Allison M. Meyer

Early life, or the neonatal period, is perhaps the most challenging time for ruminant livestock, as they adapt to the extra-uterine environment, undergo important physiological maturation, and navigate harsh ambient conditions. Maternal influences during gestation, especially energy and protein nutrition in late pregnancy, can alter many processes that affect the neonatal period. These processes include fetal growth and development, gestation length, difficulty of parturition, and maternal behavior, which interact to affect offspring vigor at birth. Moreover, colostrum and early milk production and composition are affected by gestational nutrition, and these along with the previous factors affect the neonate’s ability to obtain transfer of passive immunity, thermoregulate, perform basal metabolism, and ultimately survive to weaning. Often, the long-term effects of maternal nutrition during gestation on offspring are attributed solely to the prenatal environment, but it is critical to also consider influences of early life on later productivity and health. More research is needed to integrate these neonatal outcomes with prenatal and postnatal mechanisms as well as later ruminant livestock performance. Better understanding of the maternal environment’s effects on the neonatal period provides opportunity for improved management of ruminant livestock dams and offspring.

To investigate impacts of late gestational nutrient restriction in first-parity beef females on prenatal nutrient availability to calves, fall-calving heifers [body weight (BW): 472 ± 33 (SD) kg; body condition score (BCS): 5.4 ± 0.5] were individually-fed 100% (control; CON; n = 13) or 70% (nutrient restricted; NR; n = 13) of metabolizable energy and metabolizable protein requirements for maintenance, pregnancy, and growth from day 160 of gestation to calving. Maternal measures were determined every 21 d (BW and metabolites) or 42 d (BCS, backfat, and longissimus muscle area) during gestation and post-calving. Doppler ultrasonography of both uterine arteries was conducted every 21 d until day 244 of gestation. At birth, calf BW and size were measured, and expelled placentas were dissected and dried. Targeted messenger ribonucleic acid (mRNA) expression was determined for cotyledons. Data were analyzed with nutritional plane, treatment initiation date, heifer’s sire, and calf sex (when P < 0.25) as fixed effects. Metabolites and uterine blood flow included day and nutritional plane × day as repeated measures. Circulating glucose was less (P = 0.05) for NR dams than CON. Circulating urea N and triglycerides were less (P ≤ 0.05), but non-esterified fatty acids were greater (P ≤ 0.05), for NR at most timepoints after treatment initiation. Post-calving, NR dams were 62.1 kg and 2.1 BCS less (P < 0.01) than CON. Moreover, NR had less (P < 0.01) backfat and longissimus muscle area, but similar (P = 0.72) shoulder height compared with CON. Heart rate was less (P < 0.01) for NR dams than CON after treatment initiation. Nutritional plane did not affect (P ≥ 0.15) ipsilateral, contralateral, or total uterine artery blood flow. Number of cotyledons was greater (P = 0.03), average cotyledon weight was less (P = 0.04), and total placental weight tended to be less (P = 0.10) for NR than CON. Cotyledonary relative mRNA expression of GLUT1 was greater (P = 0.04) and SNAT2 tended to be greater (P = 0.07) for NR, but other nutrient transporters, angiogenic factors, and PAG2 were not affected (P ≥ 0.13). Calves born to NR dams weighed 14.4% (P = 0.03) less at birth and had smaller (P ≤ 0.03) heart girth and volume than CON. Despite catabolizing maternal tissue stores, beef heifers experiencing late gestational nutrient restriction had altered circulating metabolites and smaller placentas, independent of a reduction in uterine blood flow, which compromised fetal growth.

Fall-calving primiparous beef females [body weight (BW): 451 ± 28 (SD) kg; body condition score (BCS): 5.4 ± 0.7] were individually-fed either 100% (control; CON; n = 13) or 70% (nutrient restricted; NR; n = 13) of metabolizable energy and metabolizable protein requirements for maintenance, pregnancy, and growth from day 160 of gestation to parturition. Doppler ultrasonography of both uterine arteries was conducted pre-treatment and every 21 d from days 181 to 265 of gestation. Expelled placentas were collected, and ipsilateral cotyledonary tissue was sampled to assess relative messenger ribonucleic acid (mRNA) expression. Placentas were separated into ipsilateral and contralateral sides, dissected (cotyledonary vs. intercotyledonary), and dried. Data were analyzed with nutritional plane, treatment initiation date, and calf sex (when P < 0.25) as fixed effects. Uterine blood flow included day and nutritional plane × day as repeated measures. We previously reported that post-calving, NR dams weighed 64 kg less and were 2.0 BCS lower than CON, but calf birth weight was not affected. Maternal heart rate was less (P < 0.001) for NR dams than CON after nutritional planes began. Nutritional plane did not affect (P ≥ 0.20) uterine artery hemodynamics, but all variables were affected (P ≤ 0.04) by day. Contralateral cotyledonary and placental weight were less (P ≤ 0.04) and contralateral intercotyledonary weight and number of cotyledons tended to be less (P ≤ 0.10) for NR dams than CON, but ipsilateral and whole placental weights were not affected (P ≥ 0.13). Ipsilateral placental weight as a percentage of total placental weight was greater (P = 0.03) for NR dams than CON. Whole placental cotyledonary: intercotyledonary weight was less (P = 0.01) for NR dams than CON. Placental efficiency was not affected (P = 0.89) by nutritional plane. Cotyledonary relative mRNA expression of GLUT3 and SNAT2 was greater (P ≤ 0.05) and relative expression of GLUT1, GLUT4, and NOS3 tended to be greater (P ≤ 0.07) for NR dams than CON. Nutritional plane did not affect (P ≥ 0.13) relative mRNA expression of GLUT5, 4F2hc, CAT1, LAT1, LAT2, VEGFA, FLT1, KDR, GUCY1B3, and PAG2. Despite less contralateral placental growth, beef heifers experiencing late gestational nutrient restriction maintained uterine artery blood flow and total placental mass and had 4 nutrient transporters and 1 angiogenic factor upregulated in cotyledons, all of which likely contributed to conserving fetal growth. Nutrient requirements increase substantially during late gestation in the beef female to support fetal, uteroplacental, and mammary growth, and in the still-growing heifer, nutrients are also needed for maternal tissue growth. During pregnancy, the placenta serves as the interface for the metabolism and transport of nutrients, gases, and wastes between maternal and fetal circulations. Inadequate gestational nutrition can result in placental insufficiency and intrauterine growth restriction of the offspring, resulting in postnatal consequences. How late gestational undernutrition in beef heifers impacts the factors involved in placental nutrient transport capacity is poorly understood. We observed that late gestational nutrient restriction in heifers decreased maternal heart rate, but total uterine artery blood flow was not affected. Additionally, total placental mass was maintained, while cotyledonary expression of 4 nutrient transporters and an angiogenic factor were upregulated in placentas of nutrient restricted dams, which collectively allowed for similar calf birth weights as adequately-fed dams.

Fall-calving primiparous beef females [body weight (BW): 451 ± 28 (SD) kg; body condition score (BCS): 5.4 ± 0.7] were individually-fed 100% (control; CON; n = 13) or 70% (nutrient restricted; NR; n = 13) of estimated metabolizable energy and metabolizable protein requirements from day 160 of gestation to calving. Post-calving, all dams were individually-fed tall fescue hay supplemented to meet estimated nutrient requirements for maintenance, growth, and lactation in Calan gates until day 149 of lactation, which limited calves to milk only. From day 150 of lactation until weaning at day 243, dams and calves were group-fed in drylots. Dam BW and metabolic status were determined every 21 d, and BCS and backfat (BF) were determined every 42 d of lactation until weaning. Pre-weaning calf BW, size, and metabolic status were determined every 21 d. Data were analyzed with nutritional plane, calving date, and calf sex (when P < 0.25) as fixed effects. Circulating metabolites included day and nutritional plane × day as repeated measures. We previously reported that post-calving, NR dams were 64 kg and 2.0 BCS less than CON, but calf BW and size at birth were not affected. During the first 147 d of lactation, NR dams gained more (P < 0.01) BW than CON and increased (P < 0.01) BCS, while CON decreased (P ≤ 0.01) BCS and BF. Previously, NR dams had lower (P < 0.01) circulating triglycerides on day 1 of lactation, tended to have lower (P = 0.08) triglycerides on day 21, and had lower (P ≤ 0.04) non-esterified fatty acids (NEFA) on days 21 and 243 than CON. Maternal glucose and urea N were not affected (P ≥ 0.73). At weaning, NR dams weighed 17 kg less (P = 0.15), were 0.67 BCS lower (P < 0.01), and tended to have less (P = 0.06) BF. Calves born to NR dams weighed less (P = 0.02) than CON by day 42 of age and were 13% smaller (P < 0.01) at weaning. Calf girth measures diverged (P ≤ 0.05) by day 21 of age, and skeletal size measures were less (P ≤ 0.08) for calves born to NR dams at most timepoints after day 63 of age. Calves born to NR dams tended to have lower (P = 0.09) circulating urea N pre-weaning than CON, but glucose, triglycerides, and NEFA were not affected (P ≥ 0.16). In summary, first-parity beef females that were nutrient restricted during late gestation experienced compensatory growth and gained body condition during lactation but were still thinner at weaning. Nutrient restriction reduced pre-weaning calf growth, likely due to decreased milk production. Nutrient requirements increase substantially during late gestation in the beef female; however, poor forage nutrient availability can result in undernutrition. For heifers, the added nutrient requirements needed to continue growing during their first pregnancy and lactation pose an even greater challenge. It is plausible that lingering effects of late gestational nutrient restriction may exist for the dam and calf pre-weaning. We report that first-parity beef females that were nutrient restricted during late gestation and then fed to meet estimated nutrient requirements during lactation recovered quickly metabolically and experienced compensatory growth, but still had less body condition at weaning than controls. Late gestational nutrient restriction did not affect calf size at birth but resulted in calf body weight and size measures diverging early in life. Ultimately, nutrient restriction resulted in a 13% decrease in weaning weight, which was likely due to decreased milk production (in a companion paper). Despite this, metabolic status of calves born to nutrient restricted dams was not greatly altered. In summary, first-parity beef females that were nutrient restricted during late gestation prioritized partitioning nutrients to maternal growth and energy reserves over milk production during lactation, but dams were thinner at weaning, and pre-weaning calf growth was slowed.

Fall-calving primiparous beef females [body weight (BW): 451 ± 28 (SD) kg; body condition score (BCS): 5.4 ± 0.7] were individually-fed 100% (control; CON; n = 13) or 70% (nutrient restricted; NR; n = 13) of estimated metabolizable energy and metabolizable protein requirements from day 160 of gestation to calving. Post-calving, all dams were individually-fed tall fescue hay supplemented to meet estimated nutrient requirements for maintenance, growth, and lactation until day 149 of lactation. Four-hour milk yields were collected on days 21, 42, 63, 84, 105, and 147 of lactation, and milk nutrient composition was determined. Doppler ultrasonography of both pudendoepigastric arterial trunks was conducted every 21 d from days 24 to 108 of lactation. Total mammary blood flow was calculated, and hemodynamics from both sides were averaged. Data were analyzed as repeated measures with nutritional plane, day of lactation, their interaction, calving date, and calf sex (if P < 0.25) as fixed effects. We previously reported that post-calving, NR dams weighed 64 kg less and were 2.0 BCS lower than CON, but calf birth weight was not affected. Milk weight and volume were 15% less (P = 0.04) for NR dams than CON. Milk protein concentration was lower (P = 0.008) for NR dams than CON, but triglyceride and lactose concentrations were not affected (P ≥ 0.20) by nutritional plane. Milk urea N concentration of NR dams tended to be greater (P = 0.07) on day 42 but was lower (P = 0.01) on day 147 of lactation than CON. Total milk protein, triglyceride, and lactose yields were less (P ≤ 0.05) for NR dams than CON. Total milk urea N yield was less (P ≤ 0.03) for NR dams than CON on days 21, 63, and 147 of lactation. Maternal heart rate was greater (P = 0.008), but pudendoepigastric arterial trunk peak systolic velocity, resistance index, and cross-sectional area were less (P ≤ 0.04) and pulsatility index tended to be less (P = 0.06) for NR dams than CON. Mammary blood flow was 19% less (P = 0.004) for NR dams than CON, but mammary blood flow relative to milk weight or dam BW was not affected (P ≥ 0.14) by nutritional plane. Most milk yield, milk nutrient composition, and mammary blood flow variables were affected (P ≤ 0.04) by day of lactation. In summary, first-parity beef females that were nutrient restricted during late gestation and then fed to meet estimated nutrient requirements during lactation had decreased milk nutrient yield and a similar reduction in mammary blood flow. Mammary development in preparation for lactation is largely complete at the time of calving, and final prepartum mammary growth and differentiation are occurring for the first time in heifers. Nutrient requirements increase substantially during late gestation, resulting in competition for nutrient use among maternal growth, fetal growth, and mammary growth in primiparous beef females. Undernutrition during late gestation can occur due to poor forage nutrient availability or drought, potentially impacting mammary gland development and subsequent milk production. We report that first-parity beef females that were nutrient restricted during late gestation and then fed to meet estimated nutrient requirements during lactation had 15% lower milk yield, reduced milk protein concentration, and less total milk protein, triglycerides, lactose, and urea N available for their calves than controls. Additionally, previously nutrient restricted dams had 19% less total mammary blood flow, and the major arteries supplying the mammary gland were smaller.

To determine the effects of late gestational nutrient restriction on expelled cotyledonary mRNA expression, fall-calving Hereford × Simmental-Angus heifers [single-sired; BW: 451 ± 28 (SD) kg; BCS: 5.4 ± 0.7] bred to a single sire were allocated by fetal sex and expected calving date to receive either 100% (control; CON; n = 12) or 70% (nutrient restricted; NR; n = 11) of NASEM metabolizable energy and metabolizable protein requirements for maintenance, pregnancy, and growth. Beginning on d 160 of gestation, heifers were individually fed chopped sorghum sudan hay (1.74 Mcal ME/kg, 6.69% CP, 72.0% NDF; DM basis) and based on individual intakes, supplemented to meet targeted nutritional planes. Calving was observed for all females and placentas were collected at natural expulsion. Immediately after collection (average: 4.5 h post-calving; range: 2.3 to 12.1 h post-calving), 2 representative cotyledons on the ipsilateral side were cleaned with phosphate buffered saline, and cotyledonary tissue was flash frozen. Total RNA was extracted, and RT-qPCR was performed for mRNA expression of glucose transporters (SLC2A1, SLC2A3, SLC2A4), a fructose transporter (SLC2A5), amino acid transporters (SLC1A1, SLC3A2, SLC7A1, SLC7A5, SLC7A8, SLC38A2), angiogenic factors/receptors (VEGFA, NOS3, FLT1, KDR, GUCY1B3), and pregnancy-associated glycoprotein 2 (PAG2). Relative mRNA expression was analyzed with nutritional plane, treatment initiation date, time to cotyledon sampling, and calf sex (when P < 0.25) as fixed effects. We have previously reported that total uterine blood flow during late gestation was not affected (P ≥ 0.20) by nutritional plane or the nutritional plane × day of gestation interaction. Post-calving, NR dams were 64 kg BW less (P < 0.001) and 2.0 BCS less (P < 0.001), but calf birth weight was not affected (P = 0.72) by nutritional plane. Contralateral placental dry weight tended to be less (P = 0.07) in NR dams, but total placental weight was not affected (P = 0.86) by late gestational nutritional plane. Cotyledonary relative mRNA expression of SLC2A3 and SLC38A2 was greater (P ≤ 0.05) and relative expression of SLC2A1, SLC2A4, and NOS3 tended to be greater (P ≤ 0.07) in NR dams compared with CON. Late gestational nutritional plane did not affect (P ≥ 0.13) cotyledonary relative mRNA expression of FLT1, GUCY1B3, KDR, PAG2, SLC1A1, SLC2A5, SLC3A2, SLC7A1, SLC7A5, SLC7A8, and VEGFA. In summary, first parity beef females who experienced late gestational nutrient restriction maintained uterine blood flow and placental mass and had 4 nutrient transporters and 1 angiogenic factor upregulated in cotyledons, all which likely contributed to their conservation of fetal growth.

Fall-calving primiparous beef females [body weight: 451 ± 28 (SD) kg; body condition score: 5.4 ± 0.7] were individually-fed either 100% (control; CON; n = 13) or 70% (nutrient restricted; NR; n = 13) of metabolizable energy and metabolizable protein requirements for maintenance, pregnancy, and growth from day 160 of gestation to parturition. Calves were reared naturally by their dams and monitored for latency times from birth to first sternal recumbency, attempt to stand, and stand; vigor scores were assigned at 2, 5, 10, and 20 min of age. Rectal temperatures and jugular blood were obtained at 0 (pre-suckling), 6, 12, 24, and 48 h of age, and blood chemistry, hematology, cortisol, and insulin were determined. Data were analyzed with fixed effects of late gestational nutritional plane (single data point) or nutritional plane, hour, and their interaction (data over time, repeated measures). Calving date was a fixed effect; calf sex was included when P < 0.25. We previously reported that late gestational nutritional plane did not affect gestation length or calf size at birth, but calving assistance and fetal malpresentation occurred more often in NR. Nutritional plane did not affect (P = 0.65) duration of parturition, but calves born to NR dams had slower times to attempt to stand (P = 0.09), slower times to stand (P = 0.02), and poorer 20 min vigor scores (P = 0.05). Serum immunoglobulin G and A concentrations at 48 h were greater (P ≤ 0.03) for NR calves. Rectal temperature of NR calves was less (P = 0.02) at 0 h, but greater (P = 0.04) at 24 h compared with CON. Circulating glucose, non-esterified fatty acids, triglycerides, cortisol, and insulin were not affected by nutritional plane (P ≥ 0.18). Total protein and globulin from 6 to 48 h were greater (P ≤ 0.02) in NR calves. Calves from NR dams had greater (P ≤ 0.08) gamma-glutamyl transferase at 6, 12, and 48 h. Serum aspartate aminotransferase was greater (P ≤ 0.07) from 0 to 24 h and creatine kinase was greater (P ≤ 0.04) from 6 to 24 h in NR calves. At 0 h, potassium was greater (P = 0.03) in NR calves. Calves born to CON had greater chloride (P = 0.08; main effect), sodium (P ≤ 0.09) from 0 to 48 h, and anion gap (P = 0.02) at 6 h. Hematocrit from 6 to 24 h and red blood cells and hemoglobin at 6 and 12 h were greater (P ≤ 0.09) in CON calves. These data indicate that nutrient restriction during late gestation resulted in less vigorous calves with more indicators of trauma in early life. Early life after birth is challenging for beef calves, but a successful transition from the uterus to the outside environment is necessary for calves to survive and thrive pre-weaning. Despite this, limited research has investigated how maternal undernutrition during pregnancy affects the vigor and metabolism of beef calves during early life. Beef females in their first pregnancy were fed to meet their energy and protein requirements, or were fed 70% of these requirements, during late pregnancy. Their calves were monitored closely, and data were collected intensively during early life. Calves born to nutrient restricted females were less vigorous at birth, taking longer to attempt to stand and stand successfully for the first time, and having lower vigor scores at 20 min of age. These calves also showed more signs of calving trauma and stress (elevated aspartate aminotransferase, creatine kinase, and creatinine) and had less red blood cells, but they had successful transfer of passive immunity and minimally affected energy metabolism. Overall, the effects of nutrient restriction observed may have decreased neonatal calf health and survival in a normal production environment, but not through reduced passive transfer.