Ontogenic changes in the digestive enzyme patterns and characterization of proteases in Indian major carp Cirrhinus mrigala

Digestive enzymes of Cirrhinus mrigala (Ham.) were studied during ontogenic development. Specific amylase activity was detected in first feeding fish. The enzyme activity decreased up to day‐18 and then it increased with the age of fish to reach the highest level on day‐34. Protease activity was 28.61 ± 8.90 mU mg protein^?1 min^?1 on day‐4 and increased with the age throughout the study period. Trypsin activity was 31.86 ± 1.12 mU mg protein^?1 min^?1 on day‐4. The activity decreased up to day‐10 and from day‐12 onwards increased up to day‐26. Chymotrypsin activity was 14.56 ± 2.74 mU mg protein^?1 min^?1on day‐4 and constantly increased up to day‐26. A significant increase in lipase activity was observed between days‐24 and 34. SDS‐PAGE and substrate SDS‐PAGE showed the diversity of protein (17.4–127.8 kDa) and protease activity bands (16.6–88.8 kDa) during ontogenesis. Soybean trypsin inhibitor, phenyl methyl sulphonyl fluoride, N‐α‐p‐tosyl‐l ‐lysine chloromethylketone and N‐tosyl‐l ‐phenylalanine chloromethylketone inhibited the protease activity up to 79.72–97.21, 65.55–94.83, 45.41–75.31 and 40.78–64.72%, respectively. Inhibition study in substrate SDS‐PAGE revealed the abundance of serine proteases and the presence of isoforms of trypsin and chymotrypsin. Ethylenediamine‐tetraacetate showed 5.56–22.78% inhibition of metal ion‐specific enzyme activity.     

梭鱼早期发育过程中生长及主要消化酶活性的研究

选取梭鱼(Liza haematocheila)受精卵及出膜后3~28日龄鱼苗为研究对象,研究了主要消化酶活性的变化以及鱼苗的生长特点。结果表明,梭鱼苗全长、体重与日龄均呈指数相关;全长与体重呈幂函数增长相关,且为异速生长。梭鱼受精卵及鱼苗可检测到胰蛋白酶、胃蛋白酶、淀粉酶、碱性磷酸酶和酸性磷酸酶活性,且在受精卵中活性较低,未检测到脂肪酶活性。3~16日龄,胃蛋白酶活性逐渐降低至最小值,胰蛋白酶活性先升高,最大值出现在10日龄,之后降低,16日龄后,两种酶活性均显著升高(P0.05),均在28日龄达到最大;淀粉酶活性分别在3~10日龄和22~28日龄不断升高,最大值出现在28日龄;3~7日龄,碱性磷酸酶和酸性磷酸酶均显著升高(P0.05),7~22日龄碱性磷酸酶不断降低,酸性磷酸酶保持稳定,22~28日龄,两种磷酸酶活性均显著升高(P0.05),碱性磷酸酶和酸性磷酸酶活性的最大值分别在7日龄和28日龄。研究表明,梭鱼仔鱼消化酶并非外源性饵料所致,其活性与梭鱼发育阶段密切相关,亦在不同程度上受到外界环境和食性等因素的影响。     

鲢、鳙滤食及消化器官的发育、构造与食性的相互关系

本文通过大体解剖、显微解剖、组织切片及扫描电镜等方法研究鲢、鳙不同发育阶段滤食及消化器官与食性的相互关系。鳃耙左右侧对称,从外向内面积递减。鲢全长26mm时,分离的鳃耙间开始出现桥形横联;全长 85mm,开始形成外鳃耙网。鳙鳃耙的分化发生于全长 41—99mm。成鱼鳃耙均有许多粘液孔;仔鱼期鳃耙端部无粘液孔,鳃耙沟中有少量粘液孔,鳃耙基部侧缘有侧突的雏型。腭褶表面均有次级腭褶及微腭褶,微腭褶表面有味蕾分布。腭褶内部、咽上器官周围及每一鳃耙管外围均有与滤食有关的肌纤维群。鲢、鳙肠管的长度及直径发育早期相同,后期鳙的肠管短于鲢,管径大于鲢。滤食下限规格,鲢全长7.8mm 时为 25,全长56mm 以后为 10μ;鳙终生为 25μ。着重探讨了进食过程及有关器官在进食过程中的作用。     

Uptake and clearance of exogenous estradiol-17β and testosterone during the early development of coho salmon (Oncorhynchus kisutch), including eggs,alevins and fry

The uptake and clearance of estradiol-17β (E₂) and testosterone (T) were examined during the initial stages of development of coho salmon (Oncorhynchus kisutch), including eyed-eggs, newly hatched alevins and first feeding fry. Radiolabeled steroids were administered through the water in tracer amounts with or without their nonradioactive form at 400 μg l⁻¹. Regardless of developmental stage, saturation levels were invariably attained earlier for T than for E₂, thus resulting in a higher incorporation of E₂. However, both steroids had similar clearance patterns. Uptake and clearance was clearly stage-dependent, being fastest in fry, intermediate in alevins and slowest in eggs. Furthermore, combined uptake and clearance patterns showed that exposure to steroid was also higher for E₂ than for T and stage-dependent, but always markedly highest in alevins. Subsequently, based on the observed elimination of the estrogen, a double immersion in E₂ at 400 μg 1⁻¹, administered 2 days apart to maximize exposure during the alevin stage, was assayed for its effect on sex reversal and found to induce the production of 100% females. We suggest that the yolk, which is present in substantial amounts during the initial stages of development in salmonids, can retain the exogenously administered liposoluble steroids, thus providing developing embryos with an extended supply of, and exposure to, these steroids well after the treatment is finished. Together, these findings help to explain the previously observed high effectiveness of sex steroids administered during early development in regulating gonadal differentiation in salmonids, the higher effectiveness of E₂ compared to T, and clarify the localization of the most sensitive period to the action of exogenous steroids at the alevin stage in the coho salmon.     

Thermal requirements for the embryonic development and life cycle of Meloidogyne hispanica

The life cycle of a Portuguese Meloidogyne hispanica isolate on susceptible cv. Easypeel and resistant (Mi‐1.2 gene) cv. Rossol tomato plants was studied in growth chambers at constant temperatures (10–35°C). The development within the egg and hatching were compared to those of a Portuguese M. arenaria isolate. The base temperature was 10·11 and 8·31°C with 179·5 and 235·3 thermal units for M. hispanica and M. arenaria, respectively, suggesting better potential adaptation to low temperatures by M. arenaria than M. hispanica. No egg development occurred at 10 or 35°C. An increase in invasion of tomato roots by M. hispanica second‐stage juveniles (J2s) was correlated with an increase in temperature on both tomato cultivars. Tomato cv. Rossol limited M. hispanica development at 20, 25 and 30°C, but not at 35°C, indicating that these high temperatures blocked the resistance mechanism provided by the Mi‐1.2 gene. At 15°C, J2s penetrated tomato cv. Rossol roots, but failed to develop and establish feeding sites. On tomato cv. Easypeel, nematode development and reproduction occurred at 20, 25 and 30°C, but at 20°C the life cycle was 1·5 and 2·0 times longer than at 25 and 30°C, respectively. No egg production was observed at 15°C. The results of this study showed that M. hispanica is most suited to soil temperatures around 25°C. Predicted climate change might favour the spread of this nematode species into southern Europe and northwards. The thermal requirements for M. hispanica development are analysed and compared with those of M. arenaria, M. hapla, M. incognita and M. javanica.     

鲈鱼人工繁殖和早期个体发育研究

使用鲤鱼垂体ＰＧ和ＨＣＧ混合激素诱导催产，干法人工授精，鲈鱼受精率为８７％。受精卵为浮性圆球形，卵径１．２５～１．４７ｍｍ，在水温７～１６℃，盐度２２％的水中，经７９．５ｈ孵化出仔鱼，孵化率为６５％。初出膜仔鱼长３．１３ｍｍ，卵黄囊大，体上布满分枝状色素，肌节３５对左右。出膜后８天，口通体外，并开始摄食微型单细胞藻体，至第１９天，仔鱼成活率为３５％。     

Metabolic enzyme activities in larvae of the African catfish,Clarias gariepinus: changes in relation to age and nutrition

The influence of ontogeny and nutrition on metabolic enzyme activities in larvae of the African catfish, Clarias gariepinus, was studied. After start of exogenous feeding, the larvae were reared for 10 days under three different nutritional conditions: Artemia nauplii, a dry starter diet, and starvation. The live feed gave the best growth (96 mg within 10 days) whereas the dry diet resulted in low growth (33 mg). This growth difference was reflected in larval RNA and DNA concentrations, but not in the levels of soluble protein. Enzymes representing the following aspects of metabolism have been analysed: NADPH generation (G6PDH, ME), glycolysis (PFK, PK), gluconeogenesis (FDPase), amino acid catabolism (GOT, GPT) and oxidative catabolism (CS). All enzymes were present from the start of exogenous feeding onwards, but their maximum specific activities displayed different developmental patterns. In catfish larvae fed on Artemia, G6PDH and ME activities steadily increased with age and weight of the larvae. CS levels remained, after an immediate enhancement upon onset of exogenous feeding, on a rather stable plateau. The amino acid-degrading enzymes GOT and GPT showed maximum levels at days 3–5 of feeding or at a body weight of 10–20 mg, but decreased thereafter. Activities of PFK, PK and FDPase showed low initial levels, and increased significantly with age and size. Based on the ontogenetic patterns of metabolic enzymes, in C. gariepinus larvae an early and a late developmental phase can be distinguished. During the early phase, the glycolytic and gluconeogenetic capacities are low, whereas they are enforced during the later phase. The oxidative capacity is high both during the early and the late phase. The metabolic changes in catfish development coincide with other major ontogenetic events, e.g., alterations of muscle organization, gill morphology, respiration and stomach structure and function. Rearing catfish larvae on a dry diet instead of Artemia partly altered the developmental pattern described: The ontogenetic elevation of CS, PFK and FDPase was delayed and the early peak in GOT and GPT activities was not realized. Particularly during the early developmental phase, the enzyme behaviour of the larvae fed on dry food was similar to that of starved larvae.Abbreviations CS citrate synthase - FDPase fructose-1,6-diphosphatase - GOT glutamate oxaloacetate transaminase - GPT glutamate pyruvate transaminase - G6PDH glucose-6-phosphate dehydrogenase - ME malic enzyme - PFK phosphofructokinase - PK pyruvate kinase     

单环刺螠消化道的发生和分化

用组织学、组织化学和电镜技术对单环刺螠（Urechis unicinctus）消化道的发生及分化进行研究。结果表明，单环刺螠消化道起源于原肠胚期内陷而成的原肠腔；至早期担轮幼虫时期，消化道连通，开始摄食，此时据管腔直径不同消化道可分为口、食道、胃、中肠及后肠。附着变态后，其消化道在结构及功能上都有了明显的发育与分化。至蠕虫状幼虫时期。食道、胃及后肠已出现黏膜下层及肌层。嗉囊与砂囊可明显区分。中肠已产生肌纤维的分化，呼吸肠由中肠分化而来，其胞体细长，且核质比较小，可与中肠明显区分。至幼螠时期，食道前端产生了咽部的分化，至此消化道各部位在结构及功能上都与成体有了很大的相似性。即由口、咽、食道、嗉囊、砂囊、胃、中肠、呼吸肠、后肠和肛门组成。组织化学研究结果表明，随着发育，消化道各部位分别呈现不同的碱性磷酸酶、酸性磷酸酶、脂酶、ATP酶、非特异性酯酶及多糖类物质活性。     

Using Hemispherical Radiation Measurements to Predict Weight-Related Growth Traits in Oilseed Rape (Brassica napus L.) and Barley (Hordeum vulgare L.) Canopies

Methods for determining growth traits in the field are mostly work‐intensive and/or often erroneous. Further development of multi‐temporal techniques of growth analyses provided by fast and non‐destructive tools for monitoring canopy properties may result in a substantial gain for practical use. Among them, the LAI‐2000 Canopy Analyzer can be used to determine the canopy area index (CAI) resulting from radiation measurements. In plant breeding, single‐date measurements were usually made across a wide number of genoptypes. In contrast, multi‐temporal information on canopy status is needed in agronomy. In our study, data of the CAI based on hemispherical radiation measurements with the LAI‐2000 were used to estimate developmental courses of weight‐related growth traits. For this purpose, seasonal courses of shoot fresh matter, shoot dry matter, shoot N and water content and CAI of winter oilseed rape (Brassica napus L.) and spring barley (Hordeum vulgare L.) were investigated simultaneously in field experiments from 2002 to 2006 in northern Germany with and without nitrogen (N) fertilization. Courses of 2003 and 2004 were used to calculate relationships between CAI and growth traits. Results show that each of the weight‐related growth trait is related to the CAI. In this mechanistic approach relationships were found gathered across 2 years, two N‐treatments and several growth stages being unaffected by N‐fertilization, cultivar and experimental year. These relationships were verified using independent data sets of the years 2005 (winter oilseed rape and spring barley) and 2006 (winter oilseed rape). The results of the current study show that the use of LAI‐2000 and its rapid and non‐destructive determination of weight‐related growth traits over the entire growth period will substantially improve the determination of weight‐related growth traits in agriculture.     

斑马鱼发育过程中DANA相关基因的定量分析

DANA是广泛分布于斑马鱼基因组中的一个短散在核重复序列家族,通过实时荧光定量PCR相对定量方法,检测与DANA相关基因mul1a、er-Gicp2在斑马鱼整个个体发育过程中的相对表达量。结果显示:mul1a基因在1细胞期到1 000细胞期,即早期卵裂期内表达量差异不明显,从sphere时期到成体时期,其表达量显著性下调,并在shield时期降到最小值,为1细胞时期的0.009倍;er-Gicp2基因从2细胞到成体时期持续下调,到成体时期其表达量只有1细胞时期的0.023倍。