Plasma cortisol and glucose concentrations in the striped mullet (Mugil cephalus L.) subjected to intense handling stress

The plasma cortisol and glucose concentrations were determined in mature female striped mullet (Mugil cephalus L.) subjected to short term intense handling stress. The results indicated that plasma cortisol levels reached a peak 20 min after stress and declined gradually afterwards. The highest concentration of plasma glucose was observed 30 min after stress. The present study showed that the rise of plasma glucose was associated with the plasma cortisol levels.     

ANNUAL CHANGES IN PLASMA LEVELS OF CORTISOL AND SEX STEROID HORMONES IN MALE RAINBOW TROUT, ONCORHYNCHUS MYKISS

The profiles of cortisol, testosterone, ll-ketotestosterone and 17α， 20β- dihydroxy-4-pregnene-3-one in mule rainbow trout reared under constant water temperature and natural photoperiod were determined by radioimmunoassay. Gonads of male rainbow trout reached maturity when the fish were two years old. Changes in the plasma levels of both sex steroid hormones and cortisol were closely related to the GSI. Plasma levels of testosterone, 11-ketotestostemne and 17a, 2013-dihydroxy 4-pregnene-3-one showed a clear peak in the annual breeding season, when the GSI reached their maxima. Plasma cortisol levels also showed clearly seasonal changes in both two- and three-year-old fish. The results suggest that the elevated plasma levels of cortisol may not just be due to stresses during the breeding season but have certain physiological functions in the reproduction of rainbow trout.     

Low temperature stress on the hematological parameters and <Emphasis Type="Italic">HSP</Emphasis> gene expression in the turbot <Emphasis Type="Italic">Scophthalmus maximus</Emphasis>

To study the effect of low temperature stress on hematological parameters and HSP gene expression in the turbot (Scophthalmus maximus), water temperature was lowered rapidly from 18 to 1°C. During the cooling process, three individuals were removed from culture tanks at 18, 13, 8, 5, 3, and 1°C. Blood samples and tissues were taken from each individual, hematological indices and HSP gene expression in tissues were measured. The red blood cell count, white blood cell count, and hemoglobin concentration decreased significantly (P < 0.05) as temperature decreased. Enzyme activities of plasma alanine transaminase and creatine kinase increased as temperature decreased, whereas aspartic transaminase and γ-glutamyl transpeptidase activities displayed no obvious changes above 1°C and lactate dehydrogenase activity increased first and then decreased. Blood urea nitrogen and uric acid levels were highest at 8°C, and creatinine concentration was highest at 3°C. The concentrations of plasma cortisol, cholesterol, and triglyceride all increased significantly (P < 0.05) as temperature decreased. The serum glucose concentration increased first and then decreased to the initial level. The HSP70 mRNA expression showed various patterns in different tissues, whereas HSP90 mRNA expression showed the same tendency in all tissues. Overall, these results indicate that temperature decreases in the range of 8 to 5°C may induce a stress response in S. maximus and that temperature should be kept above 8°C in the aquaculture setting to avoid damage to the fish.     

低氧胁迫对珍珠龙胆石斑鱼氧化应激及能量代谢的影响

【目的】探究保活运输中低氧胁迫对珍珠龙胆石斑鱼(Epinephelus fuscoguttatus♀×Epinephelus lanceolatus♂)氧化应激的影响。【方法】诱导体质量为(498.2±27.3)g的杂交石斑鱼进入休眠状态,并放置在溶解氧为(1.74±0.21)mg·L^?1的低氧环境下,低氧胁迫0、6、12、24、48、72 h后,取样测定血清应激指标,肌肉、肝脏、鳃组织氧化应激指标及肌肉、肝脏能量利用指标。【结果】低氧胁迫过程中,杂交石斑鱼血糖含量在胁迫6 h时最大,72 h时恢复胁迫前水平;皮质醇含量在胁迫过程均显著高于胁迫前;肌肉超氧化物歧化酶(SOD)活性随胁迫时间的增加而上升,肝脏、鳃SOD活性先上升后下降;肌肉、肝脏过氧化氢酶(CAT)活性呈先升后降的趋势,鳃CAT活性在胁迫12、72 h显著低于胁迫前;肌肉、肝脏丙二醛(MDA)含量在胁迫过程中均增加,鳃MDA含量先降后升;肌肉谷胱甘肽还原酶(GSH)含量在胁迫12 h达到最大值,肝脏GSH含量呈先上升后下降的趋势,鳃中GSH含量在12 h时显著高于胁迫前,之后逐渐升高;总抗氧化能力(T-AOC)在肌肉、肝脏、鳃中均呈上升的趋势;肌肉和肝脏中糖原的含量显著减少,肌肉乳酸脱氢酶(LDH)活性显著增加,肝脏LDH先降后升。显著性水平均为0.05。【结论】低氧胁迫会刺激珍珠龙胆石斑鱼产生应激反应,影响体内正常的氧化应激系统,并对不同组织造成不同程度的氧化损伤。     

Mechanical Stress Induces Neuroendocrine and Immune Responses of Sea Cucumber(Apostichopus japonicus)

Grading procedure in routine sea cucumber hatchery production is thought to affect juvenile sea cucumber immunological response. The present study investigated the impact of a 3-min mechanical perturbation mimicking the grading procedure on neuroendocrine and immune parameters of the sea cucumber Apostichopus japonicus. During the application of stress, concentrations of noradrenaline and dopamine in coelomic fluid increased significantly, indicating that the mechanical perturbation resulted in a transient state of stress in sea cucumbers. Coelomocytes concentration in coelomic fluid increased transiently after the beginning of stressing, and reached the maximum in 1 h. Whereas, coelomocytes phagocytosis at 3 min, superoxide anion production from 3 min to 0.5 h, acid phosphatase activity at 0.5 h, and phenoloxidase activity from 3 min to 0.5 h were all significantly down-regulated. All of the immune parameters recovered to baseline levels after the experiment was conducted for 8 h, and an immunostimulation occurred after the stress considering the phagocytosis and acid phosphatase activity. The results suggested that, as in other marine invertebrates, neuroendocrine/immune connections exist in sea cucumber A. japonicus. Mechanical stress can elicit a profound influence on sea cucumber neuroendocrine system. Neuroendocrine messengers act in turn to modulate the immunity functions. Therefore, these effects should be considered for developing better husbandry procedures.     

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.     

急冷与空气暴露联合胁迫对凡纳滨对虾生理代谢影响

【目的】探明无水保活流通过程中凡纳滨对虾(Litopenaeus vannamei)响应急冷(acute cold,AC)与空气暴露(waterless duration,WD)联合胁迫的生理代谢调节规律。【方法】研究模拟产业运输实际,分析联合胁迫(AC+WD)进程中凡纳滨对虾血液和肝胰腺典型生化指标,以及肝胰腺组织病理变化。【结果】AC胁迫后,凡纳滨对虾生理应激指标血清皮质醇含量由正常组(normal control,NC)的(326.90±4.61)pg/mL显著增加到AC组的(427.16±7.92)pg/mL(P<0.05),AC+WD组则随时间延长而逐渐降低。血清葡萄糖含量在联合胁迫6 h后达到最大值(26.31±1.05)mg/dL,乳酸在9 h达到峰值(7.90±0.11)mmol/L。AC及AC+WD组中血蓝蛋白、尿素氮和肌酐含量较NC组均发生不同程度的增加,而总蛋白含量先升后降,总胆固醇和甘油三酯含量呈胁迫时间依赖性下降。血清谷草转氨酶、谷丙转氨酶、酸性和碱性磷酸酶活力呈上升趋势,并在9 h达到峰值。以上生化指标在胁迫复苏组有所恢复,表明AC+WD双重胁迫引起凡纳滨对虾物质代谢紊乱,复苏后机体生理应激和代谢系统发生一系列代偿性调节,且胁迫进程中肝胰腺组织病理损伤特征进一步解释了凡纳滨对虾响应环境联合胁迫的结构基础。【结论】无水保活流通过程中双重胁迫时间依赖性改变凡纳滨对虾生理应激水平和物质代谢,在9 h时可明显造成肝胰腺组织结构损伤,进而影响机体存活。     

基于S变换的唐山四分量钻孔应力时频特征分析

以2016-02～11期间唐山四分量深孔地应力连续观测数据为基础,使用S变换技术获得区域构造信息的时频特征,讨论其与近场小震及远场大震事件的关联性。研究发现,唐山台近场25 km范围内5次小震事件发生前的数日内,其地应力连续观测数据的S变换结果在720～1 440 min周期范围内均表现为高能量簇异常,且与潮汐因子的日常形态存在显著差异;2016-04-15日本九州M7.0级地震的同震效应主要对地应力S变换结果中周期小于240 min的频段具有较大影响,因此,唐山地应力高能量簇特征可能来自于区域构造应力场动态调整,而非远场大震的同震效应,说明地震事件前后的构造应力时频演化特征与近场地震活动可能存在较高的前兆关联性。     

STUDY ON KINETICS OF GLUCOSE UPTAKE BY SOME SPECIES OF PLANKTON

The. rates of glucose uptake by some species of plankton were determined by 3H-glucose tracer method. Experimental results indicated that the observed glucose uptake at natural sea water concentrations by Platymonas subcordiformis and Brachionus plicatilis was principally a metabolic process fitted with the Michaelis-Menten equation in the range of adaptive temperatures. Heterotrophic uptake by Platymonas subcordiformis was mainly dependent on diffusion at high glucose levels. The uptake by Brachionus plicatilis showed active transport even at high glucose levels, indicating its high heterotrophic activity. The uptake rate by Anemia salina was lower, and its Vm/K ratio was lower than those of the other two species of plankton.     

CO2协同低温有水对卵形鲳鲹麻醉保活效果的影响

【目的】探究二氧化碳CO₂麻醉协同低温对卵形鲳鲹有水保活的影响,延长卵形鲳鲹(Trachinotus ovatus)的保活时间。【方法】研究保活过程中CO₂质量浓度、麻醉时间、麻醉温度和鱼质量水比对卵形鲳鲹有水保活时长的影响;在最佳麻醉保活条件下,测定其血清生化指标、肌肉及肝脏氧化应激指标及代谢指标。【结果】卵形鲳鲹在16℃、100 mg/L CO₂水溶液中麻醉3 min其麻醉效果最佳。在15℃低温下进行有水保活,鱼水质量比为2∶1,存活时间最长,为(359±2)min,此条件下复苏率达100%。经过CO₂麻醉协同低温有水保活后的卵形鲳鲹血清谷草转氨酶(GOT)活性显著升高(P<0.05)、尿素氮(BUN)和甘油三酯(TG)含量上升,血糖含量下降。肌肉中的乳酸脱氢酶(LDH)活性显著上升(P<0.05),乳酸(LD)和糖原含量上升,过氧化氢酶(CAT)活性和丙二醛(MDA)含量下降;肝脏中的LDH活性显著上升(P<0.05)、CAT活性和LD的含量上升,MDA和糖原的含量下降。【结论】CO₂协同低温有水对卵形鲳鲹有较好的麻醉效果,有助于保活运输。在保活过程中卵形鲳鲹通过提升能量代谢水平和抗氧化能力应答环境应激,在一定程度上影响卵形鲳鲹保活过程的存活时间。     

A new simple screening method for the detection of paralytic shellfish poisoning toxins

The current testing for paralytic shellfish poisoning(PSP) in shellfish is based on the mouse bioassay(MBA).To alleviate animal welfare concerns,we evaluated the utility of using sublethal indicators of toxicity as an alternative to measuring time to death.Live mice were injected with a PSP congener and the changes in neurotransmitter levels were measured 60,90,and 120 min after injection.Acetylcholine(ACh) was the most sensitive marker for PSP toxicity.The changes in neurotransmitter levels were most pronounced in the blood.Thus,measurement of Ach levels in the blood may serve as a sensitive predictor for PSP that would not require sacrifice of the mice.This method was relatively simple,sensitive(less than 1 μg/kg weight,equivalent to 20 ng/mL),low maintenance,and rapid(less than 60 min).     

热应激对肉鸡血液生化指标及内分泌机能的影响

通过检测生产性能及血液生化指标和内分泌指标,研究了热应激对肉鸡的影响。结果表明：肉鸡体重、平均日增重、饲料转化率及血清总蛋白、白蛋白、钙、磷、甲状腺激素（Ｔ３、Ｔ４）含量显著下降（Ｐ＜０．０５或Ｐ＜０．０１）,血清钾则仅在３０日龄显著下降,但血清尿素氰、氯、皮质醇含量及血清乳酸脱氢酶、肌酸磷酸激酶活性显著升高（Ｐ＜０．０５或Ｐ＜０．０１）,热应激对血清钠和葡萄糖含量影响不大。     

Effect of dissolved oxygen on swimming ability and physiological response to swimming fatigue of whiteleg shrimp (Litopenaeus vannamei)

The swimming endurance of whiteleg shrimp(Litopenaeus vannamei, 87.66 mm ± 0.25 mm, 7.73 g ± 0.06 g) was examined at various concentrations of dissolved oxygen(DO, 1.9, 3.8, 6.8 and 13.6 mg L-1) in a swimming channel against one of the five flow velocities(v1, v2, v3, v4 and v5). Metabolite contents in the plasma, hepatopancreas and pleopods muscle of the shrimp were quantified before and after swimming fatigue. The results revealed that the swimming speed and DO concentration were significant factors that affected the swimming endurance of L. vannamei. The relationship between swimming endurance and swimming speed at various DO concentrations can be described by the power model(ν·tb = a). The relationship between DO concentration(mg L-1) and the swimming ability index(SA∫ 9000I), defined as SAI =vdt( cm), can be described as SAI = 27.947 DO0.137(R2 = 0.9312). The 0level of DO concentration directly affected the physiology of shrimp, and exposure to low concentrations of DO led to the increases in lactate and energetic substrate content in the shrimp. In responding to the low DO concentration at 1.9 mg L-1 and the swimming stress, L. vannamei exhibited a mix of aerobic and anaerobic metabolism to satisfy the energetic demand, mainly characterized by the utilization of total protein and glycogen and the production of lactate and glucose. Fatigue from swimming led to severe loss of plasma triglyceride at v1, v2, and v3 with 1.9 mg L-1 DO, and at v-11 with 3.8, 6.8 and 13.6 mg L DO, whereas the plasma glucose content increased significantly at v3, v4 and v5 with 3.8 and 6.8 mg L-1 DO, and at v5 with 13.6 mg L-1 DO. The plasma total protein and hepatopancreas glycogen were highly depleted in shrimp by swimming fatigue at various DO concentrations, whereas the plasma lactate accumulated at high levels after swimming fatigue at different velocities. These results were of particular value to understanding the locomotory ability of whiteleg shrimp and its physiological changes, further contributing to the improvement of capture and rearing technique.     

不同温度下马氏珠母贝干露耐受能力及其免疫相关基因表达的变化

【目的】探究马氏珠母贝(Pinctada fucata martensii)对干露胁迫的响应,为马氏珠母贝健康养殖和遗传育种提供基础资料。【方法】在不同温度下对马氏珠母贝进行干露胁迫,探究温度对马氏珠母贝干露耐受的影响;筛选出马氏珠母贝干露胁迫敏感组和耐受组,比较两组的免疫相关基因表达量和植核后休养期存活率。【结果】(1)22~34℃条件下,马氏珠母贝干露耐受能力随温度升高及干露时间延长而降低;(2)22℃下,S组和T组的脯氨酸羟化酶(PHD)、葡萄糖转运蛋白(GLUT)、髓样分化因子(MYD88)、热休克蛋白(Hsp70、Hsp20)和凋亡抑制因子(cIAP)6个基因在干露过程中表达量基本呈上升趋势,组间差异显著(P<0.05);(3)植核试验结果表明耐受组植核后休养期的存活率显著高于敏感组(P<0.05)。【结论】马氏珠母贝在高温环境下干露耐受能力降低,在适温条件下耐受组比敏感组表现出较高的存活率。     

环境胁迫对栉孔扇贝(Chlamys farreri)血浆蛋白及其抗菌活性的影响

采集采自山东某养殖场的栉孔扇贝(Chlamys farreri)健康贝和病贝的血淋巴,测定其血浆蛋白浓度、血浆蛋白图谱和相对分子质量,以及血浆抗菌活性、凝集素活性等几种重要的体液免疫因素,研究环境胁迫对血浆蛋白及其抗菌活性的影响。结果表明:栉孔扇贝(C.farreri)血浆蛋白质量浓度约1 260~1 650μg/mL;血浆蛋白SDS-PAGE图谱显示3条蛋白带,相对分子质量分别为34 000、25 000和14 000;健康贝血浆抗菌活性指数BI为0.34,病贝BI为0.21;凝集素活力水平为1.602~2.204。氨氮胁迫下血浆蛋白浓度下降约780μg/mL,凝集素活力下降为1.301;高温胁迫下血浆蛋白质量浓度下降约为900μg/mL,凝集素活力为1.903;高盐胁迫下血浆蛋白浓度下降约为990μg/mL,凝集素活力为2.204。     

Pharmacokinetics and tissue behavior of enrofloxacin and its metabolite ciprofloxacin in turbot Scophthalmus maximus at two water temperatures

Turbot Scophthalmus maximus, an important aquaculture species in China, currently suffers from epizootic diseases because of high density aquaculture. Enrofloxacin has been used to treat various systemic bacterial fish infections. However, studies concerning the pharmacokinetics of enrofloxacin in turbot are limited. In this study, the pharmacokinetics of enrofloxacin and its metabolite ciprofloxacin, were investigated in the turbot following intravenous and oral administration at 10 mg enrofloxacin/kg body weight, at 16°C and 10°C water temperatures. The concentrations of enrofloxacin and ciprofloxacin in the main tissues (plasma, muscle, liver and kidney) were detected by HPLC. The results show that the plasma concentration-time data for enrofloxacin were best described as a two-compartment open model after intravenous and oral administration. Three pharmacokinetic equations were established between the concentrations and temperatures. The kinetic profile of enrofloxacin was temperature dependent. The absorption half-life of enrofloxacin was 1.99 h and 2.17 h after oral administration, whereas the elimination half-life of the drug was 98.63 h and 136.59 h at 16°C and 10°C, respectively. The peak concentration of enrofloxacin in plasma and tissues was higher at 16°C than that at 10°C, and the peak plasma concentration time in the liver was the shortest at both temperatures among those of other tissues. The plasma C max /MIC ratio varied between 11.08 and 5 540.00 at 16°C; and between 7.92 and 3 960.00 at 10°C. The AUC/MIC ratio was 467.82-280 690.00 at 16°C, and 359.48-215 690.00 at 10°C. These ratios indicate that it is possible to obtain therapeutic efficacy. Very low levels of ciprofloxacin were detected. The AUC ratios of ciprofloxacin and enrofloxacin in plasma suggest that plasma ciprofloxacin might play a minor role in enrofloxacin treatment for turbot.     

Changes of serum thyroid hormone and plasma catecholamine of 16 th and 17 th Chinese Expeditioners in Antarctic environment

The serum thyroid hormone and plasma catecholamine were examined in 18 male and 2 female members of the Chinese Antarctic Expedition (who spent the 2000 or 2001 austral winter at the Great Wall Station) . The changes of serum thyroid hormone i. e. total thyroxine (TT4) and free T4 (FT4) , total triodothyronine (TT3) and freeT3 ( FT3 ) , thyroid stimulating hormone ( TSH ) and plasma catecholamine, including norepinephrine (NE) , epinephrine ( E) and dopamine ( DA ) , were investigated by Chemoluminescence Immunoassay (CLIA) and High Performance Liquid Chromatography with electrochemical detection (HPLC-ECD) . Samples were taken at different time; (1)1 day before departure to Antarctica (16th expedition 1999/12/ 09; 17th expedition 2000/12/06). (2) 1 day after returned to China after living 54 weeks in Antarctica ( 16th expedition 2000/12/25 ; 17th expedition 2001/12/25 ). Comparing the data of before departure and returned, results showed that there was a significant decrease in the contents of TT4 (P <     

Acclimation-dependent expression of heat shock protein 70 in Pacific abalone (Haliotis discus hannai Ino) and its acute response to thermal exposure

Heat shock protein 70 (Hsp70) is one important member of heat shock protein (Hsp) family that is responsible for various stresses, especially thermal stress. Here we examined the response of Hsp70 gene to both chronic and acute thermal exposure in Pacific abalone (Haliotis discus hannai Ino). For the chronic exposure, abalones were maintained at 8, 12, 20, and 30°C for four months and their mRNA levels were measured. The highest mRNA level of Hsp70 gene relative to actin gene was detected in the 30°C-acclimated group, followed by the 8°C-acclimated group and then the 12°C- and 20°C-acclimated groups. After the long-term acclimation, gills from each of the above acclimation groups were dissected and exposed to different temperatures between 8°C and 38°C for 30 min. Hsp70 expression in gills acclimated to different temperatures responded differentially to the same temperature exposure. The incubation temperature that induced maximum Hsp70 mRNA expression was higher in the higher temperature acclimation groups than lower temperature groups. Pacific abalones could alter the expression pattern of Hsp70 gene according to environmental thermal conditions, through which they deal with the stress of thermal variations.     

贝类精子质量检测与评价方法的研究Ⅴ:栉江珧精子线粒体活性的荧光检测

采用罗丹明123(Rh123)和碘化丙啶(PI)两种荧光染料对栉江珧(Atrina pectinata)精子线粒体活性进行检测,比较不同染液浓度和染色时间的检测效果,并用该方法检测和比较超低温冷冻保存对线粒体活性的影响。结果显示:Rh123和PI染色的最适染液浓度均为10μg/mL,染色时间控制在10 min之内;染色后具有线粒体活性的精子发出绿色荧光,死精子发出红色荧光,质膜受损但线粒体还未受损的精子发出红绿色荧光,表明Rh123和PI双荧光染色检测栉江珧精子线粒体活性是可行的;精液经冷冻后,线粒体活性为61.9%、质膜完整率56.8%,显著低于新鲜精液组(95.3%、93.5%),表明冷冻对精子结构会造成不同程度的损伤,Rh123和PI双荧光染色法可用来检测栉江珧精子质量。     

禁食暂养对卵形鲳鲹有水保活生理响应的影响

【目的】探究禁食暂养对卵形鲳鲹有水保活生理响应的影响,为卵形鲳鲹的保活运输奠定基础。【方法】通过测定暂养过程中水体的总氨氮和pH值、卵形鲳鲹血液生理生化指标、肌肉氧化应激指标等的变化,分析确定卵形鲳鲹有水保活的禁食暂养条件。【结果与结论】禁食暂养过程中,随着暂养时间的延长,水体总氨氮含量与pH值显著上升,但在禁食暂养时间为6 h时,其总氨氮质量浓度为3.22 mg/L和pH值为7.80;禁食暂养6 h可以降低或减少卵形鲳鲹的应激,使血液中乳酸脱氢酶(LDH)和谷草转氨酶(AST)活性以及皮质醇(Cortisol)、血糖(GLU)、尿素氮(BUN)和总胆固醇(T-CHO)含量处于较低水平,分别为2785.53 U/L、19.04 U/L、158.72 ng/mL、2.98 mmol/L、4.35 mmol/L和4.96 mmol/L;同时减轻了应激带来的肌肉组织氧化损失,使过氧化氢酶(CAT)活性和丙二醛(MDA)含量维持在较低水平,分别为6.34 U/L和5.86 mmol/L;卵形鲳鲹适宜的暂养条件为在鱼水比m鱼∶m水=1∶20和(25±1)℃的洁净海水中暂养6 h。