2.4. Feeding Protocol and Analysis of Human Milk
Powder breast milk fortifier, human milk collection
GROUP B
Following enrollment, infants were randomly assigned using a sequentially numbered sealed opaque envelope system with a ratio 1:2 to be fed with MOM supplemented, if required, either with PDM (Group A) or with TDM (Group B). If MOM was completely unavailable, the infant was fed with pasteurized PDM or TDM following randomization. Neither the attending physicians nor the nurses were aware of the type of donor milk. Only the principal investigator (not involved in the care of enrolled infants) knew the type of donor milk provided.Pasteurized PDM or TDM, as supplementary to MOM or for exclusive feeding, was provided to the study population for a three-week period after birth (defined as donor milk period) during which time the milk was analyzed regularly for protein and calorie content. After the donor milk period and until discharge, increasing demands were met with formula supplementation if MOM was not available or the milk bank was unable to provide TDM.Parenteral nutrition was started immediately after birth for all infants. Feeding of unfortified ΜOΜ or pasteurized DM from the hospital’s milk bank was initiated within the first three days of life. Powder breast milk fortifier (BMF) was added to human milk when milk intake reached 50–100 mL/kg/day. In all cases, a standard fortification with BMF (Nutricia Human Milk Fortifier) was applied (two sachets BMF per 100 mL of milk) according to the manufacturer’s instructions. Each sachet of fortifier (2.2 g) provided 0.6 g of protein in the form of extensively hydrolyzed bovine whey protein and 8 kcal of energy. The fortifier was introduced at half dose (1 sachet per 100 mL) for two to three days and then increased to full dose (2 sachets per 100 mL).Since sampling and pooling of milk samples after a full breast expression at each feed across a 24-hour period is considered the “gold standard” for human milk collection [