Effects of delayed first feeding on the survival and growth of tiger grouper,Epinephelus fuscoguttatus (Forsskål, 1775), larvae

The effects of delayed first feeding on the survival and growth of tiger grouper, Epinephelus fuscoguttatus (Forsskål 1775), larvae were examined under controlled conditions. The total length, yolk sac volume, oil globule volume, yolk sac absorption time and nutritional transition period (NTP) of the larvae fed at different first times (0, 6, 12, 18 and 24 h after the mouth opening stage; h AMO) were compared. Larval first feed intake was observed at 54 h after hatching (h AH) at 27.5 ± 0.5 °C. The yolk sac was consumed more rapidly with an increase in delayed first feeding and was significantly different among treatments (P<0.05). Larvae first fed at 0 h AMO had the longest yolk sac absorption (72 h AH) and NTP (20 h) times and had the highest survival and growth rates at the end of the experimental period (360 h AH), being significantly higher (P<0.05) than the other treatments. First mortality was observed at 69 h AH, approximately 2 h after point of no return (PNR) occurred. This study suggests that first feeding of tiger grouper larvae should commence at 0 h AMO for enhancement of larval survival and growth.     

Assessment of Photochemical Reflectance Index as a Tool for Evaluation of Chlorophyll Fluorescence Parameters in Cotton and Peanut Cultivars Under Water Stress Condition

The relationships between photochemical reflectance index （PRI） and the chlorophyll fluorescence parameters were examined to assess suitability of PRI as a remote-sensing tool for the chlorophyll fluorescence parameters. A greenhouse experiment was conducted using cotton and peanut crops under water stress condition. Five cotton and six peanut cultivars were grown using Andosole soil in pots maintained at two water levels; the control and water stress treatment （WS） of 100 and 50% of the daily transpiration, respectively. Higher non-photochemical quenching （NPQ） was exhibited by peanut than that of cotton by the water stress. On the other hand, the decreases of the actual quantum yield of photosystem II （△F/F＇m） and PRI by the water stress in cotton were larger than those in peanut. There were positively significant correlation coefficients between PRI and △F/F＇m in cotton at noon and in the afternoon including the control and WS. The correlations of PRI with NPQ were negatively significant at noon and in the afternoon for cotton, and in the afternoon for peanut. No clear relationship was found among these parameters in the morning probably due to the diurnal increase in global solar radiation. It was concluded that there would be a possibility to detect the effects of water stress on △F/F＇m and NPQ by PRI with some exceptions, although PRI could not note varietals differences in △F/F＇m and NPQ for each treatment.     

Novel nitric oxide and superoxide generation inhibitors, persenone A and B, from avocado fruit

One known, (2R)-(12Z,15Z)-2-hydroxy-4-oxoheneicosa-12,15-dien+ ++-1-yl acetate (1), and two novel compounds, persenone A (2) and B (3), have been isolated from avocado fruit (Persea americana P. Mill), as inhibitors of superoxide (O(2)(-)) and nitric oxide (NO) generation in cell culture systems. They showed marked inhibitory activities toward NO generation induced by lipopolysaccharide in combination with interferon-gamma in mouse macrophage RAW 264.7 cells. Their inhibitory potencies of NO generation (1, IC(50) = 3.6; 2, IC(50) = 1.2; and 3, IC(50) = 3.5 microM) were comparable to or higher than that of a natural NO generation inhibitor, docosahexaenoic acid (DHA; IC(50) = 4.3 microM). Furthermore, compounds 1-3 and DHA markedly suppressed tumor promoter 12-O-tetradecanoylphorbol-13-acetate-induced O(2)(-) generation in differentiated human promyelocytic HL-60 cells (1, IC(50) = 33.7; 2, IC(50) = 1.4; 3, IC(50) = 1.8; and DHA, IC(50) = 10.3 microM). It is notable that they were found to be suppressors of both NO- and O(2)(-)-generating biochemical pathways but not to be radical scavengers. The results indicate that these compounds are unique antioxidants, preferentially suppressing radical generation, and thus may be promising as effective chemopreventive agent candidates in inflammation-associated carcinogenesis.     

Promotion of initial swimbladder inflation in Pacific bluefin tuna,Thunnus orientalis (Temminck and Schlegel), larvae

Initial swimbladder inflation (ISI) of Pacific bluefin tuna (PBT), Thunnus orientalis, larvae was studied to increase the survival of cultured larvae. Experiment 1 was conducted to explore promotion and inhibition of ISI under different water surface conditions; including the use of surface skimmer to remove autogenous surface substances (SS), covering the water surface with liquid‐paraffin‐layer (LP) and oil film (OF), and a control (non‐treatment, NT). Significantly higher inflation frequency was observed in SS (62.2%) than NT (11.9%), LP (2.7%) and OF (3.9%). This indicates that ISI in PBT larvae can be promoted by removal of surface substances on rearing water which inhibit larval air gulping. Experiment 2 aimed to elucidate proper day of larval age to start skimming for promoting ISI with four different periods of oil film removal: from 3 to 8 (SF3D), 4 to 8 (SF4D), 5 to 8 (SF5D), 6 to 8 (SF6D) days‐post‐hatch (dph). Significant improvement in ISI frequency was observed in SF3D (80.2%) but the frequency was very poor in SF4D, SF5D, and SF6D (17.8–7.5%). This implies the need of oil film removal without missing a narrow window, 1 day of 3 dph, to promote ISI in practical PBT larviculture.     

Detection and sequence analysis of DNA polymerase and major envelope protein genes in koi herpesviruses derived from Cyprinus carpio in Gunma prefecture, Japan

Koi herpesvirus (KHV), which is believed to be an emerging virus, causes fatal diseases in carps. Since the 1990s, the presence of KHV has been confirmed in several countries. In Japan, from 2003 to 2004, large outbreaks of KHV infection in farmed carps resulted in the death of numerous fishes. From April to May 2004, we collected 43 dead or dying carps exhibiting typical symptoms of KHV infection in Gunma prefecture. To conduct a molecular epidemiologic study of KHV in our prefecture, we amplified DNA polymerase and the major envelope protein genes of KHV derived from carp gills using newly designed primers. We also performed sequence analysis of both genes of KHV. Sensitivity of our PCR method for amplification of DNA polymerase and the major envelope protein genes of KHV was 3 x 10(2) (100 fg) and 3 x 10(3) (1000 fg) copies of KHV genome, respectively. We detected both DNA polymerase and major envelope protein genes in 37 of 43 carps (86%). No mutation was found in both the genes sequenced from 11 strains, which included two foreign strains and one domestic strain. The results suggested that KHV strains derived from carps in our prefecture were closely related genetically to the other KHV strains.     

Hybrid aspen with a transgene for fungal manganese peroxidase is a potential contributor to phytoremediation of the environment contaminated with bisphenol A

To assess the possible utility of a fungal gene for manganese-dependent peroxidase (MnP) produced by a transgenic plant in phytoremediation, we transformed hybrid aspen with a chimeric gene for MnP. Our gene construct allowed expression of the gene for MnP in plants and relatively high MnP activity was detected in the hydroponic medium in which roots of plants that expressed the transgene had been cultured. Some of our transgenic plants were able to remove bisphenol A from the medium more efficiently than wild-type plants. Our results demonstrate that, without any modification of the coding sequence, a chimeric gene for fungal MnP can be expressed in a woody plant, with secretion of active MnP from roots into the rhizosphere. Our strategy suggests new options using woody plants for phytoremediation.     

Effect of liming using a partial mixing technique on reductions in the seed cadmium levels for soybeans (Glycine max (L.) Merr.) under field conditions

To reduce the availability of soil cadmium (Cd) to soybeans (Glycine max (L.) Merr.), we employed a liming by partial mixing (PM) technique in two drained paddy fields on Gray Lowland soils, which had 0.1 mol L^–1 hydrochloric acid-extractable Cd concentrations as high as 1.08 and 1.40 mg kg^–1. Among the different application methods tested, PM application (PM2) using a width of 20 cm and a depth of 20 cm was found to be most appropriate for reducing the seed Cd concentration and to obtain the optimum yield at Site A. Under PM2, a liming rate of 38% of that for broadcast incorporated into the surface 15 cm layer (Bc) was suitable to reduce the seed Cd concentration at Site A, whereas the lime rate with PM2 was set at 50% of that for Bc (PM2-50) at Site B due to the higher availability of soil Cd. The root system was limited within the range of lime and fertilizer application for PM2 as well as PM2-50; thus, the lime and fertilizer were supplied successfully to the rooting zone. The soil pH value was lower under PM2 at Site A and PM2-50 at Site B compared with Bc, whereas the seed Cd concentration was lower for PM2 and PM2-50. This may be explained by the intensive uptake of calcium and magnesium with PM2 as well as PM2-50. The seed Cd concentration in the cultivar “Ryuhou” at the target pH of 6.5 was approximately 30% lower with PM2-50 than Bc at Site B. In addition, the average seed Cd concentrations in one cultivar and two lines, characterized by the lower Cd uptake with higher retention in roots and higher accumulation in leaves, were approximately 40% lower compared with “Ryuhou.” Thus, the combination of liming with PM2-50 at the target pH of 6.5 and a low-Cd cultivar (or lines) minimized the seed Cd concentration. The highest seed Cd concentration was found in the first year of soybean cultivation, which was considered to be caused by the release of Cd from organic nitrogen compounds during the nitrogen mineralization process.     

Aeration rate adjustment at night to prevent sinking syndrome‐related death in the tiger grouper Epinephelus fuscoguttatus (Perciformes:Serranidae) larvae

The effects of different aeration rates at night to prevent sinking syndrome‐related death (SSRD) of the tiger grouper, Epinephelus fuscoguttatus were examined. The aeration rates were fixed at 300 mL min^?1 at daytime (07:00–19:00 hours) and regulated to 0, 300 and 900 mL min^?1 at night (19:00–07:00 hours). Larval survival, growth, feeding intake, sinking velocity, distribution and behaviour, stress level, surface tension‐related death (STRD) and flow velocity distribution were assessed. The occurrence of SSRD in the tiger grouper was observed through the accelerated sinking velocity (Vl) (from 0.15 ± 0.09 cm s^?1 at 4 days AH to 0.41 ± 0.09 cm s^?1 at 12 days AH) coupled with larval passive swimming behaviour at night‐time. On the final day of experiment (15 days AH), larvae reared in 900 mL min^?1 at night had attained significantly higher (P < 0.05) survival (34.4 ± 5.5%), growth (5.8 ± 0.5 mm) and feeding intake (60.46 ± 6.98 ind. larva^?1). A favourable flow field for the tiger grouper was produced in 900 mL min^?1 at night‐time, in which larvae were transported 15–25 cm above the tank bottom and 1.0 cm beneath the water surface. Under these night‐time rearing conditions, larval stress level and number of STRD reared in 900 mL min^?1 compared with those observed in 300 mL min^?1 remained insignificant, indicating that strong turbulence of flow velocity was not detrimental for larvae. Our findings recommend aeration at 900 mL min^?1 at night as this could improve larval survival by reducing SSRD.     

Association between bacterial community structures and mortality of fish larvae in intensive rearing systems

Bacterial community structures were analyzed in water used for rearing fish larvae by fluorescence in situ hybridization. In Experiment 1, red sea bream Pagrus major larvae were reared in two commercial seed production tanks. The survival rate in Tank 1 was higher than in Tank 2, even though phytoplankton, Nannochloropsis sp., was added to both tanks. In Tank 2, γ-proteobacteria became dominant (∼70% of total bacteria) on day 13, there after heavy larval mortalities occurred. In Tank 1, however, α-proteobacteria and the Cytophaga-Flavobacterium cluster were predominant from day − 1 until day 13; no significant mortality was recorded. In Experiment 2, marble goby Oxyeleotris marmoratus larvae were cultured with or without Nannochloropsis sp. At the end of the experiment, larval survival rates in aquaria with Nannochloropsis sp. were significantly (P <0.05) higher than those without. In rearing water without Nannochloropsis sp., γ-proteobacteria increased during rearing. In rearing water with Nannochloropsis sp., α-prote obacteria and the Cytophaga-Flavobacterium cluster were predominant at the beginning of the experiments and the relative abundance of γ-proteobacteria was maintained at a lower level throughout the experiments. The predominance of α-proteobacteria and the Cytophaga-Flavobacterium cluster appears to be a good indicator of successful larval production.     

Flow field control via aeration adjustment for the enhancement of larval survival of the kelp grouper Epinephelus bruneus (Perciformes:Serranidae)

Flow field control via aeration adjustment for the enhancement of larval survival of the kelp grouper Epinephelus bruneus was examined. Aeration rate of 300 mL min^?1 was introduced during daylight (07:00–19:00 hours) and adjusted to 0, 300 and 900 mL min^?1 at night (19:00–07:00 hours). Larval sinking velocity±SD increased from 0.08 ± 0.05 to 0.26 ± 0.24 cm sec^?1 from 4 to 12 days after hatching (DAH), indicating their susceptibility to sink. Larvae reared in 300 mLmin^?1 attained the highest survival rate at 24.9 ± 3.4%, but remained significantly smaller in growth: 4.54 ± 0.56 mm compared with 4.82 ± 0.53 mm in 900 mL min^?1. The flow field in 300 and 900 mL min^?1 was at 10–20 and 15–25 cm above the bottom of the tank and 8.0 and 1.0 cm beneath the water surface. A favourable rearing condition was observed in 300 mL min^?1 as larvae were away from the bottom and surface areas, thus preventing them from dying due to sinking and surface tension‐related death (STRD). Although sinking death was decreased with an increasing aeration rate, the stronger flow had increased larval susceptibility to STRD. Our findings suggest that aeration at 300 mL min^?1 could enhance larval survival by reducing both sinking death and STRD.