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May | June 2013 An effective source of dietary methionine for the turbot Psetta maxima

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An effective source of dietary methionine for the turbot Psetta maxima
by Rui Ma1, Huapeng Hou1, Wenbing Zhang1, Anant Bharadwaj2, Craig Browdy2 and Kangsen Mai1 he turbot Psetta maxima is an important cultured flatfish species in Europe and now increasingly in China. Dietary formulations for this species typically have been dependent on high levels of fishmeal inclusion. Increasing costs and the decreasing availability of fishmeal have necessitated lowering fishmeal levels and increasing plant proteins in feed formulations. In such diets methionine can become one of the first limiting amino acids and supplementation is frequently necessary to balance diets and achieve optimum performance. The hydroxy analog of methionine, 2-hydroxy4-methylthio butanoic acid (HMTBa) is a safe and effective source of methionine that has been used to supplement methionine deficient diets for livestock and aquaculture species. HMTBa is structurally different from L-methionine in that it has a hydroxyl group instead of an amino group at the α-carbon position, potentially reducing feed nitrogen inputs into grow out systems. It is passively absorbed and is converted to L-methionine by D-hydroxy acid dehydrogenase and L-hydroxy acid oxidase enzymes which have been confirmed in tissues of shrimp and fish. A two-part study was carried out at the Ocean University of China, China to evaluate 1. The response of turbot fed diets supplemented with either HMTBa (2-hydroxy4-methylthio-butanoic acid) or L-methionine, and 2. The dynamics of absorption of HMTBa and L-methionine.


Materials and methods
A 75-day growth trial was carried out to evaluate the effects of HMTBa and L-methionine as dietary methionine sources

on the growth of juvenile turbot (initial weight 5.6 g. N=5 tanks per treatment). Five levels (0.3, 0.6, 0.9, 1.2 and 1.5% dry matter) of HMTBa (added as Mera™Met – 84 percent HMTBa; Novus International Inc., USA) and L-methionine were added evaluate the absorption of HMTBa and respectively to a practical basal diet, that was L-methionine from diets containing these limiting in methionine (0.59% methionine; methionine sources. The basal diet used in 0.42% cystine; 1.01% total sulfur amino acids). the second trial was generally similar to that This basal diet served as the control diet used in the growth trial (0.75% methionine and contained 48 percent crude protein and and 0.45% cystine; total sulfur amino acid concentration of 1.20%). The basal diet was approximately 12.5% crude lipid. A crystalline L-amino acid premix, which was devoid of sulfur amino acids was added to the basal diet to approximate the whole body composition of the turbot. Different levels of either HMTBa or L-methionine were added to the basal diet at the expense of glutamic acid to give total methionine concentrations ranging from 0.59 - 2.09 and total sulfur amino acid concentrations ranging from 1.01 to 2.51%. A second Figure 1: Weight gain and specific growth rate in juvenile study was turbot fed either HMTBa or L-methionine conducted to
38 | InternatIonal AquAFeed | May-June 2013



NUTRACEUTICALS AND PHYTOBIOTICS FOR AQUACULTURE Growth promoters Anti-parasites Attractants Hepatoprotectors
Figure 2a: Serum free-methionine concentrations in postfed turbot fed the control, L-methionine and HMTBa diets Figure 2b: Serum HMTBa concentrations in fish fed diets containing HMTBa

Antioxidants Detoxifiers Chelated minerals

supplemented with either 0.75% HMTBa or L-methionine. In this trial larger fish (65 g; 45 fish/tank; N=3) were used to facilitate blood sampling at the end of the trial. The basal control diet and the two experimental diets were fed to the turbot for a period of 14 days twice daily to apparent satiation. Fish were fasted 12 h prior to final feeding. Following feeding, fish were sampled at 0, 0.25, 0.5, 1, 2, 3, 4, 6, 9, 12, 18 and 24 h respectively and their blood collected. Blood was centrifuged and serum collected and stored prior to analysis. Serum was analyzed using HPLC for HMTBa and free methionine.

There were no significant differences in survival between treatments. Fish fed the

basal diet (red bar) showed significantly lower weight gain than all the other treatments (Figure 1). Weight gain in fish fed both methionine sources increased in a quadratic fashion with increasing dietary concentration. In fish fed HMTBa (blue bars) maximum response was observed at 0.9% supplementation and was significantly

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May-June 2013 | InternatIonal AquAFeed | 39


data suggest that HMTBa is absorbed into circulation in fish shortly after feeding and also show that the maximum serum concentrations of HMTBa in HMTBa-fed fish are similar to serum free-methionine concentrations in L-methionine fed fish.

The results of these trials confirm that HMTBa is a safe and available source of methionine in practical diets for the turbot Psetta maxima. The HMTBa was absorbed efficiently into circulation at rates that are similar or better than L-methionine. Dynamics confirmed direct absorption demonstrated in previous livestock studies. Mera™Met can provide a cost effective methionine source in reduced fishmeal formulations with 100 percent bioavailability. HMTBa has been shown in previous studies to improve feed attractability, providing an effective alternative for optimizing performance and allowing for higher cost efficiencies through the replacement of fishmeal by plant meals in aquatic feeds.

higher than the response seen in fish supplemented 0.9% L-methionine. Generally, at doses ranging from 0.6 to 1.5%, fish fed HMTBa showed equal or higher weight gain compared to fish fed L-methionine (green bars). Similar responses were observed for other performance parameters such as final weight, specific growth rate, feed efficiency, protein efficiency ratio and protein retention. Results from the analysis of serum show that the maximum serum concentration of free-methionine (435.13µmol/L) in L-methionine-fed fish was observed at 9 h

after feeding (Figure 2a) whereas the maximum serum HMTBa level (426.17µmol/L) in HMTBa-fed fish occurred 6 h after feeding (Figure 2b). Appreciable quantities of HMTBa were observed in the serum of HMTBa-fed fish shortly after feeding as expected based on passive diffusion of HMTBa across the gut wall in fish. Two free methionine peaks were measured in the serum of HMTBa-fed fish at 3-4 h and 12 h post-feeding. This suggests contributions from either HMTBa metabolism to L-methionine, tissue protein turnover or digestion of intact dietary proteins. These
40 | InternatIonal AquAFeed | May-June 2013

More InforMatIon:
1The Key Laboratory of Mariculture, Ocean

University of China, China Email: wzhang@ouc.edu.cn
2Novus International Inc., USA

Email: craig.browdy@novusint.com Website: www.novusint.com


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