Influence of stock density on digestive enzyme activity (trypsin), heat shock protein 70 (HSP70), and oxidative stress biomarkers of narrow clawed crayfish, Astacus leptodactylus Eschscholtz, 1823 (Decapoda, Astacidae)

in Crustaceana
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Stocking density causes changes in behaviour and physiology of aquatic animals and high density can cause oxidative damage in living cells. Therefore, experiments were designed to investigate the effects of stocking density on the oxidant-antioxidant response (glutathione (GSH), glutathione peroxidase (GSH-Px), catalase (CAT), and malondialdehyde (MDA)), digestive enzyme activity (trypsin), and heat shock protein 70 (HSP70) of narrow clawed crayfish (Astacus leptodactylus). The experiment was conducted to assess effects by stocking 3, 6, 9 and 12 crayfish/m2. Crayfish were stocked into 12 fiberglass tanks, three replicates per treatment and fed for 45 days. Our results indicated that activity of GSH-Px, MDA, and GSH levels increased in hepatopancreas (p<0.05) and muscle tissues (p<0.05) with increasing stock concentration while CAT activity decreased (p<0.05). Trypsin and HSP70 insignificantly increased in haemolymph. In conclusion, oxidative stress biomarkers were negatively influenced by increased quantitative changes in stocking density.

Influence of stock density on digestive enzyme activity (trypsin), heat shock protein 70 (HSP70), and oxidative stress biomarkers of narrow clawed crayfish, Astacus leptodactylus Eschscholtz, 1823 (Decapoda, Astacidae)

in Crustaceana



AebiH.1984. Catalase in vitro. Method Enzymol.105: 121-126.

AhvenharjuT.SavolainenR.TulonenJ.RuohonenK.2005. Effects of size grading on growth, survival and cheliped injuries of signal crayfish (Pacifastacus leniusculus Dana) summerlings (age 0+). Aquacult. Res.36: 857-867.

BaysaR. P.WhangchaiN.2007. Effect of culture season and stocking density on growth and production of giant freshwater prawn (Macrobrachium rosenbergii de Man) raised in northern Thailand. Mj. Int. J. Sci. Tech.1: 216-221.

BeutlerE.DuronO.KellyB. M.1963. Improved method for the determination of blood glutathione. J. Lab. Clin. Med.61: 882-888.

CabritaE.Martínez-PáramoS.GavaiaP. J.RiescoM. F.ValcarceD. G.SarasqueteC.HerráezM. P.RoblesV.2014. Factors enhancing fish sperm quality and emerging tools for sperm analysis. Aquaculture432: 389-401.

CaraB.MoyanoF. J.ZamboninoJ. L.FauvelC. H.2004. Actividad tripsina y quimotripsina como indicadores de condición larvaria: estudio de restricción alimentaria y calidad de puesta. Conferencias Científica III Congreso Iberoamericano Virtual de Acuicultura 547-556.

CastexM.LemaireP.WabeteN.ChimL.2009. Effect of dietary probiotic Pediococcus acidilactici on antioxidant defences and oxidative stress status of shrimp Litopenaeus stylirostris. Aquaculture294: 306-313.

ChenY. Y.ChenJ. C.LinY. C.YehS. T.HuangC. L.2015. White shrimp Litopenaeus vannamei that have received Gracilaria tenuistipitata extract show early recovery of immune parameters after Ammonia stressing. Drugs13: 3606-3624.

EliaA. C.DörrA. J. M.MastrangeloC.PrearoM.AbeteM. C.2006. Glutathione and antioxidant enzymes in the hepatopancreas of crayfish Procambarus clarkii (Girard, 1852) of Lake Trasimeno (Italy). Bul. Fr. Pêche Piscic.380-381: 1351-1361.

FarhadiA.JensenM. A.2016. Effects of photoperiod and stocking density on survival, growth and physiological responses of narrow clawed crayfish (Astacus leptodactylus). Aquacult. Res.47: 2518-2527.

García-GascaaA.GalavizM. A.GutiérrezJ. N.García-OrtegaA.2006. Development of the digestive tract, trypsin activity and gene expression in eggs and larvae of the bullseye puffer fish Sphoeroides annulatus. Aquaculture251: 366-376.

GonzalezR.CeladaJ. D.GonzalezA.GarciaV.CarralJ. M.Saez-RoyuelaM.2010. Stocking density for the intensive rearing of juvenile crayfish, Pacifastacus leniusculus (Astacidae) using Artemia nauplii to supplement a dry diet from the onset of exogenous feeding. Aquacult. Int.18: 371-378.

GoyertJ. C.AvaultJ. W.1978. Effects of stocking density and substrate on growth and survival of crawfish (Procambarus clarkii) grown in a recirculating culture system. Aquaculture9: 731-735.

HarlioğluM. M.KutluyerF.GürS.2012. An investigation on the sperm number and reproductive parameters of males in wild caught freshwater crayfish (Astacus leptodactylus Eschscholtz). Anim. Biol.62: 409-418.

IwamaG. K.ThomasP.VijayanM. M.ForsythR.1998. Stress protein expression in fish. Fish Biol. Fish.8: 35-56.

JensenM. A.CarterC. G.AdamsL. R.FitzgibbonQ. P.2013. Growth and biochemistry of the spiny lobster Sagmariasus verreauxi cultured at low and high density from hatch to puerulus. Aquaculture376-379: 162-170.

JonesC. M.RuscoeI. M.2000. Assessment of stocking size and density in the production of redclaw crayfish, Cherax quadricarinatus (von Martens) (Decapoda: Parastacidae), cultured under earthen pond conditions. Aquaculture189: 63-71.

KalmarB.GreensmithL.2009. Induction of heat shock proteins for protection against oxidative stress. Adv. Drug Deliv. Rev.61: 310-318.

KimH. P.WangX.ZhangJ.SuhG. Y.BenjaminI. J.RyterS. W.ChoiA. M.2005. Heat shock protein-70 mediates the cytoprotective effect of carbon monoxide: involvement of p38 beta MAPK and heat shock factor-1. J. Immunol.175: 2622-2629.

KleinB.MoullacL. G.SellosD.Van WormhoutA.1996. Molecular cloning and sequencing of trypsin cDNAs from Penaeus vannamei (Crustacea, Decapoda): use in 11 assessing gene expression during the moult cycle. J. Biochem. Cell Biol.28: 551-563.

KöksalG.1988. Astacus leptodactylus in Europe. In: Freshwater crayfish: biology management and exploitation: 365-400. (Croom HelmLondon, United Kingdom).

KutluyerF.BenzerF.ErişirM.ÖğretmenF.İnananB. E.2016. The in vitro effect of Cypermethrin on quality and oxidative stress indices of rainbow trout Oncorhynchu smykiss spermatozoa. Pest. Biochem. Physiol.128: 63-67.

LawrenceR. A.BurkR. F.1976. Glutathione peroxidase activity in selenium-deficient rat liver. Biochem. Biophys. Res. Commun.71: 952-958.

LutzC. G.WoltersW. R.1986. The effect of five stocking densities on growth and yield of red swamp crawfish Procambarus clarkii. J. World Aquac. Soc.17: 33-36.

MazlumY.2007. Effect of stocking density affects the growth, survival, and cheliped injuries of third instars of narrow-clawed crayfish, Astacus leptodactylus Eschscholtz, 1823 juveniles. Crustaceana80: 803-815.

MikamiS.1995. Larviculture of Thenus (Decapoda Scyllaridae) the Moreton Bay Bugs. Ph.D. Thesis University of Queensland Brisbane QLD.

MillsB.McCloudP. I.1983. Effects of stocking and feeding rates on experimental pond production of the crayfish Cherax destructor Clark (Decapoda: Parastacidae). Aquaculture34: 51-72.

MinagawaM.MuranoM.1993. Effects of prey density on survival, feeding rate and development of zoeas of the red frog crab Ranina ranina (Crustacea, Decapoda, Raninidae). Aquaculture113: 91-100.

MorrissyN. M.1979. Experimental pond production of marron Cherax tenuimanus (Smith) [Decapoda: Parastacidae]. Aquaculture16: 319-344.

MourenteG.Diaz-SalvagoE.BellJ. G.TocherD. R.2002. Increased activities of hepatic antioxidant defence enzymes in juvenile gilthead sea bream (Sparus aurata L.) fed dietary oxidised oil: attenuation by dietary vitamin E. Aquaculture214: 343-361.

Naranjo-ParamoJ.Hernandez-LlamasA.VillarrealH.2004. Effect of stocking density on growth, survival and yield of juvenile redclaw crayfish Cherax quadricarinatus (Decapoda: Parastacidae) in gravel-lined commercial nursery ponds. Aquaculture242: 197-206.

PelhamH. R. B.1986. Speculations on the functions of the major heat shock and glucose regulated proteins. Cell46: 959-961.

PlacerZ. A.CusmanL. L.JohnsonB. C.1966. Estimation of product of lipid peroxidation (malondialdehyde) in biochemical systems. Anal. Biochem.16: 359-364.

RajeshkumarS.MiniJ.MunuswamyN.2013. Effects of heavy metals on antioxidants and expression of HSP70 in different tissues of Milk fish (Chanos chanos) of Kaattuppalli Island, Chennai, India. Ecotoxicol. Environ. Safety98: 8-18.

RavagnanL.GurbuxaniS.SusinS. A.MaisseC.DaugasE.ZamzamiN.MakT.JaattelaM.PenningerJ. M.GarridoC.KroemerG.2001. Heat-shock protein 70 antagonizes apoptosis-inducing factor. Nature Cell Biol.3: 839-843.

RodgersL. J.SaoudP. I.RouseD. B.2006. The effects of monosex culture and stocking density on survival, growth and yield of redclaw crayfish (Cherax quadricarinatus) in earthen ponds. Aquaculture259: 164-168.

Sainz HernándezJ. C.Córdova MuruetaJ. H.2009. Activity of trypsin from Litopenaeus vannamei. Aquaculture290: 190-195.

SainzJ. C.GarcíaC. F.SierraB. A.HernandezC. P.2004. Trypsin syntesis and storage as zymogen in the midgut gland of the shrimp Litopenaeus vannamei. J. Crustacean Biol.24: 266-273.

SantacruzH.VrizS.AngelierN.1997. Molecular characterization of a heat shock cognate cDNA of zeabrafish, HSC70, and developmental expression of the corresponding transcripts. Dev. Comp. Immunol.21: 223-233.

SavolainenR.RuohonenK.RailoE.2004. Effect of stocking density on growth, survival and cheliped injuries of stage 2 juvenile signal crayfish Pasifastacus leniusculus Dana. Aquaculture231: 237-248.

SelvakumarS.GeraldineP.ShanjuS.JayakumarT.2005. Stressor-specific induction of heat shock protein 70 in the freshwater prawn Macrobrachium malcolmsonii (H. Milne Edwards) exposed to the pesticides endosulfan and carbaryl. Pestic. Biochem. Phys.82: 125-132.

SmithG. G.RitaraJ.2006. The influence of animal density and water turbulence on growth and survival of cultured spiny lobster (Jasus edwardsii) larvae. Aquaculture258: 404-411.

VijayanM. M.PereiraC.KruzynskiG.IwamaG. K.1998. Sublethal concentrations of contaminant induce the expression of hepatic heat shock proteins 70 in two salmonids. Aquat. Toxicol.40: 101-108.

VillagranE. R.1993. Effects of stocking density and supplemental feeding on production of red swamp crawfish in pools. Master’s thesis. Louisiana State University Baton Rouge LA.

VoznesenskyM.LerzD. M.Spanings-PierrotC.TowleD. H.2004. Genomic approaches for detecting thermal stress in Calanus finmarchicus (Copepoda: Calanoida). J. Exp. Mar. Biol. Ecol.311: 37-46.


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    Effect of stocking density on (a) MDA, (b) GSH, (c) GSH-Px and (d) CAT of hepatopancreas in Astacus leptodactylus Eschscholtz, 1823 (n=6). Data are presented as mean ± SD. Superscript letters indicate significant differences among treatments (p<0.05).

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    Effect of stocking density on (a) MDA, (b) GSH, (c) GSH-Px and (d) CAT of muscle in Astacus leptodactylus Eschscholtz, 1823 (n=6). Data are presented as mean ± SD. Superscript letters indicate significant differences among treatments (p<0.05).

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