms could be the use of the young beef as donors to be tested after the milk production of their daughters, which would enormously increase the precision of the selection of bull mothers meant for use in the the I.A. The selection accuracy would increase as the phenotype value measured is no longer necessary to be corrected by heritability (Vintil, I., 1992). In March 1999, an E.T. program was carried out in Romania, with frozen embryons from milk and meat breeds, by dr. Edwin Robertson from the Company Harrogate Genetics Internationalof USA, who disposes of an advanced technology for embryopn freezing. This program was repeated in the years 2000; 2001, and the donated embryons were transferred through the non-surgical technique in several locations in the country. (Tob G.F.2000 ;2010;2011).

The results of this MOET program are given in table no.1.

Table 1. Results of MOET application with frozen embryons from USA transferred in Romania Locations Transferred embryons Recipient young beef Gestated- % 7 different locations 68-HF 68-BNR 28 - 41,17 Otopeni-Ilfov 12-AA;BNA;B.A. 12-BNR 7 - 58,33 Ardud-Satu Mare 15-AA 15-BNR 9 - 60 TOTAL 95 95 44 - 46,31

The specialty literature mentions the fact that, in Genus MOET, ( McGuirk, 1992) 95% frozen embryons and 5% fresh embryons were used, as compared to the incubation farm which may work only with fresh embryons (in order to reduce costs and increase the rate of successful transplantation), it is understood that, in this case, only frozen embryons must be worked with. Starting from the idea of E.T. closed circuit incubation farms, M.Th.Paraschivescu proposes a Project study for an open circuit MOET farm (M.Th.Paraschivescu, 2010) which


allows the valorification, for the commercial farms, of the previous stages of selection and improvement of the Holstein-Friesien breed, without disposing of advanced knowledge on the production and processing of transferrable embryons. The open circuit MOET farm is a MOET service version hiring only operations of transplantations of frozen embryons. The MOET program has bio-economic significances by the biodiversity of the metabolic and hormonal profile of embryon donor and recipient female and allowed the access to animal genetic biotechnologies, with low costs and a minimum risk of introducing diseases towards imports with live animals.

It is to retain the fact that the open circuit MOET farm executes operations regarding the production of embryons or selection operations or production control only in the extent in which the knowledge of the main recipient performances raises their selling price. The valorification of the MOET technology through open circuit farms can be extremely valuable in less developed countries, which do not have contemporary methods of genetic improvement and sophisticated systems of embryons collection and preservation. The new program was based on the donations of embryons made by the H-F Association of USA for the multiplication of the bovine population in Romania for the production of E.T. young beef resulted from recipients raised in Romania.

(a) (b)

Fig. 1. Products obtained from E.T.: (a) E.T. products (H.F.) IBNA (b) E.T. products (AA) - Ardud

2. Experiences regarding the reduction of the timeframe between two embryon collection on the BNR breed young beef

As a new experimental work approached within the postdoctoral preparation of the postdoctoral researcher dr.ing. M.Th.Paraschivescu, a research on the reduction of the timeframe between two embryon samplings. The work was carried out within the Biobase of the ICDCB Institute Baloteti, with a number of 4 young beef, with ages between 10 -12 months. Several young beef from the Blat cu Negru Romneasc (B.N.R.) breed were clinically and transrectally examined and 2 young beef were retained for follicular simulation through poliovulation (reg. no 3618 and 1092) which had a yellow body, well formed, proving the installation of puberty and the ovar was in the second half of the estrous cycle. The induction and synchronization of the estrus was provoked by the administration of an injection with PGF2. (Reglandin), and the entry into the estrus took place within 52-60 hours and the 0 day of the estrous cycle was thus obtained. (G.F Tob, 2010,2011)

The induction of the poliovulation was made starting with the 11th day of the estrous cycle, administrating a 20 ml thinner flask, containing 500 ui FSH/LH Pluset/ donor young beef every 12 hours according to the following chart:

Day 11: 7:00 a.m. 4 ml of FSH were injected intramuscular 19:00 p.m. 4 ml of FSH were injected intramuscular

Day 12: 7:00 a.m. 3 ml of FSH were injected intramuscular 19:00 p.m. 3 ml of FSH were injected intramuscular

Day 13: 7:00 a.m. 3 ml of FSH were injected intramuscular 19:00 p.m. 3 ml FSH + 2 ml PGF2 (Reglandin) were injected intramuscular

Day 14: The entry into estrus was monitored for its recording.


Day 15: Each donor was recorded when going into heat and the event time and at 17:00 the donors were administrated, intramuscular, 3 ml de Receptal (GnRH) each and, 2 hours later, at 19:00, I-IA was inseminated.

Day 16: The second IA insemination was repeated at 7:00 a.m. from the first I.A.

The embryon sampling was executed non-surgical in the 7th day following the insemination. Before the sampling, each donor young beef was examined transrectal and luteal bodies (LB) were identified and

counted on both ovars, for more safetybeing also used the ultrasound images, taken by dr. M.Ochea and presented in the photograms below.

Fig. 2. Ultrasound images D.3818

The sampling environment was PBS Dulbeco modified and the identification of the sampled embryonary

formations was made at Stereolupa Olymous, (Fig. 3 (a)) They were also evaluated according to the quality standards of the International Embryo Transfer Manual and those embryons in the blastocyst stafe D.3818. (Fig. 3(b)) were packed in yellow spangles with ethylene glycol and frozen. After sampling, PGF2alfa+GnRH was administrated to retake the estrous cyclic activity.

(a) (b)

Fig.3 Stereo-microscopic examination of embryons: (a) preparation (b) embryons in blastocyst stage-D.3818


Fig. 4 The embryon freezing diagram

It was decided that the repetition of the treatment for the multiplication of successive ovulations after the first

going into heat consecutive to the embryon sampling, Day 0, for the following poliovulation, following to obtain, by a PGF2 injection in the diestrus phase of the estrous cycle initiated by the spontaneous heat. This procedure was selected in order to reduce the effect of the medicine intervention in the ovar functioning. The collected embryons had to be frozen in white glycerol spnagles (GLY) this time being the cryoprotectant used. These results of the repeated poliovulations from the same donor young beef are illustrated in table no.2. Table 2.The results of multiple poliovulation in BNR young beef Reg. No. Poliovulation date No. of luteal bodies No. of embryons BNR-3818- 15L I - 30.05.2012 7 4 - EG 3818 II- 22.07.2012 7 7 - GLY BNR 1092- 17L 1092 TOTAL

I- 30.05.2012 II- 22.07.2012

4 9

27

0 1 - GLY

12

Following the application of the MOET program within these researches, major risk genetic resources preservation banks were realized, for the Pinzgau (G.F.Tob, 2009,2010) and Sura de Step breeds (G.F.Tob, 2007) in Tara Haegului- Retezat (N.Mang,2011), Dancu-Iasi and C.S.C.B.A.S.-Bucharest, through the management of the reduction of the disparition risk for these animal breeds ( M.Th. Paraschivescu, A.T. Bogdan, 2010). On the Romanian Buffalo, the postdoctoral researches were performed by CS III, primary vet.med.dr. Bneanu F. (2010).

We also mention the contribution of the Researchers team within C.S.C.B.A. Acad.David Davidescu in the establishment and realization of the Animal Genetic Resource Banks (A.G.R.B.), within the Durable Development Center Haegului-Retezat County, through the Project Bio and geodiversity preservation as a support of the durable development and economic and social growth in the Haeg-Retezat area, project financed by the European Economic Area (EEA) mechanism, by Norway, Island and Liechtenstein. In the inauguration ceremony participated acad. Ionel Haiduc, the president of the Romanian Academy, acad. Pun Ion Otiman, secretary general of the Romanian Academy, the projects coordinator, E.S. ystein Hovdkinn, the ambassador of the Norway Kingdom in Bucharest, Cristian Vigne, the grandson of general Berthelot, lt. col. Thierry Le Guillou, attache of the embassy of France in Bucharest, representatives of the local administration. ( Pun Ion Otiman 2010) .


Table 3. Animal Genetic Resource Banks (A.G.R.B.)

LOCATION BREED No of msc doses No.of frozen embryons SCDCB - Dancu Sur de Step. 280 29 EG + 21 GLY=50 CSCBAS- Bucharest B.N.R. 0 5 EG + 7 GLY=12 HAEG- RETEZAT TOTAL

B.R. S.S..

Pz. Female buffalo

B.G B.

0 0.................................

30 0

20 30

5 EG 5 5 GLY..5

4 EG.4 5- GLY.5 7- GLY.7 0

Fig.5. Animal Genetic Resource Bank- Haeg Conclusions:

1. The embryon transfer came to practice, in particular on taurine and it is about to be practiced also in other farm animal species. Through MOET, the embryons obtained following repeated supraovulations, the zootechnical biodiversity can be preserved. The farm animal breeds, being in critical status from the point of view of their disparition risk can be quickly consolidated through the amplification and balancing of their component family heads. The affirmation is also valid for the populations under constitution or reconstitution.

2. The bio-economy of an area can be quickly modified by MOET as animal populations can be directly transfered from one place to another, with all the zootechnical and sanitary veterinary advantages, without appealing to absorption cross- breeding.

3. The MOET biotechnology fully satisfies the sanitary veterinary and ecosanogenesis requirements as the embryons, through their processing, according to the I.E.T.S. Manual the transmission of different diseases is reduced to minimum.


Acknowledgements

This work represents the completion of the theme The bio-economic impact of the MOET European program over the safety and security of vulnerable and major risk zootechnical resources in Romania within the Interdisciplinary program for the prevention of major risk phenomena, at a national scale, a fundamental program of the Romanian Academy - INCE -coordinator Acad. Florin Gheorghe Filip

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