The following gross anatomical deficiency signs have been reported in fish fed vitamin deficient diets:
|
Vitamin/fish |
Deficiency signs 1/ |
|
RIBOFLAVIN |
|
|
Salmonids |
Anorexia, poor growth, corneal vascularisation, cloudy lens, snout erosion, spinal deformities, increased mortality rate, severe fin erosion, fin haemorrhage, rapid opercular movement, apparent muscular weakness, light or dark pigmentation, striated constrictions of abdominal wall, photophobia, incoordination, lethargy, anaemia (1-9) |
|
C. carpio |
Anorexia, poor growth, high mortality rate, haemorrhage of skin and fins, nervousness, photophobia (12,13) |
|
I. punctatus |
Short body dwarf ism, anorexia, poor growth, cataract (10,11) |
|
P. major |
Poor growth (14) |
|
A. japonica |
Fin haemorrhage, photophobia, poor growth, anorexia, lethargy (15) |
|
PANTOTHENIC ACID |
|
|
Salmonids |
Anorexia, reduced growth, gill necrosis/clubbing, anaemia, mucous covered gills, sluggish, opercules distended (1-4,16) |
|
C. carpio |
Anorexia, reduced growth, sluggishness, anaemia, skin haemorrhage, exopthalmia (13) |
|
I. punctatus |
Anorexia, clubbed gills, eroded skin, lower jaws and head; anaemia (10,17,18) |
|
P. major |
Poor growth, mortality (14) |
|
A. japonica |
Poor growth, abnormal swimming behaviour, skin lesions (15) |
|
NIACIN |
|
|
Salmonids |
Anorexia, poor growth, reduced food conversion efficiency, dark colouration, erratic swimming, muscle spasms while resting, oedema of stomach, susceptibility to sunburn (1-3,19) |
|
C. carpio |
Skin haemorrhage, high mortality (20) |
|
I. punctatus |
Haemorrhage and lesions of skin/fin, deformed jaws, anaemia, exopthalmia, high mortality (10,21) |
|
P. major |
Poor growth (14) |
|
A. japonica |
Haemorrhage and lesions of skin, reduced growth, ataxia (abnormal swimming), dark colouration (15) |
|
THIAMINE |
|
|
Salmonids |
Anorexia, poor growth, nervous disorders, impaired carbohydrate metabolism, increased sensitivity to shock by physical blow to container or from light flashes (1-4,16) |
|
C. carpio |
Fin congestion/haemorrhage, nervousness, fading of body colour, anorexia, poor growth (22) |
|
I. punctatus |
Anorexia, poor growth, dark colouration, mortality (10,23) |
|
P. major |
Anorexia, poor growth (14) |
|
A. japonica |
Anorexia, poor growth, ataxia, trunk winding syndrome, fin haemorrhage (15,24) |
|
PYRIDOXINE |
|
|
Salmonids |
Nervous disorders, hyperirritability, anorexia, indifference to light, rapid onset of rigor mortis, ataxia, oedema of peritoneal cavity, excessive flexing of opercules, erratic and rapid swimming, greenish-blue colouration of skin, anaemia, rapid and gasping breathing (1-4,16,25-28) |
|
C. carpio |
Anorexia, poor growth, nervous disorders (29) |
|
I. punctatus |
Anorexia, nervous disorders, erratic swimming, opercule extension, tetany, blue-green colouration of dorsal surface (10,30) |
|
P. major |
Poor growth (14) |
|
A. japonica |
Anorexia, poor growth, nervous disorders (15) |
|
S. maximus |
Reduced growth (31) |
|
Sparus auratus |
Anorexia, poor growth, high mortality, hyper-irritability, erratic swimming, poor food conversion efficiency (32) |
|
S. quinqueradiata |
Reduced growth (33) |
|
Channa punctata |
Reduced growth, ataxia, hyperirritability, muscular spasms, anorexia, erratic swimming, scale loss, oedema, abnormal pigmentation, lens opacity and blindness (34) |
|
BIOTIN |
|
|
Salmonids |
Anorexia, reduced growth, increased mortality, poor food conversion efficiency, blue-slime disease (brook trout only), lesions in the colon, muscle atrophy, spastic convulsions, thick gill lamellae, pale gills (2-4,16,35-39) |
|
C. carpio |
Reduced growth, reduced activity (40) |
|
I. punctatus |
Depigmentation, anaemia, anorexia, reduced growth, hypersensitivity (41,42) |
|
P. major |
No deficiency signs detected (14) |
|
A. japonica |
Poor growth, dark colouration, abnormal swimming behaviour (15) |
|
FOLIC ACID |
|
|
Salmonids |
Macrocytic normochromic anaemia, poor growth, anorexia, lethargy, dark colouration, pale gills, exopthalmia, distended abdomen with ascites fluid (1-4,16) |
|
A: japonica |
Anorexia, poor growth, dark colouration (15) |
|
C. carpio |
No deficiency signs detected (Aoe et al. 1967a) |
|
P. major |
No deficiency signs detected (14) |
|
Labeo rohita |
Reduced growth and haematocrit (43) |
|
I. punctatus |
Anorexia, increased mortality, lethargy (10) |
|
VITAMIN B12 |
|
|
Salmonids |
Anorexia, reduced growth, microcytic hypochromic anaemia, fragmented erthrocytes, poor food conversion efficiency, dark pigmentation (3,44) |
|
C. carpio |
None detected (45,46) |
|
I. punctatus |
Reduced growth, low haematocrit (10,47) |
|
A. japonica |
Poor growth (15) |
|
P. major |
Poor growth (14) |
|
L. rohita |
Reduced growth, low haematocrit, megaloblastic anaemia (43) |
|
CHOLINE |
|
|
Salmonids |
Reduced growth, fatty liver, poor food conversion efficiency, haemorrhagic kidney and intestine (1-4.16,48) |
|
C. carpio |
Reduced growth, fatty liver (49) |
|
I. punctatus |
Reduced growth, enlarged liver, haemorrhagic kidney and intestine (10) |
|
P. major |
Reduced growth, mortality (14) |
|
A. japonica |
Anorexia, reduced growth, grey-white intestine (15) |
|
INOSITOL |
|
|
Salmonids |
Reduced growth, distended abdomen, dark colour, increased gastric emptying time (1-3,16) |
|
C. carpio |
Reduced growth, skin and fin lesions/haemorrhage, loss of skin mucosa (50) |
|
I. punctatus |
None detected (51) |
|
P. major |
Reduced growth (14) |
|
A. japonica |
Reduced growth, grey-white intestine (15) |
|
ASCORBIC ACID (VITAMIN C) |
|
|
Salmonids |
Reduced growth, impaired collagen formation, scoliosis, lordosis, internal haemorrhage, fin haemorrhage, distorted and twisted gill filaments, low haematocrit, poor wound repair, increased mortality rate (1,52-56) |
|
I. punctatus |
Reduced growth, scoliosis, lordosis, increased disease susceptibility, reduced bone collagen level, broken back syndrome, internal and external haemorrhage, fin erosion, dark skin colour, anorexia (57-62) |
|
P. major |
Reduced growth (14) |
|
A. japonica |
Reduced growth, fin/head erosion, lower jaw erosion (15) |
|
C. punctatus |
Scoliosis, lordosis, anaemia, distorted gill filaments (63) |
|
O. niloticus |
Scoliosis, lordosis, reduced growth and wound repair, internal and external haemorrhage (64) |
|
Cirrhina mrigala |
Reduced growth, increased mortality, scoliosis, lordosis, hypochromic macrocytic anaemia (65) |
|
VITAMIN A |
|
|
Salmonids |
Reduced growth, exopthalmia, depigmentation, clouding and thickening of corneal epithelium, degeneration of retina (4,16,66) |
|
C. carpio |
Anorexia, faded body colour, fin and skin haemorrhage, exopthalmia, abnormal/warped gill operculae (67) |
|
I. punctatus |
Depigmentation, opaque and protruding eyes (exopthalmia), atrophy, oedema, kidney haemorrhage, increased mortality (10) |
|
VITAMIN D |
|
|
Salmonids |
Reduced growth and food conversion efficiency, anorexia, tetany, increased muscle/liver lipid content, increased plasma T3 levels (68-69) |
|
I. punctatus |
Reduced growth, low body ash, P and Ca content (70-71) |
|
VITAMIN K |
|
|
Salmonids |
Increased blood clotting time, anaemia, haemorrhagic gills, eyes, vascular tissue (44,72,73) |
|
I. punctatus |
Skin haemorrhage (10,74) |
|
VITAMIN E |
|
|
Salmonids |
Reduced growth, exopthalmia, ascites, anaemia, clubbed gills, epicarditis, ceroid deposition in spleen, increased mortality, pale gills, increased carcass fat and moisture content, erthrocyte fragility, muscle damage/degeneration, reduced egg hatching rate/spawning efficiency (75-78) |
|
C. carpio |
Muscular dystrophy, mortality, exopthalmia (79-80) |
|
I. punctatus |
Reduced growth, poor food conversion efficiency, exuadative diathesis, muscular dystrophy, depigmentation, fatty liver, anaemia, atrophy of pancreatic tissue, mortality, ceroid deposition in liver blood vessels, splenic hemosiderosis (10,81-83) |
1/ 1-McLaren et al. (1947); 2-Phillips and Brockway (1957); 3-Halver (1957); 4-Kitamura et al. (1967); 5-Poston et al. (1977); 6-Takeuchi, Takeuchi and Ogino (1980); 7-Hughes, Rumsey and Nickum (1981); 8-Woodward (1982); 9-Woodward (1985);10-Dupree (1966); 11-Murai and Andrews (1978); 12-Aoe et al. (1967); 13-Ogino (1967); 14-Yone (1975); 15-Arai, Nose and Hashimoto (1972); 16-Coates and Halver (1958); 17-Murai and Andrews (1979); 18-Wilson, Bowser and Poe (1983); 19-Poston and Di Lorenzo (1973); 20-Aoe, Masuda and Takada (1967); 21-Andrews and Murai (1978); 22-Aoe et al., (1969); 23-Murai and Andrews (1978a); 24-Hashimoto, Arai and Nose (1970); 25-Smith, Brin and Halver (1974); 26-Jürss and Jonas (1981); 27-Jürss (1978); 28-Hardy, Halver and Brannon (1979); 29-Ogino (1965); 30-Andrews and Murai (1979); 31-Adron, Knox and Cowey (1978); 32-Kissil et al. (1981); 33-Sakaguchi, Takeda and Tange (1969); 34-Agrawal and Mahajan (1983); 35-Walton, Cowey and Adron (1984a); 36-Poston and McCartney (1974); 37-Poston (1976); 38-Castledine et al. (1978); 39-Poston and Page (1982); 40-Ogino et al. (1970); 41-Robinson and Lovell (1978); 42-Lovell and Buston (1984); 43-John and Mahajan (1979); 44-Phillips et al. (1963); 45-Kashiwada and Teshima (1966); 46-Kashiwada, Teshima and Kanazawa (1970); 47-Limsuwan and Lovell (1981); 48-Ketola (1976); 49-Ogino et al. (1970a); 50-Aoe and Masuda (1967); 51-Burtle (1981); 52-Kitamura et al. (1965); 53-Hilton, Cho and Slinger (1978); 54-Sato, Yoshinaka and Ikeda (1978); 55-Poston (1967); 56-Halver, Ashley and Smith (1969); 57-Lovell (1973); 58-Andrews and Murai (1974); 59-Lovell and Lim (1978); 60-Wilson and Poe (1973); 61-Lim and Lovell (1978); 62-Li and Lovell (1985); 63-Mahajan and Agrawal (1979); 64-A. Soliman (unpublished data); 65-Agrawal and Mahajan (1980); 66-Poston et al. (1977); 67-Aoe et al. (1968); 68-Barnett, Cho and Slinger (1979); 69-Leatherland et al. (1980); 70-Lovell and Li (1978); 71-Andrews, Murai and Page (1980); 72-Poston (1964); 73-Poston (1976a); 74-Murai and Andrews (1977); 75-Woodall et al. (1964); 76-Poston (1965); 77-Poston, Combs and Leibovitz (1976); 78-Cowey et al. (1984); 79-Watanabe et al. (1970); 80-Watanabe and Takashima (1977); 81-Murai and Andrews (1974); 82-Lovell, Miyazaki and Rabegnator (1984); 83-Wilson, Bowser and Poe (1984).
Under intensive culture conditions, and in the absence of natural food organisms, dietary vitamin deficiencies may arise through:
Feed processing and storage
(i) Riboflavin: used in the form of a spray dried powder or a dry-dilution product, riboflavin is generally stable in dry multivitamin premixes. Processing losses of 26% have been reported for extruded pet foods (NRC, 1983). Feeds containing riboflavin should be protected from intense light/UV (liable to oxidation) and alkaline conditions.(ii) Pantothenic acid: used in the form of calcium d-pantothenate (92% activity) or calcium dl-pantothenate (46% activity), pantothenic acid is generally stable in dry multivitamin premixes. Processing losses during pelleting or extrusion have been reported to be as high as 10% (Slinger, Razzaque and Cho, 1979).
(iii) Niacin: used in the form of nicotinic acid or niacinamide and added as a dry-dilution product, niacin is generally stable in dry multivitamin premixes. Processing losses of 20% have been reported for extruded pet foods (NRC, 1983). Stability of niacin is good only if the feed is kept cool and dry.
(iv) Thiamine: used in the form of thiamine mononitrate (91.88% activity), thiamine is stable in dry multivitamin premixes that contain no added choline or trace elements. Thiamine is rapidly destroyed under alkaline conditions or in the presence of sulphide. Processing (pelleting/extrusion) and storage (7 months, room temperature) losses have been reported to be 0-10% and 11-12%, respectively (Slinger, Razzaque and Cho, 1979).
(v) Pyridoxine: used in the form of pyridoxine hydrochloride in dry-dilution, pyridoxine is stable in dry multivitamin premixes that contain no added trace elements. Prepared feeds that contain pyridoxine need protection from sunlight (UV), heat and moisture. Processing and storage (10 months) losses have been reported to be 7-10% (Slinger, Razzaque and Cho, 1979).
(vi) Biotin: used in the form of d-biotin in dry dilution, biotin is generally stable in dry multivitamin premixes. Processing losses in extruded pet foods have been reported to be 10% (NRC, 1983).
(vii) Folic acid: used in the crystalline form and in dry-dilution folic acid can be lost during the storage of dry multi-vitamin premixes particularly at elevated temperatures (43% activity lost after 3 months at room temperature). Processing and storage losses have been reported to be 3-10% (Slinger, Razzaque and Cho, 1979). Folic acid is liable to oxidation on storage at elevated temperatures and on exposure to sunlight.
(viii) Vitamin B12: used in the crystalline form and in dry-dilution, vitamin B12 stability in multivitamin premixes is dependent on storage temperature; elevated temperatures reduce activity, particularly in presence of mild acid conditions.
(ix) Choline: used as a 70% choline chloride solution or as a dry powder (25-60% activity), choline chloride is stable in multivitamin premixes but can decrease the stability of other vitamins present. Relatively stable on processing and storage (NRC, 1983).
(x) Vitamin C: used as L ascorbic acid, ethylcellulose or fat coated (to improve stability) and generally not added to dry multivitamin premixes because of its poor stability. Readily oxidized in the presence of moisture, trace elements, elevated temperatures, light and oxidation products (rancid oils). Stability is dependent on form of product used and method of processing. Losses of up to 90% have been reported on 'cold' pelleting and drying (original L-ascorbic acid content-400 mg/kg; Hilton, Cho and Slinger (1977). These authors even report a 70% loss in vitamin C activity on the addition of water to the feed mixture, prior to pelleting. Processing and storage losses for practical fish feeds have been reported to be as high as 95% for uncoated vitamin C (Slinger, Razzaque and Cho, 1979). Similarly, G. Reinitz reports that thawing Oregon Moist pellets for 16 h at 2°C or 25°C resulted in 53.7% and 97.2% loss of vitamin C respectively (unpublished data). However, these difficulties may be obviated by using more stable forms of vitamin C, such as ascorbate 2-sulphate (Halver et al. 1975) or ascorbyl palmitate.
(xi) Vitamin A; used as the acetate, palmitate or propionate ester, usually in beadlet form with Vitamin D3. Vitamin A is stable in dry multivitamin premixes. However, vitamin A is readily oxidized at elevated storage temperatures and in the presence of oxidation products (rancid oils). Processing losses of 20% have been reported for extruded pet foods (NRC, 1983). Similarly, six months storage at room temperature resulted in a 53% loss (NRC, 1983). Stability can be increased by suitable antioxidant protection and spraying on to the outside of the pellet in a lipid medium (NRC, 1983).
(xii) Vitamin D: used as vitamin D3 - cholecaliferol, and normally added in beadlet form with vitamin A or as a spray dried/drum dried powder. Stability is generally high.
(xiii) Vitamin K: used in the form of a menadione (K3) salt, either as menadione sodium bisulphite (50% K3 activity) or menadione sodium bisulphite complex (33% K3 activity). Stability in multivitamin premixes is good in the absence of trace elements (Frye, 1978). Under processing conditions heat, moisture, alkaline pH and trace elements accelerate the destruction of menadione salts (NRC, 1983). Prepared feeds should be protected from sunlight to obviate further oxidative losses.
(xiv) Vitamin E: used in the form of dl-a tocopherol acetate, either spray dried or absorbed, vitamin E is stable in dry multivitamin premixes stored at below room temperature. Stability is increased when used in the acetate form, but is prone to oxidation on storage at high temperatures and in the presence of oxidation products (rancid oils).
Leaching of water soluble vitamins
In contrast to the fat soluble vitamins (A, D, E and K) the water soluble vitamins can readily be 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 the 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 and 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 (1.18-2.36 mm diam. pellet). In the same study, Slinger, Razzaque and Cho (1979), 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 diets for penaeid shrimps reported water soluble vitamin losses of 97% (thiamine), 94% (pantothenic acid), 93% (pyridoxine), 90% (vitamin C),86Z (riboflavin), 50% (inositol) and 45% (choline) after a one hour immersion period in sea water (Cuzon, Hew and Cognie, 1982).
Deficiencies due to the presence of dietary anti-vitamin factors
(i) Avidin - heat labile anti-biotin factor present in raw egg white. Readily destroyed by heat treatment.(ii) Thiaminase - heat labile anti-thiamine factor present in raw fish, shellfish, rice polishings, Indian mustard seed, mung bean (green gram) and linseed (Liener, 1980). Dietary thiamine deficiency may be overcome by heat processing the contaminated raw material so as to deactivate the thiaminase, or by the use of supplemental dibenzoyl-thiamine (DBT) as a thiaminase resistant form of dietary thiamine.
(iii) Anti-vitamin A, E, D and B12 factors present in raw soybean. May be deactivated by heat treatment (Liener, 1980).
(iv) Anti-pyridoxine factor present in linseed; treatment as above.
Deficiencies due to dietary antibiotic addition
The use of feed antibiotics to treat disease outbreaks may destroy the vitamin synthesising capacity of the gut micro-flora of fish, which in omnivorous/herbivorous fish species is thought to greatly contribute toward the vitamin requirements of the fish (carp, tilapia, channel catfish -vitamin B12, folic acid and possibly biotin, thiamine and vitamin K).
In contrast to the water soluble vitamins, fish accumulate fat-soluble vitamins (A,D,E & K) under conditions where dietary intake 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. Toxicity signs which have been reported include:
|
VITAMIN A | |
|
Salmonids |
Reduced growth and haematocrit, severe necrosis/erosion of anal, caudal, pelvic and pectoral fins, scoliosis, lordosis, increased mortality, pale yellow fragile livers, reduced body fat (1,2) |
|
VITAMIN D | |
|
Salmonids |
Reduced growth, lethargy, dark colouration (3) |
|
I. punctatus |
Reduced weight gain, poor feed conversion efficiency (4) |
|
VITAMIN E | |
|
General |
Poor growth, toxic liver reaction, mortality (3) |
1-Hilton (1983); 2-Poston (1971); 3-Halyer (1980); 4-Andrews, Murai and Page (1980)
