Use of water turnover method to measure mother’s milk flow in a rat model: Application to dams receiving a low protein diet during gestation and lactation

Assessment of milk production is of utmost relevance for pediatricians and scientists interested in early life nutrition. The weight-suckle-weight (WSW) method, which consists of weighing babies before and after they suckle their mother, uses the difference in body weight as an estimate of milk intake. However, this is prone to many sources of error. In the current study, we used for the first time the water turnover method and compartmental analysis with deuterated water (D2O) as a non-toxic tracer to quantify in vivo milk production in a rat model. We assessed the effect of a nutritional intervention presumed to affect milk production, a maternal dietary protein restriction during gestation and lactation, which results in the birth of pups with intrauterine growth restriction. The specific aim of this study was to determine milk production with the body water turnover method in rat dams receiving during gestation and lactation, either a control diet (NP) or an iso-caloric low-protein diet (LP). In NP dams, mass of dam's total body water, output flow constant from dam to litter (K21) and median milk flow, calculated between days 11 to 14 after pup birth, were 282.1 g, 0.0122 h-1 and 3.30 g/h for NP dams, respectively. Maternal dietary protein restriction (-59%) during perinatal period led to a 34% reduction in milk flow (NP versus LP). With the WSW method, milk flow varied from 1.96 g/h to 2.37 g/h between days 11 to 14 for NP dams. The main advantage of the D20 method compared to the WSW method stems from its higher precision, as attested by the narrowest range of measured values of milk flow ([2.90; 3.75] and [0.98; 6.85] g/h, respectively) for NP group. This method could be suitable for testing the effectiveness of candidate galactologue molecules presumed to enhance milk production in the lactating rat model.