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.     

Effect of dietary components on the gut microbiota of aquatic animals. A never‐ending story?

It is well known that healthy gut microbiota is essential to promote host health and well‐being. The intestinal microbiota of endothermic animals as well as fish are classified as autochthonous or indigenous, when they are able to colonize the host's epithelial surface or are associated with the microvilli, or as allochthonous or transient (associated with digesta or are present in the lumen). Furthermore, the gut microbiota of aquatic animals is more fluidic than that of terrestrial vertebrates and is highly sensitive to dietary changes. In fish, it is demonstrated that [a] dietary form (live feeds or pelleted diets), [b] dietary lipid (lipid levels, lipid sources and polyunsaturated fatty acids), [c] protein sources (soybean meal, krill meal and other meal products), [d] functional glycomic ingredients (chitin and cellulose), [e] nutraceuticals (probiotics, prebiotics, synbiotics and immunostimulants), [f] antibiotics, [g] dietary iron and [h] chromic oxide affect the gut microbiota. Furthermore, some information is available on bacterial colonization of the gut enterocyte surface as a result of dietary manipulation which indicates that changes in indigenous microbial populations may have repercussion on secondary host–microbe interactions. The effect of dietary components on the gut microbiota is important to investigate, as the gastrointestinal tract has been suggested as one of the major routes of infection in fish. Possible interactions between dietary components and the protective microbiota colonizing the digestive tract are discussed.     

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.     

Apparent digestibility coefficients of selected protein feed ingredients for loach Misgurnus anguillicaudatus

Apparent digestibility coefficients of dry matter, crude protein, lipid and energy, and amino acids availability in white fish meal, brown fish meal, meat meal, fermented soybean meal, soybean meal and rapeseed meal were determined for loach (Misgurnus anguillicaudatus) (12.05 ± 0.21 g), using a reference diet with 5 g kg^?1 chromic oxide and test diets that contained 700 g kg^?1 reference diet, by weight, and 300 g kg^?1 of the test feed ingredients. The juvenile loach was held in 300‐l tanks at a density of 30 fish per tank. White fish meal, brown fish meal, meat meal and fermented soybean meal had highest apparent digestibility coefficients (ADCs) of dry matter, crude protein and gross energy among ingredients tested, ranged from 50.4% to 60.9% for dry matter, from 64.6% to 88.4% for crude protein and from 57.9% to 79.0% for gross energy. The apparent digestibility coefficients of dry matter ranged from 61.0% to 66.9% for animal products and 50.4% to 60.7% for plant products. For crude protein, apparent digestibility coefficients of protein exceeding 80% were observed for white fish meal, brown fish meal, meat meal and fermented soybean meal, and the apparent digestibility coefficients of protein in rapeseed meal were the lowest among all the treatments. Lipids from both animal and plant feedstuffs were poorly digested by loach, ranging from 64.0% to 77.6%. The apparent digestibility coefficients of energy were similar to those of dry matter and protein, and the highest and lowest ADCs of energy were found in WFM and RM, respectively. The loach used dietary phosphorus from the animal feedstuffs more efficiently than from plant feedstuffs (soybean meal and rapeseed meal), with ADC‐values ranging from 42.3% to 53.1% and from 25.1% to 32.7%, respectively. For the animal products, the availabilities of amino acids in white fish meal and brown fish meal were higher than that in meat meal, expect for Met, Asp, Pro, Gly, and Cys. Among all the plant products, the availabilities of amino acids in fermented soybean meal were higher than in soybean meal and rapeseed meal, and thus had a greater potential to be used as a dietary replacement of fish meal in loach diets.     

Gene Expression in Black Tiger Prawns Following Intramuscular Injection of β-gal Plasmid

Gene expression in black tiger prawns (Penaeus monodon) was studied following intra-muscular injection of CMVGal plasmid into the second abdominal segment. We used an in situ staining technique to detect -gal expression in one- and three-month-old injected prawns. We found that only one of the three-month-old prawns expressed the marker gene (2 days after injection), and the site of expression was confined to the sixth abdominal segment away from the injection site. We repeated the experiment on a new batch of three-month-old prawns, but using fluorometric determination technique. This time we found that -gal expression was detected (6/42) at the site of injection after 2, 7, and 14 days. In two other test samples, transgene expression was detected in the sixth abdominal segment only, further confirming the possibility of injected DNA dispersal. The results of the study also suggest that direct gene transfer is a feasible technique in black tiger prawns.     

Apparent digestibility coefficients of selected feed ingredients for juvenile snakehead,Ophiocephalus argus

Apparent digestibility coefficients (ADCs) of dry matter, crude protein, crude lipid, gross energy, phosphorus and amino acids in Peruvian fish meal, poultry by‐product meal, meat and bone meal, spray‐dried blood meal, hydrolysed feather meal, corn gluten meal, soybean meal, peanut meal, cottonseed meal and rapeseed meal were determined for juvenile snakehead (Ophiocephalus argus) with initial mean body weight of 78.1 g. A reference diet and test diets that consisted of a 70 : 30 mixture of the reference diet to test ingredient were used with 5 g kg^?1 Cr₂O₃ as an external indicator. Fish meal, poultry by‐product meal and corn gluten meal had higher ADCs of dry matter, crude protein, and gross energy among ingredients tested. Dry matter ADCs ranged 61.9–81.5% for animal ingredients and corn gluten meal and ranged 52.2–68.0% for soybean meal, peanut meal, cottonseed meal and rapeseed meal. Energy ADCs of ingredients followed similar trends to differences in dry matter digestibility. Protein ADCs of animal and plant ingredients ranged 73.6–92.8% and 75.3–85.6%, respectively. Amino acid ADCs generally reflected protein digestibility. Lipid ADCs were relatively high for the ingredients tested. Phosphorus ADCs of animal and plant ingredients ranged 39.5–65.2% and 38.7–57.1%, respectively.     

Biochemical,Physical, Chemical,and Microbiological Assessment of Blue Shrimp (Litopenaeus stylirostris) Stored in Ice

Postmortem changes in blue shrimp (Litopenaeus stylirostris) muscle were studied on the basis of biochemical, chemical, physical, and microbiological changes during an 18 day storage period at 0°C. Adenosine 5′-triphosphate (ATP) content, breakdown products, K-value, pH, trimethylamine nitrogen (TMA-N), total volatile basic nitrogen (TVB-N), water holding capacity (WHC), color, and texture (shear force) changes were examined. Also, total mesophilic and psychrophilic bacterial counts were measured. K-value increased linearly (r² = 0.98) from an initial value of 1.37 ± 0.59 to 59.42 ± 6.05% at Day 18. Spoilage indicators TVB-N and TMA-N increased from 29.56 ± 1.33 and 0.69 ± 0.25 to 39.04 and 2.04 ± 0.59 mg of N/100 g at Day 18, respectively; meanwhile, the total viable counts of mesophilic and psychrophilic bacteria increased from 3.48 ± 0.44 and 2.61 ± 0.29 log CFU/g to 6.27 ± 0.21 and 7.14 ± 0.39 log CFU/g, respectively, which indicated spoilage at the end of the storage period. The pH, texture, WHC, and color were affected (p < 0.05) during the storage period. Overall, results indicate that blue shrimp muscle quality was maintained for 12 days of storage in ice.     

Dietary α‐linolenic acid affects lipid metabolism and tissue fatty acid profile and induces apoptosis in intraperitoneal adipose tissue of juvenile grass carp (Ctenopharyngodon idella)

A 56‐day feeding trial was conducted to elucidate the effects and mechanism action of dietary α‐linolenic acid (ALA, 18:3n‐3) on lipid accumulation and fatty acid profile of muscle, hepatopancreas and intraperitoneal fat (IPF) in juvenile grass carp using three isonitrogenous and isoenergetic semi‐purified diets containing 0.0% (control group), 1.0% and 2.0% ALA, respectively. The lowest intraperitoneal fat (IPF) ratio was found in 2.0% group. In the muscle, hepatopancreas and IPF, docosahexaenoic acid (DHA, 22:6n‐3) and eicosapentaenoic acid (EPA, 20:5n‐3) contents increased with the increase in dietary ALA. In the IPF, caspase 3, caspase 8 and caspase 9 showed the highest activities in 2.0% group, while the value of Bcl‐2/Bax (B‐cell leukaemia 2/Bcl‐2‐associated X protein) reached the lowest. Meanwhile, swelling of the IPF mitochondria was observed in 2.0% group. The gene expressions of fatty acid desaturase (FAD) and fatty acid elongase (ELO) in the hepatopancreas and muscle showed significantly higher levels in the treatment groups, whereas an opposite trend was existed in the IPF. Fatty acid synthase (FAS), sterol regulatory element binding protein‐1c (SREBP‐1c) in the IPF and hepatopancreas reached the lowest in 2.0% group. Overall, dietary ALA could promote n‐3 highly unsaturated fatty acids (HUFAs) synthesis and suppress the accumulation of lipid by decreasing the expression of related genes and promoting the apoptosis in IPF.     

Effects of dietary live or heat‐inactivated autochthonous Bacillus pumilus SE5 on growth performance,immune responses and immune gene expression in grouper Epinephelus coioides

To evaluate the possible dietary application of live and heat‐inactivated probiotic Bacillus pumilus SE5 in grouper Epinephelus coioides, juveniles (14.6 ± 0.2 g) were fed either a basal control diet (without probiotic) or the basal diet supplemented with 1.0 × 10⁸ CFU g^?1 live (T1) and heat‐inactivated B. pumilus SE5 (T2). The heat‐inactivated probiotic significantly improved the final weight, weight gain (WG) and specific growth rate (SGR) at day 60 and significantly decreased the feed conversion ratio (FCR) at day 30 and 60, while the viable probiotic significantly decreased the FCR at day 60 (P < 0.05). Phagocytic activity, serum complement C3 and IgM levels as well as SOD activity elevated significantly in fish fed the heat‐inactivated probiotic for 60 days (P < 0.05). Furthermore, the heat‐inactivated probiotic remarkably up‐regulated expression of TLR2 and pro‐inflammatory cytokines (IL‐8 and IL‐1β) in head kidney (P < 0.05), but the viable probiotic failed to do so. These results indicated that heat‐inactivated B. pumilus SE5 can effectively improve the growth performance and immune responses of E. coioides.     

A SCAR marker linked to the N gene for resistance to root knot nematodes (Meloidogyne spp.) in pepper (Capsicum annuum L.)

The root-knot nematodes (Meloidogyne spp.) are important nematode pests and cause serious diseases in pepper in the world. No molecular markers linked to the nematodes resistance N gene have been reported. In this paper, ‘Carolina Wonder’ (Capsicum annuum L.), a sweet pepper line resistant to root-knot nematode with N gene, ‘20080-5-29’ (C. annuum L.), an inbred line susceptible to root-knot nematode with good horticultural characteristics, and their F₂ progeny with 320 individuals were used as materials. Evaluation of resistance and susceptibility of parental lines, F₁ and F₂ progeny inoculated with root-knot nematodes (Meloidogyne incognita) were carried out. ‘Bulked segregant analysis’ method was used to search for polymorphic markers from 512 pairs of AFLP primers. Based on the assessment of resistance and susceptibility and polymorphism of the AFLP marker in F₂ population, the genetic linkage distance between the AFLP marker and the N gene was estimated. One AFLP marker E39/M41-339 was obtained and transferred to a SCAR marker amplifying a 315 bp DNA fragment linked to the N resistant allele and a 331 bp fragment linked to the N⁺ susceptible allele. The distance between the molecular marker and the nematodes resistance N gene is 6.3 cM. This research delivered a valuable tool for the marker assisted selection of nematodes resistance in pepper.