Login| Sign Up| Help| Contact|

Patent Searching and Data


Title:
NICOTINAMIDE RIBOSIDE TREATMENTS OF DOMESTICATED MEAT ANIMALS
Document Type and Number:
WIPO Patent Application WO/2019/133815
Kind Code:
A1
Abstract:
Described herein are methods of increasing meat quantity and/or improving meat quality of domesticated meat animals using treatments of nicotinamide riboside. The nicotinamide riboside may be in the form of a chloride salt mixed or dissolved in a biologically-acceptable carrier. The treatments may be provided as an in ovo injection or orally administered to the live domesticated meat animal. The methods described herein advantageously increase the size and weight of the domesticated meat animals, reduce the incidences of transportation fatigue, and decrease meat discoloration over time.

Inventors:
GONZALEZ, John Michael (1271 Latham Drive, Watkinsville, Georgia, 30677, US)
KRUGER, Stephanie Rene (4801 NW Leedy Rd, Topeka, Kansas, 66618, US)
PAULK, Chad Bennett (5524 Turkey Foot Ln, Manhattan, Kansas, 66503, US)
WECKER, Haley Kay (815 Ratone Street, Manhattan, Kansas, 66502, US)
DUNMIRE, Kara (501 Dornoch Way, Apt. 202Manhattan, Kansas, 66502, US)
Application Number:
US2018/067865
Publication Date:
July 04, 2019
Filing Date:
December 28, 2018
Export Citation:
Click for automatic bibliography generation   Help
Assignee:
KANSAS STATE UNIVERSITY RESEARCH FOUNDATION (2005 Research Park Circle, Suite 105Manhattan, Kansas, 66502, US)
International Classes:
C07H19/048
Domestic Patent References:
WO2017109195A12017-06-29
WO2017062311A12017-04-13
Foreign References:
US20150056274A12015-02-26
CN105393976A2016-03-16
US20170204131A12017-07-20
Attorney, Agent or Firm:
SKOCH, Gregory J. (Hovey Williams, LLP10801 Mastin Blvd, Suite 1000,84 Corporate Wood, Overland Park Kansas, 66210, US)
Download PDF:
Claims:
Claims:

1. A method of increasing meat quantity and/or improving meat quality in a domesticated meat animal, the method comprising providing to the domesticated meat animal or to an embryo of the domesticated meat animal an effective amount of nicotinamide riboside.

2. The method of claim 1, wherein the nicotinamide riboside is provided as nicotinamide riboside chloride.

3. The method of claim 1, wherein the domesticated meat animal is a chicken or a pig.

4. The method of claim 3, wherein the meat animal is a chicken.

5. The method of claim 4, wherein the providing comprises injecting a quantity of nicotinamide riboside into a fertilized chicken egg.

6. The method of claim 4, wherein nicotinamide riboside is injected into the chicken egg at a concentration of at least about 2.5 mM in about 1 mΐ to about 1,000 mΐ of solution.

7. The method of claim 5 or 6, wherein the nicotinamide riboside is injected into a yolk of the chicken egg.

8. The method of claim 3, wherein the meat animal is a pig.

9. The method of claim 8, wherein the providing comprises orally administering a quantity of nicotinamide riboside to the pig.

10. The method of claim 8, wherein the nicotinamide riboside is administered at a dose of at least about 15 mg per kg of body weight.

11. The method of claim 8, wherein the nicotinamide riboside is administered at a dose of at about 15 mg per kg of body weight to about 30 mg per kg of body weight.

12. The method of claim 11, wherein the nicotinamide riboside is administered at a dose of about 30 mg per kg of bodyweight.

Description:
NICOTIN AMIDE RIBOSIDE TREATMENTS

OF DOMESTICATED MEAT ANIMALS

CROSS-REFERENCE TO RELATED APPLICATION This application claims the benefit of U.S. Provisional Application No. 62/611,087, filed December 28, 2017, entitled NICOTINAMIDE RIBOSIDE TREATMENTS OF DOMESTICATED MEAT ANIMALS, which is incorporated by reference herein.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

This invention was made with government support under Contract No. NC-1184 and Contract No. NA/1006677, both awarded by the U.S. Department of Agriculture. The government has certain rights in the invention. BACKGROUND OF THE INVENTION

Field of the Invention

The invention is generally directed to methods of increasing meat quantity and/or improving meat quality of domesticated meat animals using treatments of nicotinamide riboside.

Description of the Prior Art

Nicotinamide riboside (NR) is a pyridine-nucleoside form of vitamin B 3 that functions as a precursor to nicotinamide adenine dinucleotide, which can produce NAD+. Previous studies have found that NR enhances oxidative metabolism and protects against high-fat diet-induced obesity. Other studies have explored its effects on energy metabolism and neuroprotection. Other studies have explored the effect of NAD+ on slowing stem cell loss and aging. However, while previous studies have focused on the use of nicotinamide riboside for improving health, no one has explored its use in increasing meat quantity and/or improving meat quality in domesticated meat animals. Notably, no studies have been conducted that examine the effect of nicotinamide riboside on chicken myogenesis (muscle development) in utero or pig growth and meat quality. SUMMARY OF THE INVENTION

Embodiments of the present invention demonstrate that an in ovo injection of nicotinamide riboside in developing chicken embryos increased the body weight, and weight, length, and depth of the Pectoralis major muscle of the chickens, and particularly of the chicks immediately after hatching. This increases the efficiency and weight of birds produced for meat production. Other embodiments of the present invention demonstrate that nicotinamide riboside consumption by pigs increases growth, improves meat quality, and increases muscle NAD+ content. This increases the efficiency of growth, lengthens the time of retail meat sales, and may be used as a counter measure to delay the onset of transportation fatigue.

According to one embodiment, therefore, there is provided herein a method of increasing meat quantity and/or improving meat quality in a domesticated meat animal. The method comprises providing to the domesticated meat animal or to an embryo of the domesticated meat animal an effective amount of nicotinamide riboside.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure (Fig.) 1 is a series of graphs showing the effects on overall chick weight, and Pectoralis major length, weight, and depth of nicotinamide riboside treatments in accordance with embodiments of the present invention;

Fig. 2 is a graph showing muscle fiber cross-sectional area (CSA) during in ovo myogenesis;

Fig. 3 is a graph showing average daily weight gain of pigs treated with nicotinamide riboside in accordance with embodiments of the present invention;

Fig. 4 is a graph showing loin eye area of pigs treated with nicotinamide riboside in accordance with embodiments of the present invention;

Fig. 5 is a graph showing meat surface metmyoglobin % over time of pigs treated with nicotinamide riboside in accordance with embodiments of the present invention; Fig. 6 is a graph showing meat panelists surface discoloration % scores over time of pigs treated with nicotinamide riboside in accordance with embodiments of the present invention;

Fig. 7 is a graph showing meat metmyoglobin reducing ability % of pigs treated with nicotinamide riboside in accordance with embodiments of the present invention;

Fig. 8 is a graph showing meat muscle fiber distribution % of pigs treated with nicotinamide riboside in accordance with embodiments of the present invention;

Fig. 9 is a graph showing meat muscle fiber SDH mean intensity of pigs treated with nicotinamide riboside in accordance with embodiments of the present invention;

Fig. 10 is a graph showing Objective loin chop surface oxy- and metmyoglobin accumulation from pigs fed 0 or 30 mg/kg of nicotinamide riboside (NR) for the final 10 days of feeding;

Fig. 11 is a graph showing trained panelists loin chop discoloration scores from pigs fed 0 or 30 mg/kg of nicotinamide riboside (NR) for the final 10 days of feeding;

Fig. 12 is a graph showing oxygen consumption rate and metmyoglobin reducing ability of pigs fed 0 or 30 mg/kg of nicotinamide riboside (NR) for the final 10 days of feeding;

Fig. 13 is a graph showing the effect of nicotinamide riboside on the onset of subjective fatigue; and

Fig. 14 is a graph showing the effect of nicotinamide riboside on Semitendinosus nicotinamide adenine dinucleotide (NAD+) concentration.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

In one or more embodiments, there is provided a method of increasing meat quantity and/or improving the quality in a domesticated meat animal. As used herein, the term“domesticated meat animal” refers to animals that have been raised in captivity for consumption by people. Exemplary domesticated meat animals include birds, pigs, bovines, and the like. In certain preferred embodiments, the domesticated meat animal is a chicken (e.g., a broiler chicken) or a pig. Methods in accordance with embodiments of the present invention generally comprise providing to the domesticated meat animal or to an embryo of the domesticated meat animal an effective amount of nicotinamide riboside. Nicotinamide riboside (NR) is a pyridine-nucleoside form of vitamin B 3 that functions as a precursor to nicotinamide adenine dinucleotide, which can produce NAD+. The nicotinamide riboside is preferably provided to the domesticated meat animal as the salt nicotinamide riboside chloride. However, the nicotinamide riboside may also be provided in another biologically-acceptable form. For example, nicotinamide riboside may also be provided as nicotinamide riboside oxide, nicotinamide riboside sulfate, or combined with amino acid complexes.

In a particular embodiment, there is provided a method of increasing meat quantity of a chicken. The method comprises providing a chicken embryo with an effective amount of nicotinamide riboside. The nicotinamide riboside can be provided to the chicken embryo by injecting the nicotinamide riboside into a fertilized chicken egg during the incubation period. The injection is generally made about 10 days or more after the egg is laid and preferably about 10 to about 12 days after the egg is laid. The injection is preferably made into the yolk of the fertilized chicken egg. Injecting the nicotinamide riboside into the yolk can advantageously result in increased meat quantity of hatched chicks compared to injecting into other portions of the chicken egg, such as the albumen. The injection generally comprises a quantity of nicotinamide riboside dissolved in a biologically-acceptable liquid carrier. For example, the liquid carrier can be a sterile saline solution. The amount of nicotinamide riboside injected into the fertilized egg can be varied. However, the injection generally comprises about 1 mΐ to about 1000 mΐ, preferably about 25 mΐ to about 500 mΐ, and more preferably about 50 mΐ to about 200 mΐ of solution, wherein the solution concentration is about 0.1 mM to about 10 mM, preferably about 1 mM to about 5 mM, and more preferably about 2 mM to about 3 mM of nicotinamide riboside. However, in certain embodiments, doses providing greater amounts of nicotinamide riboside may also be used. In certain embodiments, the concentration of nicotinamide riboside in the solution is at least about 2.5 mM, at least about 5 mM, at least about 7.5 mM, or at least about 10 mM. After injection, the fertilized chicken eggs are generally incubated under natural conditions (i.e., broody hen) or in an artificial environment. Regardless, the fertilized chicken eggs are incubated at a temperature of about 30°C to about 40°C and preferably about 34 °C to about 40 °C, and a relative humidity of about 30% to about 50% and preferably about 38% to about 42%. In the final about 3 to about 5 days of incubation before hatching, the humidity may be increased to about 50% to about 70% and preferably about 58% to about 62%. The methods described herein can advantageously result in chicks having increased overall weight, as well as increased Pectoralis major muscle weight, length, and depth compared to untreated chicks.

In another particular embodiment, there is provided a method of increasing meat quantity and/or improving meat quality of a pig. The method comprises providing a pig with an effective amount of nicotinamide riboside. The pig may be an adult pig or piglet. The nicotinamide riboside can be provided to the pig by orally administering a quantity of nicotinamide riboside to the pig, although other methods of administering may also be used. The oral administration treatment comprises a quantity of nicotinamide riboside mixed with a biologically-acceptable carrier. The carrier is preferably in the form of a liquid or solid drink or foodstuff that has a desirable flavor to the pig. For example, the carrier may be corn syrup, such as Karo® dark syrup. Regardless the carrier, the nicotinamide riboside is generally administered to the pig at a dose of about 15 mg to about 30 mg of nicotinamide riboside per kg of body weight of the pig. In particularly preferred embodiments, the nicotinamide riboside is administered to the pig at a dose of about 30 mg of nicotinamide riboside per kg of body weight of the pig. However, in other preferred embodiments, the nicotinamide riboside is administered to the pig at a dose of about 15 mg of nicotinamide riboside per kg of body weight of the pig. In certain embodiments, the nicotinamide riboside is administered to the pig at a dose of at least about 5 mg, at least about 10 mg, at least about 15 mg, or at least about 30 mg of nicotinamide riboside per kg of body weight of the pig. Treatment doses providing greater amounts of nicotinamide riboside than those listed above may also be used. The dose may be administered daily for at least about 3 days, preferably at least about 5 days, and more preferably at least about 7 days. Other than the treatment doses, the pig may otherwise be given standard nursing/growing/fmishing diet and water. The methods described herein advantageously result in pigs having better average daily gain (ADG), larger loin eyes, less Longissimus lumborum (LL) surface metmyoglobin accumulation, less meat visual panelists’ surface discoloration, more metmyoglobin reducing ability (MRA), and greater NAD+ content than untreated pigs. Additionally, the methods and treatments described herein increase semitendinosus muscle NAD+ levels, providing more energy for movement, and is therefore useful as a nutritional countermeasure to reduce the incidence of transport fatigue.

Additional advantages of the various embodiments of the invention will be apparent to those skilled in the art upon review of the disclosure herein and the working examples below. It will be appreciated that the various embodiments described herein are not necessarily mutually exclusive unless otherwise indicated herein. For example, a feature described or depicted in one embodiment may also be included in other embodiments, but is not necessarily included. Thus, the present invention encompasses a variety of combinations and/or integrations of the specific embodiments described herein.

As used herein, the phrase“and/or,” when used in a list of two or more items, means that any one of the listed items can be employed by itself or any combination of two or more of the listed items can be employed. For example, if a composition is described as containing or excluding components A, B, and/or C, the composition can contain or exclude A alone; B alone; C alone; A and B in combination; A and C in combination; B and C in combination; or A, B, and C in combination.

The present description also uses numerical ranges to quantify certain parameters relating to various embodiments of the invention. It should be understood that when numerical ranges are provided, such ranges are to be construed as providing literal support for claim limitations that only recite the lower value of the range as well as claim limitations that only recite the upper value of the range. For example, a disclosed numerical range of about 10 to about 100 provides literal support for a claim reciting“greater than about 10” (with no upper bounds) and a claim reciting“less than about 100” (with no lower bounds).

EXAMPLES

The following examples set forth studies related to the treatment of domesticated meat animals with nicotinamide riboside. It is to be understood, however, that these examples are provided by way of illustration and nothing therein should be taken as a limitation upon the overall scope of the invention. EXAMPLE I

Objective

The objective of this study was to examine the effects of nicotinamide riboside (NR) on avian embryonic myogenesis.

Methods

At 11 -days of incubation, 60 fertilized broiler eggs were randomly assigned to completely randomized design with a 2 c 2 factorial arrangement. Factor 1 comprised treatment, with eggs injected with 100 mΐ of 0.9% sterile saline solution (NRNeg) or 2.5 mM NR in sterile saline (NRPos). Factor 2 comprised injection location, with treatments injected into the yolk or albumen. Eggs were incubated at 37 ± 3°C and a relative humidity of 40 ± 2% for 7 days. Humidity was increased to 60 ± 2% at the same temperature for the final 3 days of incubation. Twenty-four hours after hatching, chicks were euthanized by exposure to CO2 and decapitation. Measurements including chick weight and left Pectoralis major weight, length, and depth were taken for analysis.

Results

There were treatment x injection location interactions for chick weight and Pectoralis major weight and length (P < 0.02). In all measures, there were no differences between NRNeg and NRPos chicks when treatments were injected into the albumen ( P > 0.14). However, when treatments were injected into the yolk, NRPos chicks tended to weigh more (P = 0.07) and their Pectoralis major muscles weighed more and were longer than NRNeg chicks (P < 0.01). There was no treatment x injection location interaction (P = 0.30) for Pectoralis major depth. Treatment did affect (P = 0.03) Pectoralis major depth, with NRPos chicks having thicker muscles that NRNeg chicks. (Fig. 1).

Conclusion

Inj ection of NR into broiler eggs at day 11 of incubation increases chick weight and improves Pectoralis major development. Injecting NR into the yolk of the developing embryo has greater positive effects on chick development when compared to injecting NR into the albumen.

Immunohistochemical Analysis Immunohistochemical analysis was conducted to determine if NR supplementation increased muscle fiber cross-sectional area (CSA) during in ovo myogenesis. There were no Treatment c Location or Treatment and Location main effects for muscle fiber CSA (P > 0.06). See Fig. 2. Because muscle fiber CSA did not increase due to NR supplementation but whole muscle morphometries did, it is believed that NR increased muscle morphometries by increasing the number of muscle fibers formed during myogenesis. This is desired heavily in meat producing animals.

EXAMPLE II

Objective

The objective of this study was to further examine the effect of nicotinamide riboside (NR) concentration on avian embryonic myogenesis.

Materials and Methods

Fertilized broiler eggs ( n = 60; Cobb 500) were randomly assigned to 1 of 4 treatments: 0.0, 2.5, 5.0, or 10.0 mM NR in sterile saline. At day 10 of incubation, 100 pl of treatment solution was injected into the egg yolk. Eggs were incubated at 37 ± 3°C and a relative humidity of 40 ± 2%. At day 19 of incubation, embryos were euthanized by prolonged exposure to CO2 gas and decapitation. Measurements including: embryo weight; crown-rump length; chest circumference (CC); and left Pectoralis major (PM) weight, length, width, and depth were collected.

Results

There was no treatment effect for embryo weight (P = 0.99). Embryos treated with 10.0 mM NR had longer crown-rump measurements than all other treatments ( P < 0.05), which did not differ from each other ( P > 0.36). Embryos from the 5.0 and 10.0 mM treatments had larger CC and PM weight and width than 0.0 mM embryos ( P < 0.04), but did not differ ( P > 0.38) from each other. Embryos injected with 2.5 mM of NR did not differ in CC or PM weight and width when compared to other treatments ( P > 0.06). All NR treatments had longer PM muscles than the saline treatment ( P < 0.01), but did not differ from each other ( P < 0.41). Embryos treated with 10.0 and 2.5 mM NR had thicker PM muscles than saline injected embryos ( P < 0.04), but did not differ from each other ( P = 0.58). Embryos injected with 5.0 mM NR did not differ in PM thickness compared to other treatments ( P > 0.06). See Table 1.

Conclusion

Increasing the concentration of in ovo injected nicotinamide riboside has a quadratic influence on avian myogenesis. Injecting up to 5.0 mM NR into the yolk of the developing embryo had no effect on body weight but increased PM measures; thus, indicating NR influenced avian myogenesis.

Table 1. Effects of increasing nicotinamide riboside supplementation on avian in ovo myogenesis

_ Nicotinamide riboside dose, mM _

Item 0.0 2.5 5.0 10.0 SEM E- value

Whole body morphometries

Body weight, g 46.22 46.42 46.25 45.97 1.15 0.99 Crown to rump length, mm 84.0l a 83.69 a 85.14 3 88.20 b 1.16 0.02 Head width, mm 19.51 15.42 15.49 15.69 1.87 0.33 Head length, mm 18.25 17.70 17.60 17.98 0.28 0.36 Head circumference, cm 5 3 a 5 3 a 5 6 b 5.7 b 0.1 0.01 Chest circumference, cm 5 33 g Qa,b 6.6 b 0.3 <0.01

Pectoralis major morphometries

Weight, g 0.1 l a 0.l3 a,b 0.16 b 0.15 b 0.01 0.03 Length, mm 14.14 3 l6.73 b 17.5 l b l7.44 b 0.69 <0.01 Width, mm 4.49 a 4 90 a b 5 . 32 b 5.6 l b 0.29 0.03

Maximum thickness, mm 2.23 a 2.56 b 2.36 a,b 2.64 b 0.11 0.04

Organ weight

Heart, g 0.23 0.23 0.23 0.24 0.01 0.88 Liver, g 0.69 0.74 0.72 0.67 0.04 0.54

EXAMPLE III

Objective

The objective of this study was to examine the effects of nicotinamide riboside (NR) on pigs on Longissimus lumborum (LL) average daily gain (ADG), loin eye area, NAD+ content, and fresh meat color characteristics.

Methods

Nine growing pigs were blocked by bodyweight (BW) and placed in metabolism crates. After a 3-day acclimation period, pigs within each BW block were randomly assigned to a treatment (n = 3 pigs/treatment). Treatments comprised pigs supplemented 0, 15, or 30 mg/kg BW NR for 7 days. Pigs were allowed ad libitum access to a standard growing diet and water. Treatments were administered daily at 8 a.m. by mixing NR in 20 mL of Karo® dark syrup and orally drenching the pigs. On day 0 and 3 of the trial, muscle biopsies of the LL were taken for NAD+ analysis using standard procedures. At day 7, pigs were transported to the Kansas State LTniversity Meats Laboratory and euthanized by a captive penetrating bolt to the brain, followed by exsanguination. Following harvest procedures, 3 g of the LL were collected for NAD+ analysis. Twenty-four hours after harvest, the whole LL not sampled at harvest was removed from the carcass and aged 10 days. After aging, the LL was cut into 6 chops for retail color stability analysis, including 6-day surface oxy- and metmyoglobin percentage, 6-day objective and subjective color panel evaluation, and day 4 and 6 metmyoglobin reducing ability (MRA).

Results

The data indicates a trend for pigs in the 30 mg/kg treatment to numerically have 4.4% better ADG when compared to pigs in the other two treatments, which did not differ (Fig. 3). Treatment tended to affect ( P = 0.06) LEA, with pigs fed 30 mg/kg having 52% and 48% larger loin eyes than pigs fed 0 and 15 mg/kg NR, respectively (P < 0.03; Fig. 4). Pigs in the 0 and 15 mg/kg treatments did not differ ( P > 0.15) in LEA.

There was a treatment effect ( P = 0.04) for surface metmyoglobin accumulation (Fig. 5). Over the entire 6-day study, LL chops from the 30 mg/kg treatment had 14% less ( P = 0.02) surface metmyoglobin than 0 mg/kg chops. When compared to chops from the 0 mg/kg treatment, chops from the 15 mg/kg treatment tended to have 9% less ( P = 0.06) surface metmyoglobin accumulation. Chops from the 15 and 30 mg/kg treatments did not differ ( P = 0.21) in surface metmyoglobin accumulation.

There was a Treatment c Day interaction (P = 0.03) for visual panel discoloration scores (Fig. 6). On days 1, 4, and 5 of display, treatment discoloration scores did not differ from each other ( P > 0.15). On days 2 and 3 of display, 0 mg/kg chops tended to have more surface discoloration than the other two treatments ( P < 0.12), which did not differ ( P = 0.83) from each other. On day 6 of display, 0 mg/kg chops had 41 and 39% greater discoloration scores than 15 and 30 mg/kg chops, respectively ( P < 0.01). Discoloration scores for 15 and 30 mg/kg chops did not differ ( P = 0.82) on this day. There was a treatment effect ( P = 0.04) for discoloration scores. Over the entire 6-day study, LL chops from the 30 mg/kg treatment had 30% less ( P = 0.04) surface discoloration than 0 mg/kg chops. When compared to chops from the 0 mg/kg treatment, chops from the 15 mg/kg treatment tended to have 27% less ( P = 0.06) discoloration. Chops from the 15 and 30 mg/kg treatments did not differ ( P > 0.15) in surface discoloration.

There was a treatment effect ( P = 0.05) for MRA (Fig. 7). Over the entire study,

LL chops from the 30 mg/kg treatment had 64% more ( P = 0.03) MRA than 0 mg/kg chops. When compared to chops from the 0 mg/kg treatment, chops from the 15 mg/kg treatment tended to have 42% more (P = 0.08) MRA. Chops from the 15 and 30 mg/kg treatments did not differ ( P = 0.32) in surface discoloration. Muscle fiber distribution data indicates LL from the 30 mg/kg treatment numerically had less type 2B (glycolytic) fibers and more type 1 and 2A (oxidative) fibers than 0 mg/kg LL (Fig. 8). Succinate dehydrogenase staining indicated LL from the 30 mg/kg treatment numerically had more intense staining than LL from the other two treatments in type 2B fibers (Fig. 9).

At days 3 and 7 of NR. supplementation, there were treatment effects for the total and percent LL NAD+ change ( P < 0.05). At days 3 and 7 of supplementation, supplementing 15 mg/kg increased NAD+ on a total and percent change basis when compared to the 0 mg/kg treatment ( P = 0.02). The 30 mg/kg treatment only tended to have a greater ( P < 0.14) total change in NAD+ content when compared to the 0 mg/kg treatment. Results of the NAD+ effects of the treatments are shown in Table 2.

Conclusion In conclusion, supplementing NR at 30 mg/kg BW daily for 7 days appears to numerically increase ADG compared to 0 and 15 mg/kg pigs, which resulted in a tendency for LEA from these pigs to be bigger. When displayed under retail conditions, supplementing 15 or 30 mg/kg NR delayed surface accumulation of surface metmyoglobin when compared to 0 mg/kg chops, which was also seen by visual panelists. This improvement in color stability was most likely due to an improvement in MRA for both NR treatments compared to control. This improvement can partially be explained for the 30 mg/kg chops by what appears to be an increase in the number of oxidative fibers in the LL and an increase in SDH staining (oxidative ability) in type 2B fibers. Finally, supplementing NR increased NAD+ content in the LL of both NR treatments compared to control. However, the increase was greater in the 15 mg/kg treatment. It is hypothesized this may have occurred due to 30 mg/kg pigs utilizing more NAD+ for growth.

EXAMPLE IV

Objective

The objective of this study was to examine the effect of oral supplementation of nicotinamide riboside (NR) on pig performance, carcass characteristics, and loin chop color stability.

Materials and Methods

Seven days prior to the beginning of the experiment, 10 finishing barrows (initial BW 111.9 ± 1.6 kg) were assigned to individual pens located at the East Finisher facility of the Kansas State University Swine Teaching and Research Center (Manhattan, KS). Each pen was 7.4 m 2 with a slatted floor, contained a nipple waterer and an individual dry feeder that allowed ad libitum access to food and water. Barrows were randomly assigned to 1 of 2 NR treatments, 0 or 30 mg/kg daily of NR mixed in Karo® syrup and administered by oral gavage. Barrows were administered their assigned treatment for 10 days, after which they were harvested under USD A inspection. Twenty -four hours after harvest, carcass measurements were taken by trained personnel, carcasses were fabricated into the 5 major wholesale cuts, and whole-boneless loins were vacuum packaged and aged for 10 days. Loin were cut into 3 chops with chop 1 being used for day-0 metmyoglobin reducing ability (MRA) analysis, chop 2 being used for day-4 MRA and oxygen consumption rate (OCR) analyses, and chop 3 for 8-day objective/subjective color evaluation and MRA and OCR analyses.

Results

There were no treatment effects for any of the performance measures ( P > 0.257; Table 3). This was most likely due to the low numbers of barrows used in the pilot study. Numerically, NR supplementation increased average daily gain 8% when supplemented over 10 days.

There were no treatment effects for carcass measures and carcass fabrications measures, except for tendencies for NR to increase color score (make loin color more red) and decrease boneless loin weight ( P < 0.08), and NR decreased ( P = 0.03) Boston butt weight (Table 4). If one examines numerical trends, it appears NR increases fat deposition in all subcutaneous fat measures and marbling, but this comes at the expense of muscle deposition. This is seen with most wholesale cut weights being lower and the belly, which is mainly fat, increasing due to NR treatment.

There were no Treatment c Day interactions for all objective and subjective color measurements ( P > 0.77). Day of display affected all measures consistent with the discoloration of meat ( P < 0.02), except L* (lightness; 0 = black and 100 = white) value which was not affected ( P = 0.14; Table 5). Fig. 10 shows objective loin chop surface oxy- and metmyoglobin accumulation from pigs fed 0 or 30 mg/kg of nicotinamide riboside (NR) for the final 10 days of feeding. Chops were displayed under simulated retail display for 8 days and percent of surface oxymyoglobin and metmyoglobin were calculated using the equations of Krzywicki (1979). Objective measures indicated NR chops had greater a* (redness; -60 = green and 60 = red), greater surface oxymyoglobin, and less surface metmyoglobin formation over the 8-day display period ( P < 0.01; Table 5 and Fig. 10). Fig. 11 shows trained panelists loin chop discoloration scores from pigs fed 0 or 30 mg/kg of nicotinamide riboside (NR) for the final 10 days of feeding. Chops were displayed under simulated retail display for 8 days and 8 to 10 panelists evaluated discoloration on line scales with the following anchors: 0 = 0% discoloration and 100 = 100% discoloration. Panelists indicated chops had less discoloration form over the entire 8-day display period ( P < 0.01; Fig. 11). Fig. 12 shows oxygen consumption rate and metmyoglobin reducing ability of pigs fed 0 or 30 mg/kg of nicotinamide riboside (NR) for the final 10 days of feeding. Chops were displayed under simulated retail display for 8 days and OCR and MRA were calculated as outlined in the American Meat Science Association’s (AMSA) Meat Color Measurement Guidelines (AMSA, 2012). While treatment did not affect OCR and MRA ( P > 0.19), NR chops did have greater OCR and MRA during the 8-day display study.

Conclusion

Feeding NR the final 10 days before harvest numerically improved ADG and carcass fat measures at the expense muscle deposition. Loin chops from NR pigs were redder and had better color stability than control chops. Feeding NR at the end of the finishing period can be a useful way to increase carcass fatness (most importantly belly weight) and increase the time pork can be sold at retail.

EXAMPLE V

Objective A study was conducted to determine the effects of nicotinamide riboside (NR) on finished market barrow fatigue and semitendinosus muscle nicotinamide adenine dinucleotide (NAD+) content.

Methods

Fourteen days prior to harvest, 20 finished market barrows (initial body weight 268 pounds) were randomly assigned to 1 of 2 treatments: 0 or 30 mg/kg body weight of nicotinamide riboside, both orally administered daily in 20 ml of Karo® Syrup. Ten days prior to harvest, pigs were restrained via a snare and treatments were administered. On day 7, barrows were subjected to a performance test where they were walked around a track at 0.88 m/s until subjective fatigue was achieved. Three days following the performance test, barrows were harvested and a portion of the semitendinosus muscle was collected within 45 minutes. These samples were analyzed for NAD+ content via HPLC methodology.

Results

Data indicated NR supplemented barrows numerically ran longer and farther by 19 and 16%, respectively (Fig. 13). There was a tendency ( P = 0.09) for NR supplementation to increase the amount of NAD+ in the semitendinosus muscle by 62% (Fig. 14).

Discussion

No previous study has been conducted that examined the ability of nicotinamide riboside chloride to delay the onset of fatigue in pigs. This study demonstrated that feeding 30 mg/kg body weight of nicotinamide riboside to finished market barrows (280-300 pounds) numerically increased the time and distance barrows moved when they were subjected to a performance test. The study also demonstrated barrows fed 30 mg/kg body weight of nicotinamide riboside had greater semitendinosus muscle nicotinamide adenine dinucleotide (NAD+) content. Therefore, these data indicate supplementing nicotinamide riboside to finished market barrows delays the onset of fatigue, possibly by increasing muscle NAD+ content, and may serve as a countermeasure to prevent transportation losses.

Conclusion

Feeding 30 mg/kg body weight of nicotinamide riboside 7 days prior to a performance test increased the time and distance barrows moved. Nicotinamide riboside also increased semitendinosus muscle NAD+ levels, which could provide more energy for movement. This product may have potential to serve as a nutritional countermeasure to reduce the incidence of transport fatigue.