Corresponding author: Italo Salgado Leu (

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1 Lt. Am. J. Aqut. Res., 43(5): , Effects 215 of protein nd crohydrte on juveniles of Smstcus spinifrons 8361 DOI: /vol43-issue5-fulltext-4 Reserch Article Effects of different protein nd crohydrte contents on growth nd survivl of juveniles of southern Chilen freshwter cryfish, Smstcus spinifrons Itlo Slgdo-Leu 1 & Alert G.J. Tcon 2 1 Escuel de Acuicultur, Universidd Ctólic de Temuco. Rudecindo Orteg 295, Temuco, Chile 2 Aqutic Frms Ltd., Kmehmeh Hwy, Kneohe, HI96744, USA Corresponding uthor: Itlo Slgdo Leu ( islgdo@uct.cl) ABSTRACT. In cultivted qutic orgnisms nutritionl requirements re criticl, not only for their impct on production techniques, ut lso, for their high incidence on production costs. There is limited knowledge on some species such s the southern Chilen freshwter cryfish, Smstcus spinifrons. In order to generte prcticl knowledge, study ws crried out to determine protein nd crohydrte content requirements. These fctors were evluted upon their effects on growth nd survivl of juveniles. For this purpose, individul weight, iomss gin, survivl, nd feed conversion prmeters were mesured. The ssy ws crried out in 42 dys, It ws conducted in flow through system, using 21 plstic tnks of 1.6 L cpcity. Ech tnk ws seeded with 2 juveniles weighing 5 mg verge ech. A 3x2 fctoril design ws proposed with three protein contents (2, 3, 4%) nd two crohydrte contents (low: from 16.3 to 23.5% nd high: from 34.6 to 35.8%). Six tretments nd three replictes were performed. Individuls were fed on pprent stition once dy. The diets formulted with 3% of protein nd the two crohydrte contents resulted in higher iomss increses, food conversion efficiencies over 26%, nd specific growth rte of.78%, ll displying significnt differences. Survivl showed highly significnt differences; in ll diets were superior to 6%, however the diets with 3% of protein surpssed 9%. Keywords: Smstcus spinifrons, Crustce, diets, protein-crohydrte content, southern Chile. Efecto de diferentes niveles de proteín y crohidrtos sore el crecimiento y sorevivenci de juveniles del cmrón de río del sur de Chile, Smstcus spinifrons RESUMEN. En orgnismos cultivdos, los requerimientos nutricionles, resultn ser de sum importnci, no sólo por sus impctos en ls técnics de producción, sino tmién por su lt incidenci en los costos de producción. Este conocimiento es escso en lguns especies como el cmrón de río del sur de Chile, Smstcus spinifrons. A fin de generr un conocimiento práctico, se definieron los requerimientos de proteíns y de crohidrtos y se evluron sus efectos en el crecimiento y supervivenci de juveniles. Pr esto, se midió peso individul, ioms gnd, supervivenci y prámetros de conversión limentici. L prte experimentl se relizó en un sistem de flujo ierto, conformdo por 21 estnques de plástico con cpciddes de 1,6 L, durnte un periodo de 42 dís. Se semrron 2 individuos en cd estnque con pesos individules de 5 mg en promedio cd uno. Se propuso un diseño fctoril de 3x2 con tres niveles de proteín (2, 3 y 4%) y dos niveles de crohidrto (jo: 16,3 23,5% y lto: 34,6 35,8%). Se desrrollron seis trtmientos con tres réplics. Los individuos fueron limentdos prente sciedd, un vez l dí. Ls diets con 3% de contenido proteínico con mos niveles de crohidrtos ofrecieron incrementos de ioms más ltos, eficienci de conversión limentici de 26% y ts de crecimiento específico de,78%, todos ellos con diferencis significtivs. L supervivenci presentó diferencis ltmente significtivs, si ien en tods ls diets fue superior l 6%, en ls diets con 3% de proteín supeó el 9%. Plrs clve: Smstcus spinifrons, Crustce, diets, niveles de proteíns y crohidrtos, sur de Chile. INTRODUCTION In the Astcide field, severl investigtions on the cultivtion of species hve een performed, the Astcus Corresponding editor: Muricio Lterç nd Austropotmoius in Europe, the Pcifstcus nd Procmrus in North Americ, nd the Cherx in Austrli nd South Americ (Celd et l., 1989; Ackefors et l., 1992; Wester et l., 1994; Jones, 1995; Ghisvnd et l., 212; Xu et l., 213).

2 2837 Ltin Americn Journl of Aqutic Reserch Chile is country locted in South Americ, sustining one of the most intensive fish frming ctivities for slmon, nd shring with Norwy the first plces in slmon production (FAO, 214). However, the country s intention is to diversify its quculture ctivities towrds other potentil species. Thus, Chile hs implemented Ntionl Aquculture Policy to fulfill this purpose. Among the species, with possiilities to diversify the quculture, four species of the fmily Prstcide reported long the Chilen cost re identified. However, most of the ttention hs een drwn towrds the southern Chilen crwfish, Smstcus spinifrons, chrcterized y its good flvor nd corporl size. The existing dt on S. spinifrons, is mostly on its iology (Rudolph, 22, 215) nd on some preliminry production tests done y Fundción Chile (Augsurger, 23). However, in order to rech n effective development of the cultured species, it is necessry to hve the se of seed production to ssure the following grow-out phse. A fundmentl spect to succeed with its culture is the knowledge of juvenile nutritionl requirements (Soud et l., 212), from wening to juvenile size, tht llow them to e plced under growing techniques. S. spinifrons psses the lrvl period in the dominl cvity of grvid femles (Rudolph, 22) s ny other stcids (Jones, 1995) which is n dvntge for culturl purposes. Protein will then e relevnt to consider in for further stges, to generte informtion on their protein requirements. Proly in similr wy done for most prcticl shrimp diets sed upon cost nd growth response (Lim, 1997). Both elements re impertive to evlute different protein dietry content nd determine the optiml content to sustin mximum growth. Tking into ccount the ove-mentioned considertions, nd the nutritionl importnce in culturing production of qutic species for juvenile S. spinifrons, the present study ws performed to estlish protein content requirement, y setting up fctoril design (3x2) formulting diets with three different protein contents (2, 3 nd 4%) nd two crohydrte contents (low nd high). MATERIALS AND METHODS The ssy ws crried out t Universidd Ctólic de Temuco (UC Temuco) Aquculture Deprtment, nd ws conducted in flow through system, using 21 rectngulr plstic tnks of 1.6 L cpcity with n open flow (Q) of.12 L min -1 with constnt ertion. PVC pipe pieces of 1.27 cm dimeter nd length of 4 cm were plced into ech tnk s shelter elements. Twenty juveniles weighing 49.6 ± 4.82 mg were seeded into ech tnk nd were mintined for 42 dys. Six diets were formulted to contin three contents of protein nd two reltive crohydrte contents. A 3 2 fctoril design with three replictes per dietry tretment ws used in this study. Diets were prepred with the ingredients indicted in Tle 1. The feed formultion model ws developed using n Excel softwre (Tle 2) with the following fctors nd contents: three protein contents (2, 3 nd 4%) nd two crohydrte contents (low: from 16.3 to 23.5% referred to s: C low nd high: from 34.6 to 35.8%, nmed s: C high ) (Tle 3). Selected ingredients nd prepred diets were nlyzed t UC Temuco Aquculture Deprtment in the Nutritionl Lortory ccording to the Officil Methods of Anlysis (AOAC, 1998). For the food preprtion, ech grounded ingredient ws pssed through 3 µm mesh size nd weighed the recommended quntity ccording to the formultion. In the mnufcture process of the experimentl diets, dry ingredients, except whet flour, were mixed during 4 min in lender mchine (Kitchen id; model K5SS). Seprtely, whet flour ws hydrted nd put under cooking process for 15 min. The resulting geltinized whet ws mixed with fish oil nd cpsule of vitmin E (ntioxidnt). Finlly, the oily geltinized whet ws comined with other dry ingredients for further 45 min. A met grinder mchine RCA (1 HP) with mtrix screen of 1.5 mm orifices ws used to mke different pelleted diets. The pellets were dried during 36 h t 55 C nd lter stored under freezing conditions (-2ºC). The juveniles were fed once per dy t sme time nd mortlity ws checked nd removed. The mount of food provided in ech diet ws registered on dily sis. The uneten diet in ech tnk ws removed every dy efore feeding with suction tue nd stored in seprte recipients into the refrigertor, depending of the experimentl unit. After seven dys, the removed uneten diet ws defrosted nd dried in 55 C oven for 24 h. The removed uneten diet ws discounted from the food plced into ech tnk nd then, the ctul food consumption for ech tnk ws determined. Individul weight ws mesured t the eginning nd t the end of the experiment (dy 42) with Srtorious nlyticl scle (cpcity of 217 g nd precision of.1 g). Growth prmeters were clculted nd evluted: individul weight gin (mg), specific growth rte (SGR), iomss gin (%), nd survivl (%). Nutritionl prmeters were lso evluted: feed conversion rtio nd feed conversion efficiency (%), ccording to De l Higuer (1987) nd Bureu et l. (22).

3 Effects of protein nd crohydrte on juveniles of Smstcus spinifrons 8383 Tle 1. Proximte composition (%) of ingredients used in the formultion of diets tested on Smstcus spinifrons juveniles. Ingredients Crude protein Ether extrct Nitrogen-free extrct Crude fier Ash Wter Fishmel Blood mel Lupine mel Lupine fier mel Crrot mel Kelp mel Whet mel Fish oil 1. Tle 2. Composition of experimentl diets (%), ccording to ingredient prticiption. CP: crude protein, C: crohydrte. Diets Ingredients 2%CP 2%CP 3%CP 3%CP 4%CP 4%CP C low C high C low C high C low C high Fish mel Blood mel Lupine mel Lupine fier mel Crrot mel Kelp mel Whet mel Fish oil Vitmins nd minerls Inorgnic Normlity, homoscedsticity nd independence were tested previously on recollected dt, then twowy nlysis of vrince ws pplied to determine its interctions. If significnt (P <.5) differences were found, Tukey tests were used to compre verges etween tretments. In ech crohydrte content, protein diet contents were compred y one-wy ANOVA followed y Tukey test. All vlues were clculted using Minit sttisticl softwre (version 16.1, Minit Stte College, PA, USA). Electric heters kept the wter temperture t pproximtely 18ºC. Dissolved oxygen (5.2 ±.6 nd ph (7. ±.2) were mesured with YSI-85 equipment. RESULTS Growth prmeters such s individul weight gin (mg), SGR, iomss gin (%), food conversion rte (FCR), food conversion efficiency (FCE-%) nd survivl rte (%) of S. spinifrons juveniles re shown in Tle 4. No significnt interctions (P >.5) were found for individul weight gin nd SGR. Biomss gin nd FCR were significntly high for diets with 3% CP followed y 2% CP in interctions with low crohydrte concentrtion contents. 3% CP+C low exhiited significntly higher FCE thn other comintions. Survivl for 3% CP + C low ws the highest vlue (95%) however, differences only showed to e significnt with 2% CP + C low. Protein content ehvior under reltive low crohydrte content, offered etter growth vlues thn reltive high crohydrte content, not only in iomss gin (Fig. 1), ut lso in ll the other prmeters mesured (individul weight gin, SGR, FCR, nd FCE). Survivl hd punctul significnt exception in 2% CP with C low (62.5%). Highest vlues were significntly recorded t 3% CP for iomss gin, FCR nd FCE. For SGR, FCR nd FCE, 3% CP ws significntly different to 2% nd 4% CP. For survivl t C low significnt differences were presented in ll CP content, eing 3% CP the est. At reltive high crohydrte content, there were no significnt differences etween protein contents. Curves for reltive high crohydrte content showed more flttened tendency form thn reltive low crohydrte content.

4 4839 Ltin Americn Journl of Aqutic Reserch Tle 3. Proximte composition nlysis (%) of experimentl diets. CP: crude protein, C: crohydrte. Diets Crude protein Ether extrct Nitrogen-free extrct Crude fier Ash Wter 2% CP+C low % CP+C high % CP+C low % CP+C high % CP+C low % CP+C high Tle 4. Smstcus spinifrons. Growth nd survivl of juveniles fter 42 dys of feeding on experimentl diets contining different crude protein (2%, 3%, 4%), nd reltive crohydrte contents (C high, C low) in diets. Different letters indicte significnt difference (P <.5). SGR: specific growth rte, FCR: food conversion rte, FCE: food conversion efficiency. Prmeters 2% 3% 4% C low C high C low C high C low C high Individul weight gin (mg) ± ± ± ± ± ± 2.68 SGR.67 ±.1.63 ±.9.83 ±.1.72 ±.9.57 ±.4.65 ±.7 Biomss gin (%) ± ± ± ± ± ± 1.41 FCR 3.18 ± ± ± ± ± ±.37 FCE (%) ± ± ± ± ± ± 2.39 Survivl (%) 62.5 ± ±. 95. ± ± ± ±. In oth crohydrte cses (reltive low nd high contents), growth prmeters incresed their performnce from 2% up to 3% CP nd then descended to 4% CP. Men iomss gin dt ws fitted to qudrtic model to estimte the protein requirement ccording to the dose-response reltionship. Optimum nd mximum protein requirements were otined y pplying the following qudrtic model: y = -18x x (determintion coefficient R 2 =.927). Clcultions indicte the corresponding optimum protein content to e 28.6% CP t low crohydrte content (Fig. 1), other prmeters modelled followed the sme pthwy. Ech curve models for corresponding prmeters cn e seen in Figure 1. DISCUSSION In the present study, 3% CP + C low, showed significntly etter results in evluting the growth prmeters such s iomss gin, FCR nd FCE. Individul weight gin nd SGR showed similr vlues with no significnt differences. Survivl ws higher thn 82% for most comintions except for 2% CP+C low (62%). Dietry protein content hve een studied for other crustcen species such s Mcrorchium rosenergii, rnging from 15 to 4 g kg -1 (Blzs & Ross, 1976; New et l., 198; D Armo & Reed, 1988; Mitr et l., 25; Teshim et l., 26). High dietry protein content (5 g kg -1 ) reported mximum growth of M. rosenergii post lrve (Heinen & Mnsi, 1991). If the dietry protein content re too low (diet with 23 g kg -1 CP) the growth rte will e reduced (Blzs & Ross, 1976); Boonyrtplin & New (1982) nd Brtlett & Enkerlin (1983) (diet with 15 g kg -1 CP) nd Zki et l. (22) (diet with g kg -1 CP). This my e due to the utiliztion of protein y M. rosenergii muscle tissue to mintin other vitl functions (God, 28). On the other side, growth depression ws reported in prwn fed diets exceeding their protein requirement due to the excess dietry protein eing metolized y prwns s source of energy nd nitrogen excreted s mmoni (Burford et l., 24). Hri & Kurup (23) suggested tht further increse over 3 g kg -1 CP in dietry protein does not hve ny dded dvntge in M. rosenergii. Moreover, for 4-5 g kg -1 CP resulted in growth depression s it cn lso e seen in the present work. These protein contents re within 3-35% CP requirements for juvenile M. rosenergii (D Armo & New, 2). Ackefors et l. (1992) suggested tht commercil diets for juvenile Astcus stcus my contin pproximtely 3-35% protein, 2-25% crohydrte nd not more thn 1% lipids for the est growth rte. Ghisvnd et l. (212) expressed tht Astcus leptodctylus cn e fed prcticl diet contining 3% protein content. Celd et l. (1989)

5 Effects of protein nd crohydrte on juveniles of Smstcus spinifrons y = x x R² =.9383 Individul weight gin (mg) Low crohydrte y = -145x 2 + 9x R² =.197 High crohydrte 1% 2% 3% 4% 1,8,6,4,2 y = x x R² =.3838 Low crohydrte SGR y = x x -.88 R² = % 2% 3% 4% High crohydrte y = x x R² =.7762 Biomss gin (%) Low crohydrte y = -18x x R² =.952 High crohydrte 1% 2% 3% 4% FCR y = x x R² =.3891 Low crohydrte y = x x R² =.9628 High crohydrte 1% 2% 3% 4% FCE y = x x R² =.729 y = -14x x R² =.972 Low crohydrte 1% 2% 3% 4% High crohydrte y = -75x x + 25 R² =.9 c Survivl (%) Low crohydrte y = -225x x R² =.9847 High crohydrte % 1% 2% 3% 4% 5% Figure 1. Protein content curves ehvior of growth prmeters nd survivl under low nd high crohydrte content in juveniles of Smstcus spinifrons. Sme letters ove points t low crohydrte curve men no significnt differences. High crohydrte curves did not present significnt differences. reported tht the est growth of juvenile Pcifstcus leniusculus ws otined with diets contining 27-33% protein from severl fresh nd rtificilly compounded diets nd Gonzlez et l. (212) expressed tht 4% of protein cn e suitle in commercil dry diets for this species. For Cherx qudricrintus, Wester et

6 6841 Ltin Americn Journl of Aqutic Reserch l. (1994) indicted tht diet formulted with 33% protein ppered to e dequte for juvenile phse, wheres Zenteno-Svin et l. (28) stted tht 31% of protein is the content to stisfy nutritionl requirements for optiml growth. Xu et l. (213) reported tht 27-3% protein diet provided the sme growth efficiency for Procmrus clrki, P. leniusculus nd P. clrki. These lst species seem to e closer to S. spinifrons in protein requirements thn Cherx, which could e explined due temperture dependence s Cherx re more tropicl thn other colder species. Severl sttisticl methods cn e used to estimte nutrient requirements in dose-response experiments sed on mthemticl nd iologicl ssumptions nd principles (Sherer, 2). Estimtes of requirements for mximum growth were otined y fitting doseresponse dt to qudrtic model. The results tht S. spinifrons juveniles required etween 28 nd 29% dietry crude protein for optimum growth s result to pply model found (y = -18x x ). In this sense, Cortes et l. (23) expressed protein requirement of 31%, clculted for C. qudricrintus from using the sme second order polynomil model thn this present study (y = -.71x x ). From nutritionl point of view, protein utiliztion indexes cn e considered s good indictors of protein spring effect (God, 28). In the present study, the diet contining protein content of 3% CP + C low, provided the highest growth indexes nd dietry protein utiliztion efficiency compred with the higher dietry crohydrte content. These results suggest tht S. spinifrons could e using dietry crohydrte s min energy source t lower contents to spre dietry protein for mximum growth. Similr results were otined y D Armo & New (2), nd lso y God (28) for M. rosenergii. Best FCR ws presented t 3% CP + C low (2.32), less thn those otined y Jones et l. (1996) for C. destructor nd Cortes et l. (23) for C. qudricrintus. Hri & Kurup (23) reported their etter FCR (2.84 nd 2.89 for 3 nd 35 g kg -1 CP diet, respectively) for M. rosenergii juveniles, comprle with results otined y Millikin et l. (198), Ashmore et l. (1985) nd Gomez et l. (1988). The decresed growth rte showed y juveniles over 4% CP cn e ttriuted to the high rte of protein ctolism (Hri & Kurup, 23). Growth depression ws lredy reported in prwn fed diets exceeding their protein requirement s result of toxicity (Zein-Eldin & Corliss, 1976). Regrding crohydrte utiliztion, Butist & Suos (1997) mentioned tht protein contents in diets improve growth when comined with crohydrte contents. This mens tht crohydrte is used s n economicl energy source. In this study, two reltive contents of crohydrte were used: low content (16.3 to 23.5%) nd high content (34.6 to 35.8%). Results showed tht low reltive content is enough to keep etter growth prmeters. In this sense, crohydrte is used to uild quitine (min component of exoskeleton) nd lso component of ftty cids nd sterols (Clifford & Brick, 1978; Kuchrski & D Silv, 1991; Cruz- Surez, 1996). Diz-Herrer et l. (1992) reported crohydrtes s min energy sources for metolic requirements in post lrve nd juveniles of M. rosenergii; lter supported y Lun et l. (27). Ross et l. (2) stted the possiility to reduce protein contents in diets y crohydrtes included s strch ingredient. Survivl for CP + Crohydrte comintions showed vlues higher thn 82%, except in 2% CP + C low (62.5%). They cn e compred with others. Survivl vlues during period up to 94 dys rnged etween 65-85% for C. destructor (Jones et l., 1996). For C. qudricrintus, Ponce et l. (1998) 6-95% of survivl rte; Wester et l. (1994) 5-71%; Thompson et l. (23) 56-8%; Thompson et l. (23) 95-1%; Jcinto et l. (23) 65-89%; nd Muzinic et l. (24) 79-98%. Mortlity my e ssocited to nutrition vritions in the diet nd experimentl conditions. A survivl rte ove 5% etween stocking nd hrvesting hs een considered to e cceptle y New & Singholk (1985) nd Vlenti (199); ut Cuzon & Guillume (1997) reported tht survivl rtes greter thn 8% ws considered good in crustcen studies. REFERENCES Ackefors, H., J.D. Cstell, L.D. Boston, P. Rty & M. Svensson Stndrd experimentl diets crustcen nutrition reserch. II. Growth nd survivl of juvenile cryfish Astcus stcus (Linné) fed diets contining vrious mounts of protein, crohydrte nd lipid. Aquculture, 14: Assocition of Anlyticl Chemist (AOAC) Officil methods of nlysis. Assocition of Anlyticl Chemist, Wshington DC, 118 pp. Ashmore, S.B., R.W. Stnley, L.B. Moore & S.R. Mlech Effect on growth nd pprent digestiility of diets vrying in grin source nd protein level in Mcrorchium rosenergii. J. World Mricult. Soc., 16: Augsurger, A. 23. L experienci de cultivo de cmrón de río del sur Smstcus spinifrons en Chile. Liro Resúmenes del Primer Seminrio Interncionl

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8 843 Ltin Americn Journl of Aqutic Reserch Prstcide): I. Development of htchery nd nursery procedures. Aquculture, 138: Jones, P.L., S.S. DeSilv & B.D. Mitchell Effects of replcement of niml protein soyen mel on growth nd crcss composition in juvenile Austrlin freshwter cryfish. Aqucult. Int., 4: Kuchrski, L. & R. D Silv Effect of diet composition on the crohydrte nd lipid metolism in n esturine cr, Chsmgnthus grnulte (Dn, 1851). Comp. Biochem. Physiol. A, 99: Lim, C Replcement of mrine niml protein with penut mel in diets for juvenile white shrimp, Peneus vnnmei. J. Appl. Aqucult., 7: Lun, M., C. Grzini, E. Villrroel, M. Lemus, C. Lodeiros & G. Slzr. 27. Evlución de tres diets con diferente contenido proteico en el cultivo de postlrvs del lngostino de río Mcrorchium rosenergii. Zootec. Trop., 25(2): Millikin, M.R., A.R. Fortner, P.H. Fir & L.V. Siek Influence of dietry protein concentrtion on growth, feed conversion nd generl metolism of juvenile prwn Mcrorchium rosenergii. Proc. World Mricult. Soc., 114: Mitr, G., D.N. Chttopdhyy & P.K. Mukhopdhyy. 25. Nutrition nd feeding in freshwter prwn Mcrorchium rosenergii frming. Aqu Feeds: Formultion nd Beyond, 2(1): Muzinic, L.A., K.R. Thompson, A. Morris, C.D. Wester, D.B. Rouse & L. Mnomitis. 24. Prtil nd totl replcement of fish mel with soyen mel nd rewer s grins with yest in prcticl diets for Austrlin red clw cryfish Cherx qudricrintus. Aquculture, 23: New, M.B., M. Boonyrtplin, P. Vorsyn & S. Intrpichte An ssessment of the performnce of Mcrorchium rosenergii fed simple nonvitmin-supplemented pelleted diets. Internl Report for UNDP/FAO Progrmmer for the Expnsion of Freshwter Prwn Frming (in Thilnd). Working Pper THA. 75.8/WP/8/16. New, M.B. & S. Singholk Freshwter prwn frming: mnul for the culture of Mrcorchium rosenergii. FAO Fish. Tech. (Rev 1), Rome, 225 pp. Ponce, P.J., F.J.L. Arredondo & R.M.A. Moreno The effects of vrying dietry protein levels on growth nd survivl of juvenile nd pre-dult redclw (Cherx qudricrintus). In: K.J.C. Von Vupel & F. Schrm (eds.). The iodiversity crisis nd crustcen. Proceedings of the Fourth Interntionl Crustcen Congress. A.A. Blkem, Amsterdm, 2: Ross, C., G. Cuzon, G. Gxiol, C. Pscul, R. Brito, M. Chiml & A. Vn Worhoudt. 2. El metolismo de los crohidrtos de Litopeneus setiferus, L. vnnmei y L. stylirostris. In: L. Cruz-Surez, D. Rique-Mrie, M. Tpi-Slzr, M. Oliver-Novo & R. Civer-Cerecedo. (eds.). Avnces en nutrición cuícol V. Memoris V Symposium Interncionl de Nutrición Acuícol, Mérid, pp Rudolph, E. 22. Sore l iologí del cmrón de río Smstcus spinifrons (Philippi, 1882) (Decpod, Prstcide). Gyn, 66(2): Rudolph, E Current stte of knowledge on Virilstcus species (Crustce, Decpod, Prstcide). Lt. Am. J. Aqut. Res., 43(5): Soud, I., A. Grz de Yt & J. Ghnwi A review of nutritionl iology nd dietry requirements of redclw cryfish Cherx qudricrintus (von Mrtens 1868). Aqucult. Nutr., 18: Sherer, K.D. 2. Experimentl design, sttisticl nlysis nd modeling of dietry nutrient requirement studies for fish: criticl review. Aqucult. Nutr., 6: Teshim, S., S. Koshio, M. Ishikw, S. Alm & L. Hernndez. 26. Protein requirements of the freshwter prwn Mcrorchium rosenergii evluted y the fctoril method. J. World Aqucult. Soc., 37(2): Thompson, K.R., L.A. Muzinic, T.D. Christin, C.D. Wester, L. Mnomitis & D.B. Rouse. 23. Lecithin requirements of juvenile Austrlin red clw cryfish Cherx qudricrintus. Aqucult. Nutr., 9: Thompson, K.R., L.A. Muzinic, T.D. Christin, C.D. Wester, L. Mnomitis & D.B. Rouse. 23. Effect on growth, survivl nd ftty cid composition of Austrlin red clw cryfish Cherx qudricrintus fed prcticl diets with nd without supplementl lecithin nd/or cholesterol. J. World Aqucult. Soc., 34: 1-1. Vlenti, W.C Crico de cmroes de gu doce Mcrorchium rosenergii. Anis d 27 Reunio Anul d Sociedde Brsileir de Zootecni e 12 Reunio d Asocico Ltino-Americn de Producto niml Cmpins, Fundco de Estudos Agrrios Luiz de Queiroz (FEALQ), Pircic, pp Wester, C.D., L.S. Goodgme-Tiu, J.H. Tidwell & D.B. Rouse Evlution of prcticl feed formultions with different protein levels for juvenile red clw cryfish (Cherx qudricrintus). Trns. Kentucky Acd. Sci., 55: Xu, W., W. Liu, M. Shen, G. Li, Y. Wng & W. Zhng Effect of different dietry protein nd lipid levels on growth performnce, ody composition of juvenile red swmp cryfish (Procmrus clrkii). Aqucult. Int., 21(3): Zki, M.A., H.A. Mrouk & A.A. Nour. 22. Optimum dietry protein levels nd stocking density for freshwter prwn Mcrorchium rosenergii juveniles rered in concrete sins. Egyp. J. Anim. Produc., 7:

9 Effects of protein nd crohydrte on juveniles of Smstcus spinifrons 8449 Zein-Eldin, Z. & J. Corliss The effect of protein levels nd sources on growth of Peneus ztecus. In: T.R.V. Pilly & J.A. Dill (eds.). Advnces in Aquculture. FAO Technicl Conference on Aquculture, Kyoto, Jpn. Fishing News Books, Frnhm, pp Zenteno-Svin, T., E. Cortes-Jcinto, J. Vsquez-Medin & H. Villrrel-Colmenres. 28. Oxidtive dmge in tissues of juvenile cryfish (Cherx qudricrintus von Mrtens, 188) fed with different levels of proteins nd lipids. Hidroiologic, 18(2): Received: 14 Novemer 214; Accepted: 1 July 215