Total Sulfur Amino Acid Requirement Research Articles

62 Sulphur Amino Acids: Assessing Requirements and Utilization in Dogs and Cats

Abstract Methionine (Met), cyst(e)ine (Cys), homocysteine and taurine are the four sulfur-containing amino acids. Met can serve as a substrate for protein synthesis or to produce homocysteine, which can be remethylated to Met or irreversibly catabolized to Cys. Unlike dogs, cats are unable to further metabolize Cys to taurine, making the latter an indispensable amino acid for the cat. Met, on the other hand, is a dietary indispensable amino acid for all mammals with Cys having the ability to spare part of the Met requirement (total sulfur amino acid requirement; Met + Cys). The amount of Met that cannot be replaced by Cys is defined as the Met requirement (NRC, 2006). Empirical determinations of the Met requirement in adult dogs and cats are still limited and the recommendations proposed by the NRC (2006) are based on studies that used insensitive techniques for mature animals. When a more robust technique, the indicator amino acid oxidation, and more practical experimental diets were used to empirically determine the Met requirement, a greater estimate was found in adult cats (Pezzali et al., 2022) and adult Labrador Retrievers and Beagles, but not in miniature Dachshunds (Mansilla et al., 2020), compared with the current recommendations proposed by the NRC (2006) and AAFCO (2023). Harrison et al. (2020), however, found a decreased estimate for the Met requirement in adult Labrador Retrievers compared with Mansilla et al. (2020), likely due to differences in experimental design and diet composition. This discrepancy highlights the need to standardize methodologies and to investigate the effect of other dietary factors on the Met and total sulfur amino acid requirements owing to their complex metabolism. Indeed, the metabolism of sulfur amino acids may differ between dog breeds, resulting in differences in Met requirements and potentially making taurine a conditionally indispensable amino acid in some breeds under specific dietary conditions. While dietary Met is usually provided in excess in commercial diets, excess of Met may lead to hyperhomocysteinemia, which has been associated with detrimental health effects in other species. Due to the complex metabolism of sulfur amino acids and the complex matrix of pet foods, determining the ideal provision of sulfur amino acids in commercial diets to maximize animal health is a challenge. More long-term studies investigating the effect of different doses and ratios (Met:Cys) of sulfur amino acids on overall metabolism are necessary, in addition to requirement studies, to optimize the dietary delivery of sulfur amino acids in dogs and cats.

P07-051-23 Total Sulfur Amino Acid Requirements Are Higher in Late Gestation Compared to Early Gestation During Healthy Human Pregnancy

P07-051-23 Total Sulfur Amino Acid Requirements Are Higher in Late Gestation Compared to Early Gestation During Healthy Human Pregnancy

The dietary requirement for total sulfur amino acids in adults aged ≥60 years appears to be higher in males than in females

BackgroundThe total sulfur amino acid (TSAA) recommendation in older adults is based on data from young adults. Physiological evidence suggests that older adults have a higher requirement than young adults. ObjectivesThe objective of this study was to determine the TSAA requirement in healthy men and women aged ≥60 y. MethodsThe TSAA requirement was determined using the indicator amino acid oxidation method with L-[1-13C]phenylalanine as the indicator. At recruitment, 15 older adults (n = 7 men and n = 8 women; BMI < 30 kg/m2) were assigned to receive 7 methionine intakes (5, 10, 15, 19, 25, 35, and 40 mg/kg/d) without dietary cysteine. Intake levels were randomly assigned to each subject. Following enrollment, 2 subjects completed 2 intakes and 3 completed 3, while the remainder completed all 7. Mean TSAA requirement was determined from oxidation of L-[1-13C]phenylalanine using a mixed-effect change-point model. The 95% CI was calculated using parametric bootstrap. To test whether breakpoints were different between men and women, the overlap in the 95% CI was calculated. ResultsThe mean TSAA requirement was 26.2 (Rm2 = 0.39, Rc2 = 0.89; P < 0.001) and 17.1 mg/kg/d (Rm2 = 0.22, Rc2 = 0.79; P < 0.001) for men and women, respectively. The requirement was significantly higher in men than in women (difference in CI: 9.1 ± 8.85). ConclusionsTo our knowledge, this is the first study to determine the TSAA requirement in older adults. The requirement in older women is similar to current recommendations but is 75% higher in older men. These findings are important given recommendations for increased plant protein consumption. They will help in the assessment of diet quality and provide the basis of dietary guidelines for older adults consuming a plant-based diet.This trial was registered at clinicaltrials.gov as NCT04595188.

Revaluating the dietary methionine requirements in an improved strain of nile tilapia (Oreochromis niloticus)

It has been shown with terrestrial animals that the genetic improvement increases the nutrient requirements of the animal, which becomes more efficient in using these nutrients to achieve their higher growth potential. This study was conducted to estimate the dietary methionine requirements of different generations of the Genetically Improved Farmed Tilapia (GIFT) strain of Nile tilapia at the juvenile stage (17.51 g–19.55 g initial body weight). To achieve this objective, a completely randomized 2 × 6 factorial design was applied, with the genetic background of fish (generation) as the first independent variable with two levels, the 16th and 17th generations, and the methionine content of the diet as the second independent variable, with 6 graded levels, 0.52, 0.62, 0.84, 0.94, 1.04 and 1.27% of methionine in the diets, respectively. At the end of the 42-day experiment, the interaction effect of the generation × diet was not significant (P > 0.05) for any of the response parameters studied. The genetic improvement led to 15% more growth in the 17th generation than the 16th generation of the GIFT strain of Nile tilapia, which was accompanied with better feed conversion ratio, protein productive value, and energy productive value in the former generation (P < 0.05). Whatever the generation considered, the optimum dietary methionine requirement for the growth of the improved GIFT strain of Nile tilapia was estimated in this study, using the broken-line model, between 0.75% and 0.80% of the diet (with a cysteine level of 0.50% of the diet), which is higher than 0.49% of the diet previously estimated for conventional Nile tilapia at the juvenile stage. Therefore, the genetic improvement applied to the GIFT strain of Nile tilapia likely led to a higher methionine and total sulfur amino acid requirement than the conventional strains. The results of this study support the need to update the existing nutrient requirement databases of fish, in order to ensure that diets are formulated to effectively satisfy the requirements of not only conventional, but also improved strains of fish, for the full expression of their accrued growth potential of the latter strains.

Effects of levels of methionine supplementations in forms of L- or DL-methionine on the performance, intestinal development, immune response, and antioxidant system in broilers challenged with Eimeria spp.

The study was conducted to investigate the effects of 2 isoforms of methionine on growth performance and intestinal health induced by methionine (Met) deficiency and Eimeria infection in broilers. A total of 720 one-day old male chicks (Cobb500) were randomly allocated to 10 groups in a 2×5 factorial arrangement (6 reps/group, 12 birds/cage) with diets and Eimeria challenge as the main factors. Hundred percent DL-Met, 100% L-Met, 80% DL-Met, and 80% L-Met diets were formulated to meet approximately 100 or 80% of the total sulfur amino acid (TSAA) requirement with DL-Met or L-Met as Met supplementation sources. The 60% TSAA basal diet (60% Met) was formulated without Met supplementation. At d14, the challenge groups were gavaged with mixed Eimeria spp. Growth performance was recorded on d7, 14, 20 (6-day postinfection [DPI]), and 26 (12 DPI). The gut permeability was measured on 5 and 11 DPI. Antioxidant status and gene expression of immune cytokines and tight junction proteins were measured on 6 and 12 DPI. Data were analyzed by 1-way and 2-way ANOVA before and after the challenge, respectively. Orthogonal polynomial contrasts were used for post hoc comparison. Overall, the Eimeria challenge and 60% Met diet significantly reduced growth performance, antioxidant status, and mRNA expression of tight junction genes and immune cytokines. For other Met treatments, the L-Met groups had significantly higher BWG and lower FCR than the DL-Met group from d 1 to 20. The L-Met groups had less gut permeability than the DL-Met groups on 5 DPI. Compared to the 80% Met groups, the 100% Met groups reduced gut permeability. At 6 DPI, the 80% Met groups showed higher ZO1 expression than the 100% Met groups. The challenge groups had higher Muc2 expression and GSH/GSSG compared to the nonchallenge groups, and SOD activity was lower in the L-Met groups compared to the DL-Met groups at 6 DPI. The 100% Met groups had higher GPx activity than the 80% Met groups at 12 DPI. In conclusion, during coccidiosis, the 100% Met groups had better gut integrity and antioxidant status. Met supplementation in the form of L-Met improved growth performance in the starter phase and gut permeability in the challenge phase.

The Total Sulphur Amino Acid Requirement Is Higher in Older Men Than Women

ObjectivesThe primary aim of this study was to determine the total sulphur amino acid (TSAA) requirement of adults > 60 years, using the indicator amino acid oxidation (IAAO) method and to compare the requirement estimates between older males and females. The secondary aim was to compare the requirement estimates in older adults to the previously derived IAAO requirement in young males.MethodsFifteen healthy older adults (n = 7 male and n = 8 female participants), were randomized to receive 2 to 7 methionine intakes: 5, 10, 15, 19, 25, 35 and 40 mg. kg−1. d−1 without dietary cysteine. The TSAA requirement was determined using the IAAO method with L-[1–13C]phenylalanine as the indicator. The mean TSAA requirement was determined using two-phase linear mixed effect model, which identified a breakpoint in the F13CO2 in response to graded methionine intakes. The parametric bootstrap method was used to determine the 95% confidence interval (CI). To assess whether the breakpoints were different between older males and females or between older adults and young males the overlap in 95% CI was calculated. The null hypothesis was accepted if the interval contained zero.ResultsThe mean TSAA requirement for older males was 26.2 mg · kg−1· d−1 and 17.1 mg · kg−1· d−1 for older females. The TSAA requirement of older males is higher than older females (Difference in CI: 9.1 ± 8.85) and higher compared to young males based on our previous IAAO estimate of 12.6 mg · kg−1· d−1 (Difference in CI: 13.6 ± 5.98).ConclusionsTo our knowledge this is the first study to experimentally derive the TSAA requirement in older adults. The results can be used to assess whether the amino acid requirements of older adults can be met on a plant-based diet, especially given the current recommendations for increased plant protein consumption.Funding SourcesThis study was funded by CIHR. First author Alyssa Paoletti funded by Restracomp Scholarship provided by the Research Training Centre at SickKids Hospital.

Evaluating the dietary taurine requirement of hybrid striped bass, (Morone chrysops x M. saxatilis)

A dietary requirement for the sulfonic acid taurine has been documented in many marine species. However, it has not been well studied in hybrid fish with freshwater and marine genes, such as the hybrid striped bass (Morone chrysops x M. saxatilis). Therefore, an 8-week feeding trial was conducted to evaluate the effects of various levels of dietary taurine on growth performance, health, and body composition of juvenile hybrid striped bass. Four diets were formulated from practical ingredients and supplemented with DL-methionine at 0.54% of dry weight to meet the hybrid striped bass' established total sulfur amino acid requirement of 1.0% of dry diet, and taurine was supplemented incrementally at 0, 0.5, 1, and 2% of dry weight. Two additional diets were prepared without DL-methionine supplementation to be marginally deficient in methionine (0.49% by weight) and either supplemented with or without taurine at 1% dry weight to evaluate the potential interactions between methionine and taurine. Diets containing increment levels of taurine were determined to show no significant (P > .05) differences in terms of weight gain, feed efficiency (FE), whole-body composition, hepatosomatic index (HSI), intraperitoneal fat ratio (IPF ratio), or muscle yield. However, hybrid striped bass fed the methionine-deficient diets had significantly (P < .05) reduced percentage weight gain and FE as well as reduced muscle yield and HSI compared to fish fed the methionine-supplemented diets. Interestingly, taurine inclusion at 1% in the diet containing 0.49% methionine increased weight gain and feed efficiency of fish compared to those fed the diet without supplemental taurine or methionine. These results indicated that growth performance and body composition of hybrid striped bass did not respond to taurine supplementation when dietary methionine was adequate but did respond when dietary methionine was limiting.

The sulfur amino acid requirements of juvenile Yellowtail Kingfish (Seriola lalandi)

The dietary methionine and cysteine requirements of Yellowtail Kingfish (YTK) are unknown. Methionine, an essential sulfur-containing amino acid, acts as a sulfur and methyl donor for key metabolites, such as cysteine and taurine. Cysteine, a conditionally essential sulfur-containing amino acid, can spare significant amounts of methionine in the total sulfur amino acid (TSAA) requirement. Both methionine and cysteine are an integral part of YTK aquafeeds, in which methionine supplementation levels are based on the requirements of the closely related Japanese Yellowtail (Seriola quinqueradiata) at approximately 11.1 g kg−1 diet. However, recent research has demonstrated better growth and feed efficiency in YTK fed diets containing more than 11 g methionine kg−1, although the precise methionine requirement and potential interactions with cysteine remain unknown. Therefore, the present study was designed to (1) elucidate the methionine requirement of YTK at low and high dietary cysteine contents and to (2) quantify cysteine's sparing capacity for methionine in the TSAA requirement. These requirements and relationships were established by feeding ten isonitrogenous and isoenergetic diets, made from common feed ingredients, to triplicate groups of 12 fish (initial 52.6 ± 1.0 g fish−1) over 54 days. The orthogonal design consisted of two levels of dietary cysteine (5.6 & 13.9 g kg−1), crossed with five levels of dietary methionine, increasing from 7.9 to 25.2 g kg−1. Non-linear regression analysis indicated an average digestible TSAA requirement of 0.70 g kgBW−1 d−1 (0.56 g Met kgBW−1 d−1 & 0.14 g Cys kgBW−1 d−1) based on feed conversion ratio, specific growth rate, and protein retention efficiency. This approximates to an average dietary sulfur amino acid specification of 24.5 g kg−1 (18.9 g Met kg−1 & 5.6 g Cys kg−1). Cysteine spared 40.4–49.2% of methionine in the TSAA requirement on an equimolar sulfur basis. Sub- and supra-optimal levels of dietary methionine and cysteine induced inferior growth and feed efficiency. Additionally, fish fed the diet lowest in dietary methionine and cysteine indicated early stages of cataract. This study provides quantitative data on the sulfur amino acid requirements of juvenile YTK and will facilitate the formulation of better diets for this species.

The effect of total sulfur amino acid levels on growth performance and bone metabolism in pullets under heat stress

A study was conducted to investigate the effects of total sulfur amino acid (TSAA) levels on performance and bone metabolism in pullets under heat stress (HS). Hy-Line W36 day-old pullets (n = 216) were randomly distributed in 3 dietary treatments (70, 85, and 100% of TSAA), with 6 replicates of 12 birds. The treatments were defined as percentages of the TSAA level recommendation (100, 85, and 70%), and 85 and 100% of TSAA were obtained by adding L-Methionine to the basal deficient diet (70% of TSAA). The birds were raised under HS (35°C/7 h/D) from 1 to 18 wk. At 6, 12, and 18 wk, growth performance was measured. At 12 and 18 wk, bone weight, ash, collagenous (ColP), and noncollagenous proteins (NColP), tissue volume (TV), bone mineral content (BMC), and mineral density from total, cortical, and trabecular bones were evaluated. The means were subjected to ANOVA and, when significant (P ≤ 0.05), were compared by Dunnett's test. Regression analyses were performed to evaluate trends of TSAA dose response. Overall, birds fed 70% of TSAA showed poor growth and feed efficiency compared with other groups. Additionally, in at least 1 phase, birds fed 70% of TSAA showed lower bone ash, NColP, total BMC, and TV and higher ColP than the other treatments, whereas the cortical and trabecular TV and BMC were lower than 100% of TSAA (P < 0.04). Quadratic effects of TSAA levels on body weight gain (BWG) were found, and the level for maximum BWG was 95% of the TSAA recommendation (P < 0.03, R2 > 0.83). In conclusion, the use of a TSAA-deficient diet resulted in poor performance and delayed bone development. Additionally, the use of 100% of TSAA led to better initial structural bone development than 85% of TSAA. Therefore, the TSAA level recommended by the primary breeder guideline was enough to support growth and bone quality under HS, suggesting that HS does not alter TSAA requirement in pullets.

Ideal ratios of standardized ileal digestible methionine, threonine, and tryptophan relative to lysine for male broilers at the age of 1 to 10 days

The objective of this study was to develop ideal amino acid (AA) ratios based on the standardized ileal digestible (SID) lysine, methionine (in the presence of 4.3 g SID cysteine/kg), threonine, and tryptophan requirements, for 10-day-old male broiler chickens. Three experiments were conducted using Ross 308 male chicks fed diets based on corn, soybean meal, and additional graded crystalline AA with 228 g/kg of crude protein. Each diet contained the same amount of basal diets including corn, soybean meal, corn starch, and soybean oil, and graded AA were substituted at the expense of L-glutamic acid in order to keep all diets isonitrogenous. A total of 720 birds were allocated to 6 treatment groups, with 15 birds per cage and 8 cages per treatment using a randomized complete block design. Body weight and group feed intake were measured on day 10. The dietary SID AA levels in each experiment ranged from 2.9 to 6.9 g of methionine/kg, with increments of 0.8 g/kg (Exp. 1), 4.5–8.5 g of threonine/kg, with increments of 0.8 g/kg (Exp. 2), and 1.1–2.1 g of tryptophan/kg, with increments of 0.2 g/kg (Exp. 3), respectively. The methionine requirement was conducted in the presence of 4.5 g total cysteine/kg diet which was 4.3 g/kg diet in SID cysteine. Average daily gain and gain to feed ratio showed linear and quadratic relations to increasing levels of SID methionine, threonine, and tryptophan, respectively (P < 0.01). The SID requirements for broiler chicks at the age of 1–10 days were determined using 12.4 g/kg of SID lysine concentrations in the dietary treatments. The requirement for average daily gain and gain to feed ratios were 3.85 and 3.84 g/kg for SID methionine when the diet contained 4.3 g SID cysteine/kg diet (thus, total sulfur AA requirements were 8.15 and 8.14 g/kg, respectively), 7.11 and 7.38 g/kg for SID threonine, and 1.58 and 1.36 g/kg for SID tryptophan, respectively. By using the upper limit of the broken-line requirement, the ideal SID AA to lysine ratios of 10-day-old broiler chickens were 0.31, 0.60, and 0.13 for methionine (in the presence of 4.3 SID cysteine/kg diet), threonine, and tryptophan, respectively.

Stable isotopes to study sulfur amino acid utilization in broilers

Nutritionists have been discussing whether the dietary supplementation of cyst(e)ine is required as a part of the dietary methionine (Met) in the total sulfur amino acid (TSAA) requirement to achieve optimum performance in broilers. Part of Met is converted to cysteine (Cys) to meet the Cys requirement, especially for feather growth. The TSAA requirement has been determined by using graded levels of free Met in the diet, without supplementation of free cyst(e)ine. It has also been argued that the Met to Cys ratio (Met : Cys) changes with age and even with different Met sources. The objective of this study was to evaluate the two sources of Met, while determining the proportion of Met and Cys in total dietary TSAA that optimize the performance of broilers. A performance assay was carried out in a factorial arrangement (5 × 2) using 1080 broilers from 42 to 56 days of age fed diets having different dietary proportions of Met and Cys (44 : 56, 46 : 54, 48 : 52, 50 : 50 or 52 : 48) while maintaining the same dietary TSAA in the diets. Two synthetic Met sources (dl-Met or l-Met) were used for each of the diets with different dietary Met : Cys ratios. Twenty-one broilers of the same age were fed the diets 44 : 56, 48 : 52 and 52 : 48 by supplementing the diet with L-(15N) Met or L-(15N2) Cystine to study the metabolism of TSAA. No differences were observed between Met sources for feed intake, BW gain and feed conversion ratio (FCR; P > 0.05); however, FCR was numerically improved at 50 : 50 Met : Cys. Regarding TSAA utilization, the conversion of Met to Cys increased with increase in Met : Cys ratios, but the concentration of Met intermediates decreased. Broiler chickens responded to different dietary proportions of sulfur amino acids by altering their sulfur amino acid metabolism, and diets containing 50 : 50 Met : Cys is recommended for broilers of age 42 to 56 days.

Methionine to cystine ratio in the total sulfur amino acid requirements and sulfur amino acid metabolism using labelled amino acid approach for broilers

BackgroundAssuming that part of Methionine (Met) is converted into Cystine (Cys), but ignoring the rates with which such phenomenon occurs may lead to an excessive supply of Met in poultry diets. Such inconvenient could be easily avoided with the knowledge of the ideal Met:Cys/Total sulfur amino acids (TSAA) ratio and the rates of Met conversion into Cys.ResultsMet sources did not affect performance. Met:Cys/TSAA ideal ratio was determined using curvilinear-plateau regression model. Both optimum body weight gain and feed conversion ratio were estimated in 1007 g/day and 1.49, respectively, at 52% Met/TSAA ratio. Feed intake was not affected by Met:Cys/TSAA ratios. In the labelled amino acid assay, the rates with which Met was converted into Cys ranged from 27 to 43% in response to changes in Met:Cys/TSAA ratios, being higher at 56:44.ConclusionBased on performance outcomes, the minimum concentration of Met relative to Cys in diets for broilers from 14 to 28 d of age based on a TSAA basis, is 52% (52:48 Met:Cys/TSAA). The outcomes from labelled amino acid assay indicate that highest the Met supply in diets, the highest is its conversion into Cys.

Re-estimation of supplemented methionine as total sulfur amino acid requirement for commercial male meat-type ducks

Re-estimation of supplemented methionine as total sulfur amino acid requirement for commercial male meat-type ducks

Total Sulfur Amino Acid Requirements Are Not Altered in Children with Chronic Renal Insufficiency, but Minimum Methionine Needs Are Increased

Background: The total sulfur amino acid (TSAA) and minimum Met requirements have been previously determined in healthy children. TSAA metabolism is altered in kidney disease. Whether TSAA requirements are altered in children with chronic renal insufficiency (CRI) is unknown.Objective: We sought to determine the TSAA (Met in the absence of Cys) requirements and minimum Met (in the presence of excess Cys) requirements in children with CRI.Methods: Five children (4 boys, 1 girl) aged 10 ± 2.6 y with CRI were randomly assigned to receive graded intakes of Met (0, 5, 10, 15, 25, and 35 mg · kg-1 · d-1) with no Cys in the diet. Four of the children (3 boys, 1 girl) were then randomly assigned to receive graded dietary intakes of Met (0, 2.5, 5, 7.5, 10, and 15 mg · kg-1 · d-1) with 21 mg · kg-1 · d-1 Cys. The mean TSAA and minimum Met requirements were determined by measuring the oxidation of l-[1-13C]Phe to 13CO2 (F13CO2). A 2-phase linear-regression crossover analysis of the F13CO2 data identified a breakpoint at minimal F13CO2 Urine samples collected from all study days and from previous studies of healthy children were measured for sulfur metabolites.Results: The mean and population-safe (upper 95% CI) intakes of TSAA and minimum Met in children with CRI were determined to be 12.6 and 15.9 mg · kg-1 · d-1 and 7.3 and 10.9 mg · kg-1 · d-1, respectively. In healthy school-aged children the mean and upper 95% CI intakes of TSAA and minimum Met were determined to be 12.9 and 17.2 mg · kg-1 · d-1 and 5.8 and 7.3 mg · kg-1 · d-1, respectively. A comparison of the minimum Met requirements between healthy children and children with CRI indicated significant (P < 0.05) differences.Conclusion: These results suggest that children with CRI have a similar mean and population-safe TSAA to that of healthy children, suggesting adequate Cys synthesis via transsulfuration, but higher minimum Met requirement, suggesting reduced remethylation rates.

Evaluation of sulphur amino acid requirement of male Korean native ducklings from hatch to 21 day of age

ABSTRACT1. A dose-response experiment was conducted with male Korean native ducklings (KND) to evaluate the total sulphur amino acid (TSAA) requirement from hatch to 21 d of age.2. A completely randomised design with 7 dietary TSAA concentrations (0.62%, 0.65%, 0.68%, 0.71%, 0.74%, 0.77% and 0.80%) were used with 6 replications per treatment.3. Body weight (BW) and feed intake were measured weekly to calculate average daily gain (ADG), average daily feed intake (ADFI) and feed conversion ratio (FCR). One duckling per pen (n = 6) was killed by cervical dislocation to weigh empty body and drumsticks at the conclusion of experiment.4. BW was improved significantly with increasing TSAA content, in a non-linear manner. A significant decrease of FCR was shown with increasing TSAA contents. TSAA requirement was determined by taking a mean value after fitting the data to both a linear-plateau and a quadratic-plateau model. Estimated TSAA requirements were 0.70%, 0.70%, 0.66% and 0.70% for the maximum BW, ADG and ADFI and for the minimum FCR, respectively. Increasing TSAA content improved quantity of full body weight (FBW), empty body weight (EBW) and drumstick weight (DSW), but there was no effect on proportion of DSW in relation to EBW and proportion of EBW to FBW.5. In conclusion, the growth of male KND during 1 to 21 d of age was improved with increasing TSAA content, suggesting optimal TSAA requirements estimated by two analysis models.

Redefining the requirement for total sulfur amino acids in the diet of barramundi (Lates calcarifer) including assessment of the cystine replacement value

This study was designed to confirm a previous estimate of the methionine (Met) and total sulfur amino acid (TSAA) requirement of juvenile barramundi (Lates calcarifer) (Coloso et al., 1999) with a view for further study. Triplicate groups of fish (initial weight: 18.3g±1.5g) were fed diets with graded levels of dietary Met (7.2–12.8gkg−1 DM), centred around a previously reported requirement, and a constant dietary cystine (Cys) inclusion (5.9gkg−1 DM) over a 42day period. At the termination of the experiment, a significant linear increase (p<0.001) in %BW gain was observed in response to increasing dietary methionine, with no plateau in growth, suggesting the previous estimate of requirement may have been inadequate. A second experiment was designed to re-evaluate the Met/TSAA requirement in which a broader range of methionine inclusion levels were assessed (8.6–21.4gkg−1 diet DM Met). Triplicate groups of fish (initial weight: 36.4g±8.3g) were fed the diets for a period of 49days. A plateau and subsequent depression in growth, as well as significant (p<0.05) effects of dietary Met inclusion on %BW gain, feed conversion ratio (FCR) and protein retention efficiency (PRE) were observed at the conclusion of this experiment. The best fitting of nine nutrient response models, the Compartmental Model (R2=0.71), predicted a requirement for Met of between 10.5 (95% of maximum response) and 13.6gkg−1 (99% of maximum response) in a diet with 592g kg−1 CP and 6.6gkg−1 Cys (17.1–20.2gkg−1 TSAA; 1.8–2.3% CP Met +1.1% CP Cys). This TSAA requirement is equivalent to 43–51% of the lysine content of the diets. The applicability of this mode of expression and its relation to the ideal protein concept is discussed as is the application of different response models to the data. The impact of dietary Met:Cys ratio was also investigated with results suggesting at least 40% of dietary Met can be replaced with Cys without significantly affecting animal performance. It was concluded that disparity in the estimates of Met and TSAA requirement between this study and that of Coloso et al. (1999) was likely the result of a combination of model choice, experimental design and mode of expression of the requirements.

The cysteine, total sulfur amino acid, tyrosine, phenylalanine + tyrosine, and non-essential amino acid maintenance requirements of broiler breeders

Two hundred and fifty Cobb-Vantress broiler breeders were used to determine the maintenance requirement and efficiency of utilization of dietary Cys, Tyr, and non-essential amino acids (AA) in a 21-day experiment. The breeders were fed crystalline amino acid diets containing graded levels of Cys or Tyr representing 0, 10, 20, 30, and 40% of their suggested requirement level with all other amino acids maintained at 40% of their suggested requirement level. To determine the non-essential AA maintenance requirement, graded levels of non-essential AA were provided by glutamic acid to represent 12, 19, 26, 33, and 40% of the ideal level of glutamic acid with all other amino acids maintained at their maintenance requirement level. The total sulfur amino acid (TSAA) and Phe + Tyr requirements were calculated by combining Cys and Tyr results, respectively, with previously determined Met and Phe, respectively. The slope of Cys, Tyr, and non-essential AA accretion regression line indicated that 29% Cys, 24% TSAA, 21% Tyr, 20% Phe + Tyr, and 9% non-essential AA of crystalline amino acids were retained. The Cys requirement for zero protein accretion was calculated to be 30.48 mg/d or 17.006 mg/ kgBW0.75/d or 75.426 mg/kgCP/d. The TSAA requirement for zero accretion was calculated to be 132.25 mg/b/d, 71.48 mg/kgBW0.75/d, and 307.55 mg/kgCP/d. The Tyr requirement for zero protein accretion was calculated to be 65.907 mg/d or 37.233 mg/ kgBW0.75/d or 175.566 mg/kgCP/d. The Phe + Tyr requirement for zero protein accretion was calculated to be 352.18 mg/b/d, 190.37 mg/kgBW0.75/d, and 749.33 mg/kgCP/d. The non-essential AA requirement for zero protein accretion was calculated to be 3715.194 mg/d or 2003.155 mg/kgBW0.75/d or 9452.954 mg/kgCP/d.

Broiler responses to digestible total sulphur amino acids at different ages: a neural network approach

Three experiments were conducted with broiler chickens to evaluate the effects of digestible total sulphur amino acid (TSAA) on their performance at three different phases of starter (1–14 d), grower (15–28 d) and finisher (29–42 d). The measured traits included: average daily gain (ADG), feed intake, feed conversion ratio (FCR), carcass protein, body lipid (BL), feather weight gain, carcass plus feather protein, carcass TSAA deposition and nitrogen excretion (NE). A dilution technique was used to create seven diets (with eight replicates) increasing the TSAA content from 2.5 to 9.04 g/kg of diet for starter, 2.26 to 8.14 g/kg of diet for grower and 2.08 to 7.5 g/kg of diet for finisher. Data measured were imported to neural networks to predict the measured traits in response to dietary and intake levels of TSAA and find the optimal levels of TSAA that lead to the desired responses. Optimization results showed decreases in optimal dietary TSAA values with increasing age for all traits, while reverse was observed for intake values and requirements were increased as birds aged. The highest TSAA requirement (7.95, 7.2 and 6.6 g/kg and 283, 585 and 1150 mg/bird per d for starter, grower and finisher, respectively) were achieved for minimum BL and lowest (5.8, 5.2 and 4.9 g/kg and 201, 444 and 873 mg/bird per d for starter, grower and finisher, respectively) were suggested for minimum NE. Based on intake models, the optimal TSAA values for minimum FCR in phases 1–3 were 283, 585 and 1150 mg/bird per d while maximum ADGs were achieved with 201, 444 and 873 mg/bird per d of TSAA.

The sparing effect of cystine on methionine at a constant TSAA level in practical diets of juvenile Nile tilapiaOreochromis niloticus

The feeding experiment was conducted to evaluate the replacement effect of cystine on methionine at a constant total sulphur amino acid (TSAA) level in practical diets of juvenile Nile tilapia (Oreochromis niloticus). Soybean meal, cottonseed meal and rapeseed meal were used as the main protein sources in the diets containing 16.4 MJ kg−1 gross energy and 301 g kg−1 crude protein. Crystalline DL-methionine (Met) to L-cystine (Cys) were supplemented to fulfil various ratios of Met to Cys (60:40, 50:50, 40:60 and 30:70, based on an equimolar sulphur basis; and the analysed ratios were 53:47, 49:51, 34:66 and 31:69). Each diet was fed to triplicate groups of Nile tilapia juveniles (10.1 g mean initial weight) in a recirculation system for 8 weeks. The results showed that the growth performance and nutrient accretion were not affected by dietary methionine:cystine ratios (P > 0.05). However, the survival rates of fish tended to decrease with the decreasing methionine: cystine ratios. The only dietary treatment to achieve 100% survival was the 53:47 methionine: cystine treatment. And methionine digestibility decreased linearly with the decreasing methionine: cystine ratios (P < 0.05). Thus, it was concluded that cystine could replace up to approximately 47% of TSAA requirement on an equimolar sulphur basis without hampering growth performance in the practical diets of juvenile Nile tilapia.

Total sulphur amino acid requirement and maximum cysteine replacement value for methionine for fingerlingCatla catla(Hamilton)

Two separate 12 weeks feeding trials were performed to quantify the total sulphur amino acid (TSAA) requirement (experiment I) and cysteine replacement value for methionine (experiment II) of fingerling Catla catla. In experiment I, six casein–gelatin based (33% crude protein; 16.72 kJ g−1 gross energy) diets with graded levels of TSAA (0.56%, 0.81%, 1.06%, 1.31%, 1.56%, 1.81% dry diet) were fed to triplicate groups of fish (3.55 ± 0.06 cm; 0.65 ± 0.02 g) near to satiation. The TSAA requirement was determined by quadratic regression analysis of absolute weight gain (AWG), protein efficiency ratio (PER), feed efficiency (FE), protein gain (PG) and TSAA gain (TSAAG) against dietary TSAA concentrations at 95% maximum response. Above analysis revealed that inclusion of TSAA at 1.28% dry diet (1.22% methionine + 0.06% cysteine), corresponding to 3.87% of dietary protein is optimum. In experiment II, to determine the replacement value of cysteine for methionine, six diets containing 1.28% TSAA determined in experiment I with different ratios of l-methionine and l-cysteine (80:20, 70:30, 60:40, 50:50, 40:60, 30:70) on equimolar sulphur basis were fed to fish (3.65 ± 0.08 cm; 0.67 ± 0.04 g). Broken-line regression analysis of AWG, PER, FE, PG and TSAAG against varying methionine to cysteine ratios yielded the optimum cysteine replacement value at 40.3%. Based on above analysis, it is recommended that inclusion of 1.28% dietary TSAA, corresponding to 3.87% of dietary protein is optimum of which 40.3% could be spared by cysteine. Data generated during this study would be useful in formulating TSAA balanced, cost-effective feeds for the intensive culture of this fish.