Pet food safety testing is a comprehensive process combining ingredient toxicology, target animal data and stability of the finished product. In 2026, the use of NAMs, including in vitro and in silico models is useful for hazard identification, and nutritional adequacy can be shown via an appropriate formulation or feeding trial path within the applicable AAFCO framework. After the expiration of the FDA–AAFCO ingredient MOU in October 2024, companies wanting to develop new animal food ingredients are required to determine the regulatory pathway that applies within FDA; Animal Food Ingredient Consultation (AFIC) serves as an interim consultation route while FDA develops animal food pre-market programs.
Proving Pet-Food Safety: Emerging Toxicology and Trial Approaches Before You Launch
Dr Priya Raghavan, BVSc, PhD — Head of Pet Nutrition & Toxicology, Food Research Lab
14+ years designing pet food safety, novel-ingredient risk assessments and AAFCO / FEDIAF feeding-trial programmes across the US, UK, EU and GCC.
Quick Answer
Pet food safety testing is a comprehensive process combining ingredient toxicology, target animal data and stability of the finished product. In 2026, the use of NAMs, including in vitro and in silico models is useful for hazard identification, and nutritional adequacy can be shown via an appropriate formulation or feeding trial path within the applicable AAFCO framework. After the expiration of the FDA–AAFCO ingredient MOU in October 2024, companies wanting to develop new animal food ingredients are required to determine the regulatory pathway that applies within FDA; Animal Food Ingredient Consultation (AFIC) serves as an interim consultation route while FDA develops animal food pre-market programs.
The 2026 Safety Bar: Why Pre-Launch Testing Is More Complex
US ingredient regulation has shifted following expiration of the FDA–AAFCO Memorandum of Understanding on 1 October 2024. The FDA issued final GFI #293 for certain AAFCO-defined animal food ingredients [1] and GFI #294, along with GFI #294, creating the interim AFIC process, but continues evaluating its Food Additive Petition and GRAS Notification systems. [2] The AAFCO Official Publication remains relevant to the industry and state regulators, but not to the previous FDA scientific and technical review procedure under the MOU. Manufacturers must provide information about the ingredient identity, intended use, regulation and safety substantiation when conducting pet food regulatory compliance testing. [3]
Moreover, insect proteins, precision fermentation-derived ingredients, cell-cultured meat products and novel botanicals may raise novel exposure and hazard concerns. In the context of FSMA’s Preventive Controls for Animal Food framework, all applicable facilities are supposed to perform hazard analysis and establish preventive controls, bringing these ingredient considerations into the wider animal pet food product manufacturing system. [4]
Emerging Toxicology Approaches for Pet-Food Ingredients
New Approach Methodologies (NAMs): In Vitro and In Silico Screening
Another important technological trend in pet food toxicology studies is the application of New Approach Methodologies (NAMs), which include in vitro screening, in silico modelling, read-across and the integrated approach to weight of evidence. The methodologies help to detect potential hazards and prioritize testing, avoiding additional animal studies, while FDA’s more general NAMs activities should not be perceived as a pet-food-specific requirement to substitute animal studies. The toxicological risk assessment can include structure-based prediction, read-across, relevant in vitro tests, genotoxicity tests, and existing toxicological literature within the IATA (Integrated Approaches to Testing and Assessment) framework, to improve the pet food ingredient safety assessment while directing further testing toward genuine data gaps. [5] [6]
Tiered Risk Assessment for Novel Ingredients
Not all ingredients require the same level of effort. The tiered approach considers the relationship between the evidence and the hazard and exposure: Tier 1 includes characterization, literature review and read-across; Tier 2 comprises relevant in vitro testing and targeted target-animal studies if scientifically justified; Tier 3 reserves additional in vivo work for genuine data gaps. This keeps pet food toxicology studies proportionate and supports a defensible novel-ingredient safety assessment. [7]
Figure 1. Risk-Based Toxicological Testing Pathway for Novel Pet-Food Ingredients.
Table 1. Pre-Launch Toxicology and Ingredient Safety Testing Approaches
| Method | What it screens for | Pre-launch output |
| In silico (QSAR) | Structural alerts, predicted toxicity | Hazard prioritisation and testing rationale |
| Read-across / literature | Analogous substances, known hazards | Weight-of-evidence assessment |
| Ames + relevant in vitro assays | Mutagenicity/genotoxicity and cellular toxicity | Pass/flag for further assessment |
| Target-animal safety study | Tolerance, clinical effects, target-organ findings | Additional safety evidence where justified |
| Analytical panel | Heavy metals, mycotoxins, pathogens and other identified hazards | Raw-material and finished-product specifications |
TOXICOLOGY & INGREDIENT SCREENING
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Trial Approaches That Substantiate Your Label
Nutritional Adequacy: Feeding Trials vs Formulation
Within the relevant AAFCO regulatory system, nutritional adequacy may be demonstrated either by formulation in accordance with the relevant AAFCO Nutrient Profile or by the appropriate animal feeding trial pet food protocol. [8] Thus, the published AAFCO adult maintenance dog protocol requires at least eight healthy adult dogs, a minimum of 26 weeks’ duration for the testing, exclusive feeding of the test diet, weekly measurement of body weight and the specified clinical parameters, such as haemoglobin, packed cell volume, alkaline phosphatase and albumin, with particular criteria for completion of the study. The AAFCO feeding trials and formulation-based substantiation represent two distinct pieces of evidence, and neither should be regarded as superior; the appropriate route depends on the product, species, life stage, intended claim and applicable requirements. [9] For EU and UK markets, formulations should also be mapped against the applicable FEDIAF Nutritional Guidelines. [10]
Palatability and Digestibility Trials
Besides being nutritionally adequate, an acceptable pet food product should perform well in feeding tests. Pet food palatability testing involves either a two-bowl or single-bowl design to determine preference and consumption, while tests of digestibility quantify the apparent digestibility of nutrients using controlled feeding and faeces collection. With animal feeding trial pet food data, these studies assess product performance, tolerance and nutritional value, but do not independently prove toxicological safety. [11]
Figure 2. Integrated Pre-Launch Safety and Product Validation Pathway for Pet Food.
Building a Defensible Pre-Launch Safety Plan
Objective | Method | Output/specification |
Ingredient safety | In silico + in vitro / read across (IATA) | Safety assessment; inclusion-level rationale |
Contaminant control | Heavy metals, mycotoxins, pathogens and hazard-specific tests | Raw-material & finished-product specifications |
Nutritional adequacy | Applicable nutrient profile or appropriate feeding protocol | Nutritional adequacy substantiation |
Palatability/digestibility | Two-bowl; single-bowl; controlled digestibility methods | Acceptance and digestibility data |
Stability | Real-time + accelerated shelf-life studies | Best-before date; validated nutrient/safety margins |
Processing, storage, moisture and packaging can alter nutrient stability and microbiological or oxidative quality. Validate the final commercial format and pack, set appropriate stability-based margins, and ensure pet food regulatory compliance testing meets applicable AAFCO, FEDIAF or FSA requirements before finalising the label.
Novel-Ingredient Safety in Practice: Cultivated Chicken for Dog Food
Context: In July 2024, the UK’s Animal and Plant Health Agency (APHA) and Department for Environment, Food and Rural Affairs (Defra) approved the Meatly pet food, which contains cultured chicken. Research by the UK Food Standards Agency suggests that the product went on sale in January 2025, making the UK the first country in Europe to bring a cultivated-meat product to market.
Approach: The case illustrates the safety-assessment considerations associated with an emerging novel production technology, including characterisation of the production process and inputs, assessment of product composition and microbiological safety, and evaluation of potential chemical or biological hazards and residual process components. These are broader safety-assessment considerations identified by the UK Food Standards Agency for cell-cultivated products and should not be interpreted as a complete list of publicly disclosed tests performed specifically by Meatly.
Commercialisation:
In February 2025, Meatly announced the limited release of Chick Bites, a dog treat containing its cultivated chicken, through Pets at Home Brentford. Meatly's announcement dated 6 February 2025 states that the product would go on sale from 7 February 2025 and identifies Pets at Home as the retailer.
✓ APHA and Defra allowed the Meatly pet-food product in July 2024.
✓ The product went on sale in the UK in January 2025, according to FSA research.
✓ Meatly subsequently announced the limited retail release of Chick Bites at Pets at Home Brentford on 7 February 2025.
✓ The case provides an example of how a novel pet-food production technology progressed from regulatory assessment toward commercialisation.
Formulator's takeaway: The case illustrates a broader pre-launch principle: characterise the ingredient and production process, establish an appropriate safety evidence base, substantiate the formulation, evaluate the finished product and document the applicable regulatory pathway. [13] [14]
PRE-LAUNCH TRIALS & COMPLIANCE
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From feeding-trial design and FEDIAF substantiation to stability testing and regulatory dossiers, we build the evidence a confident launch needs.
Conclusion
In 2026, effective pet food safety testing requires integrated evidence across toxicology, nutritional adequacy and finished-product stability. Food Research Lab helps brands build launch-ready evidence through ingredient safety assessment, pet food formulation, feeding trials, stability testing and regulatory support.
Build a Safer Path From Concept to Commercialisation
Partner with Food Research Lab for end-to-end animal pet food product development, from ingredient selection and formulation to testing, regulatory compliance and pilot manufacturing.
Frequently Asked Questions
Novel pet food toxicology studies could include in silico prediction, read-across, relevant in vitro tests and genotoxicity testing in an IATA and weight-of-evidence approach. Target animal testing could be required when there are unresolved material safety issues.
No. Nutritional adequacy can be substantiated through the applicable formulation/nutrient-profile route or an appropriate animal feeding protocol, depending on the product, life stage and claim. The relevant AAFCO protocol should be followed for the specific product and life stage.
Skipping pre-launch pet food safety trials can leave gaps in ingredient safety, nutritional adequacy, contaminant control, stability or regulatory substantiation. Depending on the failure, consequences may include product withdrawal, recalls, regulatory action and animal-health concerns.
References
- S. Food and Drug Administration. (n.d.). CVM GFI #293: FDA enforcement policy for AAFCO-defined animal feed ingredients. FDA – CVM GFI #293
- S. Food and Drug Administration. (n.d.). CVM GFI #294: Animal food ingredient consultation (AFIC). FDA – CVM GFI #294
- S. Food and Drug Administration. (n.d.). Evaluation of FDA pre-market animal food programs and plan for consultations. FDA – Pre-market Animal Food Programs
- S. Food and Drug Administration. (n.d.). FDA finalizes FSMA guidance on hazard analysis and preventive controls for animal food. FDA – FSMA Guidance for Animal Food
- Rusyn, I., Boyd, K., Crincoli, C., Hanlon, P. R., & van Tongeren, T. C. A. (2026). New approach methodologies in food safety: A survey and commentary on industry adoption, barriers, and the path to regulatory adoption. Regulatory Toxicology and Pharmacology, 172, 106218. https://doi.org/10.1016/j.yrtph.2026.106218
- Reddy, N., Lynch, B., Gujral, J., & Karnik, K. (2023). Alternatives to animal testing in toxicity testing: Current status and future perspectives in food safety assessments. Food and Chemical Toxicology, 179, 113944. https://doi.org/10.1016/j.fct.2023.113944
- Organisation for Economic Co-operation and Development. (n.d.). Integrated approaches to testing and assessment. OECD – Integrated Approaches to Testing and Assessment
- Association of American Feed Control Officials. (n.d.). Reading pet food labels. AAFCO – Reading Pet Food Labels
- Association of American Feed Control Officials. (2013). Pet food report 2013: Annual appendix D. AAFCO – Pet Food Report 2013, Appendix D
- European Pet Food Industry Federation. (n.d.). Nutritional guidelines. FEDIAF – Nutritional Guidelines
- Calderón, N., White, B. L., & Seo, H. S. (2024). Measuring palatability of pet food products: Sensory components, evaluations, challenges, and opportunities. Journal of Food Science, 89(12), 8175–8196. https://doi.org/10.1111/1750-3841.17511
- S. Food and Drug Administration. (n.d.). Animal food ingredients. FDA – Animal Food Ingredients
- Food Standards Agency, Strauss, B., Kleinsorge, B., & Lotfian, P. (2025). Best practice for cell banking techniques used in cell cultivated products. FSA Research and Evidence. https://doi.org/10.46756/001c.144239
- (n.d.). Meatly launches world’s first cultivated pet food. Meatly – Cultivated Pet Food Launch
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