Molecular cloning and expression of the heat shock protein 70 gene in the Kumamoto oyster <Emphasis Type="Italic">Crassostrea sikamea</Emphasis>

Episodes of summer mortality of the Kumamoto oyster Crassostrea sikamea are a major problem for its cultivation. Expression of the heat shock protein 70 (HSP70) is induced by various environmental stresses, including heat. We cloned and sequenced hsp70 complementary DNA from C. sikamea to investigate the relationship between hsp70 expression and heat tolerance in this oyster. Quantitative real-time polymerase chain reaction was performed using gill tissue dissected from oysters before and after heat shock for 1 h. The results showed hsp70 expression was faster and greater in oysters cultured at 20–22 °C than at 10–12 °C, and survival was lower among oysters cultured at 20–22 °C than at 10–12 °C. Moreover, heat tolerance was investigated by a 1-h pre-heat treatment, followed by exposure to heat shock conditions 5 days later. Survival was higher and hsp70 expression was notably lower in oysters that received the pre-heat treatment compared with those that did not. We conclude that a pre-heat treatment of only 1 h may be useful for inducing heat tolerance in C. sikamea, and that a low level of hsp70 expression after heat shock is an important index in selecting for high heat tolerance in these oysters.     

Effects of net height of crab entangling nets on the capture of targeted economically important portunid species and non-target species

The aim of this study was to determine the effects of net height on the capture performance of crab entangling nets. Fishing trials were conducted using nets at varying net heights (1) 12 meshes down (MD), (2) 24 MD and (3) 50 MD. A total of 1290 individuals comprising 87 species belonging to 53 families were caught. One-way analysis of variance showed that net height significantly affected the various catch parameters, including catch per unit effort (CPUE) of the total and target catch, amount of non-target catch, size of catch and species richness. The use of appropriate net height is a potential technical measure for a selective but still efficient crab entangling net fishery. Lower net height significantly reduced non-target catch by up to 70%. Lower net height also decreased the CPUE of target catch such as Portunus pelagicus and Charybdis feriatus by up to 65% at 12 MD, but catch at 24 MD was not significantly different than that at 50 MD. The use of a net height of 24 MD also resulted in the capture of larger-sized P. pelagicus. The richness of the catch species decreased by up to 58% in lower nets. These results are useful to fishery managers and government institutions when developing and/or improving existing regulations towards a sustainable crab fishery, particularly blue swimming crabs.     

Growth,absorption and assimilation efficiency by mature cuttlefish (<Emphasis Type="Italic">Sepia officinalis</Emphasis>) fed with alternative and artificial diets

The effects of feeding three natural frozen diets, grass shrimp (Palaemonetes sp.), crayfish (Procambarus clarkii) and fish (Sardina pilchardus) and two semi-humid artificial diets (based on fish powder) to mature cuttlefish, Sepia officinalis, were analysed. Growth and feeding rates (GR and FR, % BW day⁻¹), food conversions (FC, %), and total protein and lipid composition of the diets were determined. Digestive gland to body weight ratio and absorption efficiency were calculated for each diet. Cuttlefish fed shrimp and crayfish grew larger (1.5 and 1.1% BW day⁻¹, respectively) compared to the other diets. Shrimp promoted the highest FC, followed by crayfish, and sardine. The highest FR was obtained for cuttlefish fed crayfish (8.4% BW day⁻¹). Although both artificial diets were accepted, none produced growth. A positive correlation (r = 0.96) between cuttlefish ingestion rate and digestive gland weight was obtained. Some cannibalism occurred among cuttlefish fed the artificial diets during the last week of the experiment. According to the results obtained, P. clarkii could be used as an alternative prey to shrimp for rearing adult mature (>50 g) S. officinalis.     

Supplemental effect of cholyltaurine and soybean lecithin to a soybean meal-based fish meal-free diet on hepatic and intestinal morphology of rainbow trout <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>

The supplemental effects of cholyltaurine and soybean lecithin to a defatted soybean meal (SBM)-based fish meal-free diet on the hepatic and intestinal morphology of rainbow trout Oncorhynchus mykiss were investigated. A fish meal-based diet (FM), three fish meal-free diets supplemented either with soybean lecithin (SL), bovine gall powder (BG) or cholyltaurine (C-tau) and an unsupplemented fish meal-free diet (NFM), were given to fish (−11 g) for 10 weeks. The growth was the lowest in fish fed diet NFM, intermediate in fish fed diet SL and the highest in fish fed diet FM, BG and C-tau. Hepatocytes of fish fed diet NFM were atrophied, and the distal intestine of these fish showed abnormal features: disintegrated microvilli and fatty degeneration in epithelial cells and increased amount of connective tissue in the submucosa. Hepatic and intestinal histological features in fish fed diets SL, BG and C-tau were similar to those in fish fed diet FM. These results indicate that soybean lecithin, bovine bile salts and cholyltaurine have similar effects on normalizing the hepatic and intestinal morphologies of rainbow trout fed the SBM-based diet, although growth promotion effect was limited in the soybean lecithin.     

Effects of triploidy on the Caspian salmon <Emphasis Type="Italic">Salmo trutta caspius</Emphasis> haematology

The aim of this study was a comparison of key haematological features of diploid (2n) and triploid (3n) Caspian salmon (Salmo trutta caspius). Morphometric indices of erythrocytes were determined on blood smears by light microscopy. Triploidy significantly (P < 0.001) increased all morphometric indices measured in the erythrocytes including cell size, cell surface area, and cell volume. The increase in cell size was larger for the major (27%) axis than for the minor (22%) axis, thus making erythrocytes of 3n Caspian salmon more ellipsoidal. The estimated increase in erythrocyte nuclear volume (87%) was bigger than the theoretical expected 50% increase. Haematological indices were measured manually by hemocytometry. Triploids had lower numbers of red blood cells (RBC: 1,120,000 cells/mL in 2n vs. 700,000 cells/mL in 3n; P < 0.001) but they were larger in size (mean erythrocytic volume [MEV]: 363.1 nm³ in 2n vs. 483.3 nm³ in 3n; P < 0.001). The decrease in RBC number was not compensated by the increase in MEV and, thus, triploidy affected the haematocrit (Hct: 38.8% in 2n vs. 33.06% in 3n; P < 0.05). Total blood hemoglobin concentration was lower in triploid fish (Hb: 9.9 g/dL in 2n vs. 8.9 g/dL in 3n; P < 0.05). In contrast, mean erythrocytic hemoglobin (MEH: 95 μg in 2n vs. 133.2 μg in 3n; P < 0.001) was higher for 3n Caspian salmon as a result of their larger erythrocytes, although MEH concentration (MEHC: 0.26 g/dL in 2n vs. 0.27 g/dL in 3n) did not significantly differ (P > 0.05). White blood cell (WBC) counts (lymphocytes and neutrophiles) were measured and WBC/RBC ratios were calculated. There were no significant differences in WBC (15,710 cells/mL in 2n vs. 12,683 cells/mL in 3n; P > 0.05), lymphocytes, and neutrophils as %WBC as well as WBC/RBC ratios between two ploidy levels (P > 0.05). Triploid Caspian salmon showed higher erythrocyte abnormalities such as ‘twisted’, ‘tailed’, and ‘anucleated’ cells as well as high portions of immature RBC in blood smears in comparison with diploids (P < 0.001).     

Aflatoxin B<Subscript>1</Subscript> (AFB<Subscript>1</Subscript>) reduces growth performance,physiological response,and disease resistance in Tra catfish (<Emphasis Type="Italic">Pangasius hypophthalmus</Emphasis>)

Triplicate groups of one hundred Tra catfish (8 g?±?0.2) were fed seven test diets containing increasing levels of AFB₁ (0, 50, 100, 250, 500, and 1000 μg AFB₁ kg^?1). Additionally Mycofix® Secure was added at 1.5% to one diet containing 500 μg AFB₁ kg^?1. Results showed that Tra catfish are sensitive to AFB₁. Reduction in weight gain (P?<?0.05) was observed for fish fed 50 μg AFB₁ kg^?1 and declined further with increasing levels of AFB₁ in the diets. Fish fed diets contaminated with 500 and 1000 μg AFB₁ kg^?1 showed increased (P?>?0.05) hepatosomatic index (HIS), while an increase in adipose somatic index (ASI) was observed in fish fed 50 μg AFB₁ kg^?1 and above when compared to the control and Mycofix® diets. After 12 weeks, blood serum analysis revealed higher alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in fish fed the 50, 100, and 250 μg AFB₁ kg^?1 suggesting occurrence of liver damage. Disease resistance of fish exposed to Edwardsiella ictaluri was also compromised by the presence of AFB₁ in the feed and was directly related to the contamination level. Seven days after Edwardsiella ictaluri exposure, survival rates were 50, 41.7, 31.7, and 8.3% for fish fed control, 50, 100, and 250 μg AFB₁ kg^?1, respectively. This trial shows that AFB₁ at a level of 50 μg AFB₁ kg^?1 and above can affect fish performance and disease resistance. Application of an effective mycotoxin management in the feed seems to be useful to prevent the negative effects of AFB₁.     

Accumulation of organotin compounds and marine birnavirus detection in Korean ascidians

ABSTRACT:   Recently, a serious disease spread extensively in aquaculture sites of the ascidian Halocynthia roretzi in Korea. To understand circumstances of ascidians in Korean aquaculture sites, residue levels of organotin compounds were analyzed, and detection of a marine birnavirus (MABV) in tissues of H. roretzi was attempted. Korean H. roretzi showed high concentrations of butyltins (mono, di, and tributyltins), especially in the gill, hepatopancreas, and digestive tract. However, there was no significant difference in the residues of butyltins in the hepatopancreas between diseased and non-diseased ascidians. The positive rate of MABV detection was high in the hepatopancreas, but also no significant difference was observed between diseased and non-diseased individuals. These observations suggest that an accumulation of tributyltin and a latency of MABV in H. roretzi tissues does not directly relate to the occurrence of the disease.     

Effect of conformation on the properties of uniaxially drawn polylacide films upon drawing temperature

Polylactide(PLA) films were drawn at various drawing temperature of 65, 90 and 120 °C. The effects of drawing temperature on structural conformation and properties of PLA films were investigated. It was confirmed that the PLA films at drawing temperature of 65 and 90 °C were composed of α′ phase crystal form. The strain-induced crystallization and molecular orientation increased with increasing the draw ratio, which result in improving the mechanical and thermal properties of α′ phase PLA films. However, at drawing temperature of 120 °C, the strain-induced crystallization and molecular orientation of PLA films were not distinctly detected. It was supposed that the rate of the chain relaxation was faster than chain orientation and strain-induced crystallization during uniaxial drawing process.     

Can facultative parthenogenesis occur in biparental mealybug species?

The existence of facultative parthenogenesis in mealybugs has been reported by different authors and questioned by others in relation to certain species. We tested the hypothesis of facultative parthenogenesis in relation to three mealybug species, Planococcus citri (Risso), Pseudococcus calceolariae (Maskell) and Pseudococcus viburni (Signoret). The results suggest that these mealybug species are obligate amphimictic.     

Selection,characterization and genetic analysis of laboratory mutants of <Emphasis Type="Italic">Botryotinia fuckeliana</Emphasis> (<Emphasis Type="Italic">Botrytis cinerea</Emphasis>) resistant to the fungicide boscalid

Resistance to the fungicide boscalid in laboratory mutants of Botryotinia fuckeliana (Botrytis cinerea) was investigated. The baseline sensitivity to boscalid was evaluated in terms of colony growth (EC₅₀ = 0.3–3 μg ml⁻¹; MIC = 10–30 μg ml⁻¹) and conidial germination (EC₅₀ = 0.03–0.1 μg ml⁻¹; MIC = 1–3 μg ml⁻¹) tests. Mutants were selected in vitro from wild-type strains of the fungus on a fungicide-amended medium containing acetate as a carbon source. Mutants showed two different levels of resistance to boscalid, distinguishable through the conidial germination tests: low (EC₅₀ ∼ 0.3 μg ml⁻¹, ranging from 0.03 to 1 μg ml⁻¹; MIC > 100 μg ml⁻¹) and high (EC₅₀ > 100 μg ml⁻¹) resistance. Analysis of meiotic progeny from crosses between resistant mutants and sensitive reference strains showed that resistant phenotypes were due to mutations in single major gene(s) inherited in a Mendelian fashion, and linked with both the Daf1 and Mbc1 genes, responsible for resistance to dicarboximide and benzimidazole fungicides, respectively. Gene sequence analysis of the four sub-units of the boscalid-target protein, the succinate dehydrogenase enzyme, revealed that single or double point mutations in the highly conserved regions of the iron-sulphur protein (Ip) gene were associated with resistance. Mutations resulted in proline to leucine or phenylalanine replacements at position 225 (P225L or P225F) in high resistant mutants, and in a histidine to tyrosine replacement at position 272 (H272Y) in low resistant mutants. Sequences of the flavoprotein and the two transmembrane sub-units of succinate dehydrogenase were never affected.