5. DISORDERS IN VITAMIN NUTRITION

5.1 Dietary vitamin deficiency

Table 8 shows the major gross anatomical deficiency signs which have been reported in fish fed vitamin deficient diets

1- Mclaren et al. (1947),
2-Phillips & Brockway (1957),
3-Halver (1957),
4-Kitamura et al.(1967),
5-Poston et al. 1977,
6-Takeuchi et al. (1980),
7-Hughes, Rumsey & Nickum(1981),
8-Woodward (1982),
9-Woodward (1985),
10-Amezaga & Knox (1990),
11-Aoe etal. (1967),
12-Ogino (1967),
13-Dupree (1966),
14-Murai & Andrews (1978a),
15-Yone(1975),
16-Arai, Nose & Hashimoto (1972),
17-Butthep, Sitasit & Boonyaratpalin (1985),
18-Boonyaratpalin & Wanakowat (1991),
19-Lim, Leamaster & Brock (1991)

¹ 1-McLaren et al. (1947),
2-Phillips & Brockway (1957),
3-Halver (1957),
4-Kitamura et al.(1967),
5-Coates & Halver (1958),
6-Masumoto, Hardy & Stickney (1991),
7-Ogino(1967),
8-Dupree (1966),
9-Murai & Andrews (1979),
10-Wilson, Bowser & Poe (1983),
11-Yone (1975),
12-Yano et al. (1988),
13-Arai, Nose & Hashimoto (1972),
14-Butthep,Sitasit & Boonyaratpalin (1985),
15-Chavez de Martinez, Escobar & Olvera-Novoa (1990),
16-Boonyaratpalin & Wanakowat (1991)

¹ 1-McLaren et al. (1947),
2-Philips & Brockway, (1957),
3-Halver (1957),
4-Poston &Di Lorenzo (1973),
5-Poston & Wolfe (1985),
6-Aoe et al. (1967),
7-Dupree (1966),
8-Andrews & Murai (1978),
9-Yone (1975),
10-Arai, Nose & Hashimoto (1972),
11-Butthep, Sitasit & Boonyaratpalin (1985),
12-Kitamura et al. (1967),
13-Coates &Halver (1958),
14-Walton et al. (1984),
15-Poston & McCartney (1974),
16-Poston(1976),
17-Castledine et al. (1978),
18-Poston & Page (1982),
19-Ogino et al.(1970),
20-Gunther & Meyer-Burgdorff (1990),
21-Robinson & Lovell (1978),
22-Lovell & Buston (1984)

1 1-McLaren et al. (1947),
2-Phillips & Brockway (1957),
3-Halver (1957),
4-Kitamura et al.(1967),
5-Coates & Halver (1958),
6-Aoe et al. (1969),
7-Dupree (1966),
8-Murai &Andrews (1978),
9-Yone (1975),
10-Arai, Nose & Hashimoto (1972),
11-Hashimoto, Arai& Nose (1970),
12-Lim, Leamaster & Brock (1991),
13-Boonyaratpalin & Wanakowat(1991),
14-Duncan & Lovell (1991),
15-John & Mahajan (1979),
16-Butthep, Sitasit &Boonyaratpalin (1985)

¹ 1-McLaren et al. (1947),
2-Phillips & Brockway (1957),
3-Halver (1957),
4-Kitamura et al.(1967),
5-Coates & Halver (1958),
6-Jurss (1978),
7-Hardy, Halver & Brannon (1979),
8-Jurss& Jonas (1981),
9-Smith, Brin & Halver (1974),
10-Herman (1985),
11-Ogino (1965),
12-Dupree (1966),
13-Andrews & Murai (1979),
14-Yone (1975),
15-Arai, Nose & Hashimoto(1972),
16-Adron, Knox & Cowey (1978),
17-Kissil et al. (1981),
18-Sakaguchi, Takeda &Tange (1969),
19-Agrawal & Mahajan (1983),
20-Butthep, Sitasit & Boonyaratpalin (1985),
21-Wanakowat et al. (1989)

¹1-McLaren et al. (1947),
2-Kitamura et al. (1965),
3-Hilton, Cho & Slinger (1978),
4-Sato,Yoshinaka & Ikeda (1978),
5-Poston (1967),
6-Halver, Ashley & Smith (1969),
7-Sandnes et al.(1984),
8-Navarre & Halver (1989),
9-Lall et al. (1989),
10-Sato, Hatano & Yoshinaka (1991),
11-Dabrowski et al. (1990),
12-Cho & Cowey (1991),
13-Lovell (1973),
14-Andrews & Murai (1974),
15-Lovell & Lim (1978),
16-Wilson & Poe (1973),
17-Lim & Lovell (1978),
18-Li & Lovell (1985),
19-Mazik, Brandt & Tomasso (1987),
20-Lovell & Naggar (1989),
21-Wilson, Poe & Robinson(1989),
22-Yone (1975),
23-Arai, Nose & Hashimoto (1972),
24-Mahajan & Agrawal (1979),
25/26-Soliman, Jauncey & Roberts (1986, 1986a),
27-Butthep, Sitasit & Boonyaratpalin (1985),
28-Agrawal & Mahajan (1980),
29-Baudin Laurencin, Messager & Stephan (1989),
30-Coustans etal. (1990),
31-Gouillou, Coustans & Guillaume (1991),
32-Rosenlund et al. (1990),
33-Boonyaratpalin, Unprasert & Buranapani it (1989),
34-Paperna (1987),
35-Chavez de Martinez(1990),
36-Gallet de Saint Aurin, Raymond & Vianas (1989),
37-Messager et al. (1986),
38-Kanazawa et al. (1992),
39-Boonyaratpalin, Boonyaratpalin & Supamataya (1992)

¹ 1-Halver (1957),
2-Philips et al. (1963),
3-Dupree (1966),
4-Limsuwan & Lovell (1981),
5-Arai,Nose & Hashimoto (1972),
6-Yone (1975),
7-John & Mahajan (1979),
8-McLaren et al. (1947),
9-Phillips & Brockway (1957),
10-Coates & Halver (1958),
11-Aoe & Masuda (1967)

¹ 1-McLaren et al. (1947),
2-Phillips & Brockway (1957),
3-Halver (1957),
4-Kitamura et al.(1967),
5-Coates & Halver (1958),
6-Ketola (1976),
7-Poston (1990),
8-Rumsey (1991),
9-Ogino et al. (1970a),
10-Dupree (1966),
11-Wilson & Poe (1988),
12-Arai, Nose &Hashimoto (1972),
13-Yone (1975),
14-Yano et al. (1988),
15-Rumsey (1991),
16-Postonet al. (1977),
17-Aoe et al. (1968),
18-Shim & Tan (1989),
19-Barnett, Cho & Slinger(1979),
20-Leatherland et al. (1980),
21-Shiau & Hwang (1992),
22-Lovell & Li (1978),
23-Andrews, Murai & Page (1980),
24-Brown (1988)

¹ 1-Woodall et al. (1964),
2-Poston (1965),
3-Poston, Combs & Leibovitz (1976),
4-Cowey et al. (1984),
5-Ndoye et al. (1989),
6-Hardie, Fletcher & Secombes (1991),
7-Watanabe et al. (1970),
8-Watanabe & Takashima (1977),
9-Dupree (1966),
10-Murai &Andrews (1974),
11-Lovell, Miyazaki & Rabegnator (1984),
12-Wilson, Browser & Poe(1984),
13-Satoh, Takeuchi & Watanabe, (1987),
14-Roem, Stickney & Kohler (1990),
15-Poston (1964),
16-Poston (1976a)

Under intensive culture conditions, and in the absence of natural food organisms, dietary vitamin deficiencies may arise through one or more of the following:

Feed processing and storage

1. Riboflavin: Slightly soluble in water and used in the form of a free flowing powder, riboflavin is generally stable in dry multivitamin premixes during extended storage and when mixed with mineral premixes and other feed ingredients. Processing losses of 26% have been reported for expanded pet foods (NRC, 1983). However, feeds containing riboflavin should be protected from intensive light/ultraviolet radiation (liable to oxidation) and alkaline conditions.

2. Pantothenic acid: Readily soluble in water and used in the form of calcium d-pantothenate (92% activity); solubility of ca. 40g/100ml) or calcium dl-pantothenate (46% activity), pantothenic acid is fairly stable in air and light if protected from humidity, but is sensitive to heat. Processing losses during pelleting or expansion have been reported to be as high as 10% within manufactured fish feeds (Slinger, Razzaque & Cho, 1979).

3. Niacin: Used in dried form as nicotinic acid or niacinamide, niacine is generally stable in dry multivitamin premixes and in the presence of air, heat and minerals. Processing losses of 20% have been reported for expanded pet foods (NRC, 1983).

4. Thiamine: Hydrochloride salt is freely soluble in water (ca. 1g/ml) whereas mononitrate salt is only sparingly soluble (ca. 2.7g/100ml). Thiamine hydrochloride is relatively stable to air if protected from light and humidity. Thiamine mononitrate is fairly stable to air if protected from light and is less sensitive to humidity than thiamine hydrochloride. Thiamine is usually stable in dry multivitamin premixes that contain no added choline or trace elements but is rapidly destroyed under alkaline conditions or in the presence of sulphide. Processing (pelleting/expansion) and storage (7 months, room temperature) losses of manufactured fish feeds have been reported to be 0–10% and 11– 12%, respectively (Slinger, Razzaque & Cho, 1979).

5. Pyridoxine: Readily soluble in water (ca. 20g/100ml) and used in dried form as pyridoxine hydrochloride. Pyridoxine is fairly stable to air and heat if protected from light and humidity, and is stable in dry multivitamin premixes that contain no added trace elements. Processing and storage (10 months) losses in manufactured fish feeds have been reported to be 7–10% (Slinger, Razzaque & Cho, 1979).

6. Biotin: Soluble in water and used in the form of d-biotin in dry dilution, biotin is fairly stable to air and heat in dry multivitamin premixes but sensitive to light and high humidity. Processing losses in expanded feeds have been reported as 10% (NRC, 1983).

7. (vii) Folic acid: Slightly soluble in water and used in dry-dilution, folic acid is fairly stable to air but is sensitive to heat and in particular to light and ultraviolet radiation. Processing and storage losses within manufactured fish feeds have been reported to be 3–10% (Slinger, Razzaque & Cho, 1979). There is some evidence to suggest that folate degrading bacteria are responsible for the destruction of folic within stored catfish feeds and that their actions are related to the occurrence of a nutritionally related anaemia in culture channel catfish (Plumb, Liu & Butterworth, 1991).

8. Vitamin B12: Moderately soluble in water and used in dry-dilution, vitamin b₁₂ stability in multivitamin premixes is good at normal storage temperatures; elevated temperatures reducing activity, particularly under mild acid conditions.

9. Choline: Used in liquid (70% activity) or dried form (25–60% activity), choline chloride is stable in multivitamin premixes but can decrease the stability of other vitamins present, and therefore should be added separately. Relatively stable on processing and storage (NRC, 1983).

10. Vitamin C: Crystalline L-ascorbic acid (AA) is readily oxidized and destroyed in the presence of oxygen, moisture, trace elements, elevated temperatures, light and oxidized lipids. It follows therefore, that considerable losses of vitamin C can occur during feed manufacture and on prolonged feed storage. For example, processing and storage losses for practical fish feeds have been reported to be as high as 95% for L-ascorbic acid (Slinger, Razzaque & Cho, 1979; Sandnes and Utne, 1982; Soliman, Jauncey & Roberts, 1987). However, the oxidation of vitamin C can be reduced by utilizing coated or ‘protected’ forms of L-ascorbic acid (ie. ethyl cellulose, silicone, gelatin, glyceride or synthetic-polymer coated ascorbic acid) or by using more stable biologically equivalent derivatives such as ascorbate-2-monophosphate (AMP) and ascorbate-2-polyphosphate (APP; Halver et al., 1975; Hilton, Cho & Slinger, 1977; Soliman, Jauncey & Roberts, 1987; Shigueno & Itoh, 1988; Grant et al, 1989; Lovell & Naggar, 1989; Skelbaek et al, 1990; Maugle, Brown & Hoffman, 1991; Halver, Felton & Palmisano, 1991; Sandnes, 1991). For example, losses of ascorbate activity from catfish diets containing AA or ethyl cellulose (EC) coated AA was reported to be 23–34% (AA) and 10–24% (ECAA) after conventional steam pelleting and 55–69% (AA) and 40–55% (ECAA) after expansion pelleting, respectively (Lovell & Lim, 1978). Although ECAA is generally regarded as being more stable than AA, Sandnes and Utne (1982) reported a 70–80% loss of ascorbate activity with ECAA supplemented diets after steam pelleting and storage at 4°C for 24 weeks, while practically no activity could be detected after 16 weeks when the feed was stored at room temperature. By contrast, a 80–100% ascorbate recovery has been reported for APP within expansion pelleted catfish feeds (Lovell & Naggar, 1989); the stability of APP within feeds at 25 or 40°C being 83 or 45 times greater than that of AA, respectively (Grant et al, 1989).

11. Vitamin A: Normally used as the acetate, palmitate or propionate ester, in powdered or stabilized beadlet form. Although vitamin A is insoluble in water and stable within dry multivitamin premixes, it is readily oxidised at elevated storage temperatures and in the presence of oxidation products (rancid oils). Processing and storage losses of 20% and 53% have been reported for expanded pet foods and after six months storage at room temperature, respectively (NRC, 1983).

12. Vitamin D₃: Insoluble in water, vitamin D₃ is usually added in a protected beadlet form with vitamin A or as a dried powder. Stability is generally high within multivitamin premixes, during feed processing, and on storage.

13. Vitamin K₃: Used in the form of the water soluble synthetic salt as menadione sodium bisulphite, the stability of vitamin K₃ in multivitamin premixes is good if protected from trace elements, heat, moisture and light (NRC, 1983).

14. Vitamin E: Insoluble in water and used in the form of dl-alpha-tocopherol acetate, either spray dried or absorbed, vitamin E is moderately stable in dry multivitamin premixes if stored below room temperature. However, vitamin E is prone to oxidation on storage in the presence of oxidation products such as rancid oils and at high ambient temperatures.

15. Inositol: Myo-inositol is fairly stable within multivitamin premixes and to normal manufacturing and storage conditions.

For a review of the effects of processing and storage on vitamin stability see Coelho (1991) and Table 9.

1 M - Moisture,
2 O - Oxidation,
3 R - Reduction,
4 - Trace minerals,
5 H - Heat
6 L - Light,
7 A - Acid Ph,
8 N - Neutral pH,
9 B - Basic pH
10 R - Resistant, MS - Mildly Sensitive, S - Sensitive, VS - Very Sensitive
11 MSBP - Menadione Sodium Bisulphite Complex, MPB - Menadione Dimethyl Pyrimidinolbisulphite

Leaching of water soluble vitamins

In contrast to the fat soluble vitamins (A, D, E and K) the water soluble vitamins can be readily lost from the feed through leaching prior to ingestion by the fish. In general, the smaller the feed particle size and the longer the feed remains uneaten in water, the greater the loss of water soluble nutrients.

L-ascorbic acid (vitamin C) has been found to be particularly prone to loss through leaching. For example, despite the excessive losses of vitamin C which occur during feed preparation and storage (90–95% loss; Slinger, Razzaque & Cho, 1979), up to 50–70% of the residual vitamin C activity was found to be lost through leaching after a 10 second immersion period in water (1.18–2.36mm diameter pellet). In the same study, the authors also reported a 5–20% loss in pantothenic acid, 0–27% loss in folic acid, 0–17% loss in thiamine and a 3–13% loss in pyridoxine activity through leaching, after a 10 second water immersion period. Murai and Andrews (1975) reported a 50% loss in pantothenic acid after a 10 second immersion of a trout pellet originally containing 500 mg/kg pantothenic acid. Similarly, water stability tests with complete pelleted diets for shrimp (Penaeus japonicus) reported dietary vitamin losses through leaching of thiamine - 98% (initial 29.5 mg/kg), pantothenic acid - 94% (initial 100 mg/kg, as calcium salt), pyridoxine - 93% (initial 14 mg/kg), ascorbic acid - 89% (initial 3089 mg/kg), riboflavin - 86% (initial 55 mg/kg), nicotinic acid - 86% (initial 120 mg/kg), inositol - 52% (initial 4000 mg/kg) and choline - 45% (initial 3368 mg/kg, as chlorohydrate) after a one hour immersion period in seawater (Cuzon, Hew & Cognie, 1982).

Deficiencies due to the presence of dietary anti-vitamin factors

1. An anti-biotin factor (avidin) is present in raw egg white, but is readily destroyed by heat.

2. Anti-thiamine factor (thiaminase enzyme) present in certain raw fish, shellfish, rice polishings, Indian mustard seed, mung bean (green gram), and linseed (Liener, 1980). The effect of thiaminase may be overcome by heat processing the raw material so as to deactivate the enzyme, or by using supplemental dibenzoylthiamine (DBT) as a thiaminase resistant form of dietary thiamine.

3. Anti-vitamin A,E,D and B₁₂ factors present in raw soybean. May be deactivated by heat treatment (Liener, 1980).

4. Anti-pyridoxine factor present in linseed; treatment as above.

Deficiencies due to dietary antibiotic addition

The use of feed antibiotics for disease treatment may reduce the vitamin synthesising capacity of the gut micro-flora of fish, which in herbivorous/omnivorous fish species is thought to significantly contribute toward the vitamin requirements of the fish (carp, tilapia, channel catfish - vitamin B₁₂, folic acid and possibly biotin, thiamine and vitamin K; Lovell & Limsuwan, 1982; Lovell & Buston, 1984; Sugita, Miyajima & Deguchi, 1990).

Vitamins - stress and immunocompetence

There is some evidence to suggest that the dietary vitamin requirement of fish may be higher under ‘stressed’ or adverse environmental conditions (ie. vitamin C-Lovell & Lim, 1978; Hilton, 1989; Sandnes, 1991) and for increased immunocompetence and disease resistance (ie. vitamins C, E and possibly vitamin B₆ , pantothenic acid and choline - Hardie, Fletcher & Secombes, 1991; Ndoye et al., 1989; Blazer, 1991; Albrektsen et al, Lall, 1991; Landolt, 1989;Yano et al, 1988; Navarre & Halver, 1989; Waagbo et al, 1991; Verlhac et al, 1991; Sandnes, Rosenlund & Waagbo, 1991). However, considerable further work is required in this newly emerging field before these results (if reconfirmed under practical farming conditions) can be applied to the commercial fish farming community.

5.2 Dietary vitamin toxicity

In contrast to the water soluble vitamins, fish accumulate fat-soluble vitamins (A, D,E & K) under conditions where dietary intakes exceeds metabolic demand. Under certain circumstances accumulation is such that a toxic condition (hypervitaminosis) may be produced. Although such a condition is unlikely to occur under practical farming conditions, hypervitaminosis has been experimentally induced in fish and the reported toxicity signs shown in Table 10.