May | June 2012 Feature title: Transfering Vitamin C from fish to embryos

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The International magazine for the aquaculture feed industry

FEATURE

from fish

Vitamin C
Transfering

to embryos
by Nagaraj G. Chatakondi PhD, National Warmwater Aquaculture Center, USA

B
Species
Tilapia

eneficial effects of ascorbic acid supplementation to broodstock of a select aquaculture species is well documented. At the present levels of feeding, dietary means of vitamin C does not meet the requirements for maturation, reproduction and needs of early life stages of larvae.

conventional methods, hence innovative approaches are needed. Mass transfer of nutrients via injection into broodstock is a novel method. Two routes of maternal transfer of vitamin C in mature channel catfish (Ictalurus punctatus) prior to hormone-induced spawning were explored as a strategy to incorporate

Table 1: Mass transfer of Vitamin C from broodfish to eggs/embryos

Mass trasfer strategy

Diet Diet

Dose
1000 mg/Kg 1250 mg/Kg

Rainbow trout Oncorhynchus mykiss Oreochromis mossambicus

ductive performance, and may subsequently improve ontogeny performance. However, the effect of vitamin C diminished with age and also in more natural conditions. Our goal was to achieve predictable fish production of robust quality for healthy, efficient, higher surviving and able to adapt to common stressors and pathogens. Improvements can be made in this area by new knowledgebased advances in nutrient delivery systems that may create large Reference improvements in terms of production, feed conversion and survival Sandness et al. (1984) in aquaculture production.

Soliman et al. (1986)

Importance of vitamin C

Ascorbic acid is an essential micronutrient in the diet of teleost Atlantic Cod Diet 500 mg/Kg Mangor-Jensen et al. (1994) fish, which do not have gulonolGadus morhua actone oxidase activity. Vitamin Rainbow trout Diet 500 mg/Kg Blom and Dabrowski (1995) C is needed for post-translatoOncorhynchus mykiss ry hydroxylation of proline and lysine moieties in collagen, minChannel catfish Diet 500 mg/Kg Zuberi et al. (2011) eral metabolism to improve stress Ictalurus punctatus response and immunity, detoxificaRainbow trout Immersion 1000 mg/L Falhatkar et al. (2006) tion reactions, steroid synthesis and Oncorhynchus mykiss vitellogenesis. Rainbow trout Immersion and Diet 1000 and 500 mg/Kg Falhatkar el.al. (2011) Egg ascorbic acid deposition levels may easily be tailored by Oncorhynchus mykiss feeding broodfish with elevated Japanese eel Injecting broodfish 50 mg/Kg Yoshikawa, 1998 levels of ascorbic acid before and Anguilla japonica after vitellogenesis. The accumulaJapanese eel Injecting broodfish 1mL* /Kg Furuita et al. 2009 tion of essential nutrients in eggs Anguilla japonica is dependent on the nutrient reserves in the female fish and Channel catfish Injecting broodfish 1 mL*/Kg Chatakondi et al. 2010 therefore on the dietary intake of Ictalurus punctatus broodfish in the period preceeding *Vitamin emulsion was prepared by dissolving Sodium Ascorbate in 0.9% NaCl Solution and during gametogenesis. Hence, broodfish nutrition consisting of In addition, this nutrient is water soluble the vitamin and to determine its effect on essential nutrients is important. The earliest steps in embryonic developand readily gets accumulated by other reproduction and progeny performance. The results of this study suggest injecting ment are dependent on and driven by materorgans before reaching the ovary. For practical reasons, it is not possible to vitamin C prior to hormone-induce spawning, nal factors deposited in the oocyte during attain the desired level of a nutrient by invokes transfer to eggs, improves repro- oogenesis. Maternal factors are stored in the
8 | InternatIonal AquAFeed | May-June 2012

FEATURE

form of specific mRNAs, proteins, hormones and other biomolecules. At egg activation and fertilization, these factors become available for embryogenesis, sometimes after a process of activation involving translation or protein modification. It has been documented that vitamin C or ascorbic acid deficiency in larval fish has been associated with hyperplasia of collagen and cartilage, scoliosis, lordosis, internal hemorrhages, resorbed opercules and abnormal support cartilage in gills, spine and fins with deformities of the jaw and snout. Based on recent research, vitamin C needs for reproduction and early life stages of fish are 10 times the recommended dose for raising young adult fish. These high levels cannot be met by dietary administration to broodfish because the nutrient is water soluble and readily absorbed / utilized by other organs during oocyte development. It has been demonstrated in several species that nutrients in broodfish diet are transferred to oocytes through uptake of extra-ovarian substances from the maternal blood. Also, there was up to a 82.4 percent loss of ascorbic acid of the prepared commercial diet. A 3.8 8.3-fold increase of vitamin C in the diet generally results in 56 to 71.9 percent increase of total ascorbic acid in the eggs respectively.

The fry produced from parents fed with elevated levels of vitamin C tend to have higher growth performance as compared with control groups. Thus, there is a need in enhance ascorbic acid in the broodfish. A diet with vitamin C content adequate for normal growth may not be sufficient for broodfish when the goal is to transfer ascorbic acids to embryos.

"Based on 2011 data, approximately 335 million pounds of catfish were processed, a reduction of over 50 percent compared to the best production in 2003

Reproduction and arval performance

Broodfish diet has a major influence on fecundity and egg quality. It has also been demonstrated that the nutritional status of broodfish can affect offspring quality. The accumulation of essential nutrients in eggs is dependent on 1) the nutrient reserves in the female fish and 2) the dietary intake preceding gonadogenesis. Vitamin C is needed for maturation, reproduction and larval metamorphosis. Beneficial effects include increased fertility, fecundity and egg quality. Nutrients in broodfish diet are trans-

ferred to oocytes through uptake of extraovarian substances from the maternal blood. Immersion enrichment of eggs is another approach to introduce compounds and nutrients into eggs. Immersion enrichment followed by feeding fry with vitamin C enhanced feed was also found to be an effective method. Injecting vitamin C in to broodfish during artificial- induced maturation improved reproduction and progeny performance. Efforts are

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"A decade ago, producing commercial quantities of hybrid catfish was believed to be unattainable. - Presently, a third of catfish farmers raise hybrid catfish in production ponds and hybrid catfish account for approximately 25% of all the total catfish processed in 2011"

furcatus male), hybridisation can be used to improve productivity immediately by producing fish that exhibit hybrid vigor. Based on numerous laboratory and field trials, hybrid catfish are superior in growth Vitamin C needs of rate, feed conversion, survival, seinability and channel catfish Channel catfish is the single largest processing traits compared to commonly aquaculture fish species cultured in the raised channel catfish. A decade ago, producing commercial quantities of hybrid catfish was believed to be unattainable. Natural hybridisation is rare and artificial spawning of channel catfish has been historically low and with no effective ovulating agents available. However, in the last 10 years, consistent and marked improvements were made in all the phases of artificial spawning and the hatchery pro"It appears that mass transfer of vitamin C duction of hybrid catfish to eggs is attained by injecting broodfish embryos. Improved production and consistent prior to hormone-induced spawning superior performance of hybrid catfish in commerto improve progeny performance" cial earthen production ponds has rejuvenated United States. Based on 2011 data, the industry with unprecedented optimism. approximately 335 million pounds of cat- Presently, a third of catfish farmers raise hybrid fish were processed, a reduction of over catfish in production ponds and hybrid catfish 50 percent compared to the best produc- account for approximately 25 percent of all tion in 2003 (2012 USDA NASS). The the total catfish processed in 2011. Our goal is to achieve predictable fish industry is currently struggling to keep pace with the increasing cost of feed, fuel, production of robust quality for healthy, production inefficiencies, foreign imports fast growing, survival and adapt to common and economy. Adopting hybrid catfish stressors and pathogens and to varying envi(channel catfish female x blue catfish, I. ronmental conditions. Improvements can be underway to develop procedures to effectively and stably accumulate vitamin C in eggs by broodstock injections (Table 1).
10 | InternatIonal AquAFeed | May-June 2012

made in this area by new knowledge-based advances in live food production or nutrient delivery systems that may create large improvements in terms of production, survival and processing yield.

Preliminary findings
Broodstock preparation is the primary requisite for hormone-induced spawning of channel catfish in the production of channel x blue hybrid catfish. Hence, broodfish management techniques must be geared towards attaining maximum production of high-quality eggs and larvae because variable egg quality is one of the limiting factors in fish hatcheries. Broodstock diet has been considered as one of the factors affecting fecundity, egg, and larval quality in fish. The accumulation of essential nutrients in eggs are dependent on the nutrient reserves in the female fish, and consequently on the dietary nutrient input of broodstock in the period preceding gonadogenesis. When eggs absorb water, it is possible to introduce compounds such as vitamins and minerals into the eggs with the water solution before water hardening. It was hypothesised that injecting female broodfish prior to hormone-induced spawning would result in mass transfer of nutrients to improve maturation, ovulation, and subsequent progeny performance. Preliminary studies confirmed accumulation of vitamin C in ovarian tissue and invoked a positive response to ovulation, fecundity and egg quality. Mass transfer of vitamin C to the eggs improved growth and reduced mortalities following Edwardseilla ictaluri disease challenge. It appears that mass transfer of vitamin C to eggs is attained by injecting broodfish prior to hormone-induced spawning to improve progeny performance.

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