NIHPrimate Concentrate Supplement, A Novel NonHuman Primate Diet Supplement

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NIH-Primate Concentrate Supplement, A Novel
Non-Human Primate Diet Supplement

• Dennis Barnard1 and Carol Clarke2
• 1 Veterinary Resources Program, Division of
  Intramural Research Services, Office of Resource
  Services
• 2 Animal Care Branch, Division of Intramural
  Research, National Institute of Allergy and
  Infectious Diseases

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Abstract
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Introduction

• The nutritional status of monkeys involved
  in biomedical research can have a significant
  influence on experimental results. Laboratory
  monkeys involved in research protocols may become
  anorexic and require dietary supplementation.
  Along with the anorexia related to research, New
  World primates such as Aotus and Callitrichidae
  are fastidious eaters that do not typically
  consume the nutritionally balanced laboratory
  primate diets available, therefore require
  dietary supplementation. A common practice in
  primate research facilities is to provide dietary
  supplements produced in-house to monkeys that are
  anorexic. The typical in-house non-human primate
  diet supplement is a combination of laboratory
  monkey diet, fruit, juice, water, and vitamin and
  mineral supplements. The ingredients are mixed
  into a mash consistency that can be formed into
  meat balls or served in a cup. To get the mash
  consistency, the major ingredient is water or
  juice, which is 60 or more of the total weight.
  On an as-is basis, this results in a dilution of
  caloric density and nutrient concentration of the
  supplement by 60 or more. Furthermore, the
  commercially available vitamin and mineral
  supplements that are used are not concentrated
  enough to overcome this dilution factor. A
  very palatable supplement that does not provide
  the nutritional requirements, and may be
  functioning as a sole-source diet, can be
  detrimental to the monkeys health and
  experimental results. The diet supplement, NIH
  Primate Concentrate (NIH-PC), has been developed
  to provide nutritional requirements to non-human
  primates that are anorexic. The NIH-PC was
  formulated so that it can be diluted up to 60,
  with whatever ingredients are required to make it
  palatable, and still meet the monkeys
  nutritional requirements. The concentrate is
  composed of highly digestible ingredients. Four
  sources of high quality protein are used to
  provide the complete amino acid requirements. The
  primary fat source is soy oil, the single source
  of dietary fat that can provide close to optimal
  essential fatty acid concentrations as well as
  fatty acid proportions. A series of palatability
  studies utilizing three flavors (cherry,
  raspberry, and cheese) of the NIH-PCS were
  performed by feeding them to healthy Aotus spp
  (owl) and Macaca mulatta (rhesus) monkeys.

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NIH- Primate Concentrate Formulation

• Ingredients Percentage by Weight Isolated
  Soy Protein 24.000
• Casein Dried 20.000
• Rice Protein Concentrate 16.000
• Soy Oil 14.000
• Calcium Phosphate Tri. 8.500
• Dryfat Plus 7.972
• Mineral Mix 3.000
• Sugar Food Product 3.000
• Vitamin Mix 2.000
• Sodium Chloride 1.500
• Tert-butylhydroquinone 0.028
• 100.000

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NIH-Primate Concentrate Proximate Analysis

• Nutrient Concentration
• Crude Protein 44.0
• Crude Fat 25.0
• Crude Fiber 0.3
• Ash 11.9

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NIH Primate Concentrate Supplement
Formulations(Ingredients measured by weight)

• Raspberry Flavor
• Primate Concentrate 40
• Monkey Diet 15
• Apple juice 24.995
• Bananas 20
• Raspberry flavor 0.5
• Cherry Flavor
• Primate Concentrate 40
• Monkey Diet 15
• Apple juice 24.995
• Bananas 20
• Cherry flavor 0.5
• Cheese Flavor
• Primate Concentrate 40
• Monkey Diet 10
• Apple juice 25
• Bananas 15

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Proximate Analysis for NIH- Primate Concentrate
Supplement

• Crude protein Minimum 20.10
• Crude fat Minimum 10.75
• Crude fiber Maximum 1.90
• Gross Energy Kcal/g Minimum 2.80

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Vitamin Concentrations for NIH- Primate
Concentrate Supplement

• Vitamin A IU/g 23.00
• Vitamin D IU/g 6.90
• Alpha-tocopherol PPM 99.00
• Thiamin PPM 29.40
• Riboflavin PPM 21.80
• Niacin PPM 120.00
• Pantothenic Acid PPM 90.00
• Choline PPM 1637.00
• Pyridoxine PPM 19.40
• Folic acid PPM 33.00
• Biotin PPM 0.40
• Vitamin B12 Mcg/kg 57.50
• Vitamin K PPM 9.50
• Vitamin C PPM 1920.00

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Mineral Concentrations for NIH- Primate
Concentrate Supplement

• Calcium 1.50
• Phosphorous 0.74
• Potassium 0.80
• Sodium 0.27
• Magnesium 0.20
• Iron PPM 203.00
• Zinc PPM 72.00
• Manganese PPM 58.00
• Copper PPM 10.30
• Cobalt PPM 0.92
• Iodine PPM 2.70

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Amino Acid Concentrations for NIH- Primate
Concentrate Supplement

• Arginine of total diet 1.25
• Lysine 1.33
• Methionine 0.68
• Cystine 0.24
• Tryptophan 0.24
• Glycine 0.35
• Histidine 0.53
• Leucine 1.73
• Isoleucine 1.11
• Phenylalanine 1.02
• Threonine 0.79
• Valine 1.25

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Equipment and Ingredients

• Batch Processor
• Balance
• Syringe
• NIH-Primate Concentrate
• Apple Juice
• Monkey Diet
• Bananas
• Fruit Flavor

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NIH-PCS Preparation Process

• Preparing to make the NIH-PCS
• Monkey biscuits after grinding in the batch
  processor
• NIH- Primate concentrate before and after being
  mixed with monkey biscuits
• NIH-PCS raspberry flavored meat balls

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Palatability Study Procedure

• Prepared NIH-PCS biweekly
• 25 Aotus monkeys were fed a 10 g NIH-PCS meat
  ball daily along with their normal diet. Uneaten
  NIH-PCS was removed from the pans and cages and
  weighed 24 hours after feeding to determine how
  much of the NIH-PCS was eaten
• 21 rhesus monkeys were fed a 25 g NIH-PCS meat
  ball daily along with their normal diet. Uneaten
  NIH-PCS was removed from the pans and cages and
  weighed 15 minutes after feeding to determine how
  much of the NIH-PCS was eaten
• The raspberry and cherry flavors were increased
  from 0.05 to 1.5 over time to determine the
  affect on palatability
• Cheese flavor was increased from 10 to 12 over
  time to determine the affect on palatability
• Daily observations were made to detect diarrhea
  and inappetence
• Monkeys were weighed regularly

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Aotus Monkeys
Rhesus Monkeys
Raspberry NIH-PCS
Raspberry NIH-PCS
Cherry NIH-PCS
Cherry NIH-PCS
Cheese NIH-PCS
Cheese NIH-PCS

• Fed each flavor for 20 to 40 days
• Recorded gastrointestinal disorders ( ie
  diarrhea)
• Recorded inappetence
• Recorded body weights

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Results of the Palatability Study

• The NIH-Primate Concentrate Supplement is highly
  palatable
• The fruit flavors were more palatable than cheese
• The monkeys did not experience diarrhea or
  constipation
• The aotus and rhesus monkeys slightly increased
  their weight during the palatability study

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PF (number of days monkeys ate more than 75 of
supplement)/ (number of days monkeys were fed
supplement)
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Discussion

• In non-human primate research facilities
  it is a common practice to feed dietary
  supplements along with the commercial primate
  diet to monkeys showing signs of inappetence or
  anorexia. There are factors that can affect
  research that must be considered when feeding
  diet supplements. Feeding diet supplements to
  monkeys on experimental protocols will introduce
  a variable into the research. To prevent
  introducing a dietary variable into the research
  it is important that there is a documented
  formulation for the supplement and that all
  modifications of the formulation are documented.
  The preparation of the diet supplement can also
  introduce variables if the ingredients are not
  measured accurately and the finished supplement
  is not a homogeneous mixture of the ingredients.
  Manual preparation is very labor intensive and
  inconsistencies in the homogeneity are probable,
  which will affect palatability and nutrient
  content. To avoid these problems the
  NIH-Primate Concentrate Supplement (NIH-PCS) was
  prepared using a syringe to measure small
  volumes, batch processor, and balance. The batch
  processor is highly recommended for primate
  research facilities because it is a labor saving
  device that can improve the ingredient
  homogeneity and nutrient consistency of the
  supplement.
• Typically, the diet supplements are
  developed in-house, from a combination of
  laboratory monkey diet, fruit, water, juice,
  vitamin and mineral supplements, and anything
  else to make it palatable. These in-house
  supplements are usually very palatable, but do
  not meet the nutritional requirements of the
  monkeys. If the diet supplement becomes the
  primary source of nutrition it can be detrimental
  to the monkeys health and affect experimental
  results. This problem is particularly true for
  New World monkeys such as the Aotus and
  callitrichids which are fastidious eaters1.
  The open formula NIH Primate Concentrate (NIH-PC)
  has been developed to provide nutritional
  requirements to non-human primates that are
  anorexic. The NIH-PC was formulated so that it
  can be diluted up to 60, with whatever
  ingredients are required to make it palatable,
  and still meet the monkeys nutritional
  requirements. The protein, fat, and gross
  energy for the NIH-Primate Concentrate Supplement
  formulations used in this study were 20.1,
  10.8, and 2.8 kcal/g, respectively. The
  vitamin, mineral, and amino acid concentrations
  met NRC requirements2. The results show that the
  formulations of the NIH-PCS used in the feeding
  trial are very palatable to Aotus spp and Macaca
  mulatta monkeys. The fruit flavors appeared to
  be slightly preferred over the cheese flavor.
  The aotus and rhesus monkeys weights increased
  slightly from the start of the palatability study
  to the end, and no gastrointestinal problems were
  observed. These results indicate that the
  NIH-PCS can be used as a dietary supplement for
  non-human primates. Further evaluation will
  determine if it can be fed as a sole source diet.

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AcknowledgementThe authors acknowledge the
contributions of Drs. Malcom Martin and Carol
Long and the VRP Primate Unit technical staff to
the concept of this project.

• References
• Barnard, D.E. and J.J. Knapka. Callitrichidae
  Nutrition, in The Cotton-Top Tamarin A Primate
  Model for the Study of Colon Carcinogenesis, (N.
  Clapp, ed.), CRC Press, 1993, Chapter 3.
• National Research Council. Nutrient Requirements
  of Nonhuman Primates, National Academy of
  Science, Washington, D. C., 1978.