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Isotopic variation of food stuffs propagates through trophic systems. But, this variation is dampened in each trophic step, due to buffering effects of metabolic and storage pools. Thus, understanding of isotopic variation in trophic systems requires knowledge of isotopic turnover. In animals, turnover is usually quantified in diet-switch experiments in controlled conditions. Such experiments usually involve changes in diet chemical composition, which may affect turnover. Furthermore, it is uncertain if diet-switch based turnover models are applicable under conditions with randomly fluctuating dietary input signals. Here, we investigate if turnover information derived from diet-switch experiments with dairy cows can predict the isotopic composition of metabolic products (milk, milk components and feces) under natural fluctuations of dietary isotope and chemical composition. First, a diet-switch from a C3-grass/maize diet to a pure C3-grass diet was used to quantify carbon turnover in whole milk, lactose, casein, milk fat and feces. Data were analyzed with a compartmental mixed effects model, which allowed for multiple pools and intra-population variability, and included a delay between feed ingestion and first tracer appearance in outputs. The delay for milk components and whole milk was ∼12 h, and that of feces ∼20 h. The half-life (t½) for carbon in the feces was 9 h, while lactose, casein and milk fat had a t½ of 10, 18 and 19 h. The 13C kinetics of whole milk revealed two pools, a fast pool with a t½ of 10 h (likely representing lactose), and a slower pool with a t½ of 21 h (likely including casein and milk fat). The diet-switch based turnover information provided a precise prediction (RMSE ∼0.2 ‰) of the natural 13C fluctuations in outputs during a 30 days-long period when cows ingested a pure C3 grass with naturally fluctuating isotope composition.

Stable isotope analysis is a fundamental tool in food origin and authenticity testing. Its use in livestock production requires knowledge of isotope discrimination between product and diet. Here, we report (13)C discrimination ((13)Δ) for milk, milk components (fat, casein and lactose) and faeces in eight lactating dairy cows, which grazed pasture or were fed fresh pasture herbage in the stall. Cows were supplemented with grain maize at 1.72 kg d(-1) (dry matter). Feed components were collected daily, and faeces, milk fat, casein, lactose and whole milk 4 times per week during an 8-week-long sampling period. Carbon isotope composition (δ(13)C) of each sample was analysed. δ(13)C was lowest in milk fat (-29.8‰) and highest in casein (-26.4‰). Compared to the diet, whole milk was depleted in (13)C ((13)Δ = 0.4‰) due to a strong (13)C-depletion of fat ((13)Δ = 2.2‰), which was not fully compensated by the (13)C-enrichment of casein ((13)Δ = -1.1‰) and lactose ((13)Δ = -0.7‰). Faeces were also depleted in (13)C ((13)Δ =1.7‰). Influences of feeding environment (stall vs. pasture) and herbage quality were minor (<0.4‰). A review of literature data shows large variation between studies. We consider that the present results are superior, as they are based on a much larger data set regarding the number of cows and milkings (total n = 256) with greater detail in analyses of diet and milk products. Also, the study covered both stall- and pasture-feeding scenarios in realistic settings with long periods of equilibration. This is the first comprehensive analysis of (13)C discrimination between diet and all main milk components (as well as faeces). Thus, the results will improve the use of stable isotope analyses in regard to authenticity testing and proof of origin.

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