2 Elk velvet antler is harvested from live elk in late spring or early summer while still in its growth phase. Antlers are appendages that grow annually from antler generating structures called pedicles located on the frontal skull bone of male elk and deer. The antler in the growing stage is a soft, cartilaginous, blood-ﬁlled tissue covered by skin with a velvet-like texture, hence the name velvet antler. Antlers grow very rapidly reaching growth rates of approxi- mately 2 cm/day. Growth and mineralization is complete at approximately 120 days after it begins. Antlers reach peak commercial value at approximately 70 to 80 days of growth; after this time, the economic value and medical usefulness of antler is gradually lost due to progressive mineralization and concomitant loss of biological activity. The soft, blood-ﬁlled cartilage tissue in the distal portions of antler will be replaced by mineralized cartilage and bone tissue advancing up from the base within this period [4–6]. In North America, the processed antler is ground and encapsulated in varying milligram amounts and sold over the counter [7, 8]. In the East, the antler is cut into wafer- thin slices and combined with other natural ingredients in prescriptions for various TCM purposes. Despite a long history and widespread use, to our knowledge, little data is available about the eﬀect of EVA on physical and neurological development of neonates particu- larly following maternal dietary supplementation with EVA. One study evaluated short-term pre- and postnatal exposure of rat pups to EVA (from the 18th day in utero to 86 days after birth) and reported no changes in normal physical, devel- opmental, or behavioral indices [2]. To add to the limited database of EVA eﬀects during postnatal development, our study was designed to comprehensively evaluate the eﬀects of long-term maternal EVA supplementation on physical, reﬂexological, and neurological development of oﬀspring and whether gender diﬀerences existed in the oﬀspring’s response to maternal supplementation. 2. Materials and Methods 2.1. Animal Diets. The control diet consisted of regular rodent chow (Prolab RMH 3000, PMI Nutrition Interna- tional, St. Louis, MO, USA) ground into powder using a laboratory mill. A 10% w/w EVA diet [2] was made using 1 : 9 EVA powder : powered regular rat chow and mixed to homogeneity. EVA powder was derived from the freeze-drying method and was supplied by Norelkco Nutraceuticals (Moosomin, SK, Canada). Brieﬂy, the velvet antler was frozen immediately after harvesting, transported to the plant where the button end (about 2 cm) was cut oﬀ in order to help control bacteria and the hide (skin) was removed. The rest of the frozen stick was shredded onto stainless steel trays to expose maximum surface area. The shred was quickly (about 12 h) dried by a blast of cold (−1◦C), dry air, which results in minimum loss of bioavailable nutrients and no pathogen buildup. The dried shred was then ground in a nitrogen-cooled grinder to a ﬁne powder. Evidence-Based Complementary and Alternative Medicine 2.2. Animals. Sixteen Wistar adult female rats weighing 200 to 250 g and 16 Wistar adult male rats weighing 400 to 450 g were obtained from Charles Rivers (Montreal, PQ, Canada). Male and female rats were randomly assigned to the two treatment groups and housed in plexiglass cages lined with sawdust. Rats were provided with water ad libitum and fed regular powdered rat chow (female and male controls) or 10% EVA powdered chow (male and female treatment groups) ad libitum for 90 consecutive days prior to mating. All animals were kept under constant temperature at 21◦C and 60% humidity and on a 12 h : 12 h light : dark schedule. Female rats were bred to males fed the same chow as the females. Female and male rats were paired for 5 days or until a vaginal mucus plug (as evidence of mating) was conﬁrmed. Females were checked twice daily until pups were born. The day of birth was designated postnatal day 0 (PND 0). During pregnancy and lactation, the female adult rats from either group continued to receive the same diet as before parturition. All females were allowed to deliver naturally and rear their young to weaning. After parturition, pups were housed with their dam for a lactation period of 21 days (PND 21), after which they were weaned and provided ad libitum with the same chow as their mothers for the remainder of the experimental period of 31 days (PND 31). In order to prevent maternal responses from playing a role in litter manipulations, eﬀorts were made to ensure the early living environment contributed by maternal behavior was the same across groups [9–11]. Pups born to dams fed EVA or regular chow were selected for study across all litters in each diet group to ensure equivalent nutritional intake and maternal care for pups. On PND 2, litters were sexed and 224 pups were assigned to one of 14 pup groups (7 control and 7 EVA groups, corresponding to the diet fed to their mothers), each group containing 8 males and 8 females. This work was approved by the University of Saskatchewan’s Animal Research Ethics Board and adhered to the Canadian Council on Animal Care guidelines for humane animal use. 2.3. Examination of Physical Development. At parturition, litter size, total litter weight, and mean birth weight of the pups were recorded for each dam. Pups were weighed every day from PND 0 to PND 21. Physical development and maturation of the pups within each dietary group was evaluated by observing the dates of onset and completion of pinna unfolding, incisor eruption, eye opening, and growth of anogenital hair [12–19]. 2.4. Examination of Neuromotor Development. Crawling, head waving, and supported and unsupported standing were examined as indicators of neuromotor development. Mean age of onset and completion of the indicators in each pup group, the number of animals in each group displaying the indicator activity from day of onset to day of completion, and the time spent in these activities were recorded. The times spent in grooming and total relative activity during a ﬁxed test period were also assessed as parameters of spontaneous activity. The equipment and detailed procedures employed in making these observations were previously described [20].