Effects of calcium and phosphate on the corpuscles of Stannius of the teleost fish,Oreochromis mossambicus

Summary Removal of the corpuscles of Stannius (CS) in Oreochromis mossambicus leads to hypercalcemia and hypophosphatemia. The effects on CS size and ultrastructure of different calcium and phosphate concentrations of the ambient water and of the food were investigated. A six-fold increase of the calcium concentration of the water leads to a four-fold increase in CS volume; this is mainly caused by an increase in the size and number of the type-1 cells. The effect of external calcium is most probably mediated by the calcium concentration of the blood plasma. Plasma ionic calcium may be the relevant factor. Changes in the calcium concentration of the food had no effect on the CS. Similarly, hyperphosphatemia or hypophosphatemia induced by high phosphate concentrations of the water or the food, or by a phosphate-deficient diet, had no noticeable effect on the CS. The results support the hypothesis that the type-1 cells produce the hypocalcemic factor of the CS. There is no evidence for the production by the CS of an endocrine factor involved in the control of phosphate metabolism.     

Evidence of steroidogenesis in postovulatory follicles of the tilapia,Oreochromis mossambicus

Summary Postovulatory follicles of the tilapia, Oreochromis mossambicus, were examined with electron microscopy and enzyme histochemistry for evidence of steroid-hormone production. Light microscopy was also used to examine changes in the ovary with time after spawning. Electron microscopy detected the presence of smooth endoplasmic reticulum, lipid droplets, and mitochondria with tubular cristae in certain cells of the theca interna. These structures are suggestive of cells that synthesize steroid hormones. Granulosa cells also contained some smooth endoplasmic reticulum, along with an augmentation of Golgi complexes, vesicles, microvilli, and microfilaments within 5–7 days after spawning. Enzyme histochemistry demonstrated an intense reaction of 5, 3-hydroxysteroid dehydrogenase (3-HSD) in variably placed thecal cells up to 7 days after spawning. At this time, the thecal cells of vitellogenic oocyte follicles also began to show strong 3-HSD activity. During the first 7 days after spawning, there was an increase in young primary oocytes and recruitment of some of these to vitellogenic oocytes. By 10 days after spawning, certain thecal cells in the follicles of these vitellogenic oocytes showed an intense 3-HSD reaction, while the postovulatory follicular tissue demonstrated a weak reaction. This arrangement continued for the lifespan of the postovulatory follicular tissue. Postovulatory follicles had a lifespan of up to 25 days after spawning in females that continued to hold the developing fry inside their mouths, i.e., mouthbrooders. At 25 days after spawning, the postovulatory follicular tissue showed signs of degeneration with the presence of vacuoles and lysosomes. In females that ate the zygotes, therefore exhibiting no parental behavior, the postovulatory follicular tissue showed signs of degeneration at l0days after spawning. In these females, the next clutch of eggs also developed at a higher rate than in mouthbrooders.     

Stomach pH,feeding rhythm and ingestion rate in Oreochromis niloticus L. (Pisces: Cichlidae) in Lake Awasa,Ethiopia

Fish were caught in February and October, 1985 with gill and trawl nets at different time intervals. The length and weight of the fish, the stomach pH and the weight of the stomach contents were determined. The feeding pattern of O. niloticus was observed to have a diel rhythm. The stomach became more acidic towards the afternoon and most of the evening. The pH of the stomach had a significant regression on stomach fullness. The feeding rate of O. niloticus in Lake Awasa was estimated from daily feeding periodicity and rate of gastric evacuation. The fish ingests 11.5% of their wet body weight daily at an average water temperature of 21.5 °C. It is believed that the fish ingests and assimilates adequate supply of energy (9.1 KJ/day) for growth, maintenance and other energy requiring activities of the fish.     

温度和盐度对吉富品系尼罗罗非鱼幼鱼鳃Na+-K+-ATPase活力的联合效应

试验采用中心组合设计(central composite face-centered design,CCF)和响应曲面法(response surface methodology,RSM)研究了温度(12—34℃)和盐度(0—26)两因素对体长为(4.36±0.105)cm,体重为(2.45±0.153)g的吉富品系尼罗罗非鱼(GIFT Nile tilapia,Oreochromis niloticus;简称吉富罗非鱼)幼鱼鳃Na+-K+-ATPase活力的联合效应。结果表明:(1)温度和盐度的一次效应和二次效应对Na+-K+-ATPase活力影响极显著(P<0.01),温度和盐度的互作效应不显著(P>0.05);(2)经响应曲面法分析,随着温度和盐度的增大,Na+-K+-ATPase活力呈先减小后增大的趋势;(3)建立了Na+-K+-ATPase活力与温度、盐度间关系的模型方程(R2=0.9829,Pred.R2=0.8550,P<0.01),并可用于预测吉富罗非鱼幼鱼鳃Na+-K+-ATPase的活力;(4)优化结果显示,温度为24.15℃,盐度为11.75时,Na+-K+-ATPase活力最小为0.62μmol无机磷.mg-1蛋白.h-1,满意度函数值高达0.961。Na+-K+-ATPase活力可以作为检测罗非鱼生长性能的指标,其活力较低时,一般反映了鱼体生存环境适宜,生长代谢旺盛,消耗于渗透调节的能量较少。     

奥利亚罗非鱼与尼罗罗非鱼rDNA内转录间隔区序列特征

核糖体DNA内转录间隔区(internal transcribed spacers,ITS)是经常被用作种和种群水平系统研究的分子序列.本文分离了奥利亚罗非鱼(Oreochromis aureus)、尼罗罗非鱼(O.niloticus)内转录间隔区,包括部分185序列,ITS1、5.8S、ITS2全序列及部分28S序列.4尾奥利亚罗非鱼的10个克隆序列分析表明,其存在长度不同的a、b两种类型ITS1.a型长为536 bp,GC含量为69.96%;b型长为520 bp,GC含量为69.04%～69.42%.4尾尼罗罗非鱼的10个克隆序列分析表明,其只存在a型ITS1,长为536～540 bp,GC含量为69.42%～70.19%.与b型ITS1相比,a型ITS1在16～31 nt有16 bp片段(GGCCCGCCTCGGCGC)的插入.奥利亚罗非鱼和尼罗罗非鱼共20条ITS序列中,5.8S长度均为157 bp,GC含量为56.69%～57.96%;ITS2为408 bp,GC含量为72.79%～74.26%.奥利亚罗非鱼和尼罗罗非鱼ITS区序列相似性高达98.2%,表明这两种罗非鱼亲缘关系很近.此外,本文对14尾奥利亚罗非鱼、15尾尼罗罗非鱼以及15尾奥尼罗非鱼[O.aureus(♂)×O.niloticus(♀)]ITS1的扩增结果显示,奥利亚罗非鱼均有a、b两种类型ITS1;15尾尼罗罗非鱼中1尾为a、b两类型ITS1,14尾为a型ITS1;15尾奥尼罗非鱼中则有6尾具有a、b两类型ITS1,9尾为单一的a型ITS1.分析表明,奥利亚罗非鱼在ITS1这个位点一致性高,但尼罗罗非鱼中有1尾混杂了奥利亚罗非鱼的基因,同时也说明分子生物学手段应用于种质鉴定比形态学手段更为精确.     

Antagonism of Aeromonas hydrophila by propolis and its effect on the performance of Nile tilapia, Oreochromis niloticus

Propolis, a resinous substance collected by Apis mellifera bees from various plant sources and mixed with secreted beeswax, is a multifunctional material used by bees in the construction, maintenance, and protection of their hives. The collected propolis sample, from High Egypt, was dark-green with olive-odor. The minimal inhibition concentration (MIC) of propolis-ethanolic-extract, against Aeromonas hydrophila, was 80 μg Propolis-ethanolic-extract and crude propolis (1%) were added to artificial basal diet with (30% crude protein) to evaluate their efficacy on the fish growth-performance, immunostimulation and resistance to A. hydrophila. Two hundred and twenty-five Oreochromis niloticus (8 ± 0.45 g/fish) were divided into three equal treatments (T) of triplet replicates. The fish of T₁ were fed on basal diet (control). The fish of T₂ were given the basal diet, containing propolis-ethanolic-extract. The fish of T₃ were given the basal diet containing crude propolis for 28 day. The fish were intraperitoneally challenged by A. hydrophila (0.2 × 10⁷ cells ml⁻¹) at the end of the feeding period and kept for 15 more days.The best growth rate and feed conversion ratio were obtained with T_2. The increase in the average daily gain, specific growth rate and feed efficiency ratio were highly significances in T₂ followed by T₃ when compared with the control group. The HCT-level and monocyte-counts were increased (T₂). No significant change, in the large lymphocytic-count was found among the three treatments (28–27–28%), while the neutrophil-count was significantly decreased (7%) with T₂ and increased (13.11%) with the control. A significant increase in serum lysozyme and serum bactericidal activities was found with T₂. The RLP against A. hydrophila was high with T₂ and T₃.The propolis-ethanolic-extract enhanced the growth, immunity and resistance of O. niloticus against A. hydrophila more than the crude propolis.     

Effects of zinc and cadmium on erythrocyte antioxidant systems of a freshwater fish Oreochromis niloticus

In this work to determine the effects of metals exposure of Oreochromis niloticus on erythrocyte antioxidant systems, fish were exposed to 5.0 mg/L Zn, 1.0 mg/L Cd, and 5.0 mg/L Zn + 1.0 mg/L Cd mixtures for 7 and 14 days and reduced glutathione (GSH) level, catalase (CAT), and glucose‐6‐phosphate dehydrogenase (G6PD) activities were investigated. In addition, Zn or Cd levels in whole blood were studied. Erythrocyte GSH level and CAT and G6PD enzyme activities increased in response to single and combined Zn and Cd exposure. The elevation observed in the CAT activity was higher in the Cd alone, and in combination with Zn, than in Zn alone. Time‐dependent alteration was not observed in all antioxidant parameters. Exposure to metals (alone and in mixture) resulted in elevatation of Zn and Cd levels in the blood. Concentration of metals in the blood of fish exposed to the Zn + Cd combination was lower than in fish exposed to the single metal. This study demonstrates that metals caused oxidative stress in fish erythrocytes, and an adaptation with an increase in CAT and G6PD activities and GSH level, which were important in the protection against metal damage, was observed. © 2010 Wiley Periodicals, Inc. J Biochem Mol Toxicol 24:223–229, 2010; View this article online at wileyonlinelibrary.com . DOI 10.1002/jbt.20327     

Identification of a sex-determining region in Nile tilapia (Oreochromis niloticus) using bulked segregant analysis

Sex determination in the Nile tilapia (Oreochromis niloticus) is thought to be an XX-XY (male heterogametic) system controlled by a major gene. We searched for DNA markers linked to this major locus using bulked segregant analysis. Ten microsatellite markers belonging to linkage group 8 were found to be linked to phenotypic sex. The putative Y-chromosome alleles correctly predict the sex of 95% of male and female individuals in two families. Our results suggest a major sex-determining locus within a few centimorgans of markers UNH995 and UNH104. A third family from the same population showed no evidence for linkage of this region with phenotypic sex, indicating that additional genetic and/or environmental factors regulate sex determination in some families. These markers have immediate utility for studying the strength of different Y chromosome alleles, and for identifying broodstock carrying one or more copies of the Y haplotype.     

The sulfide tolerance of milkfish and tilapia in relation to fish kills in farms and natural waters in the Philippines

Fish kills of milkfish Chanos chanos and tilapia Oreochromis spp. now occur frequently in brackish, marine, and freshwater farms (ponds, pens, and cages) in the Philippines. Aquafarms with high organic load, limited water exchange and circulation, no aeration, and high stocking and feeding rates can become oxygen-depleted and allow sulfide from the sediments to appear in the water column and poison free-swimming fish. The sulfide tolerance of 2–5 g milkfish and 5–8 g O. mossambicus was determined in 25-liter aquaria with flow-through sea water (100 ml min-1) at 26–30 °C and sulfide stock solutions pumped in at 1ml min-1. Total sulfide concentrations in the aquaria were measured by the methylene blue method and used in the regression against the probits of % survival. Four experiments showed that the two species have similar sulfide tolerance. In sea water of pH 8–8.5, about 163 ± 68 μM or 5.2 ± 2.2 mg l-1 total sulfide (mean ± 2 se) or 10 μM or 313 μg l-1 H2S was lethal to 50% of the fish in 4–8 h, and 61 ± 3 μM total sulfide or 4 μM H2S in 24–96 h (to convert all sulfide concentrations: 1 μM = 32 μg l-1). Earthen pond bottoms had 0–382 μM total dissolved sulfide (mean ± sd = 54 ± 79 μM, n = 76); a tenth of the samples had >200 μM. The water column may have such sulfide levels under hypoxic or anoxic conditions. To simulate some of the conditions during fish kills, 5–12 g milkfish were exposed to an abrupt increase in sulfide, alone or in combination with progressive respiratory hypoxia and decreasing pH. The tests were done in the same flow-through set-up but with sulfide pumped in at 25 ml min-1. The lethal concentration for 50% of the fish was 197 μM total sulfide or 12 μM H2S at 2 h, but 28–53 μM sulfide allowed fish to survive 6–10 h. Milkfish in aquaria with no aeration nor flow-through sea water died of respiratory hypoxia in 5–8 h when oxygen dropped from 6 to 1 mg l-1. Under respiratory hypoxia with 30–115 μM sulfide, the fish died in 2.5–4 h. Tests with low pH were done by pumping a weak sulfuric acid solution at 25 ml min-1 into aquaria with flow-through sea water such that the pH dropped from 8 to 4 in 5 h. Under these conditions, milkfish died in 7–9 h when the pH was 3.5. When 30–93 μM sulfide was pumped in with the acid, the fish died in 2–6 h when the pH was still 4.5–6.3. Thus, sulfide, hypoxia, and low pH are each toxic to milkfish at particular levels and aggravate each other's toxicity. Aquafarms must be well oxygenated to prevent sulfide toxicity and fish kills. This revised version was published online in July 2006 with corrections to the Cover Date.     

Salinity tolerance of two tropical fishes, Oreochromis aureus and O. niloticus. I. Biochemical and morphological changes in the gill epithelium

Cichlids of the genus Oreochromis are fish of economic importance in African countries. They tolerate brackish water, however, with great variations between species. In this work, two species, both from the Ivory Coast but of different origins, O. niloticus (field and laboratory strains) and O. aureus (field strain) were compared during osmotic challenges (10, 20 and 30%o salinity) in order to provide physiological support for their specific behaviour when confronted with natural hypertonic environments. Tolerance to salinity was assessed by correlated observations on gill structure, plasma sodium levels and gill Na⁺/K⁺ ATPase activity. In fresh water (FW), all fish presented a gill epithelium structure characteristic of FW stenohaline fish: no chloride cells (CC) on the lamellae and few CC on the filaments. An increase in external salinity induced the proliferation of CC on filaments, a feature typical of seawater teleosts. This change in gill structure was accompanied by an increase of gill Na⁺/K⁺ ATPase activity. In the most tolerant strains, plasma Na⁺ did not change, indicating successful ion regulation in the hypertonic media. With regard to potential interest of field strains in fish culture, O. aureus acclimated more easily to brackish water than O. niloticus . Interestingly, O. niloticus , kept for several generations in the laboratory, performed best in our challenge studies. Plasma Na⁺ levels and gill CC proliferation upon transfer to an isotonic medium may be the parameters of choice when testing these fish for their response to a salinity change.