Aquaculture Research, 2006, 37, 1378^1380

doi:10.1111/j.1365-2109.2006.01576.x

SHORT COMMUNICATION Effect of thyroxine immersion on larval survival, growth and postlarvae production of freshwater prawn,

Macrobrachium rosenbergii (de Man)

Paymon Roustaian1,2 & Lim Ai Gaik1
1

Department of Fisheries and Aquaculture, Faculty of Agrotechnology and Food Science, Kolej Universiti Sains dan Teknologi Department of Fisheries, Faculty of Marine Resources, Khoramshahr University of Marine Science and Technology (KUMST),

Malaysia (KUSTEM), Kuala Terengganu, Malaysia

2

Khoramshahr, Khozestan, Iran

Correspondence: P Roustaian, Department of Biology, University of New Brunswick, PO Box 5050, Saint John, New Brunswick, Canada E2L 4L5. E-mail: proustai@unbsj.ca

A growing body of evidence indicates that thyroid hormones (which are customarily regarded as vertebrate hormones) and their derivatives may have a role to play in the early life histories of a variety of invertebrates. For example, it has been reported that thyroid hormones are synthesized and accelerate metamorphosis in sand dollar (Saito, Seki, Amemiya,Yamasu, Suyemitsu & Ishihara 1998) and ascidians (Patricolo, Commarata & D gati 2001). These hormones have A also been shown to accelerate larval development in several sea urchins (Chino, Saito,Yamasu, Suyemitsu & Ishihara 1994), crown of thorn starsh (Johnson & Cartwright 1996) and abalone (Fukazawa, Hirai, Hori, Roberts, Nukaya, Ishida & Tsuji 2001). However, the available literature on crustacean endocrinology points to the lack of biosynthesis and/or the presence of thyroid hormones and their derivatives in this taxa (Quackenbush 1986; Huberman 2000). In light of their beneciary role in promoting larval growth and induction of metamorphosis in both invertebrates and vertebrates, it is interesting to note that no information is available on application of thyroid hormones on the larval period of the decapode crustaceans. The objective of this short communication is to examine the growth-promoting eects of exogenous thyroxine (T4) application on larviculture of freshwater prawn Macrobrachium rosenbergii. Five-day-old M. rosenbergii larvae (stage II^III) obtained from wild-caught females were stocked into eighteen 53 L aged (not new) blue plastic tanks

(experimental units), each holding 40 L of brackish water (salinity 511 1g L 1 and temperature 5 24.1^24.3 1C) with a constant gentle aeration at a density of 40 larvae L 1. They were fed solely on newly hatched Artemia nauplii (5 nauplii mL 1) once daily. Hormone treatments were performed by immersion of larvae in hormone baths once a week for 2 h. Oroxine tablets (Glaxo, Middlesex, UK) each containing 0.1mg of synthetic L -thyroxine sodium was extracted as described by Lam (1980) to prepare four hormone baths, each holding 3 L of the following hormone concentrations in triplicate: 0.01, 0.10, 1.00, 2.00 mg L 1 and control (0.00 mg L 1) (5 3 baths). Larvae from respective experimental units were gently scooped out and immersed in the respective bath (small aquarium holding 3 L of respective hormone concentration with a mild aeration) for 2 h, after which the hormone solutions containing the larvae were poured back gently into their respective tanks. Eighty per cent of water for each tank was changed weekly. Water quality parameters that were checked included pH, temperature and salinity. The water quality parameters were relatively consistent throughout the whole course of experiment; the temperature uctuated from 24 to 29 1C, the salinity was 11.011.0 g L 1 and pH ranged between 7.55 and 7.91. No abnormal larval behaviour (e.g. not exhibiting positive phototatic response, accumulation at the bottom of the tank) and lack of pigmentation were observed during the rearing period. It is worth noting

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Aquaculture Research, 2006, 37, 1378^1380

Eects of thyroxine on larval freshwater prawn P Roustaian & L A Gaik

Table 1 Growth expressed as mean developmental stage for Macrobrachium rosenbergii larvae grown under dierent thyroxine treatments,w
Age of larvae (day) Thyroxine treatments (mg L
Control 0.01 0.10 1.00 2.00
1

5
3.0a 3.0a 3.0a 3.0a 3.0a

8
4.3a 4.2ab 4.5ab 4.5ab 4.6b

11
5.5a 5.8a 5.8a 6.0a 6.1a

15
6.8a 7.2a 6.9ab 7.3ab 7.6b

18
7.5a 8.0ab 8.3bc 8.4bc 8.6c

22
8.7a 9.1ab 9.3ab 9.4ab 9.7b

25
9.4a 9.8ab 9.7bc 10.1bc 10.1c

Harvest
11.9a 11.8a 11.8a 11.9a 11.9a

Values are expressed as the mean of three replicates. wMeans in the same column followed by dierent letters are signicantly dierent among treatments within the same day according to Tukeys HSD test.

Table 2 Performance measures for Macrobrachium rosenbergii larvae grown under dierent thyroxine treatments at the termination of the experiment,w
Performance measures Thyroxine treatments (mg L 1)
Control 0.01 0.10 1.00 2.00

% survival
37.9 46.0 42.7 59.6 68.2 3.68 2.0b 6.9ab 4.8c 1.8d
a

Mean developmental stage

11.9 11.8 11.8 11.9 11.9 0.15 0.04a 0.03a 0.06a 0.03a
a

PL L 1
13.7 16.2 15.2 21.8 25.5 0.66 1.0a 2.3a 1.3b 0.8c
a

Total PL production
1639 1939 1822 2621 3062

Dry weight (mg)

0.5 1.0 1.4 1.3 1.9 0.25a 0.32ab 0.25ab 0.53ab 0.27b

Values are expressed as mean of three replicates followed by standard deviation, except for total PL production, which is expressed as

the sum of PL in all replicates for that treatment. wMeans in the same column followed by dierent letters are signicantly dierent among treatments according to Tukeys HSD test. PL, postlarvae.

that the water became murky the day after hormone treatments. This was especially obvious for 1.00 and 2.00 mg L 1 treatments. However, the murkiness disappeared after a day. No record of such an observation was found in our literature survey. The murkiness may suggest that the thyroxine did not go completely into the solution. The experiment was terminated for each replicate when postlarvae (PL) production reached 80%. The performance measures used in this study were survival, growth [mean developmental stage (MDS) as described by Lovett & Felder 1988], PL production and dry weight (after drying in oven at 60 1C until a constant weight was achieved). The eects of the hormone treatments on the performance measures were evaluated by one-way analysis of variance (signicance level set at Po0.05), and if applicable, followed by Tukeys multiple-range test using SPSS 6.0 software. To ensure normal distribution, all per cent values were arcsine-square root transformed before analysis (Zar 1999).

No signicant dierences (P40.05) in survival were observed among treatments during the course of the experiment, except for the nal survival at termination of the trial. However, the MDS values were signicantly dierent (Po0.05) among treatments, with the 2.00 mg L 1 thyroxine giving the highest growth rate (Table 1). There was a signicant dierence (Po0.05) in the time taken for PL production to reach 80%. All control and hormone replicates for 0.01 and 0.1mg L 1 treatments were harvested within 34^35 days, while 1.00 and 2.00 mg L 1 treatments were harvested at 38^39 days of culture. Table 2 represents the performance measures at the termination of experiment. Per cent survival and PL L 1 were signicantly (Po0.05) higher for larvae exposed to the 2.00 mg L 1 treatment. The ndings of the present study showed that T4 treatments (as described above) enhanced the growth, survival and PL production but did not promote early metamorphosis to shorten M. rosenbergii larviculture period. Little is known of the specic

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Eects of thyroxine on larval freshwater prawn P Roustaian & L A Gaik

Aquaculture Research, 2006, 37, 1378^1380

actions of thyroid hormones during the early life histories of invertebrates as the criteria in most of these studies, including this one, are generalized (e.g. growth, survival, induction of metamorphosis). Some specic actions of exogenous application of T4 during the early life histories of insects, the most studied arthropods, point to its inuential eects on protein, nucleic acids, glycogen, and amino acid metabolism, ecdysteroid content and activity of membrane enzymes (Thyagaraja, Kelly, Masler & Borkovec 1991; Reddy, Chaudhuri & Sukumar 1994; Reddy, Chaudhuri & Thangavelu 1994). Thyroid hormones and their derivatives have also been suggested to act as vitamins (obtained through the food chain) in various stages of the life cycle in invertebrates, including their early life histories (Eales1997). Based on the eects of thyroid hormones on the growth and dierentiation of larval teleost, the growth-promoting eects observed for thyroxine-treated M. rosenbergii larvae may be explained by mechanisms such as reduced cannibalism and aggressive behaviour, dierentiation of the cardiovascular system, the skeleto-muscular system and digestive system as reviewed by Lam (1994) and Power, Llewellyn, Faustino, Nowell, Bjornsson, Einarsdottir, Canario and Sweeney (2001). In general, the growth-promoting eects of thyroid hormones in vertebrates have been attributed to mechanisms such as induction of growth hormone synthesis and secretion, biosynthesis of biological membranes and increased mitotic rate (Hulbert 2000). Lack of early metamorphosis upon exogenous application of T4 observed in this study may have resulted from life history stage, dose or duration of the hormone application. It is also possible that exogenous T4 may have no inuence on the metamorphosis of M. rosenbergii. The dependency of the T4 eects on such factors such as age, developmental stage, temperature, salinity, nutritional and dietary factors, dose, treatment duration and method of application would certainly require more research to elucidate fully the growth-promoting eects of exogenous T4 in this economically valuable species. References
Chino Y., Saito M., Yamasu K., Suyemitsu T. & Ishihara K. (1994) Formation of the adult rudiment of sea urchins is inuenced by thyroid hormones. Developmental Biology 161,1^11. Eales J.G. (1997) Iodine metabolism and thyroid-related functions in organisms lacking thyroid follicles: are thyroid hormones also vitamins. Proceedings of the Society for Experimental Biology and Medicine 214, 302^317.

Fukazawa H., Hirai H., Hori H., Roberts R.D., Nukaya H., Ishida H. & Tsuji K. (2001) Induction of abalone larval metamorphosis by thyroid hormones. Fisheries Science 67, 985^988. Huberman A. (2000) Shrimp endocrinology. A review. Aquaculture 191,191^208. Hulbert A.J. (2000) Thyroid hormones and their eects: a new perspective. Biological Reviews 75, 519^631. Johnson L.G. & Cartwright C.M. (1996) Thyroxine accelerated larval development in the crown-of-thorns starsh, Acanthaster planci. Biology Bulletin 190, 299^301. Lam T.J. (1980) Thyroxine enhances larval development and survival in Sarotherodon (Tilapia) mossambicus Ruppell. Aquaculture 21, 287^291. Lam T.J. (1994) Hormones and egg/larval quality in sh. Journal of theWorld Aquaculture Society 25, 2^12. Lovett D.L. & Felder D.L. (1988) Evaluation of the rotifer Brachionus plicatilis as a substitute forArtemia in feeding larvae of Macrobrachium rosenbergii. Aquaculture 71, 331^338. Patricolo E., Commarata M. & D gati P. (2001) Presence of A thyroid hormones in ascidian larvae and their involvement in metamorphosis. Journal of Experimental Zoology 290, 426^430. Power D.M., Llewellyn L., Faustino M., Nowell M.A., Bjornsson B.Th., Einarstottir L.E., CanarioA.V & Sweeney G.E. .M. (2001) Thyroid hormones in growth and development of sh. Comparative Biochemistry and Physiology Part C 130, 447^459. Quackenbush L.S. (1986) Crustacean endocrinology, a review. Canadian Journal of Fisheries and Aquatic Science 43, 2271^2282. Reddy K.D., Chaudhuri A. & Sukumar K. (1994) Enrichment of ion specic adenosine triphosphatase activities by throxine in dierent tissues of the silkworm, Bombyx mori L during development. Insect Biochemistry and Molecular Biology 24, 243^248. Reddy K.D., Chaudhuri A. & Thangavelu K. (1994) L-thyroxine (T4) elevates the free amino acid pool of hemolymph plasma of tasar silkworm, Antheraea mylitta Drury (Lepidoptera, Saturniidae). Hormone and Metabolic Research 26, 570^573. Saito M., Seki M., Amemiya S., Yamasu K., Suyemitsu T. & Ishihara K. (1998) Induction of metamorphosis in the sand, dollar Peronella japonica by thyroid hormones. Development Growth and Dierentiation 40, 307^312. Thyagaraja B.S., Kelly T.J., Masler E.P. & Borkovec A.B. (1991) Thyroxine-induced hemolymph protein and ecdysteriod increases in the silkworm, Bombyx mori L: eect on larval growth and silk production. Journal of Insect Physiology 37, 153^159. Zar J.H. (1999) Biostatistical Analysis, 4th edn. Prentice Hall, Upper Saddle River, NJ, USA.

Keywords: Macrobrachium rosenbergii, thyroxine, thyroid hormones, larvae, invertebrate

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