This section contains the minimum testing methods for the substantiation of nutritional adequacy claims, calorie content claims, and procedures for establishing

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Proposed Revisions Edited per Comments for 2014 Official P ublication 1 AAFCO METHODS FOR SUBSTANTIATING NUTRITIONAL ADEQUACY OF DOG AND CAT FOODS This section contains the minimum testing methods for the substantiation of nutritional adequacy claims , calorie content claims , and procedures for establishing pet food product f amilies referenced in AAFCO Model Pet Food and Specialty Pet Food Regulations PF2, 4, 7, 8, 9 and /or 10. These methods represent minimum requirements . C ompanies may choose , or may need , to perform additional tes ting to substantiate their claims. AAFCO D og and Cat Food Nutrient Profiles Introduction The original Canine and Feline Nutrition Expert Subcommittees convened in 199 0 were charged by the chair of the AAFCO Pet Food Committee to establish practical nutrient profiles for both dog and cat foods based on commonly used ingredients. These subcommittees established the “AAFCO Dog Food Nutrient Profiles” and the “AAFCO Cat Food N utrient Profiles” that appeared in the Official Publication of the AAFCO in 1992 and 1993, respectiv ely . The profiles were reviewed in 1994/95 and updates to the maximum concentrations for vitamin A in dog foods were implemented in 1996 . The National Research Council (NRC) in 2006 updated its published Nutrient Requirements of Dogs and Nutrient Requirem ents of Cats in a single publication that combined recommendations for both specie s. 1 In 2007 the AAFCO Pet Food Committee again formed Canine and Feline Nutrition Expert Subcommittees and charged these subcommittees with the task of revising the AAFCO Nut rient Profiles in consideration of the i nformation in the 2006 NRC Nutrient Requirements of Dogs and Cat s (2006 NRC) . In addition, the subcommittees considered information in the NRC Mineral Tolerance of Animals Second Revised Edition, 2005 (2005 Mineral Tolerance of Animals ).2 Finally, the subcommittees also reviewed and considered the recommended nutrient concentrations for dog and cat food products as published in February 2008 by the European Pet Food Industry Federation ( Federation Europeenne de l™In dustrie des Alimentis pour Animaux Familiers (FEDIAF)) , titled F.E.D.I.A.F. Nutritional Guidelines for Complete and Compleme ntary P et Food fo r Cats and Dogs , (FEDIAF Guidelines) that are roughly the European -equivalent to the AAFCO Dog and Cat Food Nutrien t Profiles .3 The AAFCO Dog and Cat Food Nutrient Profiles were designed to establish pra ctical minimum and some maximum nutrient concentrations for dog and cat foods, formulated from commonly used, non -purified, complex ingredients. The concentrations dif fer from minimum nutrient requirements traditionally deve loped by the NRC Committee on Animal Nutrition. Many of the NRC minimum nutrient requirements are based on research with purified diets and/or highly bioavailable nutrient sources that are not pract ical to use in commercial dog and cat foods . Therefore, u nlike the previous NRC publications Nutrient Requirements of Dogs in 19854 and Nutrient Requirements of Cats in 1986 ,5 the Nutrient Requirements of Dogs and Cats in 2006 contained two additional list ings of nutrient concentrations for adequate intake and recommended allowance (RA) in addition to minimum requirements. The concentration s for RA™s of nutrients in the 2006 NRC are at least equal to, or greater than , concentrations for adequate intakes an d minimum requirements , respectively, and are defined as fithe concentration or amount of a nutrient in a diet formulated to support a given physiological state.fl When appropriate,

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Proposed Revisions Edited per Comments for 2014 Official P ublication 2 the RA takes into consideration the bioavailability of the nutrient. Thus , the Canine and Feline Nutrit ion Expert Subcommittees of 2007 primarily used the RA in the 2006 Nutrient Requirements of Dogs and Cats in evaluating whether revision was needed to one or more of the minimum recommended concentrations in the profiles. Values for specific nutrient concentrations were added or modified where indicated and supported by recent scientific publications, practical experience, or unpublished data. The AAFCO Dog and Cat Food Nutrient Profiles have been criticized and faulted for not explicitly indicating the apparent nutrient digestibility , sometimes called nutrient availability or bioavailability, required to make the listed concentrations adequate for meeting the animal™s daily requirements. When a minimum requirement has been establ ished for a particular nutrient, the expected apparent digestibility to meet the minimum requirement for that nutrient at the recommended concentration listed in an AAFCO Nutrient Profile can be calculated using the formula : ((minimum requirement) x (its apparent digestibility in the diet(s) used to establish the minimum requirement) / (recommended concentration in the AAFCO Profile) ) x 100 . In th e above formula, the minimum requirement is expressed in the same units as in the AAFCO Nutrient Profile and digestibility is expressed in decimal equivalents . As an example , the NRC list s the minimum crude protein requirement for puppies to be met by formulas containing 18% crude protein on a dry matter basis with the digestibility of the protein sources estim ated to be near 100% . The 2014 AAFCO Dog Food Nutrient Profile for Growth and Reproduction recommends the minimum crude protein concentration of dry matter to be 22.5%. Therefore , the expected apparent digestibility for crude protein in a diet formulated to meet the AAFCO Dog Food Nutrient Profile for Growth and Reproduction is at least 80% [(18 x (1.00) /22.5) x 100]. For nutrients known to be essential , but that lack sufficient data to establish a minimum requirement , the typical digestibility for the n utrient in ingredients and food matri ces similar to those used to establish the apparent amount to fulfill the animal ™s need for the nutrient should be ensured. The 2006 Nutrient Requirements of Dogs and Cats discusses average or typical apparent digestib ility for such nutr ients when explaining how a RA was set. As an example, for adult dogs there is no established minimum requirement for iron, alth ough iron is considered essential for adult dogs. In setting the RA of 30 mg/kg in dietary dry matter for a dult maintenance, the NRC subcommittee considered the apparent digestibility of iron to be 20%. However , the explanatory text in the publication note s that measured apparent digestibility of iron in the scientific literature has ranged from close to 100% to les s than 10% , and is affected by numerous factors such as the specific source of iron , the concentration of other specific minerals or other ingredients in the diet , as well as the iron status of the animal . The specific example for iron can be general ized to most essential minerals, and demonstrates th e impossib ility that any list of concentrations can invariably ensure that all nutrient requirements are fulfilled in all diet formulas without additional considerations. As stated for the previous editi ons of the AAFCO Dog and Cat Food Nutrient Profiles, formulating a product according to the Pr ofiles is only one part of a nutritionally sound, scientific development that must consider all other aspects of the product. The fact that a dog or cat food is formulated to meet a specific AAFCO Profile should not deter or discourage the manufacturer from co nducting appropriate feeding trials to further confirm and ensure the diet is nutritional ly adeq uate for its intended use .

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Proposed Revisions Edited per Comments for 2014 Official P ublication 3 Indications regarding expected nut rient availability from some ingredient source s are given in footnotes . It is important to read the footnotes to the tables as they contain information critical to many of the recommend ed concentrations . Additionally, manufacturers must make allo wances to nutrient concentrations prior to processing to account for losses during processing and subsequent storage. The recommended concentrations in the Profiles are those expected to be present at the time the formula is consum ed by the animal. The established profiles are the fi AAFCO Dog Food Nutrient Profiles fl and fiAAFCO Cat Food Nutrient Profiles fl as the terms are applied in AAFCO model pet food regulations referring to nutritional adequacy. Under these model regulations, dog and cat foods substantiated for nutritional adequacy by reference to the AAFCO Dog and Cat Food Nutrient Profiles for a designated life stage(s) must be formulated to contain at least the minimum concentrations of nutr ients specified in the P rofiles , and , for some nutrient s, not more tha n any maximum concentration listed for that specific nutrient in the P rofiles as shown in this section. Products with their nutritional adequacy substantiated by AAFCO Feeding Protocols are not mandated to meet the minimum or maximum concentrations listed in the Profiles. Additionally, snacks, treats or products intended for intermittent or su pplemental feeding only are not mandated to meet the concentrations in the Profiles unless their labeling references the Profiles . The AAFCO Dog and Cat Food Nutrient Profiles and the AAFCO Feeding Protocols are the only methods recognized by AAFCO for substantiating the nutritional adequacy of “complete and balanced” dog or cat foods. If a product is substantiated by a feeding trial and does not meet the AAFCO Dog or Cat Food N utrient Profiles, the label cannot reference the Profiles. An unqualified reference to an AAFCO Dog or Cat Food Nutrient Profile is an implied guarantee that the product contains the minimum concentrations for all nutrients in the profile and n o more than any maximum concentration listed for a specific nutrient in the profile. Minimum and some maximum nutrient concentrations were established in the Profiles for two categories ; growth and reproduction (gestation/lactation), and adult maintenance. Maximum nutrient concentrations were established for nutrients where the potential for ove ruse or toxicit y is of concern and likely to occur if attention is not paid to the concentration s of those nutrients . The absence of a maximum concentration should not be interpreted to mean that nutrients without a specific maximum content are safe at any concentration . Rather, it reflects the lack of info rmation in dogs and cats on toxic concentrations of that nutrient. Establishing a maximum concentration impli es safety below that concentration for long term consumption and to set a maximum arbitrarily might prove worse than no maximum at all. The nutrient concentrations are expressed on a dry matter (DM) basis and at a specified caloric density. Diets should b e corrected for caloric density as indicated below . Reference to the concentrations of nutrients on a product label in the guaranteed analysis must be expressed in the same units and order as given in the AAFCO Dog or Cat Food Nutrient Profiles. For the purposes of determining metabolizable energy (ME) , use the methods specified in Model Regulation PF9. AAFCO DOG FOOD NUTR IENT PROFILES BASED ON DRY MATTER a Nutrients Units DM Basis Growth & Reprodu ction Minimum Adult Maint enance Minimum b Maximum

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Proposed Revisions Edited per Comments for 2014 Official P ublication 4 Crude Protein % 22.5 18.0 Arginine % 1.0 0.51 Histidine % 0.44 0.19 Isoleucine % 0.71 0.38 Leucine % 1.29 0.68 Lysine % 0.90 0.63 Methionine % 0.35 0.33 Methionine -cystine % 0.70 0.65 Phenylalanine % 0.83 0.45 Phenylalanine -tyrosine % 1.30 0.74 Threonine % 1.04 0.48 Trytophan % 0.2 0 0.16 Valine % 0.68 0.49 Crude Fat c % 8.5 5.5 Linoleic acid % 1.3 1.1 alpha -Linolenic acid % 0.08 NDd Eicosapentaenoic + Docosahexaenoic acid % 0.05 NDd (Linoleic + Arachidonic):(alpha -Linolenic + Eicosapentaenoic + Docosahexaenoic) acid Ratio 30:1 Minerals Calcium % 1.2 0.5 1.8 Phosphorus % 1.0 0.4 1.6 Ca:P ratio 1:1 1:1 2:1 Potassium % 0.6 0.6 Sodium % 0.3 0.08 Chloride % 0.45 0.12 Magnesium % 0.06 0.06 Iron e mg/kg 88 40 Copper f mg/kg 12.4 7.3 Manganese mg/kg 7.2 5.0 Zinc mg/kg 100 80 Iodine mg/kg 1.0 1.0 11 Selenium mg/kg 0.35 0.35 2 Vitamins & Other Vitamin A IU/k g 5000 5000 250000 Vitamin D IU/kg 500 500 3000 Vitamin E g IU/kg 50 50 Thiamine h mg/kg 2.25 2.25 Riboflavin mg/kg 5.2 5.2 Pantothenic acid mg/kg 12 12

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Proposed Revisions Edited per Comments for 2014 Official P ublication 5 Niacin mg/kg 13.6 13.6 Pyridoxine mg/kg 1.5 1.5 Folic acid mg/kg 0.216 0.216 Vitamin B 12 mg/kg 0.02 8 0.02 8 Choline mg/kg 1360 1360 a Presumes a caloric density of 4000 kcal ME/kg, as determined in accordance with Model Regulation PF9. Formulation s greater than 4000 kcal ME/kg must be corrected for e nergy density ; formulations less than 4000 kcal ME/kg need not be corrected for energy. Formulations of low -energy density should not be considered adequate for r eproductive needs based on comparison to the Profiles alone. b Recommended concentrations for maintenance of body weight at an average caloric intake for dogs of a given optim um weight. c Although a true requirement for crude fat per se has not been established, the minimum concentration was based on recognition of crude fat as a source of essential fatty acids, as a carrier o f fat -soluble vitamins, to enhance palatability, and to supply an adequate caloric density. d ND ΠNot Determined. While a minimum requirement has not been determined, sufficient amounts of omega -3 fatty acids are necessary to meet the maximum omega -6:om ega -3 fatty acid ratio. e Average apparent digestibility for iron associated with recommended minimums is 20% of that consumed. Because of very poor apparent digestibility, iron from carbonate or oxide sources that are added to the diet should not be cons idered in determining the minimum nutrient concentration for iron. f Because of very poor apparent digestibility, copper from oxide sources that are added to the diet should not be considered in determining the minimum nutrient concentration for copper. g It is recommended that the ratio of IU of vitamin E to grams of polyunsaturated fatty acids (PUFA) be > 0.6:1. A diet containing 50 IU of vitamin E will have a ratio of > 0.6:1 when the PUFA content is 83 grams or less. D iets containing more than 83 gram s of PUFA should contain an additional 0.6 IU of vitamin E for every gram of PUFA. h Because processing may destroy up to 90% of the thiamine in the diet, allowances in formulation should be made to ensure the minimum nutrient concentration for thiamine is met after processing. AAFCO DOG FOOD NUTRIENT PROFILES BASED ON CALORIE CONTENT Nutrients Units per 1000 kcal ME Growth & Reproduction Minimum Adult Maint enance Minimum a Maximum Crude Protein g 56.3 45.0 Arginine g 2.50 1.28 Histidine g 1.10 0.48 Isoleucine g 1.78 0.95 Leucine g 3.23 1.70 Lysine g 2.25 1.58 Methionine g 0.88 0.83 Methionine -cystine g 1.75 1.63

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Proposed Revisions Edited per Comments for 2014 Official P ublication 6 Phenylalanine g 2.08 1.13 Phenylalanine -tyrosine g 3.25 1.85 Threonine g 2.60 1.20 Tryptophan g 0.5 0 0.40 Valine g 1.70 1.23 Crude Fat b g 21.3 13.8 Linoleic acid g 3.3 2.8 alpha -Linolenic g 0.2 NDc Eicosapentaenoic + Docosahexaenoic acid g 0.1 NDc (Linoleic +Arachidonic) :(alpha -Linolenic +Eicosapentaenoic+ Docosahexaenoic) acid Rati o 30:1 Minerals Calcium g 3.0 1.25 4.5 Phosphorus g 2.5 1.00 4.0 Ca:P Ratio 1:1 1:1 2:1 Potassium g 1.5 1.5 Sodium g 0.80 0.20 Chloride g 1.10 0.30 Magnesium g 0.10 0.15 Iron d mg 22 10 Copper e mg 3.1 1.83 Mang anese mg 1.8 1.25 Zinc mg 25 20 Iodine mg 0.25 0.25 2.75 Selenium mg 0.09 0.08 0.5 Vitamins & Others Vitamin A IU 1250 1250 62500 Vitamin D IU 125 125 750 Vitamin E f IU 12.5 12.5 Thiamine g mg 0.56 0.56 Riboflavin mg 1.3 1.3 Pantothenic acid mg 3.0 3.0 Niacin mg 3.4 3.4 Pyridoxine mg 0.38 0.38 Folic acid mg 0.0 54 0.0 54 Vitamin B 12 mg 0.007 0.007 Choline mg 340 340 a Recommended concentrations for maintenance of body weight at an average caloric intake for dogs of a given optim um weight. b Although a true requirement for crude fat per se has not been established, the minimum concentration was based on recognition of crude fat as a source of essential fatty acids, as a carrier of fat -soluble vitamins, to enha nce palatability, and to supply an adequate caloric density.

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Proposed Revisions Edited per Comments for 2014 Official P ublication 8 and phenylalanine plus tyrosine. The CNES felt it prudent to include spec ific minimums for methionine and phenylalanine because although some, or all, of the requirement for cystine and tyrosine can be met from excess methionine and phenylalanine, respectively, the reverse is not true. Some of the previous recommendations for dietary concentrations of essential amino acids in the Dog Food Nutrient Profile for Adult Maintenance (i.e., histidine, lysine, threonine and tryptophan) were greater than the corresponding RA in the 2006 NRC and the CNES elected to retain the previously recommended amounts for these amino acids in the current Dog Food Nutrient Profile for Adult Maintenance. Minimum concentrations of some essential amino acids in the Dog Food Nutrient Profile for Growth and Reproduction were increased, usually to match t he NRC RA for growth (i.e., arginine, leucine, methionine, methionine -cystine, phenylalanine -tyrosine and valine). Although the NRC RA for total crude protein during lactation is essentially identical to the RA for growth (22.0% versus 22.5%), several of the RA for essential amino acids during lactation are greater than the RA for growth. In some cases (i.e., histidine, isoleucine , lysine, phenylalanine, and threonine) the difference was small and the CNES elected to set the recommended amount in the Grow th and Reproduction Profile at the larger NRC RA for lactation. For other essential amino acids (i.e., leucine and valine) the RA proposed by the NRC for lactation is substantially more than the RA for growth, and in the case of leucine and valine the con centrations are equal to, or greater than, the corresponding RA for the cat during lactation, an obligate carnivore with protein requirements generally greater than those for the dog. The NRC ad hoc committee indicated that it set the RA based on, filowest concentrations of each of the essential amino acids from digestible protein in commercial dry expanded diets that have been shown to sustain normal gestation and lactation for bitches.fl 1 The CNES chose not to increase the recommended concentrations for leucine and valine to those of the NRC RA for lactation based on lack of documented problems with the previous concentrations in the AAFCO Dog Food Nutrient Profile for Growth and Reproduction and the relative disparity in the RA between canine versus felin e protein requirements. The CNES did not elect to change the tryptophan concentration in the Dog Food Nutrient Profile for Growth and Reproduction for two reasons. The CNES had access to feeding studies and a publication showing that the minimum requirem ent for t ryptophan in Labrador retriever puppies was less than the current concentration in AAFCO Dog Food Nutrient Profile for Growth and Reproduction and th at the tryptophan concentration of 0.2% DM already provided approximately a 25% safety margin. 6 The CNES was also aware that it was nearly impossible to formulate a product at the minimum protein concentration to contain more than 0.2% tryptophan on a DM basis from typical ingredients without including crystalline tryptophan in the formula. Insufficie nt data w ere available to demonstrate detrimental effects of high protein intake in the normal dog to allow for any definitive maximum concentrations for protein or amino acids to be established. The CNES is aware of the findings regarding excess lysine a t some concentration between 2.0% and 4.0% lysine/kg DM to produce depression in growth of puppies and clinical signs associated with arginine deficiency when arginine is present at 0.4% DM, and that FEDIAF has established a concentration of 2.8% lysine in DM as a maximum. 3,7 However, this information was available prior to the establishment of the original AAFCO Nutrient Profiles and did not result in a maximum lysine content being established by the 1990 Expert Subcommittee. Furthermore, the 2007 CNES notes that the minimum recommended arginine content for growth and reproduction is 2.5 times the concentration of 0.4% arginine/kg DM required to produce the noted adverse effects in combination with lysine at more than 2.0%/kg DM.

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Proposed Revisions Edited per Comments for 2014 Official P ublication 9 FAT/FATTY ACIDS The CNES increased the minimum recommended amount for total fat in the AAFCO Dog Food Nutrient Profiles by 0.5% to 8.5% for Growth and Reproduction and 5.5% for Adult Maintenance. These concentrations are consistent with the RA for total fat in the 2006 NRC and t he FEDIAF Guidelines . The CNES also increased the minimum recommended linoleic acid concentration in the Growth and Reproduction Profile from 1.0% to 1.3% and in the Adult Maintenance Profile from 1.0% to 1.1%, again consistent with the RA in the 2006 NRC . The CNES did not set a minimum recommended concentration for arachidonic acid in either profile, but did establish minimum recommended concentrations for some fatty acids in the n -3 (omega -3) series in the Growth and Reproduction Profile, specifically, alpha -linolenic acid at 0.08%, and the combination of eicosapentaenoic plus docosahexaenoic acids at 0.05%, of DM. Because the scientific evidence to date indicates that these n -3 fatty acids are needed for the development of the nervous and visual system s during fetal and neonatal life stages, the CNES did not feel there was scientific justification for setting a specific minimum recommended concentrations for n -3 fatty acids for adult maintenance . A recommendation in a comment to list quantities of alph a-linolenic acid and eicosapentaenoic plus docosahexaenoic acids for adult maintenance as being not determined (ND) was accepted by the AAFCO Pet Food Committee . The CNES did not establish maximum concentrations for fat or fatty acids despite the NRC listi ng a safe upper limit (SUL) for total crude fat, linoleic acid, and the combination of eicosapentaenoic plus docosahexaenoic acids. The CNES felt it likely that insufficiencies in other nutrients will occur in a conventional formula before an inclusion of 33% crude fat in DM is reached. Also , although some differences in delayed hypersensitivity reactions were noted in studies cited by the NRC as the basis for setting the SUL for eicosapentaenoic plus docosahexaenoic acids, the 2007 CNES noted that those differences are not unequivocally undesirable or detrimental. 8,9 The CNES did elect to set a maximum for the ratio of the sum of linoleic plus arachidonic acids to the sum of alpha -linolenic , eicosapentaenoic , and docosahexaenoic acids at 30:1 given the m odulating effects of n -3 fatty acids on n -6 metabolism and the predominant contribution of these fatty acids to the n -6 and n -3 fatty acid content s, respectively, in conventional dog food formulas. CALCIUM & PHOSPHORUS The CNES decreased the recommended m inimum concentration of calcium and phosphorus in the Adult Maintenance Profile by 0.1% to 0.5% and 0.4%, respectively. The current recommended minimum concentrations are 0.1% more than the RA for calcium and phosphorus on a DM basis for adult maintenance in the 2006 NRC b ut consistent with the concentrations in the FEDIAF Guidelines. The CNES increased the minimum calcium and phosphorus concentrations in the Growth and Reproduction Profile to 1.2% and 1.0%, respectively, consistent with the 2006 NRC RA a nd FEDIAF Guidelines. The CNES recommended that the calcium and phosphorus in growth formulas for the large -bred or large -size dog s be allowed to decrease to 0.9% and 0.75%, respectively, while still being judged to meet the G rowth and Re production Nutrie nt Profile. However, based on comments and a publication 10 demonstrating that some diets containing 0.88 % to 1.04% Ca on a DM basis (2.2 to 2.6 g Ca/1000 kcal ME) when fed to medium – or large -breed puppies produced inhibited growth in 10 -week growth studi es compared to diets containing between 1.3 to 1.8% Ca, the AAFCO Pet Food Committee elected to keep the minimum recommended calcium and phosphorus concentrations in the Growth and Reproduction Nutrient Profile at 1.2% and 1.0%, respectively, for all dog f ood products that substantiate nutritional adequacy

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Proposed Revisions Edited per Comments for 2014 Official P ublication 10 based on being formulated to meet the nutrient content of the Dog Food Nutrient Profile for Growth and Reproduction. Because of concerns for excess calcium to produce detrimental effects in growing dogs of large and giant breeds ,10-13 the 2007 CNES deemed that additional restriction to the maximum limit for calcium was warranted and lowered the maximum calcium concentration to 1.8% DM. The CNES did not believe it necessary to decrease the previous maxim um calcium concentration of 2.5% for adult dogs or growing dogs of small or moderate size breeds . However, the AAFCO Pet Food Committee felt that only one maximum value should be established , especially to ensure all life stage products were properly form ulated. Thus , the AAFCO Pet Food Committee elected to set the maximum calcium for all dog foods formulated to meet the Dog Food Nutrient Profiles at 1.8% DM. The CNES retained the maximum phosphorus concentration of 1.6% DM for both profiles, as well as the minimum and maximum values of 1:1 and 2:1, respectively, for the calcium to phosphorus ratio. OTHER MACROMINERALS POTASSIUM The 2007 CNES elected to retain the recommended minimum potassium concentration at 0.6% DM for both Profiles. Although the RA in the 2006 NRC and some concentrations in the FEDIAF Guidelines are less than 0.6% DM for potassium, the CNES felt that the potassium concentration did not warrant changing especially given that pote ntial toxicosis of potassium was not a practical concern . Thus, a maximum concentration for potassium was not established. SODIUM & CHLORIDE The 2007 CNES did not change the minimum recommendation for sodium or chloride in the Growth and Reproduction Nutrient Profile as the values are slightly above the 2006 N RC RA. The 2007 CNES made an editorial increase in the recommended minimum concentrations for sodium and chloride in the Adult Maintenance Nutrient Profile to match the 2006 NRC RA. For sodium the increase was from 0.06% to 0.08% DM and for chloride from 0.09 to 0.12% DM. The recommended minimum concentrations for sodium and chloride in both dog food nutrient profiles continue to reflect the 1:1.5 sodium to chloride ratio of salt previously used by the 1990 CNES to justify recommended chloride concentrat ions. As noted by the 1990 CNES , because palatability and food consumption would decline due to excess sodium before adverse health effects were observed, setting a maximum concentration for sodium was not of practical concern. MAGNESIUM The 2007 CNES inc reased the minimum recommended concentration for magnesium from 0.04 to 0.06% in Adult Maintenance and Growth and Reproduction Nutrient Profiles to match the 2006 NRC RA for adult maintenance and peak lactation, respectively. The 2007 CNES deleted the max imum recommended concentration for magnesium due to lack of data specific to dogs in both the 2006 NRC and the 2005 Mineral Tolerances of Animals . The only comment regarding maximum magnesium content in the 2006 NRC was that a SUL for magnesium in the die ts of dogs was greater than 1.7% DM. MICROMINERALS IRON The 2007 CNES made an editorial change to the minimum concentration for iron in the Growth and Reproduction Nutrient Profile to make the concentration consistent with a presumed caloric density of 40 00 kcal ME/kg DM which makes the recommended concentration consistent with the RA from the 2006 NRC and the

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Proposed Revisions Edited per Comments for 2014 Official P ublication 11 FEDIAF Guidelines for same life stages. The 2007 CNES decreased the recommendation for adult maintenance from 80 to 40 mg/kg DM based on considerat ions that the RA of the 2006 NRC was 30 mg/kg DM and the FEDIAF Guidelines concentration was 36 mg/kg DM. The 2007 CNES deleted the maximum concentration for iron based on one scientific and one practical regulatory consideration. First, the 2006 NRC ind icated th at appropriate data for setting a SUL for iron in dog foods are not available. The previous maximum concentration was stated to be based on tolerance data in swine. The 2005 Mineral Tolerance of Animals indicated that the listed tolerance of 300 0 mg/kg DM for swine needed to be confirmed by long -term studies and all other tolerances for iron listed in that publication are 6 times less than 3000 mg/kg DM. Second , the implied safety of a maximum concentration presumes some amount of apparent diges tibility and, as noted above, the apparent digestibility of iron in any given diet or combination of ingredients can vary from less than 10% to near 100%. Some sources of iron are considered unavailable and used for their technical effects (i.e., color) o n the product and not for their nutrient contribution of iron to the animal. Such unavailable sources will still contribute iron to an analytical result for determining product content, and thus a maximum concentration set for available sources of iron mi ght prohibit use of unavailable sources for coloring, whereas a maximum concentration set for unavailable colorants might permit use of unsafe amounts of available sources on the basis of analytical content. Thus, the 2007 CNES elected to delete the previ ous maximum of 3000 mg/kg DM and not list any other value as a maximum for iron. Manufacturers should note that iro n is toxic at some amount greater than the recommended quantities, but the ex act amount is unknown for dogs. COPPER The minimum concentratio n for copper in the Adult Maintenance Nutrient Profile was not changed from the previous amount of 7.3 mg/kg DM, the concentration being consistent with that of the FEDIAF Guidelines and slightly more than the 2006 NRC RA of 6.0 mg/kg. The 2007 CNES incre ased the minimum recommended concentration in the Growth and Reproduction Nutrient Profile to 12.4 mg/kg DM, consistent with the 2006 NRC RA for peak lactation and slightly more than FEDIAF Guidelines and the NRC RA for growth. Because of poor bioavailabi lity, the use of copper oxide as a nutritional source is excluded. 15 The 2007 CNES deleted the copper max imum concentration for many of the same science -based reason s cited above for deleting the maximum for iron content. MANGANESE The minimum concentrat ion for manganese in the Adult Maintenance Nutrient Profile was not changed from the previous amount of 5.0 mg/kg DM, the amount being slightly more than the 2006 NRC RA of 4.8 and slightly less than the FEDIAF Guidelines of 5.6 mg/kg DM. The 2007 CNES increased the minimum recommended concentration in the Growth and Reproduction Nutrient Profile to 7.2 mg/kg DM, consistent with the 2006 NRC RA for peak lactation and slightly more than FEDIAF Guidelines concentrations and NRC RA for growth. ZINC The 2006 N RC RA for zinc in growth, reproduction, and adult maintenance formulations was less than the previous concentration in the Dog Food Nutrient Profiles of 120 mg/kg DM and the 2007 CNES decreased the recommended minimum concentration to 100 mg/kg DM in the G rowth and Reproduction Nutrient Profile and to 80 mg/kg DM in the Adult Maintenance Nutrient Profile consistent with the 2006 NRC RA and FEDIAF Guidelines concentrations . Both the 2005 Mineral Tolerance of Animals and the 2006 Nutrient Requirements of Dog s and Cats state there is not enough data available to set a tolerance or SUL for zinc in dog food s. The 2007 CNES

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