Non-specific immunity parameters and the formation of advanced glycosylation end-products (AGE) in rainbow trout, Oncorhynchus mykiss (Walbaum), fed high levels of dietary carbohydrates

The effects of long-term hyperglycaemia (5 months), through feeding high levels of dietary carbohydrates, on the non-specific immunity parameters of rainbow trout, Oncorhynchus mykiss (Walbaum), was assessed. Fish were fed one of three diets composed of a basal diet containing ≈ 14% digestible carbohydrate which was progressively diluted with gelatinized potato starch in the following ratios of basal diet to supplemental carbohydrate (gelatinized potato starch): 65:00, 65:20, 65:35. The three diets were pair-fed based on the feed intake of the fish fed the diet containing the highest level of starch (65:35) and representing 100% intake. The other diets were then fed at different levels in a manner that allowed all the groups of fish in the same block to receive the same amount of the basal diet [e.g. the basal diet (undiluted, 65:00) was fed at 65% of the dietary intake of the 65:35 diet]. Blood glucose concentrations and relative liver-to-body size increased with increased dietary carbohydrate intake. Feeding supplemental carbohydrates resulted in a small increase in weight gain of the fish at both supplemental levels. Pronephros tissue lysozyme activity and pronephros macrophage superoxide production were not affected by the dietary treatments. The results suggest the presence of advanced glycosylation end-products in muscle tissue collagen, but were not significantly different between treatments. No substantial effect of long-term feeding of a high carbohydrate diet on the non-specific immunity of rainbow trout was observed. However, the results suggest that dietary carbohydrates may have a slight stimulatory effect on phagocytosis at low–moderate levels.     

Incorporation of composted and dried water hyacinth in pelleted feed for the tilapia Oreochromis niloticus (Peters)

Abstract. Eight diets were formulated using either composted or dried water hyacinth, Eichornia crassipes (Mart.) Solms., incorporated into a conventional pelleted tilapia feed in a simple displacement procedure: 100. 75, 50 and 25% composted water hyacinth: 100, 75 and 25% dried water hyacinth; and a control diet with 0% displacement of the conventional feed. The experimental diets were fed in duplicate treatments to Oreochromis niloticus in a series of 16 outdoor 4-m³ tanks stocked with eight fish/tank for 84 days. Good growth and feed utilization efficiencies were obtained with diets containing up to 75% composted water hyacinth with no significant reduction in fish performance compared to the control diet. Fish performed better with diets containing composted than with diets containing dried water hyacinth, except for the 100% composted water hyacinth diet in which the pellet was exceptionally hard. The better performance with diets containing composted compared to dried water hyacinth may have been due to the lower levels of crude fibre in the former diets, although significant fish growth was achieved even with the diet containing 100% dried water hyacinth. Some nutrition must have been obtained by the fish indirectly from the plankton in the static water experimental system but it was concluded that the fish also obtained a significant amount of their nutrition directly from the diets, including those with a high percentage of composted water hyacinth which in some treatments had relatively low mean phytoplankton biomass levels in the water. An experimental protocol is recommended for the evaluation of the nutritional value of non-conventional fish feeds in which determination of fish growth without concern for underlying mechanisms precedes studies to assess the relative contribution of the direct and indirect nutritional value of the feed.     

The effects of less-expensive plant protein sources replaced with soybean meal in the juvenile diet of grass carp (Ctenopharyngodon idella): growth, nutrient utilization and body composition

In this study, the effects of less-expensive protein sources replaced with soybean meal on growth performance, nutrient utilization and body composition of juvenile grass carp were investigated. For this purpose, a control diet containing 42% soybean meal was prepared. Cotton seed meal (CSM), sunflower meal (SFM) and corn meal (CM) were added to the experimental diets by replacing 100% of the soybean meal protein used in the control diet. All diets were iso-nitrogenous (40% crude protein) and iso-caloric (19.5?MJ?kg^?1 gross energy). Each treatment had three replicates, and 20 juvenile grass carps (3.5?±?0.1?g, initial weight) were located in per replicate. The fish were fed to satiety three times in a day during 3?months at 26°C of water temperature in glass aquarium (45 l). The results of the study showed that the weight gain (308.6?C448.6%), specific growth rate (1.56?C1.89%/day), feed conversion rate (1.2?C1.9), protein efficiency ratio (1.32?C2.08), and apparent digestibility coefficients of protein (80.5?C89%) or sum of amino acids (80.5?C88.9%) were not significantly affected by the experimental diets, except diet CM. All of these values obtained for the diet CM were significantly lower than those of fish fed on the control, CSM and SFM diets. However, there were no significant differences between the lipid digestibility of the diets, and crude protein, lipid, and gross energy contents in the whole body of fish in experiments. In conclusion, CSM and SFM diets were highly utilized by juvenile grass carp, but not CM diet. These results combined with the remarkable acceptability of diets containing high levels of plant protein ingredients with identical growth performances of juvenile grass carp show clearly that dietary soybean meal level can be considerably reduced without any adverse consequence in terms of somatic growth and nitrogen utilization.     

Infectious haematopoietic necrosis virus from salmonids cultured in Korea

Abstract. Four isolates of infectious haematopoietic necrosis virus (IHNV) were obtained from rainbow trout, Oncorhynchus mykiss (Walbaum), and masu salmon, Oncorhynchus masou (Walbaum), fry during epizootics at hatcheries in Korea. The four isolates of IHNV were compared with three from salmonids in the USA (SRCV, OSV and RB-76) by analysis of virion proteins in sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and neutralization tests, with two monoclonal antibodies raised against SRCV (MAb SRCV/A4) and RB-76 (MAb RB/B5). Based on the antigenicity and the size of the structural proteins, one Korean isolate from masu salmon (SCS) is similar to RB-76 and is an electropherotype 1. The other three isolates from rainbow trout (PRT, YRT and MRT) were identical to each other in the mobilities of their virion proteins in SDS-PAGE, and although their nucleocapsid (N) proteins comigrated with that of the RB-76 isolate, they differed from RB-76 in the smaller matrix 2 (M2) protein they contained. In addition, the three Korean isolates (PRT, YRT and MRT) could be divided into two groups by reactivity with MAb RB/B5. While the YRT isolate was neutralized by this MAb, the PRT and MRT isolates were not, suggesting that there are at least two neutralizing antigenic variants of IHNV in Korea.     

Gonadal sex differentiation and sex control in red sea bream, Pagrus major

The gonadal sex differentiation in red sea bream, Pagrus major, which is one of the most important species for aquaculture in Japan, was revealed histologically. The suitable conditions for induction of all-male groups in the fish were investigated, and functional males were induced by the conditions of oral administration of 17α-methyltestosterone. The sex determination of this fish was also discussed.     

Differentiation and development of Leydig cell, and induction of spermatogenesis during testicular differentiation in the Japanese eel, Anguilla japonica

The initial appearance and the development of Leydig cells (LCs), the sites of steroid hormone production in the testis, were investigated ultrastructurally during testicular differentiation in the Japanese eel, Anguilla japonica. In addition, the effects of a single injection of human chorionic gonadotropin (HCG; 5 IU g body weight^-1) on histological changes of the testes and serum 11-ketotestosterone (11-KT) were examined at various stages (15–18, 20–23, 26–29, 32–35, 38–41 and 46–50 cm body length (BL)) of testicular differentiation. Testicular differentiation was morphologically characterized by the development of loose connective tissue on the medial side in animals 18–29 cm in BL. Ultrastructurally, LCs were first identified in the loose connective tissue of the testis of the 23 cm fish. In the testes of fish over 32 cm, clusters of LCs were distributed throughout the interstitial region accompanying the increase in number of spermatogonia. In fish larger than 32 cm, spermatogenesis was induced by administration of HCG; serum 11-KT levels were also raised. On the other hand, there was no effect on spermatogenesis or serum 11-KT levels in fish less than 29 cm, or in the controls. These result suggests that morphological differentiation of LCs occurs in testis of the 23 cm eel, and subsequently, the testes of eels of BL more than 32 cm acquire the capability to produce steroid hormones.     

Methods of assessing extinction risk in marine fishes

The decline and disappearance of species from large parts of their former geographical range has become an important issue in fisheries ecology. There is a need to identify which species are at risk of extinction. The available approaches have been subject to considerable debate – particularly when applied to commercially exploited species. Here we have compiled methods that have been used or may be used for assessing threat status of marine organisms. We organize the methods according to the availability of data on the natural history, ecology and population biology of species. There are three general approaches to inferring or assessing extinction risk: (i) correlative approaches based on knowledge of life histories and ecology; (ii) time‐series approaches that examine changes in abundance; and (iii) demographic approaches based on age‐ or stage‐based schedules of vital rates and fisheries reference points. Many methods are well suited to species that are highly catchable and/or have relatively low productivity, but theory is less well developed for assessing extinction risk in species exhibiting narrow geographical distributions or ecological specialization. There is considerable variation in both definitions of extinction risk and the precision and defensibility of the available risk assessment methods, so we suggest a two‐tiered approach for defining and assessing extinction risk. First, simple methods requiring a few easily estimated parameters are used to triage or rapidly assess large numbers of populations and species to identify potentially vulnerable populations or species. Second, the populations and species identified as vulnerable by this process can then be subject to more detailed and rigorous population analysis explicitly considering sources of error and uncertainty.     

Dominant red colour morphology used to detect paternal contamination in batches of Oreochromis mossambicus (Peters) gynogens

Abstract. Gynogenetic (meiotic) Oreochromis mossambicus (Peters) lines can be produced easily with the simple ultraviolet (UV) spermatozoan irradiation technique detailed in this study. One hundred per cent haploid gynogens were achieved by eggs fertilized with UV-irradiated (254 nm, 4.2 W/m² for 7 min) red tilapia spermatozoa. Survival of the haploid gynogens were 5% and all haploid fry were deformed. Hybridization between female O. mossambicus and male red tilapia produced 100% red offspring. Thus the red colour can be used as a marker to identify fish that are not gynogens. Activation of eggs with 7-min UV-irradiated spermatozoa from red tilapia and subsequent heat shocking at 42⁰C for 3 min resulted in 100% diploid gynogens (black).     

Ultrastructural morphogenesis of salmonid alphavirus 1

Studies on the ultrastructural morphogenesis of viruses give an insight into how the host cell mechanisms are utilized for new virion synthesis. A time course examining salmonid alphavirus 1 (SAV 1) assembly was performed by culturing the virus on Chinook salmon embryo cells (CHSE‐214). Different stages of viral replication were observed under electron microscopy. Virus‐like particles were observed inside membrane‐bound vesicles as early as 1 h following contact of the virus with the cells. Membrane‐dependent replication complexes were observed in the cytoplasm of the cells, with spherules found at the periphery of late endosome‐like vacuoles. The use of intracellular membranes for RNA replication is similar to other positive‐sense single‐stranded RNA (+ssRNA) viruses. The number of Golgi apparatus and associated vacuoles characterized by ‘fuzzy’‐coated membranes was greater in virus‐infected cells. The mature enveloped virions started to bud out from the cells at approximately 24 h post‐infection. These observations suggest that the pathway used by SAV 1 for the generation of new virus particles in vitro is comparable to viral replication observed with mammalian alphaviruses but with some interesting differences.     

Assessment of clonal fidelity of micropropagated gerbera plants by ISSR markers

True-to-type clonal fidelity is one of the most important pre-requisites in micropropagation of crop species. Genetic fidelity of in vitro raised 45 plants of gerbera (Gerbera jamesonii Bolus) derived from three different explants, viz., capitulum, leaf and shoot tips, was assessed by 32 ISSR markers, for their genetic stability. Out of 32 ISSR markers, 15 markers produced clear, distinct and scorable bands with an average of 5.47 bands per marker. The markers designed from AG motif amplified more number of bands. The markers anchored at 3′ ends produced high number of consistent bands than unanchored markers. Fifteen ISSR markers generated a total of 3773 bands, out of which 3770 were monomorphic among all the clones. The Jaccard's similarity coefficient revealed that out of 45 clones derived from different explants, 44 were grouped into a single large cluster alongwith the mother plant with a similarity coefficient value of 1.00, whereas one clone (C38) remained ungrouped. The clones derived from capitulum and shoot tip explants did not show any genetic variation, whereas, one of the leaf-derived clones exhibited some degree of variation.