Name: in ovo feeding of commercial broiler eggs: an accurate and reproducible method to affect muscle development and growth
Uploaded: 2021-09-20T15:00:00
Description: Watch this Scientific Journal Video about In Ovo Feeding of Commercial Broiler Eggs: An Accurate and Reproducible Method to Affect Muscle Development and Growth at JoVE.com

The authors would like to thank Cobb Vantress, Inc. for donation of the eggs and providing technical assistance on egg incubation. The authors would like to thank ChromaDex, Inc. for nicotinamide riboside technical assistance.

All methodologies were approved by the University of Georgia Institutional Animal Care and Use Committee.
1. Egg incubation and treatment administration
<ol>
	<li>Egg procurement and treatment assignment
	<ol>
		<li>Obtain unincubated, fertilized high-yield broiler eggs and transport them to the laboratory.</li>
		<li>Inspect and discard eggs deemed poor quality. 
		NOTE: Eliminate misshapen eggs (round, elongated, slab-sided), cracked, dirty/stained, thin-shelled, and wrinkled. This is...

<ol>
	<li>Biressi, S., Molinaro, M., Cossu, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cellular+heterogeneity+during+vertebrate+skeletal+muscle+development.">Cellular heterogeneity during vertebrate skeletal muscle development.</a> Developmental Biology. 308 (2), 281-293 (2007).</li><li>Chal, J., Pourquie, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Making+muscle:+Skeletal+myogenesis+in+vivo+and+in+vitro.">Making muscle: Skeletal myogenesis in vivo and in vitro.</a> Development. 144 (12), 2104-2122 (2017).</li><li>Sharma, J., Burmester, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Resistance+of+Marek's+disease+at+hatching+in+chickens+vaccinated+as+embryos+with+the+Turkey+herpesvirus.">Resistance of Marek's disease at hatching in chickens vaccinated as embryos with the Turkey herpesvirus.</a> Avian Diseases. 26 (1), 134-149 (1982).</li><li>Al-Murrani, W. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+injecting+amino+acids+into+the+egg+on+embryonic+and+subsequent+growth+in+the+domestic+fowl.">Effect of injecting amino acids into the egg on embryonic and subsequent growth in the domestic fowl.</a> British Poultry Science. 23 (2), 171-174 (1982).</li><li>Ohta, Y., Kidd, M. T., Ishibashi, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Embryo+growth+and+amino+acid+concentration+profiles+of+broiler+breeder+eggs,+embryos,+and+chicks+after+in+ovo+administration+of+amino+acids.">Embryo growth and amino acid concentration profiles of broiler breeder eggs, embryos, and chicks after in ovo administration of amino acids.</a> Poultry Science. 80 (10), 1430-1436 (2001).</li><li>Zhao, M. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+ovo+feeding+of+creatie+pyruvate+increases+hatching+weight,+growth+performance,+and+muscle+growth+but+has+no+effect+on+meat+quality+in+broiler+chickens.">In ovo feeding of creatie pyruvate increases hatching weight, growth performance, and muscle growth but has no effect on meat quality in broiler chickens.</a> Livestock Science. 206, 59-64 (2017).</li><li>Bieganowski, P., Brenner, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Discoveries+of+nicotinamide+riboside+as+a+nutrient+and+conserved+NRK+genes+establish+a+Preiss+Handler+independent+route+to+NAD1+in+fungi+and+humans.">Discoveries of nicotinamide riboside as a nutrient and conserved NRK genes establish a Preiss Handler independent route to NAD1 in fungi and humans.</a> Cell. 117 (4), 495-502 (2004).</li><li>Chi, Y., Sauve, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nicotinamide+riboside,+a+trace+nutrient+in+foods,+is+a+Vitamin+B3+with+effects+on+energy+metabolism+and+neuroprotection.">Nicotinamide riboside, a trace nutrient in foods, is a Vitamin B3 with effects on energy metabolism and neuroprotection.</a> Current Opinion in Clinical Nutrition and Metabolic Care. 16 (6), 657-661 (2013).</li><li>Gonzalez, J. M., Jackson, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+ovo+feeding+of+nicotinamide+riboside+affects+pectoralis+major+muscle+development.">In ovo feeding of nicotinamide riboside affects pectoralis major muscle development.</a> Translational Animal Science. 4 (3), 1-7 (2020).</li><li>Xu, X., Jackson, A. R., Gonzalez, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+in+ovo+nicotinamide+riboside+dose+on+broiler+myogenesis.">The effects of in ovo nicotinamide riboside dose on broiler myogenesis.</a> Poultry Science. 100 (3), 100926 (2021).</li></ol>

To date, the Gonzalez Laboratory at the University of Georgia is the only group to demonstrate positive effects of nicotinamide riboside in ovo feeding on pectoralis major muscle&#160;development and growth. The first study found that in ovo feeding of 250 mM nicotinamide riboside increased muscle weight and dimensions when injected in the yolk sac9. In the follow-up study, injecting increasing nicotinamide riboside dose into the yolk, similar to the doses tested in the ...

Since 1960, the United States&#39; per capita consumption of meat from poultry has risen at an astounding rate, while other primary protein sources have remained stagnant, declined, or minimally increased. The poultry industry invested considerable time and research efforts optimizing nutrition and genetics to produce an efficient bird to keep up with demand. Because the poultry industry&#39;s main objective is to produce muscle for conversion to meat, their efforts have drastically changed the bird&#39;s ultimate muscle mass at harvest.
Like most species, poultry develops muscle in a biphasic manner. Primary myogenesis utilizes mesenchymal stem cells to produce primary muscle fibers, which serve as the scaffold for the second wave of muscle fiber development1. In poultry, primary myogenesis occurs during embryonic days 3 to 8, and secondary myogenesis happens from days 8 to 212. Once developed, primary and secondary muscle fibers serve as the basis for all future muscle growth through cellular hypertrophy. Therefore, scientists and industry spent considerable effort attempting to manipulate primary and secondary myogenesis in all meat-producing species to maximize meat yield.
One technology explored in poultry, called in ovo feeding, involves feeding compounds through injection. In ovo feeding, a technology employed by the poultry industry for almost 40 years, was initially developed for vaccine administration3. The literature documents that in ovo feeding of various compounds and nutrients at different developmental periods and locations within the egg positively affected in ovo muscle development and growth4,5,6. To date, the Gonzalez Laboratory at the University of Georgia is the pioneer in utilizing nicotinamide riboside in ovo feeding to manipulate poultry muscle development.
Nicotinamide riboside, a pyridine-nucleoside analog of vitamin B3, produces NAD+ through the salvage pathway7. Since this pathway utilizes fewer enzymatic steps to produce NAD+, production is the most efficient8. Gonzalez and Jackson9 demonstrated that supplementation of developing broiler embryo yolk sac with nicotinamide riboside increased hatched chick pectoralis major muscle weight and muscle fiber density. This was later confirmed by Xu et al.10, who found that increasing nicotinamide riboside dose increased muscle weight and increased muscle fiber density. These first two studies were conducted in a commercial yield broiler. Because high yield broilers possess a more significant genetic potential for ultimate muscle mass size, the study&#39;s objective was to determine the effects of nicotinamide riboside dose on high yield broiler hatched chick pectoralis major muscle development and growth at hatch.

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			<td>VWR</td>
			<td>VWR-214B2</td>
			<td>https://us.vwr.com/store/product/20970740/vwr-b2-series-analytical-and-precision-balances</td>
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			<td>Complete Dissection Set</td>
			<td>DOCAZON</td>
			<td>DK1001</td>
			<td>https://www.amazon.com/DOCAZON-Complete-Dissection-Set-Dissecting/dp/B07VBHKSW3</td>
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			<td>Fisherbrand&#8482; Isotemp&#8482; General Purpose Deluxe Water Baths</td>
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			<td>https://www.fishersci.com/shop/products/isotemp-general-purpose-water-baths/p-6448020</td>
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			<td>Fisherbrand&#8482; Sterile Syringes for Single Use</td>
			<td>Fisher Scientific</td>
			<td>14-955-464</td>
			<td>https://www.fishersci.com/shop/products/sterile-syringes-single-use-12/p-7114739#?keyword=</td>
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			<td>HIGH INTENSITY EGG CANDLER</td>
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			<td></td>
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			<td>Microsoft Excel</td>
			<td>Microsoft</td>
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			<td>Neiko Tools Digital Caliper</td>
			<td>Neiko Tools</td>
			<td>01408A</td>
			<td>https://www.amazon.com/Neiko-01407A-Electronic-Digital-Stainless/dp/B000NEA0P8?th=1</td>
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			<td>Nexcare Absolute Waterproof Tape</td>
			<td>Nexcare Brand</td>
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			<td>https://www.nexcare.com/3M/en_US/nexcare/products/catalog/~/Nexcare-Absolute-Waterproof-Tape/?N=4326+3294529207+3294631805 
			&#38;rt=rud</td>
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			<td>Pen Size Temperature and Humidity USB Data Logger with Display</td>
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			<td>SAS 9.4 for Windows</td>
			<td>SAS Institute</td>
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			<td>https://www.sas.com/en_us/home.html</td>
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			<td>Sportsman 1502 Incubator</td>
			<td>GQF Manufacturing</td>
			<td>1502</td>
			<td>https://www.gqfmfg.com/item/1502-digital-sportsman/</td>
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			<td>Tru Niagen (Nicotinamide riboside)</td>
			<td>ChromaDex, Inc.</td>
			<td>N/A</td>
			<td>https://www.truniagen.com/truniagen-300mg/ - note, contact company for pure product</td>
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			<td>Wood Craft Sticks</td>
			<td>Creatology</td>
			<td>M20001547</td>
			<td>https://www.michaels.com/wood-craft-sticks-by-creatology/M20001547.html</td>
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in ovo feeding of commercial broiler eggs: an accurate and reproducible method to affect muscle development and growth

Within the past three decades, red meat and poultry scientists focused on developing strategies and technologies to manipulate muscle development during embryonic and fetal development. This area continues to be an area of focus because muscle fiber number is established during this time and determines the basis for all future growth. In poultry, numerous studies demonstrated in ovo feeding of growth factors, vitamins, or other nutrients improved chick embryonic muscle and intestinal development. Improving in ovo muscle development could benefit the poultry industry by possibly influencing meat yield, growth rate, or myopathy conditions. During the past five years, the Gonzalez Laboratory at the University of Georgia developed a nicotinamide riboside in ovo feeding methodology for broiler-chicken embryos, which altered&#160;muscle development. When injected into a developing embryo&#39;s yolk sac, nicotinamide riboside increased pectoralis major muscle weight and muscle fiber density at hatch. This protocol will demonstrate a methodology to accurately and reproducibly conduct in ovo feeding studies utilizing commercial standard- and high-yielding broiler embryos. These data and methods will allow other research groups to perform in ovo feeding studies with much success and reproducibility.

There were no DOS effects for the bodyweight of day-18 embryos and hatched chicks (P &#62; 0.52; Figure 2). There were no DOS effects for all day-18 embryo pectoralis major muscle measurements (P &#62; 0.24; Figure 3). There were no DOS effects for hatched chick pectoralis major muscle length and width measurements (P &#62; 0.26); however, DOS did affect muscle weight and depth (P &#60; 0.03; <strong class="x...

Watch this Scientific Journal Video about In Ovo Feeding of Commercial Broiler Eggs: An Accurate and Reproducible Method to Affect Muscle Development and Growth at JoVE.com

In Ovo Feeding of Commercial Broiler Eggs: An Accurate and Reproducible Method to Affect Muscle Development and Growth

A robust methodology has been developed for conducting in ovo feeding research trials utilizing unincubated commercial broiler eggs to test the ability of natural and synthetic compounds, in this case, nicotinamide riboside, to influence muscle development and growth.

In Ovo Feeding of Commercial Broiler Eggs: An Accurate and Reproducible Method to Affect Muscle Development and Growth

Within the past three decades, red meat and poultry scientists focused on developing strategies and technologies to manipulate muscle development during ...

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