From September 18 to October 17, 2025, we conducted a 30-day public consultation on the proposed amended description for 25-hydroxyvitamin D3 monohydrate from fermentation process.
Summary of feedback received
During the consultation period, we received 3 sets of comments from stakeholders.
Comments received and our response
A stakeholder highlighted to us that the European Food Safety Authority (EFSA) had some concerns regarding the production microorganism, Pseudonocardia autotrophica, used to produce 25-hydroxyvitamin D3. These concerns included the potential presence of viable cells of the production microorganism in the ingredient, uncertainties with regards to genotoxicity, and the purity due to the presence of impurities including 1α,25-dihydroxycholecalciferol (1,25-dihydroxyvitamin D3). The same stakeholder also asked whether 1,25-dihydroxyvitamin D3 was allowed as feed additive or ingredient in Canada.
- we cannot comment on the data provided to EFSA and their evaluation, but the data provided to our Animal Feed Program (AFP) demonstrated that this source of this part 2 single ingredient feed (SIF) is safe and efficacious for livestock, humans consuming products from livestock fed the product, and individuals handling the product
- based on the data and the weight of evidence:
- no viable cells or DNA from the production microorganism were detected
- genotoxicity was not observed in the provided studies for the SIF
- the source of this part 2 SIF is not crystallized; therefore, some impurities from the fermentation process might be present in the final SIF, such as 1,25-dihydroxyvitamin D3 and vitamin D3
- the glycosylated form of 1,25-dihydroxyvitamin D3 (a metabolite of this active form of vitamin D3) is found in the waxy-leaf nightshade leaf powder (2-504-010) which is an approved SIF set out in part 2 of the Canadian Feed Ingredients Table (CFIT)
- this SIF can be safely and efficaciously fed to broiler chickens where, in vivo, deglycosylation occurs to produce the active form,1,25-dihydroxyvitamin D3
A second stakeholder also referred to the opinion from EFSA and had concerns about the purity of the final SIF product, the presence of impurities and the inconsistencies in the "potency" of the final SIF product. A third stakeholder expressed concerns regarding the differences in the relative potency of the different forms of vitamin D3 and questioned if the conversion factor between International Units (IUs) of vitamin D3 and 25-hydroxyvitamin D3 was appropriate. In other words, if the IU equivalency statement in the SIF description was relevant for the active form of vitamin D3.
- we have reviewed the data for the source of this part 2 SIF, which included considering the purity and the potential for any substances/impurities originating from the fermentation process, and determined that it is safe and efficacious
- as a part 2 SIF, each source requires registration and an evaluation of each source's specific characteristics and safety
- we removed the IU equivalency statement from the amended description
- equivalency is based on the biological activity of vitamin D, rather than the biological activity of its metabolites, and therefore expressing the biological activity in IU for 25-hydroxyvitamin D3 is not appropriate
As a result of the comments and feedback received, we proposed the following changes to:
- the purpose statement in the amended SIF description
- included the approved production stages/classes of chickens to provide clarification
- added the maximum inclusion rates to the description
- clarified that this SIF should be used in combination with vitamin D (D2 and/or D3) (and not as the only source of vitamin D activity) and included the maximum content of the combination of 25-hydroxyvitamin D3 and vitamin D in the description to ensure the maximum value for vitamin D is not being exceeded in the total diet as indicated in the Tables of Maximum Nutrient Values for Feeds
- the labelling statements
- removed the International Units (IU) equivalency statement
- the IUs are meant to express the biological activity of vitamin D rather than the biological activity of its metabolites, and the hydroxylated form of vitamin D3 may exhibit a greater biological activity than vitamin D3
- since this SIF is not intended to replace the supplementation of vitamin D, the IU equivalency statement may lead to a misinterpretation of the potency of this active form
- the IUs are meant to express the biological activity of vitamin D rather than the biological activity of its metabolites, and the hydroxylated form of vitamin D3 may exhibit a greater biological activity than vitamin D3
- updated the required labelling statement with its intended purpose to clarify the approved production stages/classes of chickens and included the maximum inclusion rates allowed in feeds and drinking water
- added the statement "Do not use in combination with sources or forms of 1,25-dihydroxyvitamin D3."
- to prevent over supplementation and to avoid the concurrent use of the two active forms of vitamin D3 (25-hydroxyvitamin D3 and glycosylated 1,25-dihydroxyvitamin D3)
- 1,25-dihydroxyvitamin D3 is a biologically active form of vitamin D3 and is not intended to replace the nutritional requirements of vitamin D in the total diet of the intended livestock species
- a similar statement was included in the amended description for waxy-leaf nightshade leaf powder
- removed the International Units (IU) equivalency statement
- the required label guarantees in the amended description
- removed the required label guarantee for equivalent minimum International Units of vitamin D activity per kilogram as discussed above
- changed minimum milligrams of 25-hydroxyvitamin D3 per kilogram to minimum micrograms of 25-hydroxyvitamin D3 per kilogram
- micrograms are commonly used to express the concentration of 25-hydroxyvitamin D3
Since the comments received during the public consultation of this amended description included some concerns on the safety of this SIF, the description was revised to address some of these comments. The proposed amendments to this description were shared with implicated stakeholders with the purpose of offering an additional opportunity for them to provide feedback on the proposed changes to this SIF description.
We received 1 set of comments during the post-consultation engagement reiterating concerns about the SIF's impurities and potential effects on certain livestock species.
- we reviewed all the documents submitted by this stakeholder, including the scientific literature provided, evaluated the information and updated our safety evaluation based on the previously assessed information and additional information provided
- based on our review, we concluded that that this source of 25-hydroxyvitamin D3 monohydrate from fermentation process is safe and efficacious for its intended purpose and intended livestock species
- all vitamin D sources, including 25-hydroxyvitamin D3 monohydrate from fermentation process are placed into part 2 of the CFIT and each source must undergo a pre-market evaluation
- this evaluation considers differences between sources which includes variability in the manufacturing process including the production microorganism, variations in purity profiles (including characterization of potential impurities) and the analytical methodology used to guarantee the active ingredient(s)
The final amended SIF description is:
- 2-504-008 25-Hydroxyvitamin D3 monohydrate from fermentation process (or calcifediol monohydrate from fermentation process or 25-hydroxycholecalciferol monohydrate from fermentation process)
- is the monohydrate form of 25-hydroxyvitamin D3, generally expressed as C27H44O2·H2O and having the CAS# 63283-36-3. It is obtained from a manufacturing process that includes a fermentation process using a non-pathogenic strain of Saccharomyces cerevisiae or Pseudonocardia autotrophica that does not contain a novel trait. The manufacturing process may include further processing (for example, chemical transformation or purification) after fermentation. The manufacturing process shall be conducted in accordance with good manufacturing practices. It shall not contain other extraneous materials except in such amounts as may occur unavoidably during good manufacturing practices.
If the product bears a name descriptive of the form (for example, crystalline), it shall correspond thereto and be indicated on the label.
This ingredient is approved for use in supplements for drinking water and complete feeds for broiler chickens and turkeys in an amount not to exceed 100 µg/kg and for laying and pre-layer chickens in an amount not to exceed 80 µg/kg. This ingredient is also approved for use in complete feeds for swine in an amount not to exceed 50 µg/kg.
This ingredient should be used in combination with vitamin D (D2 and/or D3). Maximum content of the combination of 25-hydroxyvitamin D3 with vitamin D must not exceed 87.5 µg/kg (equivalent to 3,500 International Units of vitamin D per kg) for swine, 250 µg/kg (equivalent to 10,000 International Units of vitamin D per kg) for turkeys, broiler chickens, and pre-layer chickens, and 162.5 µg/kg (equivalent to 6,500 International Units of vitamin D per kg) for laying chickens.
It may be blended with suitable carriers listed in the Canadian Feed Ingredients Table to standardize the 25-hydroxyvitamin D3 concentration.
If a carrier is used, it must be approved for use in livestock feeds, it shall be used at the approved rate, and the common name or names shall be indicated on the label.
If an antioxidant is used, it must be approved for use in livestock feeds, it shall be used at the approved rate, and the common name or names shall be indicated on the label.
It shall be labelled with the following statements:
"This ingredient is approved for use in supplements for drinking water and complete feeds for broiler chickens and turkeys in an amount not to exceed 100 µg/kg and for laying and pre-layer chickens in an amount not to exceed 80 µg/kg. This ingredient is also approved for use in complete feeds for swine in an amount not to exceed 50 µg/kg."
"This ingredient is free of antimicrobial activity and is not a source of viable microbial cells."
"Do not use in combination with sources or forms of 1,25-dihydroxvitamin D3."
It shall be labelled with guarantees for minimum micrograms of 25-hydroxyvitamin D3 per kilogram, and maximum percent moisture.
Consequential amendments to the waxy-leaf nightshade leaf powder description
Following the amendments to the description for 25-hydroxyvitamin D3 monohydrate from fermentation process, our AFP has undertaken a review of the waxy-leaf nightshade leaf powder description (2-504-010). These amendments were not associated with an application to evaluate a new source of waxy-leaf nightshade leaf powder, rather, they were made to ensure consistency with the amended description for 25-hydroxyvitamin D3 monohydrate from fermentation process and to improve scientific accuracy. Overall, the changes better reflect the nature of the SIF, its approved purpose, and its alignment with the description for 25-hydroxyvitamin D3 monohydrate from fermentation process.
This description has been amended with the following changes to:
- the purpose statement found in the SIF description
- to clarify the intended purpose, which is a source of glycosylated 1,25-dihydroxyvitamin D3 and not a source of vitamin D activity
- this SIF is not intended to replace the required amount of vitamin D (D3 or D2), but as an additional source of the active form of vitamin D3 (glycosylated 1,25-dihydroxyvitamin D3)
- the required labelling statements
- removed the International Units (IU) equivalency statement
- the IUs are meant to express the biological activity of vitamin D rather than the biological activity of its metabolites, and since this SIF is not intended to replace the supplementation of vitamin D the IU equivalency statement may lead to a misinterpretation of the potency of this active form
- updated the required labelling statement with its intended purpose as a source of glycosylated 1,25-dihydroxyvitamin D3, and not a source of vitamin D activity
- added a statement "This ingredient is not intended to replace the required amount of vitamin D in the total diet for broiler chickens."
- this statement will clarify that the purpose of this SIF is to complement the activity of vitamin D3 (and is not to replace the nutritional requirements of vitamin D3 in the total diet of the intended livestock species)
- added a statement "Do not use in combination with sources of 25-hydroxyvitamin D3."
- to prevent over supplementation and to avoid the concurrent use of the two active forms of vitamin D3 (glycosylated 1,25-dihydroxyvitamin D3 and 25-hydroxyvitamin D3)
- a similar statement was included in the amended description for 25-hydroxyvitamin D3 monohydrate from fermentation process
- removed the International Units (IU) equivalency statement
- the required label guarantees in the description
- removed the required label guarantee for equivalent minimum International Units of vitamin D3 activity per kilogram as discussed above
The final amended SIF description is:
- 2-504-010 Waxy-leaf nightshade leaf powder (or Solanum glaucophyllum leaf powder)
- is the product that consists of waxy-leaf nightshade (Solanum glaucophyllum) leaves that have been dried by thermal means and finely ground under controlled conditions. It shall contain a minimum of 50 mg/kg of glycosylated 1,25-dihydroxyvitamin D3 (glycosylated 1,25-dihydroxycholecalciferol), and it shall contain not more than 10% moisture.
This ingredient is approved for use as a source of glycosylated 1,25-dihydroxyvitamin D3 in the total diet for broiler chickens.
It may be blended with suitable carriers listed in the Canadian Feed Ingredients Table to standardize the glycosylated 1,25-dihydroxyvitamin D3 concentration.
If a carrier is used, it must be approved for use in livestock feeds, it shall be used at the approved rate, and the common name or names shall be indicated on the label.
It shall be labelled with the following statements:
"This ingredient is approved for use as a source of glycosylated 1,25-dihydroxyvitamin D3 in the total diet for broiler chickens."
"This ingredient is not intended to replace the required amount of vitamin D in the total diet for broiler chickens."
"Do not use in combination with sources of 25-hydroxyvitamin D3."
It shall be labelled with guarantees for minimum milligrams of glycosylated 1,25-dihydroxyvitamin D3 per kilogram, and maximum percent moisture.
Outcome and next steps
Following the public consultation and request for additional feedback from stakeholders on the amended descriptions, we have finalized the amended descriptions for 25-hydroxyvitamin D3 monohydrate from fermentation process and waxy-leaf nightshade leaf powder.
These amended SIFs will be added to the CFIT at the next update.
We are committed to reviewing any new scientific information on the safety of any SIF. Anyone who becomes aware of new scientific information regarding the safety of these SIFs are encouraged to contact us.
Related information
- Feeds Act
- Consultations on proposed regulatory and policy changes
- Proposed amended livestock feed ingredient – 25-Hydroxyvitamin D3 monohydrate from fermentation process
Contact us
Animal Feed Program (AFP)
Canadian Food Inspection Agency
Email: cfia.afp-paa.acia@inspection.gc.ca