Mass mortalities associated with viral nervous necrosis (VNN) disease in two species of hatchery-reared grouper, Epinephelus fuscogutatus and Epinephelus akaara (Temminck & Schlegel)

Mass mortalities of hatchery-reared juvenile groupers have occurred in southern Taiwan. The diseased fish swam in a darting, corkscrew fashion. Light microscopy revealed vacuolation in the brain tissue. Electron microscopy showed numerous non-enveloped, cytoplasmic viral particles (20–25 nm in diameter) in the brain cells, and many virions were enclosed in the membrane-bound organelles of the cells. Two structural proteins of the purified grouper virus, with molecular weights of 44 and 43 kDa, were revealed by SDS-PAGE. Moreover, the results of RT-PCR and nested PCR diagnosis using primers specific to the T2 and T4 target segments of striped jack nervous necrosis virus (SJNNV) RNA2 genes suggest that this virus is a fish nodavirus, and is designated as GNNV 9410 strain (grouper nervous necrosis virus strain 9410). This is the first case report of viral nervous necrosis among marine fish in Taiwan.     

Adaptation of the LIGNUM model for simulations of growth and light response in Jack pine

LIGNUM is a whole tree model, developed for Pinus sylvestris in Finland, that combines tree metabolism with a realistic spatial distribution of morphological parts. We hypothesize that its general concepts, which include the pipe model, functional balance, yearly carbon budget, and a set of architectural growth rules, are applicable to all trees. Adaptation of the model to Pinus banksiana, a widespread species of economic importance in North America, is demonstrated.

Conversion of the model to Jack pine entailed finding new values for 16 physiological and morphological parameters, and three growth functions. Calibration of the LIGNUM Jack pine model for open grown trees up to 15 years of age was achieved by matching crown appearance and structural parameters (height, foliage biomass, aboveground biomass) with those of real trees. A sensitivity study indicated that uncertainty in the photosynthesis and respiration parameters will primarily cause changes to the net annual carbon gain, which can be corrected through calibration of the growth rate. The effect of a decrease in light level on height, biomass, total tree branch length, and productivity were simulated and compared with field data. Additional studies yielded insight into branch pruning, carbon allocation patterns, crown structure, and carbon stress. We discuss the value of the LIGNUM model as a tool for understanding tree growth and survival dynamics in natural and managed forests.     

Flavonoids of zoysiagrass (Zoysia spp.) cultivars varying in fall armyworm (Spodoptera frugiperda) resistance

Flavonoid profiles of 12 zoysiagrass (Zoysia spp.) cultivars sampled six times in 1998 were correlated to fall armyworm (Spodoptera frugiperda J. E. Smith) larval weights and survival on replicated field-grown plant material and analyzed to determine genetic and seasonal variations of flavonoids among zoysiagrass cultivars. From multiple regression analyses and correlations, flavonoid peak 10 (luteolin-glucoside) had the greatest positive association with average fall armyworm weight; however, resistance appeared to be correlated with a number of other flavonoids. The flavonoid profiles of cultivars subjected to clustering procedures showed consistent genetic variability for five of six samplings and was used to genotype 23 cultivars. The dendrogram supported the results of the FASTCLUS procedure in clustering certain genotypes such as fall armyworm-resistant Cavalier and Zeon together, as well as J-36 and Meyer. Flavonoid evaluations measure genetic relatedness among cultivars and could be used for selective breeding of resistance to fall armyworm.     

5-Hydroxy-3,6,7,8,3',4'-hexamethoxyflavone induces apoptosis through reactive oxygen species production, growth arrest and DNA damage-inducible gene 153 expression, and caspase activation in human leukemia cells

This study examined the growth inhibitory effects of structurally related polymethoxylated flavones in human cancer cells. Here, we report that 5-hydroxy-3,6,7,8,3',4'-hexamethoxyflavone (5-OH-HxMF) induces growth inhibition of human cancer cells and induction of apoptosis in HL-60 cells through modulation of mitochondrial functions regulated by reactive oxygen species (ROS). ROS generation occurs in the early stages of 5-OH-HxMF-induced apoptosis, preceding cytochrome c release, caspase activation, and DNA fragmentation. The changes occurred after single breaks in DNA were detected, suggesting that 5-OH-HxMF induced irreparable DNA damage, which in turn triggered the process of apoptosis. Up-regulation of Bax was found in 5-OH-HxMF-treated HL-60 cells. In addition, a caspase-independent pathway indicated by endonuclease G also contributed to apoptosis caused by 5-OH-HxMF. Antioxidants suppress 5-OH-HxMF-induced apoptosis. 5-OH-HxMF markedly enhanced growth arrest DNA damage-inducible gene 153 (GADD153) protein in a time-dependent manner. N-acetylcysteine (NAC) and catalase prevented up-regulation of GADD153 expression caused by 5-OH-HxMF. These findings suggest that 5-OH-HxMF creates an oxidative cellular environment that induces DNA damage and GADD153 gene activation, which in turn helps trigger apoptosis in HL-60 cells. Meanwhile, ROS were proven an important inducer in this apoptotic process. The C-5 hydroxyl on the ring of 5-OH-HxMF was found to be essential for the antiproliferative and apoptosis-inducing activity. Our study identified the novel mechanisms of 5-OH-HxMF-induced apoptosis and indicated that these results have significant applications as potential chemopreventive and chemotherapeutic agents.     

Gallic acid induces apoptosis in 3T3-L1 pre-adipocytes via a Fas- and mitochondrial-mediated pathway

Gallic acid (3,4,5-trihydroxybenzoic acid) is a naturally abundant plant phenolic compound. Our previous studies have shown that some phenolic acids such as gallic acid inhibit cell growth and induce apoptosis in 3T3-L1 pre-adipocytes. However, the molecular mechanism of gallic acid in the induction of cell apoptosis is still unclear. In this study, we investigated the effect of gallic acid on the apoptotic pathway in 3T3-L1 pre-adipocytes. Western blot data revealed that gallic acid stimulated an increase in the protein expression of Fas, FasL, and p53. The ratio of expression levels of pro- and anti-apoptotic Bcl-2 family members was changed by gallic acid treatment. Gallic acid released mitochondrial cytochrome c into the cytosol and subsequently induced the activation of caspase-9 and caspase-3, which were followed by the cleavage of poly(ADP-ribose) polymerase. Pretreatment with a general caspase-9 inhibitor (Z-LEHD-FMK) and caspase-3 inhibitor (Z-DEVD-FMK) prevented gallic acid from inhibiting cell viability in 3T3-L1 pre-adipocytes. The data also indicated that treatment with gallic acid inhibited histone deacetylase activity in 3T3-L1 pre-adipocytes. These results demonstrate that gallic acid induces apoptosis in 3T3-L1 pre-adipocytes through the Fas and mitochondrial pathway. The induction of cell apoptosis by gallic acid may prove to be a pivotal mechanism for decreased pre-adipocyte proliferation.     

Conventional analyses of data from dietary validation studies may misestimate reporting accuracy: illustration from a study of the effect of interview modality on children's reporting accuracy

OBJECTIVE: To compare two approaches to analysing energy- and nutrient-converted data from dietary validation (and relative validation) studies - conventional analyses, in which the accuracy of reported items is not ascertained, and reporting-error-sensitive analyses, in which reported items are classified as matches (items actually eaten) or intrusions (items not actually eaten), and reported amounts are classified as corresponding or overreported. DESIGN: Subjects were observed eating school breakfast and lunch, and interviewed that evening about that day's intake. For conventional analyses, reference and reported information were converted to energy and macronutrients; then t-tests, correlation coefficients and report rates (reported/reference) were calculated. For reporting error-sensitive analyses, reported items were classified as matches or intrusions, reported amounts were classified as corresponding or overreported, and correspondence rates (corresponding amount/reference amount) and inflation ratios (overreported amount/reference amount) were calculated. SUBJECTS: Sixty-nine fourth-grade children (35 girls) from 10 elementary schools in Georgia (USA). RESULTS: For energy and each macronutrient, conventional analyses found that reported amounts were significantly less than reference amounts (every P < 0.021; paired t-tests); correlations between reported and reference amounts exceeded 0.52 (every P < 0.001); and median report rates ranged from 76% to 95%. Analyses sensitive to reporting errors found median correspondence rates between 67% and 79%, and that median inflation ratios, which ranged from 7% to 17%, differed significantly from 0 (every P < 0.0001; sign tests). CONCLUSIONS: Conventional analyses of energy and nutrient data from dietary reporting validation (and relative validation) studies may overestimate accuracy and mask the complexity of dietary reporting error.     

Treatment of hatchery wastewater by a sequencing batch reactor process

After settling pretreatment, the primary sludge wastewaters from a fish hatchery were treated in three five-litre bench-scale sequencing batch reactors. The sequencing batch reactors were operated concurrently under the same operating conditions. A cycle of 24 hours was adopted during the course of this study. Two different temperature regimes (22°C and 10°C) were employed, and a range of initial BOD₅ concentrations was treated. The results showed the occurrence of very high reductions in BOD₅ and COD concentrations, and very high removal of ammonia---nitrogen and suspended solids. The average five-day BOD₅ reduction was 98% for high influent concentrations, while the reduction for lower influent concentrations was 95%. The average five-day BOD₅ concentration for the effluents was less than 100 mg litre⁻¹. Nitrification was pronounced after 40 days of reactor operation. Denitrification occurred when the reactors were fed wastewaters with high BOD₅ concentrations, although it was not observed with low influent BOD₅ concentrations or at low temperatures (10°C). The overall treatment efficiency remained very high when the reactors were operated under a low temperature regime (10°C).     

Effect of salinity on natural community and production of Litopenaeus vannamei (Boone), within experimental zero-water exchange culture systems

Recent efforts have been made to culture marine shrimp in systems operating under low or zero‐water exchange and with decreased water salinity. The aim of this study was to investigate the impact of various salinity levels on qualitative and quantitative characteristics of the natural community and, more particularly, ciliated protozoa, and compare this information with shrimp growth and survival. Tanks with 9‰ salinity were characterized by a higher pH, but also by a significantly higher concentration of chlorophyll a (Chl a) per weight of suspended matter (1.93 ± 0.72 µg Chl a/mg TSS) than tanks with 18‰ (1.29 ± 0.68 µg Chl a/mg TSS) or 36‰ (1.37 ± 0.61 µg Chl a/mg TSS) salinity. Concentrations of ciliates (max 6000 cells mL^?1) showed considerable fluctuations over the sampling period, reflecting the impact of water salinity, dynamic interactions between ciliates and their diverse roles within the shrimp production system. There was no significant difference between survival rates of shrimp reared at 9‰, 18‰ or 36‰, but decreasing salinity from 36‰ to 9‰ led to a significant decrease in final shrimp body weight (from 13.40 ± 0.26 g to 10.23 ± 2.72 g). Future work should address the potential of ciliates as an indicator of aquaculture water quality, as is currently being done in the wastewater industry, and the contribution of ciliates as food sources.     

Characterization of molecular variants of GnRH, induction of spermiation and sex reversal using salmon GnRH-A and domperidone in the protogynous diandric fish, Synbranchus marmoratus Bloch, (Teleostei, Synbranchidae)

This paper studies the molecular variants of gonadotropin-releasing hormone (GnRH) present in the brain of the protogynous swamp eel, Synbranchus marmoratus, and the effects of the administration of salmon GnRH analogue (sGnRH-A) and the dopamine receptor antagonist, domperidone (DOM) on final maturation and gamete release in this species. Evidence for the presence of two GnRH variants, sGnRH and cIIGnRH were obtained by reverse phase high-pressure liquid chromatography (RP-HPLC) and radioimmunoassay with different antisera. The effects of treatment with sGnRH-A+DOM were checked by three ways: oocyte and milt release by stripping, histological analysis of the gonadal tissue, and androgen serum levels at different times throughout the experiment. In males, spermiation was induced after three weeks of treatment. In the female group, sGnRH-A+DOM did not induce ovulation at the end of the experiment. Histological analysis of the gonads from the female group showed evidence of sex reversal. All the treated fish had elevated androgen serum levels from the third week, with respect to control fish. In all cases, serum estradiol levels were undetectable. These results suggest that treatment with sGnRH analog and DOM induce sex reversal in female and spermiation in males. This revised version was published online in July 2006 with corrections to the Cover Date.