The effects of dietary nucleotide content on the growth performance,digestibility and immune responses of juvenile narrow clawed crayfish,Astacus leptodactylus leptodactylus Eschscholtz, 1823

A 126‐day experiment was carried out under controlled conditions to compare the effects of five levels (0.5, 1.0, 1.5, 2.0 and 2.5 g kg^?1) of dietary nucleotide (Vannagen^®) on the growth performance and biological indices of juvenile crayfish (8.25 ± 0.39 g). The protein efficiency ratio (PER, 1.69–2.17), lipid efficiency ratio (LER, 2.29–3.00), energy efficiency ratio (EER, 2.68–3.65), protein productive value (PPV, 57.01–68.95%), lipid productive value (LPV, 63.02–75.15%) and energy productive value (EPV, 68.20–88.15%) increased significantly (P < 0.05) as linear response to increased dietary nucleotide from 0.5 to 2.5 g kg^?1 in the feed. With an increase in the dietary nucleotide, the uricase activity (654.29–827.63 U g^?1) and lactobacillus count to total count ratio (1.21–2.17) of crayfish fed the experimental diets increased significantly (P < 0.05). Crayfish fed the diets containing different levels of nucleotide (from 0.5 to 2.5 g kg^?1) had higher phenoloxidase activity (1.57–2.11 U min^?1) than that of control after air exposure challenge. At the levels tested, 2 g kg^?1 nucleotide in the diet was considered optimum for growth performance, digestibility and immune responses. It can be concluded that dietary nucleotide exerted positive effects on growth performance, feed utilization and accelerate crayfish immune response against air exposure challenge.     

Antifungal effect of Origanum onites essential oil as an alternative to formalin in the artificial incubation of narrow‐clawed crayfish (Astacus leptodactylus Eschscholtz, 1823)

As alternative to formalin, the antifungal effect of a plant product [Origanum onites L. (Lamiaceae) oil] was investigated for use in the artificial incubation of narrow‐clawed crayfish (Astacus leptodactylus Eschscholtz) eggs. For this purpose, this study was conducted as two experiments. In experiment I, the eggs were artificially incubated for 40 days. In experiment II, juveniles were cultured to determine effects of O. onites oil on juveniles for 30 days. The experimental groups were as follows: formalin (3500 ppm for 15 min), O. onites oil (300 ppm for 15 min, 700 ppm for 2 min and 1000 ppm as a dip treatment 15 split‐second) and a control (no treatment). In the experiment I, the highest hatching rate (86%) and survival rate of stage II juveniles (80%) were observed in 1000 ppm dip group. These results were similar to that of formalin group (85% and 79%) respectively. The control group exhibited the lowest hatching rate (49%) and stage II rate (42%) compared with the 1000 ppm dip group and 3500 ppm formalin treatments. However, other concentrations (300 and 700 ppm) of O. onites showed toxic effects on the eggs and there was no hatching. In the experiment II, the survival rate and growth performance of the crayfish juveniles were similar in all groups. This study indicated that the 1000 ppm O. onites dip treatment could be a good alternative to formalin for improved egg hatchability in the artificial incubation of crayfish eggs.     

The effects of seasonal,sex and size on the digestive enzyme activities in freshwater crayfish (Astacus leptodactylus Eschscholtz, 1823)

The effects of seasonal, sex and size on digestive enzyme activities of Astacus leptodactylus in natural habitat were investigated in the present study. The freshwater crayfish were sampled 584 individual as seasonally from Egirdir Lake. Ten male and 10 female individuals were sampled randomly from caught crayfish in each two different size ranges (4.5–7.0, 7.1–9.9 cm) and each season. The gastrointestinal tract, gastric, intestine and midgut gland were individually dissected on ice for enzyme analysis and stomach contents. At the end of the study, amylase activities were affected by interaction of season × sex × size in intestinal, season × sex in midgut gland. However, any interaction was not determined in amylase activities of the gastric. Lipase activities were affected by interaction of season × sex × size in intestinal and gastric, season × sex in midgut gland. The protease activities in each organ were affected by interaction of season, sex and size (p ? .05). In all organs, amylase and lipase activities were similar. However, the highest protease activity was in stomach while the lowest protease activity was the intestine. The protease activities were higher than amylase and lipase activities. There is a positive correlation between protease enzyme activities in gastric with gonad maturation and active feeding period. It can be said based on data of digestive system and high protease activity that A. leptodactylus is an omnivore species in need high protein. In addition, the reason for the low levels of lipase activity may be A. leptodactylus not prefer high fat foods or not fed high fat foods.     

Effects of photoperiod and stocking density on survival,growth and physiological responses of narrow clawed crayfish (Astacus leptodactylus)

Photoperiod and stocking density are critical factors influencing the performance of decapod crustaceans in culture, however, their influence on growth; survival and biochemical physiology of crayfish broodstock have rarely been considered. Analysis of biochemical physiology in crayfish broodstock during the non‐breeding season provides information on the energy storage requirements of broodstock for increased survival and reproductive output. Growth rate, moulting frequency, survival and biochemical physiology were measured in Astacus leptodactylus broodstock that were cultured at three different photoperiods (18L:6D; 12L:12D; and 6L:18D) and three stocking densities (10, 20 and 40 individuals m^?2) during non‐breeding season. Survival of crayfish was highest at 18L:6D photophase and 10 m^?2 (100%) than other treatments. Survival in the high stocking density was high when combined with 18L:6D photophase, but weight gain (WG) and specific growth rate (SGR) were higher at shorter photophase and lower stocking density. Longer photophase (18L:6D) increased stress responses, characterized by increased haemolymph lactate and glucose levels. Stocking density did not affect proximate composition of crayfish; however, individuals cultured at 18L:6D photophase had higher lipid content than other photoperiod treatments. The study demonstrated that culturing A. leptodactylus at 18L:6D photophase and 10 m^?2 is critical for increased survival of broodstock.     

Effects of dietary onion (Allium cepa) powder on growth performance,hemolymph indices and fillet organoleptic properties of juvenile narrow‐clawed crayfish,Astacus leptodactylus leptodactylus Eschscholtz, 1823

A 18‐week feeding trial was carried out under controlled conditions to compare the effects of onion powder (OP) at six levels (5, 10, 20, 30, 40 and 50 g/kg) on the growth performance, nutritional efficiency indices, hemolymph indices and fillet organoleptic properties of juvenile (5.62 ± 0.39 g) crayfish. The significantly (p < .05) highest values of final weight (71.30 g) and SGR (2.02% per day) and the lowest FCR (1.03) were observed in the juvenile crayfish fed the diet containing 40 g OP/kg. The juvenile crayfish fed the diet containing 40 g OP/kg had the significantly (p < .05) highest THC (105.27 × 10⁵ cell/ml), HC (98.33 × 10⁵ cell/ml), SGC (38.54 × 10⁵ cell/ml) and LGC (49.51 × 10⁵ cell/ml). The crayfish fed the levels of dietary OP higher than 30 g/kg showed the significantly (p < .05) higher values of SOD (4.07–4.30 U/min) and LYZ (6.73–7.20 U/min) compared with those fed 5, 10, 20 and 30 g of dietary OP/kg and control. Polynomial regression of SGR, FCR, PPV and PER suggested that the optimum dietary OP level could be higher than 30 and <50 mg/kg in crayfish reared in culture conditions.     

Growth performance and RNA/DNA ratio of noble crayfish (Astacus astacus) and narrow‐clawed crayfish (Pontastacus leptodactylus) fed fish waste diets

New and viable species for aquaponics and integrated multi‐trophic aquaculture (IMTA) in freshwater systems can improve yields and sustainability of aquaculture. Freshwater crayfish species such as Astacus astacus and Pontastacus leptodactylus are omnivorous feeders and considered candidates for feeding on faecal matters in existing aquaculture systems. Feeding trials were conducted to determine growth response and RNA/DNA ratio in freshwater crayfish fed fish waste. Carapace length and wet weight were measured to determine the growth response. Juvenile A. astacus was fed faeces of hybrid striped bass (Morone chrysops × Morone saxatilis) and rainbow trout (Oncorhynchus mykiss), while adult P. leptodactylus was fed with two commercial pellet diets and pikeperch (Sander lucioperca) faeces. The nutritional composition of hybrid striped bass faeces was close to optimal diet composition of A. astacus, and crayfish showed significantly higher carapace growth, weight gain and weight gain per moult as the group fed rainbow trout faeces. The growth of P. leptodactylus was significantly lower in terms of weight gain and weight gain percentage per moult for crayfish fed on pikeperch faeces. Thus, this study can recommend a co‐cultivation of hybrid striped bass and A. astacus within one system, but cannot recommend co‐cultivation of P. leptodactylus with pikeperch. Additionally, this study showed controversial results of RNA/DNA ratio and weight gain of both crayfish species. Thus, RNA/DNA ratio cannot be approved for investigations on crayfish physiological status in controlled feeding experiments if animals are fed with an inadequate diet.     

Effect of in vitro selected synbiotics (galactooligosaccharide and mannanoligosaccharide with or without Enterococcus faecalis) on growth performance,immune responses and intestinal microbiota of juvenile narrow clawed crayfish,Astacus leptodactylus leptodactylus Eschscholtz, 1823

The aim of this study was to determine the best synbiotic combination (based on growth and short‐chain fatty acids production) between Enterococcus faecalis and eight prebiotics. Based on the results of in vitro studies, E. faecalis + galactooligosaccharide (EGOS) and E. faecalis + mannanoligosaccharide (EMOS) were selected as synbiotics. A 126‐day feeding trial was conducted to compare the effects of prebiotics, probiotic and synbiotics on the growth indices, In vivo ADC of nutrients, digestive enzymes, hemolymph indices and finally, biological responses against 48‐hr Aeromonas hydrophila exposure challenges of juvenile (4.13 ± 0.12 g) crayfish. The highest values of SGR (2.19% body weight day^?1), VFI (2.75% body weight day^?1), survival rate (96.67%) and the lowest FCR (2.33) were observed in the juvenile crayfish fed the EGOS‐ diet. The significantly (p < .05) highest means of in vivo ADC_OM, in vivo ADC_CP, in vivo ADC_CF and in vivo ADC_GE were measured in crayfish fed the EGOS‐ diet. The mean survival rate of Aeromonas hydrophila‐injected crayfish fed the EGOS‐ diet (56%) was significantly (p < .05) higher than those of fed the control (8.67%) and other diets (22.67–35.32%). At the levels tested, 7.86 log CFU E. faecalis g^?1 + 10 g kg^?1 GOS in the diet was considered optimum.     

Effect of dietary encapsulated organic salts (Na‐acetate,Na‐butyrate,Na‐lactate and Na‐propionate) on growth performance,haemolymph, antioxidant and digestive enzyme activities and gut microbiota of juvenile narrow clawed crayfish,Astacus leptodactylus leptodactylus Eschscholtz, 1823

A 63‐day experiment was done to study the effects of four levels (5, 10, 20 and 50 g/kg) of encapsulated organic salts (Na‐acetate, Na‐butyrate, Na‐lactate and Na‐propionate) on the growth indices and haemato‐immunological responses of crayfish Astacus leptodactylus leptodactylus (4.38 ± 0.08 g). Crayfish were distributed at 51 1,000‐L tanks (17 treatments at triplicate). The highest values of final weight (27.86 g), specific growth rate (2.94% body weight per day) and survival rate (96%) were observed in the crayfish fed the 20 g/kg of encapsulated Na‐propionate diet (p < .05). The highest activities of phenoloxidase (7.4 U/min), superoxide dismutase (7.80 U/min) and lysozyme (9.40 U/min) were observed in the gut of crayfish fed the 20 g/kg of encapsulated Na‐propionate diet (p < .05), as well as the highest activities of alkaline protease (10.70 U/mg), lipase (9.10 U/mg), amylase (9.60 U/mg) and the lactobacillus count (p < .05). Broken line regression model of SGR and phenoloxidase activity suggested that the optimum dietary levels of encapsulated Na‐acetate, Na‐butyrate, Na‐lactate and Na‐propionate could be 30.7, 31.8, 31.4 and 33.5 g/kg, respectively, in crayfish reared in culture conditions.     

Effects of dietary antioxidants on oxidative stress,antioxidant defence and growth of freshwater crayfish Astacus leptodactylus (Eschscholtz, 1823) during the reproductive period in females

In this study, we examined the effects of dietary antioxidant vitamins [vitamin E (Vit E), vitamin C (Vit C) and vitamin A (Vit A)], astaxanthin (AX) and β‐carotene (βC) supplementation on oxidative stress [malondialdehyde (MDA)], antioxidant enzyme activities [superoxide dismutase (SOD), catalase, glutathione peroxidase (GSH‐Px)] and glutathione (GSH) levels in hepatopancreas, ovarian, muscle, gill tissues and survival rates (SR), live weight gain (LWG), specific growth rate (SGR) of Astacus leptodactylus during ovarian development. The EE, EC, EA, EAX and EβC groups were formed by addingClick here to enter text. Vit E, Vit C, Vit A, AX and βC to the diet respectively. The antioxidants did not have a significant impact on the values of SR, LWG and SGR. After 137 days, the MDA levels in the Control (C) group were statistically higher in the hepatopancreas, ovarian and muscle tissues according negative control (NC), EE, EC, EA, EAX and EβC groups. The SOD activities in ovarian, muscle and gill tissues and GSH levels in ovarian and muscle tissues were lower in the C group compared with the NC group, but GSH‐Px activities in hepatopancreas and ovarian tissues and GSH levels in gill tissues were higher. The MDA levels were statistically lower in all tissues of crayfish in experimental groups according to C group. The antioxidant enzyme activities and GSH levels changed according to tissue. The results showed that Vit E and AX should be supplemented in diets to increase the reproductive performance of crayfish, because Vit E and AX were of higher potential as antioxidants than other vitamins and carotenoids in ovarian tissues.     

The effect of dietary vitamin E on the pleopodal egg number of Astacus leptodactylus (Eschscholtz, 1823)

The effect of vitamin E on the pleopodal egg number of Astacus leptodactylus (Eschscholtz, 1823) was studied. Crayfish were fed 2% of their total wet weight daily with vitamin E supplemented diets and a control for 70 days. The vitamin E content of the control diet, diet 1, diet 2 and diet 3 were 20 mg kg^–1, 40 mg kg^–1, 80 mg kg^–1 and 160 mg kg^–1 respectively on a dry weight basis. Vitamin E levels of the control and experimental diets were analysed by High Performance Liquid Chromatography. Results showed that diets containing supplemental vitamin E were associated with an increase in the number of pleopodal eggs. The best result was obtained with diet 2 containing 80 mg kg^–1 supplemental vitamin E.     

The effect of dietary n‐3 series fatty acids on the number of pleopadal egg and stage 1 juvenile in freshwater crayfish,Astacus leptodactylus Eschscholtz

The effect of dietary n‐3 series fatty acids on the number of pleopadal egg and stage 1 juvenile in freshwater crayfish, Astacus leptodactylus was investigated. Crayfish were fed with n‐3 series fatty acids supplemented diets and a control diet for 251 days. Control diet did not comprise additional n‐3 series fatty acids. However, D2, D3 and D4 groups were supplemented with n‐3 series fatty acids at 1%, 2% and 3% level respectively. Results showed the beneficial effects of dietary n‐3 series fatty acids on the production of pleopodal egg and stage 1 juvenile number. At the end of the experiment, pleopodal egg (from 177 to 234) and stage 1 juvenile number (from 167 to 225) increased significantly with increased dietary n‐3 series fatty acid level. In addition, an increase in dietary n‐3 series fatty acids led to an increase in number of pleopodal egg and stage 1 juvenile produced per gram of female crayfish weight. It can be concluded that the reproductive efficiency of A. leptodactylus (i.e., pleopodal egg and stage 1 juvenile number) can be improved in controlled hatchery conditions by adding n‐3 series fatty acid into the diet of this species.     

Effect of serotonin injection on the reproductive parameters and haemolymph methyl farnesoate level in the narrow‐clawed crayfish Pontastacus leptodactylus (Eschscholtz, 1823)

The effects of serotonin injection (1, 5 and 10 µg/g body weight) on spermatozoal production, vas deferens index (VDI), hepatosomatic index (HSI), testicular index (TI), gonado‐somatic index (GSI), ovarian egg number, total protein, methyl farnesoate (MF) levels of haemolymph, amino acid and fatty acid profiles of hepatopancreas were evaluated in Pontastacus leptodactylus. In male P. leptodactylus, the highest spermatozoal number (9.3 × 10⁶ ± 1.2) and MF level (0.66 ± 0.06 ng^?1 ml) were observed after injection of 10 µg/g of serotonin (p < .05). Serotonin injection, however, did not significantly affect the reproductive parameters in female P. leptodactylus (p > .05). It was also found that serotonin injection caused an increase in the level of MF in male P. leptodactylus but a reduction in female P. leptodactylus (p < .05). Therefore, a sex‐dependent response to serotonin could exist for MF synthesis in P. leptodactylus. Serotonin injection did not affect the total protein, amino acid and fatty acid profiles in the hepatopancreas of male P. leptodactylus (p > .05). It can be concluded that serotonin injection affects the reproductive parameters such as spermatozoal production, VDI, TI, GSI, HSI and haemolymph MF levels in P. leptodactylus.     

Predation impact of common backswimmer (Notonecta glauca L.) on juvenile narrow‐clawed crayfish (Astacus leptodactylus Esch.)

The aim of the study was to investigate the effect of the presence of common backswimmer Notonecta glauca and their potential predation, on growth, mortality, survival and cheliped loss of juvenile narrow‐clawed crayfish Astacus leptodactylus. The presence of N. glauca in tanks during the day caused a radical change in the behaviour of juvenile A. leptodactylus, which remained for more time in hiding places. At night, such differences in crayfish behaviour were not found. The negative effect of N. glauca in tanks on crayfish total body length and body weight was statistically significant (p < .05). The crayfish survival decreased with increasing the number N. glauca in tanks. The crayfish mortality in the first week was higher than in the second week. The effect of the presence of N. glauca in tanks on crayfish chelae autotomy was statistically significant (p < .05). The number of crayfish with complete sets of chelipeds decreased and the number of crayfish with loss of chelipeds increased when the number of N. glauca in tanks increased. Common backswimmers N. glauca have been effective predators not only of victims caught on the surface of water, but also of bottom‐dwelling organisms such as juvenile crayfish.     

Effect of dietary n-3 series fatty acids on sperm production in the freshwater crayfish, Astacus leptodactylus (Eschscholtz) (Astacidae)

The objective of the present work was to evaluate the effects of three different rates of long chain n-3 polyunsaturated fatty acid (LC n-3 PUFA) on the sperm number of Astacus leptodactylus. The experiment was carried out with the four following treatments: control group (not supplemented LC n-3 PUFA), D1 (1 %), D2 (2 %) and D3 (3 %). The sperm number of A. leptodactylus was counted; the reproductive system was weighted; and the macroscopical analysis of the testes and vasa deferentia (DVD) were conducted. In addition, gonado-somatic index (GI) and testes index (TI) were also calculated. The results showed that there was a significant difference between the crayfish fed LC n-3 PUFA supplemented diets (D1, D2, D3) and control in mean sperm number (P < 0.05). The highest sperm number was found for the crayfish fed D3 (6.27 ± 0.14 × 10⁸ sperm/distal vas deferens section). The results also showed that the reproductive system of the crayfish fed D3 and D2 was significantly heavier than that of the crayfish fed the control and D1 (P < 0.05). On the other hand, there was not a significant difference in the testes weight and vasa deferentia weight between the crayfish fed the control and D1, and between the crayfish fed D2 and D3 (P < 0.05). GI of the crayfish fed D3 and D2 was significantly higher than that of the crayfish fed control and D1 (P > 0.05), but there was not a significant difference in the TI of the crayfish fed control, D1, D2 and D3 (P > 0.05). In conclusion, diets containing different levels of LC n-3 PUFA significantly increase the sperm number of A. leptodactylus, and they are crucial for the production of higher number of sperm in this species.     

Effect of hydroxyproline supplementation on growth performance,body composition,amino acid profiles,blood‐biochemistry and collagen synthesis of juvenile chu's croaker (Nibea coibor)

Hydroxyproline (Hyp) may play an important role in collagen metabolism, as it is almost exclusively found in collagen. To examine this possibility, a 56‐day feeding trial was conducted to evaluate the effect of graded supplementaries of Hyp (0, 2.5, 5, 7.5, 10 and 12.5 g/kg dry diet weight) on growth performance, feed utilization, biochemical composition and collagen synthesis of chu's croaker, Nibea coibor (13.6 ± 0.28 g). The growth performance and feed utilization were improved with increased levels of dietary Hyp, reaching a maximum at 5 g/kg (diet H2) followed by a slight decrease. Polynomial regression analysis showed that the maximum specific growth rate (SGR) was attained at 7.177 g/kg dietary Hyp. Increasing levels of Hyp did not affect body composition of juvenile Nibea coibor. However, dietary Hyp supplementation significantly affected the levels of Leu, Ile, Met, Lys, Ser, Glu and Pro in muscle and the levels of Met, Lys, Glu, Pro and Gly in swim bladder. The serum calcium (SC) level decreased significantly with increasing levels of dietary Hyp. However, the triglycerides (TG) and total cholesterol (T‐CH) levels in serum and liver were relatively stable. Importantly, the acid‐soluble collagen (ASC) and total collagen (TC) levels in swim bladder increased significantly initially and then decreased after reaching a maximum. Polynomial regression analysis revealed that the maximum TC level in swim bladder was attained at 9.736 g/kg of dietary Hyp. Collectively, these results suggest that the appropriate provision of hydroxyproline is essential for maximal collagen synthesis and growth.     

The effects on some non‐enzymatic antioxidants and oxidative stress of Astacus leptodactylus (Esch., 1823) of starvation periods

This study investigated the effect of starvation (78 days) and refeeding (33 days) on the oxidative stress [malondialdehyde (MDA)] and the non‐enzymatic antioxidants [vitamin E (VE), vitamin C (VC), vitamin A (VA), beta carotene (βC) and reduced glutathione (GSH)] in the hepatopancreas, muscle and gill tissues of freshwater crayfish (Astacus leptodactylus). Crayfish were divided into three experimental groups: control (fed), starved (not fed) crayfish for 78 days and refeeding crayfish for 33 days after 78 days of starvation. The biochemical analysis of the tissues was conducted at 3, 18, 33, 48, 63 and 78 days of starvation and feeding and at 3, 18 and 33 days of refeeding. It was determined that crayfish can withstand starvation period of 78 days. In all of the periods, the MDA levels were significantly higher in the tissues of starved crayfish when compared with the control. The findings of this study demonstrate that starvation has a negative effect on the VE, VC, VA, βC and GSH levels in the crayfish. The measured parameters returned to control values after 33 days of the refeeding. Additionally, the starvation resulted in decreased levels of VE, VA and βC in the abdomen muscle of crayfish consumed by humans.     

Effect of dietary phosphorus sources and varying levels of supplemental phosphorus on survival,growth and body composition of postlarval shrimp (Litopenaeus vannamei)

Two experiments were conducted for 30 days each to investigate the effective phosphorus source and supplemental phosphorus levels for postlarval Litopenaeus vannamei. The first experiment was performed in postlarval shrimp (mean initial wet weight 2 mg) fed four isoenergic and isonitrogenous diets containing three supplemented inorganic phosphorus sources [D1: no supplemental phosphorus, D2: NaH₂PO₄·2H₂O, D3: KH₂PO₄·2H₂O, D4: Ca(H₂PO₄)₂·2H₂O]. The quantities of the three supplemental NaH₂PO₄·2H₂O, KH₂PO₄·2H₂O and Ca(H₂PO₄)₂·2H₂O were 11.6, 12.8 and 10 g kg^?1 of the diet, respectively in order to make the three diets have the same total phosphorus. Growth performance (final mean body weight, FBW; weight gain, WG; specific growth ratio, SGR) of shrimp in D3 treatment was the highest and had significant difference with the D1 treatment. The survival of shrimp in D3 treatment was the highest and had significant difference with the other treatments. The mineral concentration and body composition of shrimp were not significantly different among treatments. We could conclude that KH₂PO₄·2H₂O was the optimal phosphorus source for postlarval L. vannamei from the growth performance and survival. The second experiment was performed in postlarval shrimp (mean initial wet weight 0.88 mg) fed four isoenergic and isonitrogenous diets containing four supplemental KH₂PO₄·2H₂O levels (d1, d2, d3 and d4 with 0, 5, 10 and 20 g kg^?1, respectively). Shrimp in d2 treatment showed the highest growth performance and survival and also showed significant difference with other diet treatments. The whole body content of zinc (Zn) increased with the increase of dietary KH₂PO₄·2H₂O and significant differences were observed when dietary KH₂PO₄·2H₂O reached 5 g kg^?1, excess KH₂PO₄·2H₂O supplementation (10 and 20 g kg^?1) had a negative effect on Zn content, the Zn content significantly decreased when KH₂PO₄·2H₂O was 20 g kg^?1. We can conclude that the amount of total phosphorus in the diet should be maintained between 20.9 and 22.0 g kg^?1, the amount of supplemental KH₂PO₄·2H₂O in the diet is less than 10 g kg^?1.     

Dietary calcium requirement of red swamp crayfish (Procambarus clarkia)

To investigate dietary calcium requirement of red swamp crayfish (Procambarus clarkia), six semi‐purified diets were formulated to contain different concentrations of calcium (2.7(control group), 6.1, 11.9, 17.6, 23.5 and 29.1 g/kg calcium). Each diet was hand‐fed to triplicate of 15 crayfish with average initial body weight (6.22 ± 0.87) g for 8 weeks. The results showed that weight gain rate (WGR) significantly increased and feed conversion ratio (FCR) significantly decreased from 11.9 to 23.5 g/kg groups (p < .05). Protease activities in intestine and hepatopancreas and parathyroid hormone concentrations in serum significantly decreased with increasing dietary calcium levels (p < .05), while calcium and phosphorus contents in exoskeleton, calcium content in muscle and calcitonin concentrations in serum significantly increased (p < .05). The activities of lipase and amylase in intestine and hepatopancreas, serum alkaline phosphatase and total vitamin D concentrations in serum had significant increase as dietary calcium content increased up to 11.9–17.6 g/kg (p < .05). The inorganic phosphorus content in 29.1 g/kg group was significantly lower than those in other groups (p < .05). Broken‐line model analysis based on WGR and quadratic curve model analysis based on FCR showed that optimal dietary calcium requirement of red swamp crayfish ranged from 12.7 to 17.1 g/kg.