Reproducibility of the creamatocrit technique for the measurement of fat content in human milk.

Quantification of human milk (HM) fat is important for determining the energy intake of infants. The simplest and most rapid method is the creamatocrit method. However, the reliability of the creamatocrit has not been comprehensively investigated. The aims of this study were to test the inter- and-intra-rater reliability of: 1) HM sampling after hand- or-machine mixing methods and 2) HM fat measurement by the creamatocrit method. Inter-and-intra rater HM sampling after hand- or-machine mixing methods had high intraclass correlation coefficient (>0.91). Inter-rater reliability of measurement of HM with low fat (<2%) resulted in high variability (median coefficient of variations (CVs) > 15%). Intra- and inter-rater reliability of measurement of HM with higher fat (>3.5%) had low variability (median CVs < 10%). As the greatest variation in the creamatocrit method occurred during the measurement of HM samples with low fat, duplicate readings are necessary to reduce discrepancies in every HM fat determination.

The Effect of Caffeine and Trifluralin on Chromosome Doubling in Wheat Anther Culture.

Challenges for wheat doubled haploid (DH) production using anther culture include genotype variability in green plant regeneration and spontaneous chromosome doubling. The frequency of chromosome doubling in our program can vary from 14% to 80%. Caffeine or trifluralin was applied at the start of the induction phase to improve early genome doubling. Caffeine treatment at 0.5 mM for 24 h significantly improved green plant production in two of the six spring wheat crosses but had no effect on the other crosses. The improvements were observed in Trojan/Havoc and Lancer/LPB14-0392, where green plant numbers increased by 14% and 27% to 161 and 42 green plants per 30 anthers, respectively. Caffeine had no significant effect on chromosome doubling, despite a higher frequency of doubling in several caffeine treatments in the first experiment (67-68%) compared to the control (56%). In contrast, trifluralin significantly improved doubling following a 48 h treatment, from 38% in the control to 51% and 53% in the 1 µM and 3 µM trifluralin treatments, respectively. However, trifluralin had a significant negative effect on green plant regeneration, declining from 31.8 green plants per 20 anthers (control) to 9-25 green plants per 20 anthers in the trifluralin treatments. Further work is required to identify a treatment regime with caffeine and/or anti-mitotic herbicides that consistently increases chromosome doubling in wheat without reducing green plant regeneration.

Preterm Infant Feeding: A Mechanistic Comparison between a Vacuum Triggered Novel Teat and Breastfeeding.

The goal for preterm infants is to achieve full oral feeds quickly and ultimately progress to full breastfeeding. Supplementary oral feeds are often given when the mother is not available to breastfeed. Bottles typically deliver milk in a different fashion compared to breastfeeding, which is thought to hamper transition to full breastfeeding. The aim of this study was to compare the sucking dynamics of preterm infants fed at the breast to feeding with an experimental novel teat (NT) designed to release milk only upon the application of vacuum. Simultaneous ultrasound imaging of the infant oral cavity and measurement of intra-oral vacuum was performed during a breastfeed and a feed with the NT. Test weighs were used to measure milk intake. Linear mixed effects models were performed to investigate differences by feed type, and simultaneous linear regression was performed to investigate individual patterns. Tongue movement was not different between breastfeeding and the NT. Intra-oral vacuums (median (interquartile range: IQR)) were significantly lower with the NT (Baseline vacuum: -5.8 mmHg (-11.0, 0.1); Peak: 40.0 mmHg (-54.6, -27.1)) compared to breastfeeding (Baseline: -31.1 mmHg (-60.0, -12.7); Peak: -106.2 mmHg (-153.0, -65.5)). Milk intake was significantly higher with the NT (33 mL (22.5, 42.5)) compared to the breastfeed (12 mL (3, 15.5)). The novel teat encouraged a similar tongue action to breastfeeding, and infants transferred a greater volume of milk with the novel teat. Intra-oral vacuums were lower in strength with the novel teat compared to the breast. Use of the novel teat for the training of sucking dynamics in preterm infants has the potential to improve breastfeeding success and requires further investigation.

Relationships between Breastfeeding Patterns and Maternal and Infant Body Composition over the First 12 Months of Lactation.

Breastfeeding has been implicated in the establishment of infant appetite regulation, feeding patterns and body composition (BC). A holistic approach is required to elucidate relationships between infant and maternal BC and contributing factors, such as breastfeeding parameters. Associations between maternal and breastfed term infant BC (n = 20) and feeding parameters during first 12 months of lactation were investigated. BC was measured at 2, 5, 9 and/or 12 months postpartum with ultrasound skinfolds (US; infants only) and bioimpedance spectroscopy (infants and mothers). 24-h milk intake (MI) and feeding frequency (FFQ) were measured. Higher FFQ was associated with larger 24-h MI (p ≤ 0.003). Higher 24-h MI was associated with larger infant fat mass (FM) (US: p ≤ 0.002), greater percentage FM (US: p ≤ 0.008), greater FM index (FMI) (US: p ≤ 0.001) and lower fat-free mass index (FFMI) (US: p = 0.015). Lower FFQ was associated with both larger FFM (US: p ≤ 0.001) and FFMI (US: p < 0.001). Greater maternal adiposity was associated with smaller infant FFM measured with US (BMI: p < 0.010; %FM: p = 0.004; FMI: p < 0.011). Maternal BC was not associated with FFQ or 24-h MI. These results reinforce that early life is a critical window for infant programming and that breastfeeding may influence risk of later disease via modulation of BC.

Gastric emptying of different meal volumes of identical composition in preterm infants: a time series analysis.

BackgroundPreterm infants' meals typically progress to higher volumes over time. Knowledge of gastric emptying (GE) responses to differing meal volumes may inform enteral feeding management. We examined the effect of meal volume and composition on preterm GE.MethodsForty infants were studied at 33.3±1.4 (29.7-35.6) weeks postmenstrual age when fully enteral fed (target 150 ml/kg/day). Intraindividual comparisons of GE were made for paired meals of 100% and 75% prescribed volume and identical composition of mother's own milk (n=21) and pasteurized donor human milk (n=19). Serial stomach ultrasound images were used to calculate gastric residual volumes (GRVs) and remaining meal proportions (% meal).ResultsGE was faster in the early postprandial period and slowed over time (P<0.001). Reduced volume meals had slower GE rates and lower GRV (P<0.001). Serial postprandial % meal was similar between reduced and full volume meals (P=0.41). Higher milk casein concentration was associated with slower GE (P=0.04). Complete gastric emptying (GRV=0 ml) was more common in infants fed at 3 h intervals compared with those fed every 2 h (P=0.002).ConclusionEarly postprandial GE is more rapid for larger meal volumes. Stable preterm infants may tolerate feeding of a 3 h meal volume at shorter intervals.

Characterisation of sucking dynamics of breastfeeding preterm infants: a cross sectional study.

BACKGROUND: Full breastfeeding is the ultimate aim for preterm infants to ensure they receive the full benefits of human milk however, preterm infants face a number of challenges associated with their immaturity and associated morbidities. In order to facilitate oral feeding, it is essential to have a sound knowledge of the sucking dynamics of the breastfed infant. The aim of this study was to measure and describe the sucking dynamics of the preterm breastfeeding infant. METHODS: A prospective cross sectional observational study was carried out at King Edward Memorial Hospital, Perth. 38 mothers and their preterm infants (birth gestation age: 23.6-33.3 weeks; corrected gestation age 32.7 to 39.9 weeks) were recruited. Intra-oral vacuum levels, tongue movement and milk intake for a single breastfeed was measured. Statistical analysis employed linear regression and linear mixed effects models. RESULTS: Synchronised ultrasound and intra-oral vacuum measurements show that the preterm infant generates vacuum by lowering their tongue in a parallel fashion, without distortion of the nipple/nipple shield. Baseline (B), mean (M) and (P) peak suck burst vacuums weakened over the course of a feed (B: p = 0.015; M: p = 0.018; P: p = 0.044) and mean and peak vacuums were weaker if the mother fed with a nipple shield (M: p = 0.012; P: p = 0.021). Infant milk intakes were higher when infants sucked for longer (p = 0.002), sucked for a greater proportion of the feed (p = 0.002), or had a greater sucking efficiency (p < 0.001). CONCLUSIONS: Breastfeeding preterm infants generated intra-oral vacuum in the same manner as term infants. Nipple shields were associated with weaker intra-oral vacuums. However, vacuum strengths were not associated with milk intake rather time spent actively sucking was related to milk volumes. Further research is required to elucidate factors that influence preterm infant milk intake during breastfeeding.

Changes in Fatty Acid Composition of Human Milk in Response to Cold-Like Symptoms in the Lactating Mother and Infant.

Infants rely on their innate immune systems to protect them from infection. Human milk (HM) contains fatty acids (FAs) and monoacylglycerols that are known to exhibit antiviral and antibacterial properties in vitro. The specific fat content of HM may potentially affect the efficacy of this antimicrobial activity. This preliminary study investigates whether the proportions of FA in HM change in response to infections, leading to cold-like symptoms in the mother or infant. Milk samples were obtained from mothers (n = 26) when they and their infants were healthy, and when mother, infant, or both suffered cold-like symptoms. The milk was hydrolysed and FA proportions were measured using gas chromatography. Fifteen FAs were recorded, of which eight were detected in sufficient quantities for statistical analysis. The proportions of capric (C10:0) and lauric acids (C12:0) in HM were significantly lower, and palmitic acid (C16:0) was higher when mothers and infants were ill compared to healthy samples. Palmitoleic (C16:1, n-7) and stearic acid (C18:0) proportions were higher in HM when the infant was unwell, but were not related to maternal health. Whilst the differences detected were small (less than 0.5%), the effects may be additive and potentially have a protective function. The value of further studies is certainly indicated.

Associations between Maternal Body Composition and Appetite Hormones and Macronutrients in Human Milk.

Human milk (HM) appetite hormones and macronutrients may mediate satiety in breastfed infants. This study investigated associations between maternal adiposity and concentrations of HM leptin, adiponectin, protein and lactose, and whether these concentrations and the relationship between body mass index and percentage fat mass (%FM) in a breastfeeding population change over the first year of lactation. Lactating women (n = 59) provided milk samples (n = 283) at the 2nd, 5th, 9th and/or 12th month of lactation. Concentrations of leptin, adiponectin, total protein and lactose were measured. Maternal %FM was measured using bioimpedance spectroscopy. Higher maternal %FM was associated with higher leptin concentrations in both whole (0.006 ± 0.002 ng/mL, p = 0.008) and skim HM (0.005 ± 0.002 ng/mL, p = 0.007), and protein (0.16 ± 0.07 g/L, p = 0.028) concentrations. Adiponectin and lactose concentrations were not associated with %FM (0.01 ± 0.06 ng/mL, p = 0.81; 0.08 ± 0.11 g/L, p = 0.48, respectively). Whole milk concentrations of adiponectin and leptin did not differ significantly over the first year of lactation. These findings suggest that the level of maternal adiposity during lactation may influence the early appetite programming of breastfed infants by modulating concentrations of HM components.

The relationship of human milk leptin and macronutrients with gastric emptying in term breastfed infants.

BackgroundInfants breastfed on demand exhibit a variety of feeding patterns and self-regulate their nutrient intake, but factors influencing their gastric emptying (GE) are poorly understood. Despite research into appetite regulation properties of leptin, there is limited information about relationships between human milk leptin and infant GE.MethodsGastric volumes were calculated from ultrasound scans of infants' stomachs (n=20) taken before and after breastfeeding, and then every 12.5 min (median; range: 3-45 min) until the next feed. Skim milk leptin and macronutrient concentrations were measured and doses were calculated.ResultsThe leptin concentration was (mean±SD) 0.51±0.16 ng/ml; the leptin dose was 45.5±20.5 ng per feed. No relationships between both concentration and dose of leptin and time between the feeds (P=0.57; P=1, respectively) or residual stomach volumes before the subsequent feed (P=0.20; P=0.050) were found. Post-feed stomach volumes (GE rate) were not associated with leptin concentration (P=0.77) or dose (P=0.85).ConclusionGE in term breastfed infants was not associated with either skim milk leptin concentration or dose. Further investigation with inclusion of whole-milk leptin and other hormones that affect gastrointestinal activity is warranted.

Changes in milk composition associated with pethidine-PCEA usage after Caesarean section.

The effect of pethidine as patient-controlled epidural analgesia (PCEA) on specific biochemical components in breast milk in relation to the timing of secretory activation is not well investigated. The aim of this study was to compare biochemical timing of secretory activation between women who had a vaginal (V) or Caesarean birth with pethidine-PCEA (CBP). Several milk samples were collected daily from 36 mothers (17 V, 19 CBP) for the first 265 h post-partum. Protein and lactose concentrations and Na+ and K+ ion levels were measured. Samples were assigned to three time periods: 0-72, >72-165 and >165-265 h post-partum for statistical analyses. Data were analyzed using linear mixed effect models. In the first 72 h post-partum, the mean difference in lactose concentration was 5 gL-1 higher in group V (P < 0.05). From >72-165 h post-partum, protein and Na+ concentrations were lower in group V (P = 0.05, P = 0.02), and K+ levels were higher in group V (P < 0.001). From >165-265 h post-partum, there were no significant differences between the groups. Biochemically, secretory activation had occurred by 72 h post-partum in both groups. There were greater variations in measured biochemical components observed within group CBP initially. However, by 165 h post-partum, there were no differences in the biochemical components between the groups. This suggests that effects of pethidine-PCEA are diminished by 72 h post-partum and undetected by 165 h.

Determinants of body composition in breastfed infants using bioimpedance spectroscopy and ultrasound skinfolds-methods comparison.

BACKGROUND: Accurate, noninvasive, and inexpensive methods are required to measure infant body composition. Ultrasound (US) and bioimpedance spectroscopy (BIS) have been validated in adults and introduced in pediatric populations. The aim of this study was to evaluate the performance of both methods in determining percentage fat mass (%FM) in breastfed infants. METHODS: %FM of 2, 5, 9, and 12 mo-old healthy, breastfed term infants (n = 58) was calculated using BIS-derived total body water equations and skinfold equations then compared with reference models. Skinfolds were measured with US at two and four sites (biceps, suprailiac and/or triceps, and subscapular). RESULTS: %FM differed widely within and between methods, with the degree of variation affected by infant age/sex. Not a single method/equation was consistent with the distributions of appropriate reference values for all age/sex groups. Moderate number of matches with references values (13-24 out of 36) was seen for both types of equations. High number of matches (25-36) was seen for US skinfold-based equations. %FM values calculated from US and BIS were not significantly different (P = 0.35). CONCLUSION: Both BIS and US are practical for predicting %FM in infants. BIS calculations are highly dependent upon an appropriate set of validated age-matched equations.

Effect of Human Milk Appetite Hormones, Macronutrients, and Infant Characteristics on Gastric Emptying and Breastfeeding Patterns of Term Fully Breastfed Infants.

Human milk (HM) components influence infant feeding patterns and nutrient intake, yet it is unclear how they influence gastric emptying (GE), a key component of appetite regulation. This study analyzed GE of a single breastfeed, HM appetite hormones/macronutrients and demographics/anthropometrics/body composition of term fully breastfed infants (n = 41, 2 and/or 5 mo). Stomach volumes (SV) were calculated from pre-/post-feed ultrasound scans, then repeatedly until the next feed. Feed volume (FV) was measured by the test-weigh method. HM samples were analyzed for adiponectin, leptin, fat, lactose, total carbohydrate, lysozyme, and total/whey/casein protein. Linear regression/mixed effect models were used to determine associations between GE/feed variables and HM components/infant anthropometrics/adiposity. Higher FVs were associated with faster (-0.07 [-0.10, -0.03], p < 0.001) GE rate, higher post-feed SVs (0.82 [0.53, 1.12], p < 0.001), and longer GE times (0.24 [0.03, 0.46], p = 0.033). Higher whey protein concentration was associated with higher post-feed SVs (4.99 [0.84, 9.13], p = 0.023). Longer GE time was associated with higher adiponectin concentration (2.29 [0.92, 3.66], p = 0.002) and dose (0.02 [0.01, 0.03], p = 0.005), and lower casein:whey ratio (-65.89 [-107.13, -2.66], p = 0.003). FV and HM composition influence GE and breastfeeding patterns in term breastfed infants.

The Effects of Leptin on Breastfeeding Behaviour.

Breastfed infants have a reduced risk of becoming overweight and/or obese later in life. This protective effect has been partly attributed to leptin present in breastmilk. This study investigated 24-h variations of skim milk leptin and its relationship with breastmilk macronutrients and infant breastfeeding patterns. Exclusive breastfeeding mothers of term singletons (n = 19; age 10 ± 5 weeks) collected pre- and post-feed breastmilk samples for every breastfeed over a 24-h period and test-weighed their infants to determine milk intake at every breastfeed over a 24-h period. Samples (n = 454) were analysed for leptin, protein, lactose and fat content. Skim milk leptin concentration did not change with feeding (p = 0.184). However, larger feed volumes (>105 g) were associated with a decrease in post-feed leptin levels (p = 0.009). There was no relationship between the change in leptin levels and change in protein (p = 0.313) or lactose levels (p = 0.587) between pre- and post-feed milk, but there was a trend for a positive association with changes in milk fat content (p = 0.056). Leptin concentration significantly increased at night (p < 0.001) indicating a possible 24-h pattern. Leptin dose (ng) was not associated with the time between feeds (p = 0.232). Further research should include analysis of whole breastmilk and other breastmilk fractions to extend these findings.

Persistent Nipple Pain in Breastfeeding Mothers Associated with Abnormal Infant Tongue Movement.

BACKGROUND: Infants of breastfeeding mothers with persistent nipple pain have been shown to apply stronger vacuums to the breast and transfer less milk during one monitored feed. This may be associated with differences in the movement of the tongue. The aim was to analyse the intra-oral nipple shape and movement of the tongue of infants of mothers with and without nipple pain. METHODS: Breastfeeding infants of mothers with or without nipple pain were monitored using ultrasound and intra-oral vacuum during one breastfeed. From cine clips of the ultrasound scans measurements were made of the depth of the intra-oral space between the hard-soft palate junction (HSPJ) and the mid-tongue; the distance of the tip of the nipple to the HSPJ; and nipple diameters from the tip to the base. RESULTS: During nutritive sucking, tongue movements of infants of mothers with nipple pain resulted in a smaller intra-oral space (p = 0.040) and restricted nipple expansion compared to controls (p < 0.012). Stronger baseline and peak vacuums compared to controls were confirmed (p = 0.002). CONCLUSION: In these mothers, nipple pain was associated with restricted infant tongue movement. Ultrasound may complement measurement of intra-oral vacuum in monitoring treatment strategies in breastfeeding women experiencing nipple pain.

Gene expression in breastmilk cells is associated with maternal and infant characteristics.

Breastmilk is a rich source of cells with a heterogeneous composition comprising early-stage stem cells, progenitors and more differentiated cells. The gene expression profiles of these cells and their associations with characteristics of the breastfeeding mother and infant are poorly understood. This study investigated factors associated with the cellular dynamics of breastmilk and explored variations amongst women. Genes representing different breastmilk cell populations including mammary epithelial and myoepithelial cells, progenitors, and multi-lineage stem cells showed great variation in expression. Stem cell markers ESRRB and CK5, myoepithelial marker CK14, and lactocyte marker α-lactalbumin were amongst the genes most highly expressed across all samples tested. Genes exerting similar functions, such as either stem cell regulation or milk production, were found to be closely associated. Infant gestational age at delivery and changes in maternal bra cup size between pre-pregnancy and postpartum lactation were associated with expression of genes controlling stemness as well as milk synthesis. Additional correlations were found between genes and dyad characteristics, which may explain abnormalities related to low breastmilk supply or preterm birth. Our findings highlight the heterogeneity of breastmilk cell content and its changes associated with characteristics of the breastfeeding dyad that may reflect changing infant needs.

Impact of Measuring Milk Production by Test Weighing on Breastfeeding Confidence in Mothers of Term Infants.

AIM: The duration of exclusive breastfeeding is affected by maternal confidence and perception of milk supply, but objective measurement of milk supply is rarely used. Mothers of preterm infants have found measuring milk supply by in-home test weighing to be helpful to ascertain their infants' breastfeeding intake. This study aimed to determine if this technique affects breastfeeding confidence of mothers of term infants. MATERIALS AND METHODS: Participants filled in a survey of their breastfeeding confidence and perceptions of breastfeeding behavior prior to and after a 24-hour period of recording the times of feeds, amounts of each breastfeed or bottle feed (expressed breastmilk or formula), and amounts of breastmilk expressed. RESULTS: The breastfeeding confidence of at least 66% of mothers who were initially confident was maintained, and the breastfeeding confidence of at least 11% of mothers who were initially not confident was improved after objective measurement of milk supply and breastfeeding behavior. CONCLUSIONS: Measurement of actual milk supply and breastfeeding behavior provides objective information that can guide clinicians in the management of lactation. It can also help to maintain or improve breastfeeding confidence.

Human Milk MicroRNA and Total RNA Differ Depending on Milk Fractionation.

MicroRNA have been recently discovered in human milk signifying potentially important functions for both the lactating breast and the infant. Whilst human milk microRNA have started to be explored, little data exist on the evaluation of sample processing, and analysis to ensure that a full spectrum of microRNA can be obtained. Human milk comprises three main fractions: cells, skim milk, and lipids. Typically, the skim milk fraction has been measured in isolation despite evidence that the lipid fraction may contain more microRNA. This study aimed to standardize isolation of microRNA and total RNA from all three fractions of human milk to determine the most appropriate sampling and analysis procedure for future studies. Three different methods from eight commercially available kits were tested for their efficacy in extracting total RNA and microRNA from the lipid, skim, and cell fractions of human milk. Each fraction yielded different concentrations of RNA and microRNA, with the highest quantities found in the cell and lipid fractions, and the lowest in skim milk. The column-based phenol-free method was the most efficient extraction method for all three milk fractions. Two microRNAs were expressed and validated in the three milk fractions by qPCR using the three recommended extraction kits for each fraction. High expression levels were identified in the skim and lipid milk factions for these microRNAs. These results suggest that careful consideration of both the human milk sample preparation and extraction protocols should be made prior to embarking upon research in this area.

Gastric Emptying and Curding of Pasteurized Donor Human Milk and Mother's Own Milk in Preterm Infants.

We evaluated the effects of fortification and composition on gastric emptying and curding in un/fortified pairs of mother's own milk (MOM, n = 17) and pasteurized donor human milk (PDHM, n = 15) in preterm infants. Retained meal proportions (%) and curding were determined from sonography. Immediate and subsequent postprandial % were higher for PDHM (23%, P = 0.026; 15%, P = 0.006) and fortified meals (31.5%; 8.8%, both P < 0.001), whereas higher casein, whey, and lactose concentrations were associated with lower immediate postprandial % (all P < 0.006). Curding did not affect emptying. Influences of fortification, pasteurization, and differing breast milk compositions are small and unlikely implicated in preterm feeding intolerance.

Influences of breast milk composition on gastric emptying in preterm infants.

OBJECTIVES: The aim of the present study was to determine whether specific biochemical and energy concentrations influence gastric emptying of unfortified and fortified mother's own milk (MOM) in stable preterm infants, and whether gastric emptying differs between feeds of unfortified MOM and feeds fortified with S-26 or FM 85 human milk fortifier (HMF) when infants are fed the same volume under similar conditions. Influences of infant gestation, age, and weight, and feed characteristics were also explored. METHODS: Stomach volumes of 25 paired unfortified and fortified MOM feeds were monitored prefeed and postfeed delivery and at 30-minute intervals thereafter. For each feed, MOM samples were analyzed to determine concentrations of total protein, casein, whey, carbohydrate, lactose, fat, and energy. Fortified feed compositions were calculated by adding fortifier biochemical and energy concentrations to unfortified MOM concentrations. Ultrasound images were used to calculate infant stomach volumes. Statistical comparisons were made of paired stomach volume measurements. RESULTS: Higher feed concentrations of casein were associated with faster gastric emptying during feed delivery (P = 0.007). When compared with unfortified MOM, S-26 fortified feeds emptied similarly, whereas FM 85 fortified feeds emptied more slowly both during feed delivery and during the postprandial period (P = 0.002, <0.001, respectively). Gastric emptying was slower for 2-hourly feeds compared with that for 3-hourly feeds (P = 0.003) and in supine position compared with that in prone (P = 0.001). CONCLUSIONS: Breast milk composition influences gastric emptying in stable preterm infants, with feeds of higher casein concentration emptying faster during feeding than otherwise equivalent feeds, and FM 85 fortified MOM emptying more slowly than unfortified MOM.

Repeatability of gastric volume measurements and intragastric content using ultrasound in preterm infants.

OBJECTIVES: The aim of the present study was to determine whether serial gastric volumes and intragastric curding are repeatable within individual preterm infants when given sequential feeds of the same volume and composition. Infant and feeding characteristics that may influence differences in measured gastric volumes, echogenicity, and intragastric curding were also explored. METHODS: Ultrasound images were used to calculate gastric volumes and to rate echogenicity and intragastric curding for 20 infants. A total of 29 paired feeds of the same volume and composition were monitored prefeed and postfeed and at 30-minute intervals thereafter. Statistical comparisons of paired gastric volume measurements and agreement between echogenicity and curding ratings were made for each time point. Analyses of factors that influence discrepancies between volume measurements and between curding ratings were performed. RESULTS: Paired gastric volume measurements were repeatable (intraclass correlation coefficient [ICC] = 0.971, 0.938 < ICC < 0.987). Most (75%) discrepancies were <2 mL and increased over time, although volume differences were small. Overall moderate levels of consistency were observed for ratings of echogenicity (κ = 0.44), and curd presence (κ ≤ 0.65), density (κ = 0.41), and volume (κ = 0.47). Gastric emptying during feed delivery is influenced with infant positioning, fortification of breast milk, and feeding frequency. CONCLUSIONS: For preterm infants serial gastric volumes are repeatable and ratings of intragastric echogenicity and curding are moderately consistent when fed milk of the same volume and composition. Ultrasound has the potential to further explore factors that influence gastric emptying in the preterm infant.