Determination of allogeneic and xenogeneic markers in the genus of Tilapia. II. Identification of T. aurea,T. vulcani and T. nilotica by electrophoretic analysis of their serum proteins

Sera from Israeli stocks of T. nilotica, T. vulcani and T. aurea were analysed, by polyacrylamide gel electrophoresis, for the existence of species-specific markers. Markers situated in the fast region of the electropherogram were found to be male sex and species specific. Interspecies polymorphism in markers in the post albumin area (transferrins) permit the distinction between these related species of Tilapia. No significant polymorphism was found in the electropherogram of T. vulcani. T. aurea presented some slight variations in the transferrin zone, and T. nilotica displayed a high polymorphism everywhere along the electropherogram. Two other species specific bands were found in the prealbumin zone; bands 1 (specific for T. aurea) and 2 (specific for T. vulcani). A selection of fish stocks of T. aurea and T. vulcani is currently done now on the basis of this electrophoretic criteria and on the ability to produce a high percentage of male hybrids. It was concluded that the stock of T. nilotica which exist in Israel do not represent the pure species of T. nilotica. The failure to obtain 100% males in all paired interspecific mating (Pruginin et al., 1975) was explained on this basis.     

Physiological investigations of a neurotoxin-producing phytoflagellate, Chattonella marina (Raphidophyceae)

The effects of temperature, salinity, light intensity and pH on the growth and morphology of Chattonella marina (Subrahmanyan) Hara & Chihara were examined. Optimal growth was observed at temperatures of 20-25°C, salinities of 20-30%o, light intensities of 60-140 μE m^?2 s^_1 and pH 7.5-8.5. Growth did not occur at temperatures below 15°C or above 30°C, and at salinities below 10%o. The morphology (shape) of the cells was strongly affected by temperature. At 20°C and 25°C, the population occurred mostly in a spindle-like form, whereas at 10°C, 90% of the cells became spherical within 10 days of inoculation and stationary phase cultures consisted entirely of spherical cells. Morphology was also markedly affected at 30°C. The number of spindle-like cells was highest at 20-30%o and was less at lower salinities. Light intensity and pH did not influence morphology markedly under the range of light intensities (20-180 μE m ^?2 s-^?1) and pH (6.5-8.5) tested.     

Effect of dietary protein level on the reproductive performance of Nile tilapia, Oreochromis niloticus (L.)

The effects of five dietary protein levels (25%, 30%, 35%, 40% and 45%) on the individual spawning frequency and the egg production of 135 tagged female Nile tilapia, Oreochromis niloticus (L.), were studied from 8 May to 19 November 1996 in outdoor concrete tanks. Virtually identical spawning patterns were found in all treatments, but there was a great deal of variation in the frequency of spawning and egg production. Overall, individual spawning frequency varied from one to 14 and individual egg production from 31 to 2828 eggs per spawning. The average number of spawnings and average number of eggs per spawning for fish receiving dietary protein levels of 25%, 30%, 35%, 40% and 45% were 8.0 ±1.6, 8.4 ± 2.2, 8.5 ± 2.7, 8.4 ± 2.5 and 9.4 ± 2.1 spawnings, and 1167 ± 305, 1082 ± 410, 1288 ± 324, 1145 ± 389 and 1328 ± 311 eggs per spawning, respectively. Fish receiving 45% dietary protein spawned more frequently than fish receiving 25% dietary protein. The total number of eggs produced per female was significantly higher for females fed 45% protein feed than females receiving 25% and 30% protein feeds. No definite trend was found in the number of eggs produced per spawning and the number of eggs produced per gram in females fed at different protein levels. Based on weekly checking, the time interval between spawnings varied from 7 to 77 days, and mean spawning intervals ranged from 15.8 to 17.1 days. Sixty per cent of females spawned after an interval 14 days, 15% after 21 days, 13.6% after 7 days, 7.2% after 28 days, 1.8% after 35 days and 1.0% after 42 days, and the time interval was 49-77 days for the rest of the females (1.5%). In all treatments, maximum spawning activity was recorded from May to August, and thereafter, it gradually decreased and no spawning females were found in November.     

Ultraviolet-induced androgenesis in Nile tilapia, Oreochromis niloticus (L.), and hybrid Nile × blue tilapia, O. aureus (Steindachner)

Fertilization of ultraviolet (UV) irradiated oocytes of Nile tilapia, Oreochromis niloticus (L.), with sperm from O, niloticus or blue tilapia, O, aureus (Steindachner), and subsequent suppression of the first cleavage of fertilized eggs successfully induced androgenesis in Nile and blue tilapia. The optimal doses of UV irradiation to denucleate a female genome of Nile tilapia prior to androgenesis ranged from 5940 to 6930 erg mm^?2 for 54-63 s at a fixed intensity of 110 erg mm^?2 s^?1. Putative androgenetic fish were created from eggs which were irradiated at various times and several durations of heat-shock. Eggs which were treated for 5 min at 41.6 ^oC at 2 7.5 min after fertilization were the most successful at suppressing the first cleavage and producing viable androgenetic diploids in Nile or hybrid Nile X blue tilapia. The maximal survivals of putative androgenetic diploids in relation to the control were 1.60% and 0.90% in Nile and hybrid Nile X blue tilapia, respectively. The androgenetic offspring established exhibited active feeding and normal growth.     

Role of T-bet in commitment of TH1 cells before IL-12-dependent selection

How cytokines control differentiation of helper T (TH) cells is controversial. We show that T-bet, without apparent assistance from interleukin 12 (IL-12)/STAT4, specifies TH1 effector fate by targeting chromatin remodeling to individual interferon-gamma (IFN-gamma) alleles and by inducing IL-12 receptor beta2 expression. Subsequently, it appears that IL-12/STAT4 serves two essential functions in the development of TH1 cells: as growth signal, inducing survival and cell division; and as trans-activator, prolonging IFN-gamma synthesis through a genetic interaction with the coactivator, CREB-binding protein. These results suggest that a cytokine does not simply induce TH fate choice but instead may act as an essential secondary stimulus that mediates selective survival of a lineage.     

Cloning of bovine CD69

CD69 is rapidly inducible on various hematopoietic cells upon stimulation and is detectable as an early activation antigen. Although CD69 is well characterized in human and mouse, no information is available on bovine CD69. We report here that, bovine CD69 was cloned from a cDNA expression library prepared from activated peripheral blood lymphocytes. The full-length cDNA contained an 80bp 5' untranslated region, followed by a 600bp coding region and AU-rich motifs in a 3' untranslated region (GenBank accession number AF272828). Comparison of the bovine CD69 coding sequence reveals 69.4 and 78.2% nucleotide sequence identities with mouse and human CD69, respectively. The predicted amino acid sequence of bovine CD69 shares 56.3 and 62.3% sequence identity when compared with mouse and human CD69, respectively. Bovine CD69 has the highly conserved amino acid sequences found in the C-type lectin family, suggesting that the conserved residues may be important for conformation and binding to the, as yet unidentified ligand. In addition, the cytoplasmic tail of bovine CD69 has two casein kinase-2 (CK-2) phosphorylation sites. These data suggest that bovine CD69 plays an important role in the activation of lymphocytes.     

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.     

Effect of dietary carbohydrate‐to‐lipid ratios on growth performance,body composition,nutrient utilization and hepatic enzymes activities of herbivorous grass carp (Ctenopharyngodon idella)

Six isonitrogenous (390 g kg^?1) and isoenergetic (16.2 kJ g^?1) diets with varying carbohydrate : lipid (CHO : L) ratios (202.5–1.74), were fed to triplicate groups of 25 fish in indoor recirculation system. Over 8‐week‐growth trial, best weight gain (WG), specific growth rate, feed conversion ratio, protein efficiency ratio and protein production value (P < 0.05) were observed in fish‐fed diets with CHO : L ratio of 7.5. Fish fed either the lowest (1.7) or highest (202.5) CHO : L ratio tended to produce lower (P < 0.05) growth and feed conversion efficiencies. The values of viscerosomatic index, hepatosomatic index and intraperitoneal fat ratio increased as dietary CHO : L ratios decreased. There were no significant differences in whole body and liver crude protein among dietary treatments. Whole body and liver lipid increased as CHO : L ratios decreased. Plasma cholesterol and triacylglyceride levels increased linearly as dietary CHO : L ratios decreased. Activities of glucokinase and pyruvate kinase were stimulated by elevated levels of dietary carbohydrate; however, activities of lipase (LPS) and alkaline phosphatase were stimulated by elevated levels of dietary lipid. Based on a second‐order polynomial regression analysis of WG against dietary carbohydrate and lipid levels, 275 g kg^?1 of carbohydrate and 59 g kg^?1 of lipid, corresponding to a CHO : L ratio of 4.7, in a diet holding 390 g kg^?1 of crude protein and 16.3 kJ g^?1 of gross energy, proved to be optimal for grass carp. These results indicated that utilization of dietary lipid and carbohydrate was moderate in grass carp, but the fish were a little more capable of utilizing lipid compared with carbohydrate.     

Prevalence and different characteristics of two serotypes of Streptococcus parauberis isolated from the farmed olive flounder, Paralichthys olivaceus (Temminck and Schlegel), in Korea

The prevalence of two serotypes of Streptococcus parauberis isolated from the olive flounder, Paralichthys olivaceus, was evaluated in a total of 29 isolates between 2003 and 2010 in Korea. Streptococcus parauberis isolates were divided into two serologically distinct types (serotype 1 and serotype 2), except for one strain (S1091), using an agglutination assay with rabbit antiserum, and serotype 1 was identified as the dominant type (24 of 29 isolates) in this study. To identify the characteristics of the two serotypes of S. parauberis, we conducted a biochemical test using the API 20 Strep kit, a transmission electron microscopy (TEM) assay, sequence analysis of 16S‐23S rRNA intergenic spacer region (ISR) and a pathogenicity test. In TEM, both serotypes possessed polysaccharide capsule layers around the cell surface when bacterial cells were treated with a homologous serotype of rabbit antiserum. However, we were unable to discriminate serotype‐specific biochemical characteristics and genetic characteristics of 16S‐23S rRNA ISR between the two serotypes. In the pathogenicity test, the serotype 1 strains induced significantly higher mortality than the serotype 2 strains in olive flounder when experimentally inoculated via the intraperitoneal route.