Heat-treated colostrum feeding promotes beneficial bacteria colonization in the small intestine of neonatal calves.

The present study investigated the effect of heat-treated colostrum feeding on the bacterial colonization in calf small intestine of neonatal calves within the first 12h of life. Newborn Holstein bull calves (n=32) were assigned to 3 treatment groups and fed with either fresh colostrum (FC, n=12) or heat-treated (60°C, 60 min) colostrum (HC, n=12) soon after birth, whereas the control (NC, n=8) group did not receive colostrum or water. Small intestinal tissues and contents were collected from proximal jejunum, distal jejunum, and ileum at 6 and 12h after birth, following euthanasia. Quantitative real time-PCR was used to explore the colonization of total bacteria, Lactobacillus, Bifidobacterium, and Escherichia coli. The feeding of colostrum soon after birth increased the colonization of total bacteria in calf gut within the first 12h compared with NC. In contrast, the prevalence of Lactobacillus was lower in HC and FC compared to NC. Remarkable changes in the prevalence of small intestinal tissue-attached Bifidobacterium were observed with the feeding of HC, but not that in small intestinal contents. The prevalence of Bifidobacterium was 3.2 and 5.2 fold higher in HC than FC and NC, respectively, at 6h. Although the feeding of FC did not enhance the prevalence of tissue-attached Bifidobacterium at 6h compared with NC, it displayed a gradual increase over the time that was higher than NC, but similar to that of HC at 12h. Moreover, the colonization of E. coli was drastically reduced in HC calves compared with FC and NC. Thus, the present study suggests that the feeding of HC enhances the colonization of Bifidobacterium but lessens E. coli in the calf small intestine immediately postpartum compared with that of FC and NC. The increased colonization of beneficial bacteria along with the decreased colonization of potential pathogens in calf gut may also diminish the neonatal calf diarrhea when calves are fed heat-treated colostrum soon after birth.

Choosing healthier foods in recreational sports settings: a mixed methods investigation of the impact of nudging and an economic incentive.

BACKGROUND: Nudging is an approach to environmental change that alters social and physical environments to shift behaviors in positive, self-interested directions. Evidence indicates that eating is largely an automatic behavior governed by environmental cues, suggesting that it might be possible to nudge healthier dietary behaviors. This study assessed the comparative and additive efficacy of two nudges and an economic incentive in supporting healthy food purchases by patrons at a recreational swimming pool. METHODS: An initial pre-intervention period was followed by three successive and additive interventions that promoted sales of healthy items through: signage, taste testing, and 30% price reductions; concluding with a return to baseline conditions. Each period was 8 days in length. The primary outcome was the change in the proportion of healthy items sold in the intervention periods relative to pre- and post-intervention in the full sample, and in a subsample of patrons whose purchases were directly observed. Secondary outcomes included change in the caloric value of purchases, change in revenues and gross profits, and qualitative process observations. Data were analyzed using analysis of covariance, chi-square tests and thematic content analysis. RESULTS: Healthy items represented 41% of sales and were significantly lower than sales of unhealthy items (p < 0.0001). In the full sample, sales of healthy items did not differ across periods, whereas in the subsample, sales of healthy items increased by 30% when a signage + taste testing intervention was implemented (p < 0.01). This increase was maintained when prices of healthy items were reduced by 30%, and when all interventions were removed. When adults were alone they purchased more healthy items compared to when children were present during food purchases (p < 0.001), however parental choices were not substantially better than choices made by children alone. CONCLUSIONS: This study found mixed evidence for the efficacy of nudging in cueing healthier dietary behaviors. Moreover, price reductions appeared ineffectual in this setting. Our findings point to complex, context-specific patterns of effectiveness and suggest that nudging should not supplant the use of other strategies that have proven to promote healthier dietary behaviors.

Effect of calf starter feeding on gut microbial diversity and expression of genes involved in host immune responses and tight junctions in dairy calves during weaning transition.

Calf starters are usually offered to dairy calves to facilitate the weaning process, however, the effect of solid feed consumption on gut health has not been well studied. This study aimed to investigate the effect of calf starter feeding on the gut bacterial community and mucosal immune functions in dairy calves during weaning transition. Mucosal tissue and digesta samples were collected from rumen, jejunum, ileum, cecum, and colon upon slaughtering of calves (n=8) after feeding the experimental diets [milk replacer (MR) or milk replacer + calf starter (MR+S)] for 6 wk. Expression of toll-like receptor (TLR) 10 was downregulated along the gut, whereas TLR2 in colon and TLR6 along the gut were upregulated in MR+S-fed calves compared with MR-fed calves. Ileal TLR9 and TLR10 showed higher expression compared with the other regions regardless of the diet. Peptidoglycan recognition protein 1 demonstrated a diet- and gut-regional dependent expression pattern, whereas ß-defensin did not. The diet and gut region also affected the expression of tight junction-regulating genes claudin 4 and occludin. Bacterial diversity tended to be different between the 2 diets, whereas the bacterial density was different among gut regions and sample type. The present study revealed that changes in bacterial diversity, expression of genes encoding host mucosal immune responses, and barrier functions were associated with the MR+S diet, and suggests that solid feed consumption may alter gut microbiome and host mucosal functions during weaning transition.

Impact of feed efficiency and diet on adaptive variations in the bacterial community in the rumen fluid of cattle.

Limited knowledge of the structure and activities of the ruminal bacterial community prevents the understanding of the effect of population dynamics on functional bacterial groups and on host productivity. This study aimed to identify particular bacteria associated with host feed efficiency in steers with differing diets and residual feed intake (RFI) using culture-independent methods: PCR-denaturing gradient gel electrophoresis (DGGE) and quantitative real-time PCR analysis. PCR-DGGE profiles were generated from the ruminal fluid of 55 steers fed a low-energy-density diet and then switched to a high-energy-density diet. Bacterial profile comparisons by multivariate statistical analysis showed a trend only for RFI-related clusters on the high-energy diet. When steers (n = 19) belonging to the same RFI group under both diets were used to identify specific bacterial phylotypes related to feed efficiency traits, correlations were detected between dry matter intake, average daily gain, and copy numbers of the 16S rRNA gene of Succinivibrio sp. in low-RFI (efficient) steers, whereas correlations between Robinsoniella sp. and RFI (P < 0.05) were observed for high-RFI (inefficient) animals. Eubacterium sp. differed significantly (P < 0.05) between RFI groups that were only on the high-energy diet. Our work provides a comprehensive framework to understand how particular bacterial phylotypes contribute to differences in feed efficiency and ultimately influence host productivity, which may either depend on or be independent from diet factors.

Correlation of particular bacterial PCR-denaturing gradient gel electrophoresis patterns with bovine ruminal fermentation parameters and feed efficiency traits.

The influence of rumen microbial structure and functions on host physiology remains poorly understood. This study aimed to investigate the interaction between the ruminal microflora and the host by correlating bacterial diversity with fermentation measurements and feed efficiency traits, including dry matter intake, feed conversion ratio, average daily gain, and residual feed intake, using culture-independent methods. Universal bacterial partial 16S rRNA gene products were amplified from ruminal fluid collected from 58 steers raised under a low-energy diet and were subjected to PCR-denaturing gradient gel electrophoresis (DGGE) analysis. Multivariate statistical analysis was used to relate specific PCR-DGGE bands to various feed efficiency traits and metabolites. Analysis of volatile fatty acid profiles showed that butyrate was positively correlated with daily dry matter intake (P < 0.05) and tended to have higher concentration in inefficient animals (P = 0.10), while isovalerate was associated with residual feed intake (P < 0.05). Our results suggest that particular bacteria and their metabolism in the rumen may contribute to differences in host feed efficiency under a low-energy diet. This is the first study correlating PCR-DGGE bands representing specific bacteria to metabolites in the bovine rumen and to host feed efficiency traits.

Fermentation of barley by using Saccharomyces cerevisiae: examination of barley as a feedstock for bioethanol production and value-added products.

The objective of this study was to examine the ethanol yield potential of three barley varieties (Xena, Bold, and Fibar) in comparison to two benchmarks, corn and wheat. Very high gravity (VHG; 30% solids) fermentations using both conventional and Stargen 001 enzymes for starch hydrolysis were carried out as simultaneous saccharification and fermentation. The grains and their corresponding dried distiller's grain with solubles (DDGS) were also analyzed for nutritional and value-added characteristics. A VHG traditional fermentation approach utilizing jet-cooking fermentation revealed that both dehulled Bold and Xena barley produced ethanol concentrations higher than that produced by wheat (12.3, 12.2, and 11.9%, respectively) but lower than that produced by corn (13.8%). VHG-modified Stargen-based fermentation of dehulled Bold barley demonstrated comparable performance (14.3% ethanol) relative to that of corn (14.5%) and wheat (13.3%). Several important components were found to survive fermentation and were concentrated in DDGS. The highest yield of phenolics was detected in the DDGS (modified Stargen 001, 20% solids) of Xena (14.6 mg of gallic acid/g) and Bold (15.0 mg of gallic acid/g) when the hull was not removed before fermentation. The highest concentration of sterols in DDGS from barley was found in Xena (3.9 mg/g) when the hull was included. The DDGS recovered from corn had the highest concentration of fatty acids (72.6 and 77.5 mg/g). The DDGS recovered from VHG jet-cooking fermentations of Fibar, dehulled Bold, and corn demonstrated similar levels of tocopherols and tocotrienols. Corn DDGS was highest in crude fat but was lowest in crude protein and in vitro energy digestibility. Wheat DDGS was highest in crude protein content, similar to previous studies. The barley DDGS was the highest in in vitro energy digestibility.

Modification and validation of a Macronutrient Preference Checklist for use in North America.

A Macronutrient Preference Checklist (MPC) is a simple tool that assesses momentary macronutrient and taste preferences. The purpose of this study was to modify and validate an existing European 32-item MPC and adapt it for use in North America. A total of 160 subjects completed questionnaires (demographic survey, appetite assessment, MPC, and MPC items rated on a 9-pt hedonic scale) on two occasions (Part 1). The MPC showed good internal consistency (Cronbach's alpha values of 0.76, 0.72, 0.64, and 0.57 for the four macronutrient categories of high protein, high carbohydrate, high fat, and low energy) and high test-retest reliability, with strong correlations for all four macronutrient categories. A larger sample size (n=239) was used to explore the influences of age, gender, and appetite on macronutrient and taste preference (Part 2). Subjects completed study questionnaires once. The influences of appetite, age, and gender on macronutrient preferences assessed with the MPC concur with the literature, confirming the tool's validity. The MPC was found to be a valid and reliable tool that may be used as a simple and efficient method of assessing momentary macronutrient and taste preferences in future research and clinical settings in North America.

Evaluation of early conception factor lateral flow test to determine nonpregnancy in dairy cattle.

The early conception factor (ECF) lateral flow test was evaluated for its ability to accurately determine nonpregnant status in dairy cattle. Results of 2 field trials involving 191 cows and 832 tests indicated the probability that a cow can be correctly diagnosed as nonpregnant by using the ECF test is only about 50%. Agreement of test results between milk and serum obtained from the same cow was 57.5%. The ECF test was not consistent in identifying nonpregnancy when the same cows were tested repeatedly over a period of 4 weeks. We conclude that the ECF lateral flow test does not accurately identify nonpregnancy in dairy cattle.

Aromatic amino acid requirements in healthy men measured by indicator amino acid oxidation.

BACKGROUND: In the current literature, no agreement exists on estimates for aromatic amino acid (phenylalanine plus tyrosine) requirements as measured by stable-isotope techniques. OBJECTIVE: The goal of the present study was to determine the phenylalanine requirement in healthy men who were fed a diet without tyrosine by using the indicator amino acid oxidation method. DESIGN: Five healthy men were assigned to receive in random order diets devoid of tyrosine and with 8 graded intakes of phenylalanine (5, 10, 15, 25, 35, 45, 60, and 70 mg x kg(-1) x d(-1)). The phenylalanine requirement was measured by the rate of 13CO2 release (F13CO2) from L-[1-(13)C]lysine oxidation. RESULTS: The graded intakes of phenylalanine had no effect on lysine flux, as required for this method. The phenylalanine (ie, total aromatic amino acid) requirement, in the absence of tyrosine, was estimated to be 48 mg x kg(-1) x d(-1) by applying a two-phase linear regression crossover model to the F13CO2 data. CONCLUSIONS: In the absence of tyrosine, the mean phenylalanine requirement is higher than the current FAO/WHO/UNU (1985) and Dietary Reference Intake (2002) recommendations.

If we offer it, will children buy it? Sales of healthy foods mirrored their availability in a community sport, commercial setting in Alberta, Canada.

BACKGROUND: Community sports settings are often sources of unhealthy foods for children. Many managers in these settings are reluctant to increase availability of healthy food options because they perceive that healthy foods are not profitable. This study assessed the independent contribution of increased availability of healthy foods to their sales in a community sport, commercial context. Change in revenues per patron was also examined. METHODS: The availability of healthy items was increased from 9.1% at baseline (35 days) to 25.0% during the intervention period (40 days), returning to 9.1% postintervention (6 days). Purchases of all patrons who bought foods/beverages (n=17,262 items sold) from two concessions at an outdoor community pool were assessed from baseline to postintervention. Chi-square analyses assessed differences in the proportion of healthy and unhealthy items sold, as well as in the proportion of total revenues per patron across periods. A trained observer also recorded qualitative observations pertaining to a subset of patrons' (n=221) dietary behaviors and activities. RESULTS: Healthy items represented 7.7%, 22.7%, and 9.8% of sales during the preintervention, intervention, and postintervention periods, respectively (p<0.01). Sales of healthy beverages exceeded sales of all other product types. The proportion of total revenues per patron did not differ by period. CONCLUSIONS: Food availability was an important environmental determinant of food purchasing behaviors in this community commercial context, given that sales of healthy foods closely mirrored their availability. Increased availability of healthy foods in community and commercial settings is important because concurrent changes within multiple environments will be required to improve children's dietary behaviors.

Pretreatment with an intravenous lipid emulsion increases plasma eicosapentanoic acid and downregulates leukotriene b4, procalcitonin, and lymphocyte concentrations after open heart surgery in infants.

BACKGROUND: The effect of providing a lipid emulsion containing medium-chain triglyceride (MCT), soybean oil, and fish oil in critically ill infants is not widely studied. This study investigated lipid emulsion effects on plasma phospholipids and immune biomarkers. MATERIALS AND METHODS: Thirty-two infants undergoing cardiopulmonary bypass (CPB) and dependent on parenteral nutrition (PN) were randomized to receive either soybean oil (control, n = 16) or a 50:40:10 mixture of MCT, soybean oil, and fish oil (treatment, n = 16). PN was administered for 3 days preoperatively and 10 days postoperatively. Fatty acids, procalcitonin (PCT), leukotriene B4 (LTB4), and lymphocytes were quantified at baseline, before surgery, and days 1, 7 and 10 after surgery. RESULTS: PCT was significantly lower in the treatment vs control group 1 day postoperatively (P = .01). The treatment group exhibited a lower ω-6 to ω-3 ratio (P = .0001) and a higher ω-3 concentration at all postoperative study periods (P = .001). Treatment resulted in higher (P < .05) plasma phospholipid eicosapentaenoic acid (EPA) on days 7 and 10, while α-linolenic acid, arachidonic acid, and docosahexaenoic acid remained constant. An increase in plasma phospholipid EPA concentration was associated with a decrease in plasma phospholipid LTB4 concentration (P < .05). On postoperative day 10, treatment infants with high Pediatric Risk of Mortality III scores exhibited a 45% lower lymphocyte concentration (P < .05). CONCLUSION: These findings suggest that treating infants undergoing CPB with a lipid emulsion containing ω-3 improves fatty acid status and results in a lower inflammatory response after surgery. Overall, this alternative ω-3-enriched lipid emulsion may benefit clinical outcomes of critically ill infants after cardiac surgery.

Influence of sire breed on the interplay among rumen microbial populations inhabiting the rumen liquid of the progeny in beef cattle.

This study aimed to evaluate whether the host genetic background impact the ruminal microbial communities of the progeny of sires from three different breeds under different diets. Eighty five bacterial and twenty eight methanogen phylotypes from 49 individuals of diverging sire breed (Angus, ANG; Charolais, CHA; and Hybrid, HYB), fed high energy density (HE) and low energy density (LE) diets were determined and correlated with breed, rumen fermentation and phenotypic variables, using multivariate statistical approaches. When bacterial phylotypes were compared between diets, ANG offspring showed the lowest number of diet-associated phylotypes, whereas CHA and HYB progenies had seventeen and twenty-three diet-associated phylotypes, respectively. For the methanogen phylotypes, there were no sire breed-associated phylotypes; however, seven phylotypes were significantly different among breeds on either diet (P<0.05). Sire breed did not influence the metabolic variables measured when high energy diet was fed. A correlation matrix of all pairwise comparisons among frequencies of bacterial and methanogen phylotypes uncovered their relationships with sire breed. A cluster containing methanogen phylotypes M16 (Methanobrevibacter gottschalkii) and M20 (Methanobrevibacter smithii), and bacterial phylotype B62 (Robinsoniella sp.) in Angus offspring fed low energy diet reflected the metabolic interactions among microbial consortia. The clustering of the phylotype frequencies from the three breeds indicated that phylotypes detected in CHA and HYB progenies are more similar among them, compared to ANG animals. Our results revealed that the frequency of particular microbial phylotypes in the progeny of cattle may be influenced by the sire breed when different diets are fed and ultimately further impact host metabolic functions, such as feed efficiency.

Low energy intakes are associated with adverse outcomes in infants after open heart surgery.

BACKGROUND: Infants with congenital heart lesions who undergo open heart surgery may experience physiologic and metabolic stress in the postoperative period, leading to altered metabolism and hypercatabolism. The purpose of this study was to determine the relationship between energy intake and hospital outcomes during the first 10 days following neonatal open heart surgery. MATERIALS AND METHODS: A post hoc analysis of all patients in a prospective randomized controlled trial was performed. Nutrition intake and hospital outcomes were assessed in 32 infants (40 ± 2.2 weeks, 3.4 ± 0.5 kg) in the neonatal and pediatric intensive care units. Infants received parenteral nutrition (PN) for 1-4 days before and 10 days after open heart surgery. Infants were separated into those who received a cumulative energy intake of <689 kcal (average 63 kcal/kg/d) and those who received an intake ≥689 kcal during postoperative days 0-10. RESULTS: Lower energy intake was associated with a significantly increased duration of artificial ventilation (5 ± 1.2 days), time to chest closure (1.4 ± 0.5 days), time in intensive care (5 ± 1.8 days), and stay in the hospital (25 ± 6.4 days). Lower energy intake was also associated with a significant increase in the length of time infants required PN (8 ± 2.9 days) and longer time to achieve full enteral intake of 100 mL/kg/d (7 ± 2.2 days) and before enteral feeds could be initiated (5 ± 1.5 days). CONCLUSIONS: Providing <63 kcal/kg/d to infants after open heart surgery was associated with adverse pediatric intensive care outcomes.

Pre-treatment with an intravenous lipid emulsion containing fish oil (eicosapentaenoic and docosahexaenoic acid) decreases inflammatory markers after open-heart surgery in infants: a randomized, controlled trial.

BACKGROUND & AIMS: This study assessed the effects of administering a lipid emulsion containing eicosapentaenoic and docosahexaenoic acid before and after open-heart surgery on cytokine production and length of hospital stay in infants. METHODS: Thirty-two infants (40 ± 2.3 weeks gestational age; 10.6 days at time of surgery) undergoing open-heart surgery with cardiopulmonary bypass were randomized to receive an intravenous lipid emulsion with (treatment) or without (control) eicosapentaenoic and docosahexaenoic acid in this prospective, randomized, double-blind, controlled trial. RESULTS: Mean plasma TNF-α concentration was significantly (p = 0.003) lower in the treatment (5.9 pg/mL) compared to the control group (14.8 pg/mL). In infants without sepsis, plasma TNF-α did not differ according to treatment, however when sepsis developed, mean plasma TNF-α was 21.1 pg/mL and 1.5 pg/mL (p = 0.0007) in control and treatment groups, respectively. Plasma TNF-α was positively correlated with length of hospital stay in the control group (p = 0.01), and negatively correlated with length of stay in the treatment group (p = 0.004), with a significant time by treatment interaction (p = 0.02). CONCLUSIONS: Providing a lipid emulsion containing eicosapentaenoic and docosahexaenoic acid reduces TNF-α concentrations in infants undergoing open-heart surgery. Lipid emulsions containing eicosapentaenoic and docosahexaenoic acid may ameliorate the inflammatory response among critically ill infants.

Evaluation of value-added components of dried distiller's grain with solubles from triticale and wheat.

This study focused on the detection of value-added co-products in dried distiller's grain plus soluble (DDGS), a possibility that could open new avenues for further processing and marketing of DDGS and improving economic sustainability of ethanol industry. Varieties of triticale, wheat and two benchmarks, CPS wheat and Pioneer Hi-Bred corn, were fermented using two very high gravity (VHG) fermentation approaches: jet-cooking and raw starch processing (STARGEN fermentation). DDGS from STARGEN fermentation could be promising sources of value-added co-products. Pronghorn triticale DDGS (STARGEN fermentation) had the highest concentration of sterols (3.7 mg/g), phenolic compounds (13.61 mg GAE/g), and ß-glucan (2.07%). CDC Ptarmigan DDGS (STARGEN fermentation) had the highest concentration of tocopherols and tocotrienols (107.0 µg/g), 1.93% of ß-glucan, and 53.0mg/g of fatty acids. AC Reed DDGS (STARGEN method) showed 1.97% of ß-glucan. This study shows that proper choice of fermentation approach and feedstock for ethanol production could improve commercial quality of DDGS.

Leucine is not a good choice as an indicator amino acid for determining amino acid requirements in men.

Leucine tracer has been widely used for examining whole-body protein turnover in humans, but has not been evaluated as an indicator to be used in the indicator amino acid oxidation (IAAO) method. The goal of this study was to determine whether the L-[1-(13C)]leucine isotope is an acceptable indicator by comparing it with an established tracer, L-[1-(13C)]lysine. Healthy men (n = 7; 29.9 +/- 4.8 y old) were fed in random order a diet with 7 graded intakes of phenylalanine without tyrosine. In the first study (n = 5), subjects were administered an excess leucine intake of 65 mg/(kg.d), and in the second study (n = 5), they were given the mean requirement of 45 mg/(kg.d) to determine whether leucine intake affected the pattern of response. Previous IAAO studies using lysine and phenylalanine demonstrated a clear pattern in 13CO2 production, i.e., increasing test amino acid intake resulted in a linear decrease to plateau, with a readily discernable breakpoint indicating the requirement. This pattern of production of 13CO2, indicates clear partitioning of the indicator amino acid between oxidation and protein synthesis. This was not observed with leucine at an intake of 65 mg/(kg.d). Conversely, at the lower leucine intake of 45 mg/(kg.d), a breakpoint was seen and a total aromatic amino acid requirement estimate that did not differ from that obtain using lysine as the indicator was obtained. In conclusion, leucine may be used as the indicator in the IAAO technique only when the daily intake leucine is given at its mean requirement level and the potential metabolic effects of other variables are taken into consideration.

Parenteral and enteral routes of feeding in neonatal piglets require different ratios of branched-chain amino acids.

The requirements for total branched-chain amino acids (BCAA), isoleucine, leucine and valine, in neonatal piglets receiving parenteral and enteral nutrition was determined recently. The optimum ratio among BCAA during different routes of feeding is not yet known. In this study, the ratio of BCAA during parenteral and enteral feeding was tested using the indicator amino acid oxidation (IAAO) technique. Male Yorkshire piglets (n=24) received amino acid-based diets containing adequate nutrients for 5 d. Phenylalanine oxidation and kinetics were determined from a 4-h primed, constant infusion of L-[1-14C]-phenylalanine on d 6 and 8. On d 6, all piglets received a BCAA diet which met 75% of the total BCAA requirement, based on our previous research, with a ratio of 1:1.8:1.2 of isoleucine/leucine/valine. On d 8, the piglets were randomly assigned to receive one of the 3 test diets supplemented with isoleucine (+isoleucine), leucine (+leucine) or valine (+valine) to meet 100% of requirement, with the remaining two BCAA at 75% of requirement. The difference in phenylalanine oxidation (% of dose) between d 6 and 8 was used as an indicator of BCAA adequacy. In enterally fed piglets, the change in the percentage of the dose oxidized was minimal for all 3 test diets (mean=1.15%). In parenterally fed piglets, the difference in phenylalanine oxidation (% of dose) between d 6 and 8 was +isoleucine (12.6%), +leucine (2%) and +valine (6.6%). The ratio of 1:1.8:1.2 of isoleucine/leucine/valine is appropriate for enteral feeding, but during parenteral feeding, isoleucine was first limiting and valine was second limiting.

Phase of menstrual cycle affects lysine requirement in healthy women.

The aim of this study was to investigate whether the phases of the menstrual cycle affect lysine requirement in healthy adult females, as determined by the indicator amino acid oxidation (IAAO) method. Five healthy females with regular menstrual cycles were studied at seven graded levels of lysine intake, in random order, with an oral [13C]phenylalanine tracer protocol in both the follicular and luteal phases. A total of 14 studies were conducted for each subject. Breath and plasma samples were collected according to the standard IAAO protocol. Serum 17beta-estradiol and progesterone concentrations were measured on each IAAO study day. The rate of release of 13CO2 from [13C]phenylalanine oxidation (F13CO2) was measured, and a two-phase linear regression crossover model was applied to determine lysine requirement. F13CO2 was higher during the luteal phase (P < 0.001) and was positively associated with serum concentrations of 17beta-estradiol and progesterone. The F13CO2 data were adjusted for subjects and sex hormones and used to define breakpoints for lysine requirements. The lysine requirement of healthy females in the luteal phase was 37.7 mg.kg(-1).day(-1) and higher (P = 0.025) than that of females in the follicular phase (35.0 mg.kg(-1).day(-1)). At all lysine intake levels, plasma amino acids were lower and phenylalanine oxidation was higher in the luteal relative to the follicular phase. Therefore, we reason that the higher lysine requirement observed in the luteal phase is probably due to higher amino acid catabolism.