胶束电动色谱法快速测定鱼血液中的类固醇激素

建立了一种同时分离测定鱼血液中皮质醇、雌二醇及睾酮含量的胶束电动色谱法.研究了缓冲溶液浓度、添加剂含量、pH、分离电压及温度等条件对分离的影响,得到了最佳分离条件,即:缓冲溶液组成为12mmol/L硼砂+60 mmol/L十二烷基硫酸钠+6 mmol/Lβ-环糊精+8%乙腈,pH 9.5;检测波长为225 nm,分离电压为25 kV,分离温度为25℃;采用毛细管短端进样(毛细管有效长度仅为10 cm)、反向分离模式.3种类固醇激素在4 min内达到基线分离,且分析物在1.0～100.0μg/mL呈良好的线性关系,相关系数大于0.993 2.该方法用于鱼血液中类固醇激素的检测,回收率为84.60%～113.30%,满足生物样品检测的要求.

Persistence of malachite green and leucomalachite green in perch (Lateolabrax japonicus)

The persistence of malachite green (MG), and its metabolite leucomalachite green (LMG), in fish tissues is still unclear, leading to many trade disputes. In this research, we established and evaluated an HPLC method that could detect MG and LMG simultaneously, and then investigated the persistence of these two toxins in the tissues of juvenile perch (Lateolabrax japonicus) post sub-chronic MG exposure at 1.0 mg/L. Exposure lasted for 2 h everyday and was repeated six times. The perch were then placed in MG-free seawater for 100 d to eliminate the toxins. Results show that MG accumulated in the tissues, including the gills, liver, muscle, blood and viscera, and then was metabolized rapidly to LMG. The concentrations of these two toxins increased significantly with the accumulation process. In general, the highest concentrations of MG and LMG in all tissue exceeded 1 000 μg/kg, except for MG in the muscle. The order of accumulation levels (highest to lowest) of MG was gill>blood>liver>viscera>muscle, while that of LMG was liver>blood>gill>viscera>muscle. High levels of MG or LMG could persist for several hours but decreased rapidly during the elimination process. The concentration of LMG was much higher than that of MG during the experiment, especially in the gill, liver and blood. Therefore, the three tissues play important roles in toxin accumulation, biotransformation, and elimination. Although the MG and LMG concentrations in muscle were much lower than in other tissues, the content still exceeded the European minimum required performance limit (MRPL), even after 2 400 h (100 d) of elimination. This demonstrates that it is extremely difficult to eliminate MG and LMG from tissues of perch, and therefore use of these toxins is of concern to public health.

Effect of hypoxia on the blood of large yellow croaker (Pseudosciaena crocea)

Large areas of hypoxic water have recently been reported in the East China Sea. It is hypothesized that hypoxia may be partially responsible for the decline of some fish stocks. We evaluated the effect of hypoxia on large yellow croaker (Pseudosciaena crocea). The fish were exposed to three concentrations of dissolved oxygen (DO; 1.5 mg/L and 2.0 mg/L, and 6.5 mg/L control). We collected blood after 6, 12, 24, 48, and 96 h exposure. There was a significant increase in red blood count, hematocrit, hemoglobin concentration, and mean corpuscular hemoglobin in the group exposed to 1.5 mg/L DO after 6 h or 12 h, and a delayed increase (only elevated after 48 h and 96 h) in these indices in the group exposed to 2.0 mg/L DO. Plasma glucose concentrations increased significantly in both hypoxic groups after 24 h. Furthermore, plasma lactate and lactate dehydrogenase activity increased significantly after the first 6 h exposure in both hypoxic groups. Our results suggest that large yellow croakers could not maintain the aerobic pathway and instead use anaerobic metabolism for survival when DO levels fall below 2.0 mg/L. We conclude that the occurrence of hypoxia (<2 mg/L DO) in the East China Sea could cause metabolic stress for large yellow croakers and may be partially responsible for the population decline of this species.

Embryology of Maldives Clownfish, Amphiprion nigripes (Amphiprioninae)

This study investigated the embryonic development of Maldives clownfish Amphiprion nigripes under natural conditions (28 - 30℃) at a lagoon of Agatti Island, Lakshadweep, India. The newly deposited fish egg was capsule-shaped and orange-red, with a (0.73 ± 0.04) mm3 yolk containing 5 - 10 fat globules. The embryonic development of fertilized eggs was divided into 26 stages and the time elapsing for each stage was recorded. Results showed that the cleavage was rapid, with the first division observed 1 h 20 min after fertilization. Blastulation occurred 4 h later, followed by gastrulation 12 h after fertilization, with a yolk volume of (0.61 ± 0.06) mm3. The organogenesis process started 22 h after fertilization when the blastopores closed and notochord formation began. The embryonic stage was recorded 24 h later, with the appearance of forebrain, midbrain, hindbrain, melanophores on yolk-sac and 22 somites, and a decreased yolk volume of (0.54 ± 0.08) mm3. Other organs developed well 31 h after fertilization, whereas the heart started beating and blood circulation began 78 h later. Red pigmentation (erytrophores) appeared 96 h after fertilization, with a small yolk volume of (0.22 ± 0.02) mm3. Mouth developed well and eyes were noticeable 120 h later, with head, pectoral fin and tail frequently moving 144 h after fertilization. The embryo reached the pre-hatching stage 168 h later and started to hatch after 170 - 180 h incubation. This study first detailed the embryonic development and yolk absorption of A. nigripes under natural conditions.

Effect of Using Baker＇s Yeast and Exogenous Digestive Enzymes as Growth Promoters on Growth, Feed Utilization and Hematological Indices of Nile tilapia, Oreochromis niloticus Fingerlings

The present study was conducted to evaluate the effect of baker yeast, Saccharomyces cerevisiae （SC） and exogenous digestive enzymes （pepsin, papain and a-amylase, EDE） dietary supplementation on growth performance, feed utilization and hematological indices of Nile tilapia, Oreochromis niloticus fingerlings. A total of 630 Nile tilapia fingerlings with an average body weight of 26.4 ± 0.2 g were divided in the seven experimental net-pen treatments （three replicates each）. The experiment was conducted for 119 days. Seven isonitrogenous （26.50%） digestible protein and isocaloric （13.40 MJ kgl） digestible energy experimental diets were formulated. The control diet had no SC and EDE added. Diets 2-3 each contained SC at levels of 2 and 4 g 100 g diet-t, respectively, while diets 4-5 each contained EDE at levels of （0.64, 1.28, 0.16） and （1.28, 2.56, 0.32） g 100 gdiefI of pepsin, papain and a-amylase, respectively. Diet 6 contained mixture of SC and EDE at levels of 1 g yeast and 0.32, 0.64, 0.08 g of pepsin, papain and a-amylase, respectively 100 gdiet1 and diet D7 contained 2 g yeast and 0.64, 1.28, 0.16 g of pepsin, papain and a-amylase, respectively 100 g dietl. Growth performance and feed utilization efficiency of Nile tilapia were significantly （P 〈 0.05） higher in all treatments receiving SC and/or EDE supplemented-diets than the control diet which suggests that the addition of SC and EDE enhanced the growth performance. Red blood cells counts, hematocrit and hemoglobin were significantly （P 〈 0.05） highest in all treatments receiving mixture of SC and EDE supplemented-diets （D6 ＋ D7）. The same trend was observed for total plasma protein and total plasma globulin levels. The results of present study suggested that Nile tilapia fingerlings fed diets containing the mixture of I g yeast, SC and 0.32, 0.64, 0.08 g of pepsin, papain and a-amylase, respectively 100 gdiet^-1, for 119 days had enhanced growth performance, diet utilization efficiency and hematological indices.

Influences of Amazonian White and Black Waters on the Hematological and Biochemical Plasma Features of Arapaima gigas （Osteoglossiformes）

The Arapaima gigas, despite being an air breather, its gill structure is quite close to water breathers, especially in early stages of development. The effects of Amazonian waters is well notices in other Teleostei expose to BW （black water）, and WW （white water）. However, information about hematological adjustments and its implications to ionic regulation patters are scarce. Therefore, our aim was to analyzed A. gigas hematological parameters when exposed to BW and WW providing suitable hematological data concerning about physiological responses in Amazonian waters. Fish were acclimated in three separated ponds containing BW, WW and well water as control （C）. Blood samples were taken from the caudal vessel in order to perform measurement assays on levels of hemoglobin, hematocrit, mean corpuscular volume, corpuscular hemoglobin, corpuscular hemoglobin concentration, glucose, cholesterol and protein. Our findings corroborate the hypothesis stating that BW does interfere on fish adaptation specialy in smallfish （-100 g）. However in largefish （-1,000 g） neither WW or BW can interfere on plasma profile of analysed fish. Despite black water systems being considered a barrier constraining the dispersion of several species, this seems not to be a problem for this specie which has kept its ion-regulatory mechanisms even in black waters.