Effects of lysine and methionine supplementation on growth,body composition and digestive function of grass carp (Ctenopharyngodon idella) fed plant protein diets using high‐level canola meal

A 76‐day feeding trial was carried out to evaluate the effects of Lysine and Methionine supplementation on growth and digestive capacity of grass carp (Ctenopharyngodon idella) fed plant protein diets using high‐level canola meal (CM). Fish with initial average weight 103.9 ± 0.6 g were fed three extruded diets. Fish meal (FM) diet was formulated as the normal control with 40 g kg^?1 FM and 300 g kg^?1 CM; CM diet was prepared by replacing all FM with CM (total 340 g kg^?1) without Lys or Met supplementation; CM supplement (CMS) diet was similar to CM diet but was supplemented with essential amino acids (EAA) to ensure the levels of Lys and Met similar to those in the FM diet. Feed intake, feed efficiency and specific growth rate of the grass carp fed CMS and FM diets were similar (P > 0.05), but higher than those of the grass carp fed CM diet (P < 0.05). The hepatosomatic index, relative gut length, intestosomatic index and intestinal folds height were significantly improved in fish fed FM and CMS diets as compared to CM diet (P < 0.05). Lower activities of trypsin, lipase and amylase in hepatopancreas were observed in fish fed CM diet (P < 0.05). Three hundred and forty gram per kilogram CM without Lys or Met supplementation significantly decreased trypsin, lipase and amylase mRNA levels in hepatopancreas (P < 0.05). These results indicated that the high supply of CM (340 g kg^?1) in plant protein (200 g kg^?1 soybean meal and 100 g kg^?1 cottonseed meal) diets decreased digestive ability through decreasing digestive enzyme activities and enzyme gene's expressions of grass carp, and these side effects can be reversed by supplementing Lys and Met. Therefore, CM could be high level used in a plant protein blend‐based extruded diet for grass carp as long as EAA were supplemented.     

Apparent digestibility coefficients of animal and plant feed ingredients for juvenile Pseudobagrus ussuriensis

A study was conducted to determine apparent digestibility coefficients (ADCs) of dry matter (DM), crude protein (CP), lipid, gross energy, phosphorus and amino acids (AAs) from Peruvian fish meal (PFM), native fish meal (NFM), meat bone meal (MBM), dried daphnia meal (DDM), soybean meal (SBM), full‐fat soybean (FSBM), cottonseed meals (CSM), rapeseed meal (RSM) and corn gluten meal (CGM) for Pseudobagrus ussuriensis (7.8 ± 0.06 g). Test diets were formulated to contain a 70:30 mixture of reference diet to test ingredient with chromic oxide (5 g kg^–1) serving as the inert marker. ADCs of DM ranged from 49.94% (CGM) to 87.11% (PFM), the highest ADCs of DM were observed in PFM and the lowest was found in CGM. ADCs of CP in the test ingredients ranged from 74.92% (CGM) to 93.41% (SBM). ADCs of crude lipid ranged from 51.80% (CGM) to 94.07% (PFM) for fish. ADCs of energy ranged from 56.42% (CGM) to 90.09% (PFM). ADCs of phosphorus ranged from 21.81% (CSM) to 65.70% (PFM). AA availability for the test ingredients followed a similar pattern to ADCs of CP, in which values for PFM, NFM and MBM were generally higher (p < .05) than those of other protein ingredients. Among all plant meals, the AA availability values in SBM were higher (p < .05) than those in CGM, CSM and RSM. The availability of most AAs in CGM was the lowest (p < .05) among all tested ingredients. In conclusion, PFM and NFM are good sources of available protein and AAs. When fishmeal supply is limited, MBM and DDM are substitutes for fishmeal reduction in P. ussuriensis diets; at the same time, SBM and FSBM are also very acceptable protein feedstuffs. However, CSM, RSM and CGM could not be used at high levels in P. ussuriensis diets because of their inferior digestibilities.     

Effect of dietary protein reduction with lysine and methionnine supplementation on growth performance,body composition and total ammonia nitrogen excretion of juvenile grass carp,Ctenopharyngodon idella

A 63‐day growth trial was undertaken to estimate the effects of supplemented lysine and methionine with different dietary protein levels on growth performance and feed utilization in Grass Carp (Ctenopharyngodon idella). Six plant‐based practical diets were prepared, and 32CP, 30CP and 28CP diets were formulated to contain 320 g kg^?1, 300 g kg^?1 and 280 g kg^?1 crude protein without lysine and methionine supplementation. In the supplementary group, lysine and methionine were added to formulate 32AA, 30AA and 28AA diets with 320 g kg^?1, 300 g kg^?1 and 280 g kg^?1 dietary crude protein, respectively, according to the whole body amino acid composition of Grass Carp. In the groups without lysine and methionine supplementation, weight gain (WG, %) and specific growth rate (SGR, % day^?1) of the fish fed 32CP diet were significantly higher than that of fish fed 30CP and 28CP diets, but no significant differences were found between 30CP‐ and 28CP‐diet treatments. WG and SGR of the fish fed 32AA and 30AA diets were significantly higher than that of fish fed 28AA diets, and the performance of grass carp was also significantly improved when fed diets with lysine and methionine supplementation (P < 0.05), and the interaction between dietary protein level and amino acid supplementation was noted between WG and SGR (P < 0.05). Feed intake (FI) was significantly increased with the increase in dietary protein level and the supplementation of lysine and methionine (P < 0.05), but feed conversion ratio (FCR) showed a significant decreasing trend (P < 0.05). Two days after total ammonia nitrogen (TAN) concentration test, the values of TAN discharged by the fish 8 h after feeding were 207.1, 187.5, 170.6, 157.3, 141.3 and 128.9 mg kg^?1 body weight for fish fed 32CP, 32AA, 30CP, 30AA, 28CP and 28AA diets, respectively. TAN excretion by grass carp was reduced in plant‐based practical diets with the increase in dietary protein level and the supplementation of lysine and methionine (P < 0.05). The results indicated that lysine and methionine supplementation to the plant protein sources‐based practical diets can improve growth performance and feed utilization of grass carp, and the dietary crude protein can be reduced from 320 g kg^?1 to 300 g kg^?1 through balancing amino acids profile. The positive effect was not observed at 280 g kg^?1 crude protein level.     

Effects of dietary thiamin supplement on growth,body composition and intestinal enzyme activities of juvenile Jian carp (Cyprinus carpio var. Jian)

A total of 1050 juvenile Jian carp (Cyprinus carpio var. Jian) (8.20 ± 0.02 g) were fed diets containing seven graded levels of thiamin (0.25, 0.48, 0.79, 1.06, 1.37, 1.63 and 2.65 mg kg^?1) for 60 days to investigate the effects of thiamin on growth, body composition and digestive enzyme activities. Percent weight gain (PWG), feed intake and feed efficiency (FE) were the lowest in fish fed the basal diet (P < 0.05). Protein productive value and lipid productive value increased with increasing dietary thiamin levels up to 1.06 and 0.79 mg kg^?1 diet, respectively (P < 0.05). Body protein and lipid increased with increasing dietary thiamin levels (P < 0.05), while moisture and ash of fish carcasses decreased with the increase in dietary thiamin supplementation (P < 0.05). Intestinal folds height had a similar trend to PWG (P < 0.05). Activities of α‐amylase, lipase, trypsin, Na⁺, K⁺‐ATPase, alkaline phosphatase and gamma‐glutamyl transpeptidase in intestine were all affected by the dietary thiamin (P < 0.05). In conclusion, thiamin could improve growth and intestinal enzyme activities of juvenile Jian carp. The dietary thiamin requirement of juvenile Jian carp (8.0–60.2 g) based on PWG was 1.02 mg kg^?1 diet.     

Seminal plasma composition and its relationship with physical spermatological parameters of Grass carp (Ctenopharyngodon idella) semen: with emphasis on sperm motility

In this research, the mineral and organic composition of the seminal plasma, physical spermatological parameters and their physiological relationships were investigated in grass carp (Ctenopharyngodon idella). The seminal plasma contained 98.14±5.23 mM L^?1 (Na⁺), 380.85±25.95 mM L^?1 (K⁺), 30.25±4.96 mg dL^?1 (Ca²⁺), 19.16±1.70 mEq L^?1 (Mg²⁺), 1.36±0.11 mg dL^?1 glucose, 0.37±0.08 g dL^?1 total protein, 12.02±1.18 mg dL^?1 cholesterol, 14.85±1.50 mg dL^?1 triglyceride and 43.5±9.56 mg dL^?1 urea. The following spermatological parameters were found: sperm volume 14.44±1.16 mL, sperm motility 80.60±1.55%, movement duration 67.68±4.32 s, density 15.43±0.72 × 10⁹ mL^?1, total density 337.43+45.86 × 10⁹ and pH 7.24±0.17. The Na⁺ and Ca²⁺ ions correlated negatively with spermatozoa motility (r=?0.453, P>0.05 and r=?0.192, P>0.05) respectively. The K⁺ ion correlated positively with spermatozoa motility (r=0.545, P>0.05). But a statistically significant correlation was not observed between sperm motility and seminal plasma parameters. The following correlations were observed between mineral and organic components. The Mg²⁺ was positively correlated with glucose and cholesterol (r=0.692, P<0.05 and r=0.680, P<0.05) respectively. A highly significant positive relationship was also found between Mg²⁺ and total protein (r=0.837, P<0.01). On the other hand, a significantly negative relationship was found between Ca²⁺ and triglyceride (r=?0.639, P<0.05). These parameters should be considered when developing procedures for either artificial fertilization or for cryopreservation of grass carp sperm.     

The pathology of selenium deficiency in Cyprinus carpio L

Selenium is essential for the normal life processes, and all animals, including fish, need this inorganic element. In order to research the pathology of selenium deficiency in common carp, Cyprinus carpio L., 360 juvenile carp were allocated to three treatments. Each treatment containing 120 fish was randomly divided into four groups, fed with purified diets containing selenium at 0, 0.15, 0.30 and 0.45 mg kg^?1. The results indicated that the morbidity and mortality rates of the test groups were negatively correlated with the level of selenium in the diets. The morbidity and mortality rates were, respectively, 46.7%, 33.3%, 13.3%, 0 and 26.7% 16.7%, 6.7%, 0. The affected fish exhibited pathological changes, such as ‘thin back disease’ and lordosis. Histologically, there was no abnormality observed in muscles, liver and pancreas. Ultrastructurally, there was swelling of the mitochondria of the cardiac muscle cells, the liver cells and renal tubular epithelial cells, with disintegration and lysis of the cristae of the mitochondria which vesiculated. Serum glutamic oxaloacetic transaminase and glutamic pyruvic transaminase levels were increased and the activity of the serum glutathione peroxidase and the serum superoxide dismutase was decreased; in contrast, the serum malonaldehyde concentration was increased.     

Effects of Cu2+-exchanged montmorillonite on intestinal microflora, digestibility and digestive enzyme activities of Nile tilapia

A total of 360 Nile tilapia at an average initial body weight of 3.9 g were randomly allocated to four treatments, each of which had three replicates of 30 fish per tank and used to investigate the effects of Cu²⁺‐exchanged montmorillonite (Cu‐MMT) on intestinal microflora, digestibility and digestive enzyme activities. The dietary treatments were: (i) basal diet, (ii) basal diet + 1.5 g kg^?1 MMT; (iii) basal diet + 30 mg kg^?1 copper as CuSO₄ (equivalent to the copper in the Cu‐MMT treatment group), or (iv) basal diet + 1.5 g kg^?1 Cu‐MMT. The results showed that supplementation with Cu‐MMT significantly improved growth performance as compared with the control and fish fed with Cu‐MMT had higher growth performance than those fed with MMT or CuSO₄. Supplementation with Cu‐MMT reduced (P < 0.05) the total intestinal aerobic bacterial counts and affected the composition of intestinal microflora with Aeromonas, Vibrio, Pseudomonas, Flavobacterium, Acinetobacter, Alcaligence, Enterobacteriaceae decreasing as compared with the control. Addition of MMT improved (P < 0.05) the apparent digestibility of dry matter and crude protein, and the activities of intestinal amylase, lipase, and alkaline phosphatase as compared with the control. Addition of Cu‐MMT improved (P < 0.05) the apparent digestibilities of dry matter, crude protein, ether extract, and crude ash, and the activities of total protease, amylase, lipase, and alkaline phosphatase as compared with the control. While supplementation with CuSO₄ had no (P > 0.05) effect on intestinal microflora, digestibility, and digestive enzyme activities the results showed that Cu‐MMT exhibited antibacterial activity in vivo resulting in a positive effect on digestive enzyme activities, and then promoted the digestion and absorption of dietary nutrition.     

Cloning and expression analysis of innate immune genes from red sea bream to assess different susceptibility to megalocytivirus infection

As suggested by the Office International des Epizooties (OIE), fishes belonging to the genus Oplegnathus are more sensitive to megalocytivirus infection than other fish species including red sea bream (Pagrus major). To assess the roles of the innate immune response to these different susceptibilities, we cloned the genes encoding inflammatory factors including IL‐8 and COX‐2, and the antiviral factor like Mx from red sea bream for the first time and performed phylogenetic and structural analysis. Analysed expression levels of IL‐1β, IL‐8 and COX‐2 and the antiviral factor like Mx genes performed with in vivo challenge experiment showed no difference in inflammatory gene expression or respiratory burst activity between red sea bream and rock bream (Oplegnathus fasciatus). However, the Mx gene expression levels in red sea bream were markedly higher than those in rock bream, suggesting the importance of type I interferon (IFN)‐induced proteins, particularly Mx, during megalocytivirus infection, rather than inflammation‐related genes. The in vitro challenge experiments using embryonic primary cultures derived from both fish species showed no difference in cytopathic effects (CPE), viral replication profiles, and inflammatory and Mx gene expression pattern between the two fish species.