Availability of supplemental amino acid-chelated trace elements in diets containing tricalcium phosphate and phytate to rainbow trout, Oncorhynchus mykiss

Aquaculture 225 (2003) 431 – 444 www.elsevier.com/locate/aqua-online

Availability of supplemental amino acid-chelated trace elements in diets containing tricalcium phosphate and phytate to rainbow trout, Oncorhynchus mykiss Mary Jane S. Apines a,b, Shuichi Satoh a,*, Viswanath Kiron a, Takeshi Watanabe a, Takashi Aoki c a

Laboratory of Fish Nutrition, Department of Aquatic Biosciences, Tokyo University of Fisheries, Tokyo 108-8477, Japan b Institute of Aquaculture, College of Fisheries and Ocean Sciences, University of the Philippines in the Visayas, Miag-ao, Iloilo 5023, Philippines c Laboratory of Genetics and Biochemistry, Department of Aquatic Biosciences, Tokyo University of Fisheries, Tokyo 108-8477, Japan

Abstract This study investigated the availability of amino acid-chelated trace elements to rainbow trout fed diets containing tricalcium phosphate (TCP) and phytate (PH) that are known mineral inhibitors. Six semi-purified diets were supplemented with trace elements either from the sulfate (SF, diets 1 and 2) or amino acid-chelate (AM, diets 3 – 6). Diets 1 (SF) and 3 (AM) contained neither TCP nor PH. Diet 4 (AMPO) included TCP while Diet 5 (AMPH) contained PH alone. Diets 2 (SF++) and 6 (AM++), on the other hand, contained both inhibitors. Rainbow trout weighing 1.6 g were fed the experimental diets three times daily to near satiation for 18 weeks. Growth, whole body mineral deposition, and enzyme activity/expression were measured after the trial. Growth of fish fed the SF++ diet was significantly lower than the rest of the groups. Whole body Cu content was significantly higher in AM groups than in SF groups with or without TCP and PH. Whole body and bone Zn contents were significantly higher in fish fed the diet containing AM alone compared to the rest. Alkaline phosphatase activity was significantly higher in fish fed the diet containing AM alone than with fish fed the SF diet with or without TCP and PH. CuZnSOD and DNA polymerase were highly expressed in the AM compared to the SF groups. The results show that availability of trace elements is

* Corresponding author. Tel.: +81-3-5463-0557; fax: +81-3-5463-0553. E-mail address: [email protected] (S. Satoh). 0044-8486/03/$ - see front matter D 2003 Elsevier Science B.V. All rights reserved. doi:10.1016/S0044-8486(03)00307-7

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significantly affected by their chemical form and that amino acid-chelates are more beneficial for rainbow trout even in the presence of TCP and PH compared to the inorganic sources tested. D 2003 Elsevier Science B.V. All rights reserved. Keywords: Amino acid-chelates; Trace elements; Availability; Rainbow trout; Enzymes; Mineral inhibitors

1. Introduction Most practical diets for carnivorous fish such as rainbow trout are heavily dependent on fish meal as a protein source. However, the need for alternative plant protein sources has been recognized due to the high cost of fish meal and its high phosphorus content which contributes to increased pollution of water bodies from aquaculture effluents. Fish meal and plant protein sources contain antinutritional substances particularly tricalcium phosphate and phytate that inhibit mineral availability (Hardy and Shearer, 1985; Richardson et al., 1985; Satoh et al., 1987a,b,c; Lovell, 1989; Francis et al., 2001). Vertebral zinc content was significantly reduced with increasing levels of phytic acid in diets containing 50 mg Zn kg 1 (Satoh et al., 1989). Further, the authors observed a significant reduction in growth and feed efficiency with the inclusion of phytic acid at 2.2%. Thus, requirements of fish for minerals increase with increasing levels of inhibitors present in the diet. Channel catfish require only 5 to 10 mg zinc kg 1 when fed purified diets but when fed practical diets containing fish meal and soybean meal, the requirement increased from 88 to 200 mg zinc kg 1 of diet (Lovell, 1977; Gatlin and Wilson, 1983, 1984; Gatlin and Phillips, 1989). It has been shown that chelates and complexes have the ability to compete with dietary mineral inhibitors thus making minerals more available to animals (Ashmead, 1992). Hence, mineral sources with higher availability should be considered in feed formulation. Recently, interest in using alternative mineral sources, particularly those chelated with proteins/amino acids, has increased due to their reportedly higher availability compared to conventional (inorganic) sources (Ashmead, 1992). However, there are still contentions as to the availability of organic mineral complexes to different species. Studies with swine did not show differences in the availability of Zn between organic and inorganic sources (Hill et al., 1986; Swinkels et al., 1996). Likewise, Gomez and Kaushik (1993) did not observe significant differences between ZnSO4 and ZnMet fed to rainbow trout. Wedekind et al. (1994) observed that Zn from ZnSO4 was more available than from zinc –lysine and zinc –methionine. However, improved growth and increased bone Zn deposition were reported in chicks fed ZnMet with higher bioavailability compared to ZnO and ZnSO4 (Wedekind and Baker, 1989; Wedekind et al., 1992). In catfish also, availability of ZnMet was greater than ZnSO4 in terms of growth and bone Zn content (Paripatananont and Lovell, 1995). This was validated by our previous studies that Zn and Mn from their amino acid-chelates seem to be more available than the inorganic sources tested (Apines et al., 2001; Satoh et al., 2001).

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This study investigated the availability of amino acid-chelated trace elements to rainbow trout using semi-purified diets containing tricalcium phosphate at a level normally contained in practical diets and phytate at a higher level anticipated for the gradual replacement of fish meal with plant protein sources in rainbow trout diet. Parameters considered include growth, mineral absorption, tissue deposition, and retention. As trace elements mainly function as co-factor of numerous enzymes, activity/expression of selected metalloenzymes was also measured.

2. Materials and methods 2.1. Experimental diets and design Six semi-purified diets containing trace elements either from the sulfate (Kokusan Chemical Works, Tokyo, Japan) or amino acid-chelate (Eisai, Tokyo, Japan) sources with or without tricalcium phosphate (TCP) and phytate (PH) were formulated (Table 1). The diets contained 89.6% basal components and 0.05% trace element mix from either source. Diets 1 and 3 contained neither TCP nor PH and were designated as SF (sulfate) and AM (amino acid-chelate), respectively. In diets 4 and 5, either TCP or PH was included and designated as AMPO and AMPH. Diets 2 and 6 contained both TCP and PH, and thus coded as SF++ and AM++ (Table 2). Tricalcium phosphate was added at 4% and Ca-phytate at 1.5%. The diets were pelleted using the laboratory pelletizer (AEZ12M, Hiraga-Seikakusho, Kobe, Japan), dried in a vacuum freeze-drier (RLE-206, Kyowa Vacuum Tech., Saitama, Japan) and stored at 4 jC until used. The experimental set-up was arranged in a completely randomized design with two replications per treatment. 2.2. Fish, experimental conditions, and feeding Rainbow trout (Oncorhynchus mykiss) of about 1.63 F 0.31 g were randomly distributed in 60-l glass tanks supplied with dechlorinated tap water in a semi-recirculating

Table 1 Composition of the experimental diets (g kg

1

)

Diet code

SF

SF ++

AM

AMPO

AMPH

AM ++

Basal dieta Zn, Mn, Cu – Sulfate mix Zn, Mn, Cu – AA chelate mix Ca3(PO4)2 Ca-phytate Cellulose Total

896.0 0.5 – – – 103.5 1000

896.0 0.5 – 40.0 15.0 48.5 1000

896.0 – 0.5 – – 103.5 1000

896.0 – 0.5 40.0 – 63.5 1000

896.0 – 0.5 – 15.0 88.5 1000

896.0 – 0.5 40.0 15.0 48.5 1000

a

Basal diet (g kg 1)—Casein 450, Pregelatinized starch 200, Dextrin 50, Pollock liver oil 60, Soybean oil 60, Vitamin mix 30, Choline chloride 5, Vitamin E (50%) 1, Zn, Mn, Cu – free mineral mix 10, and NaH2PO4
2H2O 30.

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Table 2 Proximate and mineral compositions of the experimental diets* Diet code

SF

SF ++

AM

AMPO

AMPH

AM ++

Moisture Protein (%)a Lipid (%) Ash (%) Zn (Ag g 1) Cu (Ag g 1) Mn (Ag g 1) Ca (mg g 1) P (mg g 1)

9.32 F 0.05 46.3 F 0.04 8.11 F 0.85 3.44 F 0.22 32.9 F 0.69 4.70 F 0.11 13.5 F 2.34 0.11 F 0 11.4 F 0.24

9.99 F 0.12 45.1 F 0.02 7.53 F 0.18 8.17 F 0.02 30.0 F 0.64 4.62 F 0.24 13.9 F 0.29 18.3 F 0.15 21.5 F 0.12

8.99 F 0.14 45.9 F 0.01 8.29 F 0.26 3.50 F 0.06 30.5 F 0.56 4.67 F 0.09 13.7 F 0.48 0.16 F 0.05 11.6 F 0.16

9.99 F 0.07 45.0 F 0.22 8.00 F 0.11 7.43 F 0.09 31.3 F 1.12 4.91 F 0.19 13.7 F 0.04 15.6 F 0.44 20.4 F 0.18

9.74 F 0.04 46.2 F 0.20 8.21 F 0.47 4.35 F 0.02 32.4 F 0.35 4.86 F 0.05 13.4 F 0.83 2.90 F 0.02 13.5 F 0.30

9.36 F 0.13 45.2 F 0.13 8.55 F 0.34 8.17 F 0.06 33.3 F 0.83 4.69 F 0.12 13.2 F 0.03 18.2 F 0.44 22.1 F 0.54

a

Dry matter basis. * Values represent means F S.D.

system at a flow-rate of 600 –700 ml min 1 with an average daily temperature of 18.8 F 1.31 jC. The fish were fed the experimental diets three times daily to near satiation for 18 weeks. 2.3. Sample collection and chemical analyses Initial weight data were obtained at the start of the experiment and growth of fish was measured every 3 weeks thereafter. After the feeding trial, samples were collected and analyzed as described in Apines et al. (2001), except for minerals other than phosphorus which were measured using the Polarized Zeeman Atomic Absorption Spectrophotometer (Hitachi Z-5010, Tokyo, Japan). Plasma and liver samples for the analysis of alkaline phosphatase, CuZnSOD, and DNA polymerase were taken from the same fish used in bone analysis. 2.4. Enzyme assay 2.4.1. Alkaline phosphatase (ALP) Alkaline phosphatase activity was determined colorimetrically using p-nitrophenyl phosphate as substrate following the method of Snedeker and Greger (1983) with some Table 3 Growth and feed performance in rainbow trout cultured for 18 weeks* Diet code

Weight (g) Initial

Final

Wt. gain (g)

SGR (% day

SF SF ++ AM AMPO AMPH AM ++

1.66 F 0.04 1.61 F 0.03 1.62 F 0 1.64 F 0.01 1.64 F 0.02 1.64 F 0.04

33.3 F 0.04 23.4 F 0.04 33.9 F 0.04 36.8 F 0.04 36.8 F 0.04 33.4 F 0.04

31.7 F 1.61ab 21.8 F 2.18a 32.3 F 1.14b 35.2 F 0.22b 35.2 F 1.63b 31.8 F 1.49b

2.38 F 0.05 2.12 F 0.14 2.42 F 0.05 2.47 F 0 2.47 F 0.05 2.39 F 0.04

1

)

Feed FGR consumption (g fish 1)

TGC

31.4 F 1.43b 22.8 F 1.56a 34.1 F 1.52b 34.6 F 1.05b 35.3 F 1.10b 31.6 F 1.17b

0.000860 F 3 0.000712 F 6 0.000873 F 2 0.000908 F 4 0.000907 F 9 0.000863 F 3

0.99 F 0.01 1.05 F 0.05 1.06 F 0.01 0.99 F 0.04 1.01 F 0.02 1.00 F 0.02

* Values (means F S.D.) in a column not sharing the same letters are significantly different ( P < 0.05).

5b 5a 5b 5b 5b 5b

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modifications. Briefly, 15 Al of the diluted sample was added to the substrate and incubated at 20 jC for 30 min. Then, 2 ml of 3 N NaOH was added to arrest the reaction. The change in absorbance at 405 nm due to the formation of p-nitrophenol measured in a spectrophotometer (Beckman DU640, California, USA) is directly proportional to the ALP activity. 2.4.2. CuZnSOD and DNA polymerase Total RNA was extracted from the liver using Trizol (Invitrogen, California, USA) following the manufacturer’s protocol and reverse-transcribed using the AMV reverse

Fig. 1. Whole body mineral content in rainbow trout fed diets supplemented with trace element from different sources (n = pooled samples of 5 fish/tank). Bars represent means F S.D. Means with different letters are significantly different ( P < 0.05).

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transcriptase first-strand cDNA synthesis kit (Life Sciences, Florida, USA). The 25-Al cDNA synthesis reaction contained 16 Al total RNA (10 Ag), 1.0 Al pd(T)12 – 18 primer, 1.0 Al of 0.25 M DDT, 1.0 Al RNAsin, 5.0 Al 5  reaction buffer, and 1.0 Al AMV-RT. The reverse-transcribed cDNA samples were amplified using specific primers for rainbow trout CuZnSOD (GenBank Acc. No. AF469663): sense, 5V-GTGGCTAAGATCAACATCCA3V; anti-sense, 5V-CTTGTCTAAATAGGGAGTCA-3V and DNA polymerase (GenBank Acc. No. U53366): sense, 5V-TCTCAATATGCATGTCAGAG-3V; anti-sense, 5V-TCGATGATGCATCCAACAGC-3V. The following conditions were adopted for the amplification process: initial denaturation at 95 jC for 3 min, followed by 30 cycles of denaturation at 95 jC for 30 s, annealing at 49 jC (DNA Polymerase), 50 jC (CuZnSOD), and 46 jC (hactin) for 30 s, and elongation at 72 jC for 1 min, with final elongation step at 72 jC for 5 min. The PCR products were electrophoresed in 1% ethidium bromide stained-agarose gel and the signal was quantified using Atto densitograph software (Version 4.1). h-actin was used as an internal control and was also amplified using the specific primers for rainbow trout (GenBank Acc. No. AF157514): sense, 5V-ATCCTGACTCTGAAGTACCC-3V; anti-sense, 5V-GCGGTGCCCATCTCCTGCTC-3V.

Fig. 2. Bone Zn (A) and liver Cu (B) contents in rainbow trout fed diets supplemented with trace element from different sources (A: n = pooled samples of 5 fish/tank, B: n = 10 fish). Bars represent means F S.D. Means with different letters are significantly different ( P < 0.05).

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Fig. 3. Alkaline phosphatase activity in rainbow trout fed diets supplemented with trace element from different sources (n = 10 fish). Bars represent means F S.D. Means with different letters are significantly different ( P < 0.05).

2.5. Statistical analyses Results were analyzed by one-way ANOVA (Systat 8.0, SPSS, Chicago, USA). Differences between treatments were evaluated using Tukey’s test. Alkaline phosphatase activity between groups was compared using the Fisher’s Least Significant Difference Test. Values were considered significant at P < 0.05.

3. Results 3.1. Growth Growth, feed consumption, and thermal-unit growth coefficient (TGC) of the fish were significantly lower in the SF++ compared to the chelate-supplemented groups (Table 3).

Fig. 4. CuZnSOD and DNA polymerase mRNA expression in rainbow trout fed diets supplemented with trace element from different sources.

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However, specific growth rate (SGR) and feed to gain ratio (FGR) did not differ significantly among treatments. 3.2. Whole body, bone, and liver mineral content Whole body Zn content of the fish in the AM supplemented group was significantly higher than the other groups (Fig. 1). The same pattern was also observed in bone Zn content (Fig. 2A). Whole body Cu and Mn contents were significantly higher in the chelate- compared to the sulfate-supplemented groups (Fig. 1). Liver Cu content also tended to be higher in the chelate than the sulfate group (Fig. 2B).

Fig. 5. Retention of trace elements from different sources in rainbow trout. Bars represent means F S.D. Means with different letters are significantly different ( P < 0.05).

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3.3. Enzyme Alkaline phosphatase activity was significantly higher in the AM group compared to the sulfate-supplemented group with or without inhibitors (Fig. 3). Likewise, CuZnSOD and DNA polymerase were highly expressed in the chelate than the sulfate group (Fig. 4). 3.4. Retention The result showed that Zn retention in the AM group was significantly higher than the rest of the treatments (Fig. 5). Cu and Mn retention were likewise higher in the chelate than the sulfate group.

4. Discussion In this study, we were able to show the advantages of using the amino acid-chelate as a source of trace elements for rainbow trout. The higher growth exhibited by fish fed the amino acid-chelate compared to the sulfate-supplemented group validated our previous finding that amino acid complexes of Mn significantly improved the growth of rainbow trout (Satoh et al., 2001). On the other hand, this effect of the chelates on growth was not observed in similar studies using practical diets (Apines et al., 2001, unpublished). Several authors have previously reported that Zn sources have no significant influence on the growth performance of vertebrates including fish (Roth and Kirchgessner, 1983; Spears, 1989; Mohanna and Nys, 1999; Maage et al., 2001). In the present study, the significantly lower weight gain in the SF++ group might be due to the lower feed consumption in this group compared to the rest of the treatments as SGR and FGR were similar. Reduced feed consumption was detected only from the 15th week of the trial and could not be due to the palatability of the feed itself but probably due to taste dysfunction accompanying low Zn utilization as observed in rats and humans (Komai et al., 2000; Goto et al., 2001; Kitagoh et al., 2002). Prolonged feeding resulted to a reduction in feed intake and cumulative effect of the diet could have been the cause of reduced growth in the SF++ group. Previous studies showed that Zn and Cu are mainly stored in bone and liver, respectively (Keen and Graham, 1989; Linder et al., 1998; Turnlund, 1998; Peretz et al., 2001; Apines et al., unpublished) and therefore their levels in these organs are promising indices for evaluating their status. In this study, the significantly higher levels of Zn and Cu in bone and liver from the AM group compared to SF and SF++ suggests a higher availability of the elements from this source. Zinc can influence bone mineralization either directly, as a divalent cation acting on nucleation and mineral accumulation, or indirectly, as a co-factor of enzymes involved in the process like alkaline phosphatase (Gomez et al., 1999). In humans, the role of Zn in bone formation involves the activation of bone alkaline phosphatase, osteoblast tyrosine kinase and RNA synthetase (Yamaguchi and Hashizime, 1994). An in vitro study in rats demonstrated that Zn increased the protein components in the femoral-diaphyseal and metaphyseal tissues, and enhanced bone growth in collaboration with insulin-like growth factor I (Ma and Yamaguchi, 2001). On the other hand, the higher level of Cu in the liver in this study may also indicate a

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higher tissue retention of this element, with the amino acid-chelate showing a higher retention than the sulfate group, as previously observed in ruminants (Mills et al., 1985). This indicates that Cu ingested from chelates is highly retained by this tissue. Likewise, whole body trace element content clearly demonstrated the beneficial effect of chelate over the inorganic source. This result agrees with our previous finding that fish fed amino acid-chelated Mn had a significantly higher whole body Mn content compared to the group supplemented with MnSO4 (Satoh et al., 2001). A similar study with rainbow trout also reported a higher whole body Zn level in fish fed diets supplemented with Zn amino acid-chelate than those on ZnSO4 (Hardy et al., 1987). Higher availability of an element from a particular source could result in reduction in the amount needed to meet the dietary requirement of the animal for such element. For example, the requirement of channel catfish for maximum bone Zn deposition was determined to be lower with ZnMet compared with ZnS (Paripatananont and Lovell, 1995). In a corresponding observation on shellfish, Tan and Mai (2001) reported that juvenile abalone require Zn at only 16 to 18 mg Zn kg 1 diet from ZnMet while 35 mg kg 1 is required from ZnSO4. The higher activity of the Zn-requiring enzyme, alkaline phosphatase (ALP), in the chelate compared to the sulfate fed group in this study lends further support to our previous results that utilization of the element from amino acid-chelate are better than from inorganic source (Apines et al., 2001). Inadequate amount of Zn to activate the enzyme could impair the synthesis of mRNA needed for the translation of new proteins and may affect bone formation (Kfoury et al., 1968; Reinhold and Kfoury, 1969; Rose et al., 1978; Yamaguchi and Hashizime, 1994; Ma and Yamaguchi, 2001). Several studies indicate that ALP also influences the skeletal mineralization of aquatic organisms (Donachy et al., 1990; Olsen et al., 1991; Blasco et al., 1993). Therefore, as in other animals (Kfoury et al., 1968; Huber and Gershoff, 1973), ALP activity in fish can also be a good indicator of their Zn status. Numerous studies have shown that CuZnSOD, an antioxidant enzyme, is a sensitive marker of Cu status (Gatlin and Wilson, 1986; Harris, 1992; Hari et al., 1998). CuZnSOD is a product of the SOD1 gene and is largely cytosolic but based on studies in yeast, it seems to protect both the cytosolic and mitochondrial components from oxidative damage (Gralla and Kosman, 1992; Sturtz et al., 2001). It has been demonstrated that Zn did not cause any significant alteration in the activities of antioxidant enzymes such as SOD (Pathak et al., 2002). However, Cu deficiency can destabilize the enzyme structure and decrease its activity resulting in an increased rate of lipid peroxidation due to reactive oxygen species (ROS), excess of which can as well damage DNA, protein, and nucleic acids (Cerutti, 1985; Stadtman and Levine, 2000; Lee et al., 2001). In the present study, the higher CuZnSOD mRNA expression in the amino acid-chelate compared to the sulfate-supplemented group again indicates a higher availability and better retention of the elements from this group. CuZnSOD mRNA level in the chelated group seemed to be unaffected by the presence of both tricalcium phosphate and phytate in the diet. In humans, alteration in the CuZnSOD mRNA level has been observed to occur concomitantly with enzyme activity (Sugino et al., 2002) implying that the enzyme activity is associated with the mRNA level produced. CuZnSOD has been shown to protect cells against reactive free radicals by scavenging superoxide anions produced under oxidative conditions (McCord and Fridovich, 1969; Keele et al., 1971; Fridovich, 1983, 1995; Sturtz et al., 2001). When

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the enzyme is damaged and begins to release Cu2 +, it could aggravate oxidative damage (Park and Floyd, 1997; Kwon et al., 1998). Being an antioxidant, alteration in the enzyme activity could impair the cell immune function thereby making the animal more susceptible to infection. CuZnSOD is also involved in Zn homeostasis in addition to its antioxidant properties (Wei et al., 2001). DNA polymerase is another enzyme that requires the catalytic function of Zn in the replication and transcription of genetic materials (Endre et al., 1990). The largest subunit of the replication protein A (RPA) contains an evolutionarily conserved Zn finger motif that aids in DNA replication and mismatch repair (Lin et al., 1998). The higher expression of DNA polymerase mRNA in the amino acid-chelate than in the sulfatesupplemented group in this study further indicates a higher absorption of the elements from this source which can support faster growth due to the influence of the enzyme on protein synthesis. Production of the enzyme mRNA in the chelated group was shown to be higher compared to the sulfate-supplemented group even when both the inhibitors were present in the diet. Apart from Zn, Cu has also been found to have a protective role against DNA damage in rats as measured by the activity of some enzymes related to DNA repair such as DNA polymerase beta (Webster et al., 1996). Further, genetic defects mimicking Cu deficiency have been reported in humans and mice (Keen and Graham, 1989) which indicates the importance of the trace elements in the process of gene activation and protein synthesis. The higher retention of the elements in the amino acid-chelate compared to the sulfate-supplemented group supports our contention that the former is a better source of trace elements than the latter. This again substantiated our previous findings that Zn and Mn retention from amino acid-chelates at any level of supplementation were higher than from their sulfate form (Apines et al., 2001; Satoh et al., 2001). Among the trace elements evaluated, Zn has the highest retention followed by Cu and Mn which has the least retention. The same pattern was observed in our unpublished study which conforms with reports in higher animals that Mn is as efficiently absorbed as it is excreted and thus, only a small amount is retained in the tissues (O’Dell and Campbell, 1971; Wapnir, 1990). As indicated by the results, the use of amino acid-chelated trace elements as feed supplement to rainbow trout provides more advantages than the use of inorganic source of the elements tested. The amino acid-chelate showed a higher availability than the inorganic compound even in the presence of both tricalcium phosphate and phytate which are normal components of plant protein sources in practical fish diets. Further studies in this area are warranted.

Acknowledgements We wish to thank Christopher Marlowe A. Caipang for his technical inputs in the molecular analysis. The Fuji Trout Hatchery of the Shizuoka Prefectural Fisheries Experimental Station is also gratefully acknowledged for providing the experimental fish. This study was supported in part by the Ministry of Education, Culture, Sports, Science, and Technology of Japan (MONBUKAGAKUSHO).

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References Apines, M.J., Satoh, S., Kiron, V., Watanabe, T., Nasu, N., Fujita, S., 2001. Bioavailability of amino acids chelated and glass embedded zinc to rainbow trout, Oncorhynchus mykiss, fingerlings. Aquac. Nutr. 7, 221 – 228. Ashmead, H.D., 1992. The Roles of Amino Acids Chelates in Animal Nutrition. Noyes Publications, Park Ridge, NJ. 479 pp. Blasco, J., Puppo, J., Sarasquete, M.C., 1993. Acid and alkaline phosphatase activities in the clam Ruditapes philippinarum. Mar. Biol. 115, 113 – 118. Cerutti, P.A., 1985. Prooxidant states and tumor promotion. Science 227, 375 – 380. Donachy, J.E., Watabe, N., Showman, R.M., 1990. Alkaline phosphatase and carbonic anhydrase activity associated with arm generation in the seastar Asterias forbesi. Mar. Biol. 105, 471 – 476. Endre, L., Beck, F.W.J., Prasad, A.S., 1990. The role of zinc in human health. J. Trace Elem. Exp. Med. 3, 337 – 375. Francis, G., Makkar, H.P.S., Becker, K., 2001. Antinutritional factors present in plant-derived alternate fish feed ingredients and their effects in fish. Aquaculture 199, 197 – 227. Fridovich, I., 1983. Superoxide radical: an endogenous toxicant. Annu. Rev. Pharmacol. Toxicol. 23, 239 – 257. Fridovich, I., 1995. Superoxide radical and superoxide dismutases. Ann. Rev. Biochem. 64, 97 – 112. Gatlin III, D.M., Phillips, H.F., 1989. Dietary calcium, phytate and zinc interactions in channel catfish. Aquaculture 79, 259 – 266. Gatlin III, D.M., Wilson, R.P., 1983. Dietary zinc requirement of fingerling channel catfish. J. Nutr. 113, 630 – 635. Gatlin III, D.M., Wilson, R.P., 1984. Zinc supplementation of practical channel catfish diets. Aquaculture 41, 31 – 36. Gatlin III, D.M., Wilson, R.P., 1986. Dietary copper requirement of fingerling channel catfish. Aquaculture 54, 277 – 285. Gomez, E.F., Kaushik, S.J., 1993. Effect of replacement of dietary inorganic zinc by zinc methionine on vegetable and animal protein utilization by rainbow trout. In: Kaushik, S.J., Luquet, P.J. (Eds.), Fish Nutrition in Practice: Les Colloques, vol. 61. INRA, Paris, France, pp. 897 – 902. Gomez, S., Rizzo, R., Pozzi-Mucelli, M., Bonucci, E., Vittur, F., 1999. Zinc mapping in bone tissues by histochemistry and synchroton radiation-induced x-ray emission: correlation with the distribution of alkaline phosphatase. Bone 25, 33 – 38. Goto, T., Komai, M., Suzuki, H., Furukawa, Y., 2001. Long-term zinc deficiency decreases taste sensitivity in rats. J. Nutr. 131, 305 – 310. Gralla, E.B., Kosman, D.J., 1992. Molecular genetics of superoxide dismutases in yeasts and related fungi. Adv. Genet. 30, 251 – 319. Hardy, R.W., Shearer, K.D., 1985. Effect of dietary calcium phosphate and zinc supplementation on whole body zinc concentration of rainbow trout (Salmo gairdneri). Can. J. Fish. Aquat. Sci. 42, 181 – 184. Hardy, R.W., Sullivan, C.V., Koziol, A.M., 1987. Absorption, body distribution, and excretion of dietary zinc by rainbow trout (Salmo gairdneri). Fish Physiol. Biochem. 3, 133 – 143. Hari, R., Burde, V., Arking, R., 1998. Immunological confirmation of elevated levels of CuZn superoxide dismutase protein in an artificially selected long-lived strain of Drosophila melanogaster. Exp. Gerontol. 33, 227 – 237. Harris, E.D., 1992. Copper as a cofactor and regulator of copper, zinc superoxide dismutase. J. Nutr. 122, 636 – 640. Hill, D.A., Peo Jr., E.R., Lewis, A.J., Crenshaw, J.D., 1986. Zinc – amino acid complexes for swine. J. Anim. Sci. 63, 121 – 130. Huber, A.M., Gershoff, S.N., 1973. Effects of dietary zinc on zinc enzymes in the rat. J. Nutr. 103, 1175 – 1181. Keele Jr., B.B., McCord, J.M., Fridovich, I., 1971. Further characterization of bovine superoxide dismutase and its isolation from bovine heart. J. Biol. Chem. 246, 2875 – 2880. Keen, C.L., Graham, T.W., 1989. Trace elements. In: Kaneko, J.J. (Ed.), Clinical Biochemistry of Domestic Animals, 4th ed. Academic Press, San Diego, CA, pp. 753 – 795.

M.J.S. Apines et al. / Aquaculture 225 (2003) 431–444

443

Kfoury, G.A., Reinhold, J.G., Simonian, S.J., 1968. Enzyme activities in tissues of zinc-deficient rats. J. Nutr. 95, 102 – 110. Kitagoh, H., Tomita, H., Ikui, A., Ikeda, M., 2002. Course of recovery from taste receptor disturbance. Acta Otolaryngol., Suppl. 546, 83 – 93. Komai, M., Goto, T., Suzuki, H., Takeda, T., Furukawa, Y., 2000. Zinc deficiency and taste dysfunction; contribution of carbonic anhydrase, a zinc – metalloenzyme, to normal taste sensation. BioFactors 12, 65 – 70. Kwon, O.J., Lee, S.M., Floyd, R.A., Park, J.W., 1998. Thiol-dependent metal-catalyzed oxidation of copper, zinc superoxide dismutase. Biochim. Biophys. Acta 1387, 249 – 256. Lee, J.H., Choi, I.Y., Kil, I.S., Kim, S.Y., Yang, E.S., Park, J.W., 2001. Protective role of superoxide dismutases against ionizing radiation in yeast. Biochim. Biophys. Acta 1526, 191 – 198. Lin, Y.L., Shivji, M.K.K., Chen, C., Kolodner, R., Wood, R.D., Dutta, A., 1998. The evolutionarily conserved zinc finger motif in the largest subunit of human replication protein A is required for DNA replication and mismatch repair but not for nucleotide excision repair. J. Biol. Chem. 273, 1453 – 1461. Linder, M.C., Wooten, L., Cerveza, P., Cotton, S., Shulze, R., Lomeli, N., 1998. Copper transport. Am. J. Clin. Nutr. 67S, 965 – 971. Lovell, R.T., 1977. Mineral requirements. In: Strickney, R.R., Lovell, R.T. (Eds.), Nutrition and Feeding of Channel Catfish. Southern Cooperative Series Bulletin, vol. 218. Auburn University, Auburn, AL, pp. 30 – 32. Lovell, R.T., 1989. Nutrition and Feeding of Fish. Van Nostrand-Reinhold, New York. Ma, Z.J., Yamaguchi, M., 2001. Stimulatory effect of zinc and growth factor on bone protein component in newborn rats: enhancement with zinc and insulin-like growth factor I. Int. J. Mol. Med. 7, 73 – 78. Maage, A., Julshamn, K., Berge, G.E., 2001. Zinc gluconate and zinc sulphate as dietary zinc sources for Atlantic salmon. Aquac. Nutr. 7, 183 – 187. McCord, J.M., Fridovich, I., 1969. Superoxide dismutase: an enzymic function for erythrocuprein (hemocuprein). J. Biol. Chem. 244, 6049 – 6055. Mills, C.F., Bremner, I., Chester, J.K., 1985. Trace Elements in Man and Animals. Commonwealth Agricultural Bureaux, Farnham Royal, UK. 339 pp. Mohanna, C., Nys, Y., 1999. Effect of dietary zinc content and sources on the growth, body zinc deposition and retention, zinc excretion and immune response in chicken. Br. Poult. Sci. 40, 108 – 114. O’Dell, B.L., Campbell, B.J., 1971. Trace elements: metabolism and metabolic function. In: Florkin, M., Stotz, E.H. (Eds.), Metabolism of Vitamins and Trace Elements. Comp. Biochem., vol. 21, pp. 179 – 267. Olsen, R.L., Overbo, K., Myrnes, B., 1991. Alkaline phosphatase from the hepatopancreas of shrimp (Pandalus borealis): a dimeric enzyme with catalytically active subunits. Comp. Biochem. Physiol. 99B, 755 – 761. Paripatananont, T., Lovell, R.T., 1995. Chelated zinc reduces the dietary zinc requirement of channel catfish, Ictalurus punctatus. Aquaculture 133, 73 – 82. Park, J.W., Floyd, R.A., 1997. Glutathione/Fe3 +/O2-mediated DNA strand breaks and 8-hydroxydeoxyguanosine formation: enhancement by copper, zinc superoxide dismutase. Biochim. Biophys. Acta 1336, 263 – 268. Pathak, A., Mahmood, A., Pathak, R., Dhawan, D., 2002. Effect of zinc on hepatic lipid peroxidation and antioxidative enzymes in ethanol-fed rats. J. Appl. Toxicol. 22, 207 – 210. Peretz, A., Papadopoulos, T., Willems, D., Hotimsky, A., Michiels, N., Siderova, V., Bergmann, P., Neve, J., 2001. Zinc supplementation increases bone alkaline phosphatase in healthy men. J. Trace Elem. Med. Biol. 15, 175 – 178. Reinhold, J.G., Kfoury, G.A., 1969. Zinc-dependent enzymes in zinc-depleted rats; intestinal alkaline phosphatase. Am. J. Clin. Nutr. 22, 1250 – 1263. Richardson, N.L., Higgs, D.A., Beames, R.M., McBride, J.R., 1985. Influence of dietary calcium, phosphorus, zinc and sodium phytate level on cataract incidence, growth and histopathology in juvenile chinook salmon (Oncorhynchus tshawytscha). J. Nutr. 115, 553 – 567. Rose, K.M., Allen, M.S., Crawford, I.L., Jacob, S.T., 1978. Functional role of zinc in poly(A)synthesis catalysed by nuclear poly(A)polymerase. Eur. J. Biochem. 88, 29 – 36. Roth, H.P., Kirchgessner, M., 1983. Effect of different concentrations of various zinc complexes (picolinate, citrate, 8-hydroxyquinolate) in comparison with sulfate on zinc supply status in rats. Z. Ernahr.wiss. 22, 34 – 44 (in German with English abstract).

444

M.J.S. Apines et al. / Aquaculture 225 (2003) 431–444

Satoh, S., Izume, K., Takeuchi, K., Watanabe, T., 1987a. Availability to rainbow trout of zinc contained in various types of fish meals. Nippon Suisan Gakkaishi 53, 1861 – 1866. Satoh, S., Tabata, K., Izume, K., Takeuchi, K., Watanabe, T., 1987b. Effect of dietary tri-calcium phosphate on availability of zinc to rainbow trout. Nippon Suisan Gakkaishi 53, 1199 – 1205. Satoh, S., Takeuchi, K., Watanabe, T., 1987c. Availability to rainbow trout of zinc in white fish meal and of various zinc compounds. Nippon Suisan Gakkaishi 53, 595 – 599. Satoh, S., Poe, W.E., Wilson, R.P., 1989. Effect of supplemental phytate and/or tricalcium phosphate on weight gain, feed efficiency and zinc content in vertebrae of channel catfish. Aquaculture 80, 155 – 161. Satoh, S., Apines, M.J., Tsukioka, T., Kiron, V., Watanabe, T., Fujita, S., 2001. Bioavailability of amino acids chelated and glass embedded manganese to rainbow trout, Onchorhynchus mykiss, fingerlings. Aquac. Res. 32S, 18 – 25. Snedeker, S.M., Greger, J.L., 1983. Metabolism of zinc, copper and iron as affected by dietary protein, cysteine and histidine. J. Nutr. 113, 644 – 652. Spears, J.W., 1989. Zinc methionine for ruminants: relative bioavailability of zinc in lambs and effects of growth and performance of growing heifers. J. Anim. Sci. 67, 835 – 843. Stadtman, E.R., Levine, R.L., 2000. Protein oxidation. Ann. N. Y. Acad. Sci. 899, 191 – 208. Sturtz, L.A., Diekert, K., Jensen, L.T., Lill, R., Culotta, V.C., 2001. A fraction of yeast Cu,Zn-Superoxide dismutase and its metallochaperone, CCS, localize to the intermembrane space of mitochondria: a physiological role for SOD1 in guarding against mitochondrial oxidative damage. J. Biol. Chem. 276, 38084 – 38089. Sugino, N., Karube-Harada, A., Kashida, S., Takiguchi, S., Kato, H., 2002. Differential regulation of copper – zinc superoxide dismutase and manganese superoxide dismutase by progesterone withdrawal in human endometrial stromal cells. Mol. Hum. Reprod. 8, 68 – 74. Swinkels, J.W., Kornegay, E.T., Zhou, W., Lindemann, M.D., Webb Jr., K.E., Verstegen, M.W., 1996. Effectiveness of a zinc amino acid chelate and zinc sulfate in restoring serum and soft tissue zinc concentrations when fed to zinc-depleted pigs. J. Anim. Sci. 74, 2420 – 2430. Tan, B., Mai, K., 2001. Zinc methionine and zinc sulfate as sources of dietary zinc for juvenile abalone, Haliotis discus hannai Ino. Aquaculture 192, 67 – 84. Turnlund, J.R., 1998. Human whole-body copper metabolism. Am. J. Clin. Nutr. 67S, 960 – 964. Wapnir, R.A., 1990. Protein Nutrition and Mineral Absorption. CRC Press, Boca Raton, FL. Webster, R.P., Gawde, M.D., Bhattacharya, R.K., 1996. Modulation by dietary copper of aflatoxin B1-induced activity of DNA repair enzymes poly (ADP-ribose) polymerase, DNA polymerase beta and DNA ligase. In Vivo 10, 533 – 536. Wedekind, K.J., Baker, D.H., 1989. Zinc bioavailability in feed-grade zinc sources. J. Anim. Sci. 67S, 126. Wedekind, K.J., Hortin, A.E., Baker, D.H., 1992. Methodology for assessing zinc bioavailability: efficacy estimates for zinc – methionine, zinc sulfate, and zinc oxide. J. Anim. Sci. 70, 178 – 187. Wedekind, K.J., Lewis, A.J., Giesemann, M.A., Miller, P.S., 1994. Bioavailability of zinc from inorganic and organic sources for pigs fed corn – soybean meal diets. J. Anim. Sci. 72, 2681 – 2689. Wei, J.P.J., Srinivasan, C., Han, H., Valentine, J.S., Gralla, E.B., 2001. Evidence for a novel role of copper – zinc superoxide dismutase in zinc metabolism. J. Biol. Chem. 276, 44798 – 44803. Yamaguchi, M., Hashizime, M., 1994. Effect of beta-alanyl-L-histidinato zinc on protein components in osteoblastic MC3T3-E1 cells: increase in osteocalcin, insulin-like growth factor-I and transforming growth factorbeta. Mol. Cell. Biochem. 136, 163 – 169.