Journal of

Applied Ichthyology
J. Appl. Ichthyol. 26 (2010), 453455 Published 2010. This article is a US Government work and is in the public domain in the USA. ISSN 01758659 Received: November 1, 2008 Accepted: August 28, 2009 doi:10.1111/j.1439-0426.2009.01371.x

Short communication Eect of salt addition and feeding frequency on cascudo preto Rhinelepis aspera (Pisces: Loricariidae) larviculture
By R. K. Luz1 and J. C. E. dos Santos2
Laboratorio de Aquacultura da Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil; 2Estac a o de Hidrobiologia e Piscicultura de Tres Marias-CODEVASF, Tres Marias, Minas Gerais, Brazil
1

Introduction The cascudo preto Rhinelepis aspera is a detritivorous sh species that occurs in the Sa o Francisco River (Sato et al., River (Agostinho et al., 1995). This 1998) and the Parana neotropical freshwater sh is important for reservoir sheries and river ecology. It also has aquaculture potential, in particular for the international ornamental aquarium industry. However, little is known about eective larviculture of this species. Fish larval rearing is considered to be a dicult stage for many species (Santos and Luz, 2009). Determining the best feeding frequency is important to achieve better growth and survival (Rabe and Brown, 2000), and to optimize laboratory culture procedures (Luz and Portella, 2005). One potential management option during larviculture can be the addition of salt to the culture water for improvement in survival and or growth of some freshwater neotropical species (Beux and Zaniboni-Filho, 2007; Luz and Santos, 2008; Santos and Luz, 2009). Thus, rearing in dierent salinities during some days of the culture then returning to freshwater at later stages is important because the environment into which the juveniles will be released for production or for stocking of reservoirs will be freshwater. The aim of this study was to evaluate the eect of low level salt additions to the rearing water during the rst 7 days of exogenous feeding and, subsequently, determine the eect of the return to freshwater for seven more days, using two dierent feeding frequencies on growth and survival of R. aspera. Materials and methods The experiment was conducted at the Hydrobiology and Hatchery Station of Tre s Marias, Minas Gerais, Brazil. Larvae were reared under largely controlled conditions in 24 circular aquaria (2-L volume; white color; constant aeration; photoperiod 10L:14D). The light level at the water surface was 150 lux. Each day at 09.00 h, temperature and dissolved oxygen were determined in all aquaria using the YSI 55 equipment. Dissolved oxygen was 5.0 0.9 mg L)1 and the average temperature was 26.0 0.8C. Artemia nauplii were oered as food (daily prey concentrations = 400, 600 and 800 Artemia larvae)1, from 1 to 5, 6 to 10, and 11 to 14 days of feeding, respectively). These levels were divided in accordance to the feeding frequency. Aquaria were siphoned daily to

remove waste and about 50% total volume was renewed with water from prepared stock solutions. First-feeding larvae of R. aspera (age 5 days; total length = 6.23 0.32 mm; weight = 3.22 0.11 mg, n = 20) were transferred directly from freshwater to various salinities: S0 = freshwater (control); S2 = 2 g salt L)1; S4 = 4 g salt L)1; and S6 = 6 g salt L)1. The larvae were reared under these conditions during the rst 7 days of feeding. For each salinity, two feeding frequencies were used: F2 = twice a day (at 08.00 and 17.00 h), and F4 = 4 times a day (at 08.00, 11.00, 14.00 and 17.00 h). Stocking density was 10 larvae L)1 (20 larvae aquaria)1). The work was conducted with a 4 2 factorial design (four salinities, two feeding frequencies, and three replicates each). The test media of freshwater and three salinities used in the experiments were prepared and stocked in 500-L containers. Commercial salt was used to prepare the solutions (Remo Brazilian Industry). The pH values of the stock solutions were 7.15, 7.19, 7.21 and 7.27, and the respective conductivity values (lS cm)1) were 60, 3780, 6540, 9480 in freshwater, and 2, 4 and 6 g salt L)1, respectively (measured on the HORIBA U10 equipment). After the rst 7 days of feeding in dierent salinities and the control, all larvae were counted to determine survival; samples (n = 35) of larvae from each aquarium were xed in formalin 10% to measure total length (mm) with an electronic caliper (Starret), and individual weights (mg) to 0.0001 g precision. To continue larval rearing, the larvae of all treatments were transferred directly to freshwater and subsequently reared in the freshwater for 7 days. For the transfer, sh were captured with a 0.5 mm diameter mesh. After a complete water exchange, sh were returned to the same aquaria. Due to some mortality and larvae samples taken for measurement, the stocking density was adjusted to reach 7 larvae L)1 (14 larvae aquaria)1). At the end, after 7 days in freshwater (14 days of feeding), all animals were collected to determine the survival and growth (weight and total length) as describe above. The specic growth rate (SGR) of larvae was determined [SGR = 100 (ln Wtf ) ln Wti) Dt)1, with Dt the time interval (in days) between Wti (initial weight) and Wtf (nal weight)]. Survival and growth data were compared by parametric Factorial ANOVA, and means were compared using Tukeys test at a 5% probability level using the SATISTICA 7.0 program (Statsoft INC., 2004).

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454 SGR (% mg day)1)

R. K. Luz and J. C. E. dos Santos Table 1 Total length, weight, specic growth rate (SGR), and survival of Rhinelepis aspera larvae reared for rst 7 days of feeding in dierent salt concentrations and feeding frequencies

Total length (mm) F2 F4 S0 S2 S4 S6 Statistical Feeding frequency (F) Salt concentration (S) Interaction (F S) 13.4 1.4a 12.6 1.2b 14.2 13.3 13.3 11.4 0.9a 1.3a 0.4a 0.8b

Weight (mg)

Survival (%) 88.7 7.4 85.9 11.5 92.5 86.6 86.0 84.2 9.3 10.8 6.5 10.7

5.16* 10.82** 1.16ns

Means for Feeding Frequency (F) 33.6 4.5 33.5 2.0 27.7 8.3 30.3 4.4 Means for Salinity (S) 31.9 6.9 35.5 3.2 30.8 8.4 31.8 4.4 34.7 5.4 33.9 2.2 26.5 6.5 29.8 4.0 F values ns 4.44ns 3.91 ns 1.93 1.74ns 1.86ns 1.37ns

0.13ns 0.91ns 0.20ns

Means followed by same superscript letters did not dier in Tukeys test (P < 0.05); *(P < 0.05); **(P < 0.01); ns(not signicant). F2 = twice a day, and F4 = 4 times a day feeding frequency. S0 = freshwater; S2 = 2 g salt L)1; S4 = 4 g salt L)1; and S6 = 6 g salt L)1.

Total length (mm) F2 F4 S00 S20 S40 S60 Statistical Feeding frequency (F) Initial salt concentration (Si) Interaction (F Si) 21.1 1.0 20.9 0.7 21.8 21.4 20.5 20.1 0.5a 0.7a 0.4ab 0.6b

Weight (mg)

SGR (% mg day)1)

Survival (%) 97.0 7.1 95.2 10.1 94.3 7.8 95.2 11.6 95.7 9.6 100.0 0.06ns 0.50ns 0.66ns

0.43ns 6.81* 0.32ns

Means for feeding frequency (F) 121.7 14.9 18.4 2.5 117.6 20.4 20.0 4.4 Means for initial salt concentrations (Si) 125.7 14.2 19.2 3.4 126.3 25.9 20.4 4.9 119.0 9.9 17.6 2.6 107.6 5.8 18.9 2.3 F values 0.36ns 0.56ns 1.04ns 0.59ns 0.11ns 0.85ns

Table 2 Total length, weight, specic growth rate (SGR), and survival of Rhinelepis aspera larvae during 7 days of feeding in freshwater, after transfer from initial rearing in dierent salt concentrations

Means followed by same superscript letters did not dier in Tukeys test (P < 0.05); *(P < 0.05); **(P < 0.01); ns(not signicant). F2 = 2 times a day, and F4 = 4 times a day feeding frequency. S00, S20, S40, and S60 initial salt concentrations used in rst 7 days of feeding; nal rearing in freshwater (more than 7 days of feeding).

Results Table 1 shows the growth and survival of R. aspera after 7 days of initial feeding at dierent salinity regimes and in freshwater (control). Weight, SGR and survival were similar among all treatments, without pronounced eects from salinity (S), feeding frequency (F) or interaction between S F. Total length, however, was signicantly inuenced by salinity (P < 0.01) and feeding frequency (P < 0.05). The twice daily feeding frequency showed better length growth than the four times daily feeding regime, while rearing at a salinity of 6 g salt L)1 resulted in lower growth (as expressed by total length) compared to the other salt treatments. Table 2 shows the growth and survival of R. aspera reared in freshwater after being returned from exposure to dierent salt concentrations. Rearing for an additional 7 days resulted in survival rates, weight, and SGR similar to control, and were not signicantly inuenced by the initial exposure to various salt concentrations (Si) or feeding frequency (F). There also was no Si F interaction eect. In contrast, total length was signicantly aected by the initial exposure to salt concentrations. The worst eects (P < 0.05) were registered in specimens exposed to the treatment S60. Discussion Five-day-old R. aspera larvae can be transferred directly from freshwater to water containing salt (up to 6 g salt L)1) and

reared under these conditions for the rst 7 days of feeding. After this period the animals can be returned to freshwater. These osmotic changes do not aect survival. Low salinities, up to 4 g salt L)1, have been demonstrated as allowing for good survival when rearing freshwater larvae (Luz and Portella, 2002; Beux and Zaniboni-Filho, 2007; Luz and Santos, 2008; Santos and Luz, 2009). However, those studies did not evaluate the eects of returning the test animals from dierent salinities to freshwater. This tolerance to dierent salinities during the larvae initial development suggests the presence of mechanisms and structures related to the acclimation process to osmotic variations. This is an important point that can be used in the prevention of diseases and to improve juvenile production. The addition of salt (up to 4 g salt L)1) can help reduce infestations in Rhamdia quelen of Ichthyophthirius multiliis (Garcia et al., 2007), a common parasite in freshwater larviculture and which often causes high mortality. Similar growth rates in freshwater and dierent salt concentrations were veried in R. aspera larvae, except for total length. This indicates that the species presents good acclimation and tolerance capacity to salt concentration exposures up to 6 g salt L)1. However, the reduced total length registered in the sh reared at higher salinities is not well understood. Future works need to elucidate fully the treatment eects on the total length of R. aspera larvae. After transfer of the test specimens from the dierent salt concentration exposures to freshwater, the weight gain and

Management of R. aspera larviculture

455 Cho, S. H.; Lim, Y. S.; Lee, J. H.; Lee, J. K.; Park, S.; Lee, S. M., 2003: Eects of feeding rate and feeding frequency on survival, growth and body composition of Ayu post-larvae Plecoglossus altivelis. J. World Aquac. Soc. 34, 8591. Garcia, L. O.; Becker, A. G.; Copatti, C. E.; Baldisserotto, B.; Neto, J. R., 2007: Salt in the food and water as a supportive therapy for Ichthyophthirius multiliis infestation on silver catsh, Rhamdia quelen, ngerlings. J. World Aquac. Soc. 38, 111. Kikuchi, K.; Nakahiro, I.; Kawabata, T.; Yanagawa, T., 2006: Eect of feed frequency, water temperature, and stocking density on the growth of tiger puer, Takifugu rubripes. J. World Aquac. Soc. 37, 1220. Luz, R. K.; Portella, M. C., 2002: Traira o (Hoplias lacerdae) larviculture in slightly saline freshwater. Rev. Bras. Zootec. 31, 829834. Luz, R. K.; Portella, M. C., 2005: Feeding frequency throughout traira o Hoplias lacerdae larval rearing. Rev. Bras. Zootec. 34, 14421448. Luz, R. K.; Santos, J. C. E., 2008: Stocking density and water salinity on pacama larviculture. Pesqui. Agropecu. Bras. 43, 903909. Mohseni, M.; Pourkazemi, M.; Bahmani, M.; Falahatkar, B.; Pourali, H. R.; Salehpour, M., 2006: Eects of feeding rate and feeding frequency on growth performance yearling great sturgeon, Huso huso. J. Appl. Ichthyol. 22(Suppl. 1), 278282. Petkam, R.; Moodie, G. E. E., 2001: Food particle size, feeding frequency, and the use of prepared food to culture larval walking sh (Clarias macrocephalus). Aquaculture 194, 349362. Rabe, J.; Brown, J. A., 2000: A pulse feeding strategy for rearing larval sh: an experiment with yellowtail ounder. Aquaculture 191, 289302. Santos, J. C. E.; Luz, R. K., 2009: Eect of salinity and prey concentrations on Pseudoplatystoma corruscans, Prochilodus costatus and Lophiosilurus alexandri larviculture. Aquaculture 287, 324328. Sato, Y.; Verani, N. F.; Verani, J. R.; Godinho, H. P.; Sampaio, E. V., 1998: Induced reproduction and reproductive characteristics of Rhinelepis aspera Agassiz, 1829 (Ostheichthyes: Siluriformes, Loricariidae). Braz. Arch. Biol. Tech. 43, 309314. Statsoft INC., 2004: Statistica (data analysis software system). Version 7.0. C. E. dos Santos, Estac Authors address: Jose a o de Hidrobiologia e Piscicultura de Tre s Marias-CODEVASF, CEP. 39205-000, Tre s Marias, Minas Gerais, Brazil. E-mail: jose.claudio@codevasf.gov.br

SGR in freshwater were similar to the controls, conrming acclimation capacity to short-term osmotic changes. However, the lower total length in the S60 treatment was simply a consequence of the lower initial total length result from the rst 7 days of exposure to high salt concentrations. The feeding frequency did not aect R. aspera larval survival, weight gain or SGR during rearing in dierent salt concentrations; however, total length was reduced when larvae were oered food rations four times per day. After transfer from dierent salinities to freshwater, the larvae can continue being fed twice a day. Feeding frequency between twice and four times a day also did not aected survival and growth of Hoplias lacerdae (Luz and Portella, 2005), or walking catsh Clarias macrocephalus (Petkam and Moodie, 2001) larvae. For these species, twice daily feeding frequencies optimize labor for other larviculture activities. However, feeding frequencies have dierent results in dierent species (Cho et al., 2003; Kikuchi et al., 2006; Mohseni et al., 2006). Dierent feeding frequencies must still be tested for R. aspera juveniles and adults to determine optimum management regimes for rearing. Acknowledgements This research received support from CODEVASF CEMIG, ` Pesquisa do Brazil and FAPEMIG (Fundac a o de Amparo a Estado de Minas Gerais, Brazil). We thank Dr Yoshimi Sato for assistance and support during this work. References
Agostinho, A. A.; Matsuura, Y.; Okada, E. K.; Nakatani, K., 1995: The catsh, Rhinelepis aspera (Teleostei: Loricariidae), in the River: an example of population Gua ra region of the Parana estimation from catch-eort and tagging data when emigration and immigration are high. Fish. Res. 23, 333344. Beux, L. F.; Zaniboni-Filho, E., 2007: Survival and the growth of pintado (Pseudoplatystoma corruscans) post-larvae on dierent salinities. Braz. Arch. Biol. Tech. 50, 821829.