CORRELATION OF ZONA-BINDING WITH OOCYTE MATURATION AND SPERM MOTILITY IN RHESUS-MONKEYS BY HEMIZONA ASSAY

The hemizona assay (HZA) in Rhesus monkeys was employed to study the correlation of zona-binding ability with sperm motility or with naturally developing oocytes at various maturational stages. Oocytes from unstimulated ovaries were retrieved within 2 hr from monkeys sacrificed for vaccine production (in reproductive season, but with their menstrual cycles not determined). Oocytes were divided into four groups based on their morphological maturation: 1) Oocytes surrounded by more than one cumulus layer (MC); 2) Oocytes retaining intact germinal vesicle nuclei (GV); 3) Oocytes with germinal vesicle breakdown showing distinct perivitelline space (PVS); and 4) Oocytes extruding the first polar body (PB1). The mean numbers of sperm bound to hemizona for PBI, PVS, GV, and MC groups were 132.9 +/- 12.0, 71.5 +/- 10.1, 36.1 +/- 4.0, and 20.1 +/- 2.9 (Mean +/- SE), respectively. The four groups showed significant differences from each other in sperm/egg binding ability (P < 0.01). The number of bound sperm significantly increased with oocyte maturation. The present study also showed that zona-binding ability was also affected by sperm motility. For sperm with 67.7% motility and sperm with 31.2% motility, the average numbers of bound sperm were 43.5 +/- 2.2 and 25.3 +/- 2.9 (Mean +/- SE), respectively. There was significantly higher binding ability for sperm with higher motility (P < 0.01). The results suggest that: 1)The rhesus monkey model can serve as a very sensitive model for studying sperm/egg interaction by HZA; 2) Sperm motility positively correlated with sperm/egg binding; and 3) Sperm/egg binding ability increases with oocyte maturation. The binding ability is highest when oocytes matured to the PB1 stage, which is also the best opportunity for fertilization. This is strong evidence for the ''zona maturation'' hypothesis. (C) 1994 Wiley-Liss, Inc.

In vivo and in vitro storage of kutum, Rutilus frisii kutum eggs

Effects of post-ovulatory and post-stripping retention time and temperature on egg viability rates were studied in kutum (Rutilus frisii kutum). Eggs were retained inside (in vivo storage) or outside the ovarian cavity with ovarian fluid (in vitro storage) at various temperatures. Two experiments were performed: 1) Partial volumes of eggs were stripped and fertilized at 24- hour intervals for 96 hours post-ovulation (HPO) (at 11 °C) and at 12-hour intervals for 72 HPO (at 14 °C), and 2) stored eggs were fertilized after 0, 2, 4, 6, and 8 hours post-stripping (HPS) at temperatures of 4, 10, 12, and 26 °C. In the first experiment, the highest eyeing and hatching rates (76% and 60% at 11 °C; 81% and 71% at 14 °C) and the lowest eyed-egg mortalities (20% at 11 °C; 12% at 14 °C) occurred in the eggs fertilized immediately (0–24 HPO at 11 °C and 0–12 HPO at 14 °C) after ovulation. Egg viability, as shown by successful eyeing and hatching rates, was completely lost by 72–96 HPO at 11 °C, and 60–72 HPO at 14 °C. In the second experiment, the maximum eyeing (87%) and hatching (75%) rates of eggs took place at 0 HPS followed by 8 HPS (> 80% and > 70%, respectively) at 4 °C. As storage temperature increased, egg viability decreased: 80%, 70%, and 50% viable at 8 HPS at 4, 10, and 12 °C, respectively. The eggs stored at 26 °C lost their viability almost completely after 4 HPS. Eyed-egg mortality increased from 13% at 0 HPS to 48.2% at 4 HPS at 26°C. These results demonstrate that egg stripping should take place within 168 °C-hours after ovulation and that complete loss of viability of the eggs occurs by 672°C-hours after ovulation. The in vivo storage method is more effective compared to in vitro storage. Also successful in vitro storage of eggs can be used atleast within 8 hours at temperatures ranging from 4 to 12ºC.

The study of biological characteristics and determination of population dynamic parameters of sawtooth barracuda (Sphyraena putnamae) in the Persian Gulf (Hormozgun Province waters)

The biological characteristics and population dynamisms of Sphyraena putnamae, were studied in the northern Persian Gulf and Oman Sea restricted to Hormuzgan province waters within 13 months period, from November 2006 up to November 2007. Biometrical and anatomical measurements were carried out, and biological surveys were conducted on 486 specimens. On the other hand, the growth and mortality parameters were estimated by using 3096 samples. These samples were collected from 3 landings, namely Bandar Abbas, Bandar Lengeh and Bandar Jask. The measurements of the minimum and maximum Fork lengths and weights were 11.7 to 8.03 cm and 135.0 to 4140.0 g, respectively. The results indicated that this species, having the Relative Length of Gut, RLG=0.34±0.002, is strongly carnivorous (often fish-eater), proven by the fact that more than 98% of its stomach contents were fish pieces. Examining the changes in the index of stomach emptiness by the percentage of CV = 0.47% indicates that this fish is Moderate feeder. The level of feeding increased in March, before spawning and decreased in June and September, simultaneously with the spawning season. There are 2 peaks of reproduction or spawning seasons during the months of April-May and September, of which the prior is assumed as the main spawning. The sex ratio (M:F) was calculated 0.5:1.0(X2 =2.11), which did not show a significant difference with expected level of 1:1 (P>0.05). The average absolute and relative reproduction rates of Sphyraena putnamae is respectively as follows: 1866827.1±255448.9 and 1097.7±94.3. The highest and the lowest diameter of matured egg are from 200 to 750 μ, and its average diameter is 402.10 ± 0.190 μ. A parameter for Saw-tooth barracuda length measurement, Lm50, based on the Fork-length, was calculated as 54.01 cm. In other words, as far as the fisheries management is concerned, the fish whose lengths are less than 54.01 cm should not be caught. The calculated level of (R2) (correlations of total length & weight), indicated strong correlations between length and weight of this fish, and the obtained formula included W =0.007100 FL 2.9295 and reinforced this assumption. The “K” Index for this fish in 3 above mentioned landings (Jask, Bandar-Abbas and Bandar-Length) were 1.24, 0.37 and 0.46 per year, respectively and the FL index for the same landings were estimated as 129, 110 and 134 cm, respectively. The growth coefficient (MONRO) for the above mentioned regions were calculated as 3.601, 3.647 and 3.917, respectively; and in the surveyed regions there were no significant differences in populations. The Total mortality coefficient (Z) was calculated 0.76, 1.12 and 1.07 per year, the Natural mortality coefficient was 0.46, 0.63 and 0.70, and the Fishing mortality coefficient (rate) (F) was found to be 0.30, 0.49 and 0.37 per year. The value of the exploitation rate (E) is equal to 0.39 per year, indicating that this species is an under-exploited resource, and there is no excessive fishing pressure on the fish supply of this species in the afore-said regions. The highest level of exploitation was found for ‘Bandar Abbas’ fishing region and the lowest level of exploitation is in ‘Bandar Lengeh’ waters.

Effect of amino acids on cryopreservation of cynomolgus monkey (Macaca fascicularis) sperm

The effects of three amino acids (proline, glutamine, and glycine) added to the freezing medium Tes-Tris-egg yolk (TTE) for cryopreservation of cynomolgus monkey (Macaca fascicularis) spermatozoa were studied. This is the first report on the effects of amino acids on nonhuman primate sperm cryopreservation. The addition of 5 mM proline, 10 mM glutamine, and 10 or 20 mM glycine each significantly improved post-thaw sperm motility and membrane and acrosome integrity compared with the control (TTE alone). However, a significant decrease in motility and membrane/acrosome integrity was observed when amino acid concentrations increased to 60 mM for proline and glutamine, and 80 mM for glycine. The results suggest that adding a limited amount of amino acids to the freezing media is beneficial for freezing cynomolgus monkey sperm.

Cryopreservation of cynomolgus monkey (Macaca fascicularis) spermatozoa in a chemically defined extender

Aim: To establish a method for cynomolgus monkey sperm cryopreservation in a chemically defined extender. Methods: Semen samples were collected by electro-ejaculation from four sexually mature male cynomolgus monkeys. The spermatozoa were frozen in straws by liquid nitrogen vapor using egg-yolk-free Tes-Tris (mTTE) synthetic extender and glycerol as cryoprotectant. The effects of glycerol concentration (1%,3%, 5%, 10% and 15% [v/v]) and its equilibration time (10 min, 30 min, 60 min and 90 min) on post-thaw spermatozoa were examined by sperm motility and sperm head membrane integrity. Results: The post-thaw motility and head membrane integrity of spermatozoa were significantly higher (P < 0.05) for 5% glycerol (42.95 +/- 2.55 and 50.39 +/- 2.42, respectively) than those of the other groups (1%: 19.19 +/- 3.22 and 24.84 +/- 3.64; 3%: 34.23 +/- 3.43 and 41.37 +/- 3.42; 10%: 15.68 +/- 2.36 and 21.39 +/- 3.14; 15%: 7.47 +/- 1.44 and 12.90 +/- 2.18). The parameters for 30 min equilibration (42.95 2.55 and 50.39 2.42) were better (P < 0.05) than those of the other groups (10 min: 31.33 +/- 3.06 and 38. 98 +/- 3.31; 60 min: 32.49 +/- 3.86 and 40.01 +/- 4.18; 90 min: 31.16 +/- 3.66 and 38.30 +/- 3.78). Five percent glycerol and 30 min equilibration yielded the highest post-thaw sperm motility and head membrane integrity. Conclusion: Cynomolgus monkey spermatozoa can be successfully cryopreserved in a chemically defined extender, which is related to the concentration and the equilibration time of glycerol.

Evaluation and comparison of ozone, hydrogen peroxide and physical treatment on fungal infection prevention and hatching rate of Iranian sturgeon (Acipenser persicus)

Ozone due to having low half-life and devoid of environmental harmful effects is recognized as one of the most effective disinfectant and fungicide in aquaculture. The objective of this study is to consider the effects of periodicay ozonation, hydrogen peroxide treatment, and physical treatment capability in hatching rate enhancement. Three concentrations of 0.05, 0.1 and 0.15 ppm ozone (10 min) and peroxide hydrogen with dose of 500 and 1000 ppm in two procedures accompanied with physical treatment and without physical treatment were examined on hatching rate. In the first year, Egg ozonation (0.1 ppm) with physical treatment have been resulted the greatest hatching rate (81.4%). In the second year, egg treatment with 1000 ppm hydrogen peroxide with physical treatment have been showed the greatest hatching rate (78%). Average hatching rate for the blank control treatment (without disinfectin and physical treatment) was 32.7%. From the economic viewpoint, 0.05 ppm ozone with physical treatment, due to considerable minimizing at consumption energy and ozonation system retention costs, indicated as the best treatment than other ozone treatments for fungal control. Very low correlation (r=-0.14) have been observed between hatchery water temperature and fungal infection percentage in control treatment.

Survey of marine turtles ecobiology in north of Persian Gulf

Sea turtles are counted as rare and extincting species. During the last century the population of sea turtles are drastically reducd and the remaining ones are considerd either threatend or endangerd because of some factors as sea pollution, economic benefits, intense fishing effort, beach man-made structures and so on. So, in recent years, more attention to conserving and surviving them is assumed. After a preliminary studies on some of the biological specifications in northern beach of Persian Gulf, three islands ; Hormoz, Hengam and Larak for the biometry of the turtles, a total number of 179, from 1999 to 2002 (4 years) are chosen. Some of the biological attributes as weight, length and their egg-laying are recorded, these items are then analyzed using SPSS software. The observed results are as follows . The lowest weight of the turtles was 35 kg, the highest 59 kg, and the average 45.24 kg. The lowest carapace length was 64 cm. the highest 86 cm and the average 76.79. the lowest number of laying eggs was 73, the highest 126, and the average 86.79. The results are discussed in three following sections: 1-A contrastive analysis of the biometric characteristics during 1999 in the three mentiond islands. 2-A contrastive analysis of the biometric characteristics in Hormoz island from 1999 to 2002. 3-A contrastive analysis of all the biometric characteristics in the three islands and other parts of the world. The results obtained from the turtles biometry in 1999 shows that the average of the turtles weight in Larak is lower than the other mentioned ones, and the heighest average is observed in Hormoz island. The turtles of Hengm have laid more eggs than the others. The results of Hengani turtles biometry during 1999-2002 indicates that the average of their weight in 2002 is more than the other three years in the same place. The least number of eggs laid during these years are 53 and the most are 126. The most number of eggs are laid in Hormoz in 2002, but the average number does not show any significant change.

The technology of milkfish hatchery in Indonesia

The culture of milkfish (Chanos chanos), which is the oldest fish cultured in Indonesia, has spread to almost all the provinces in the country. In order to answer the demand of milkfish fry, without decreasing production and wild fry supply, artificial fry production in big and small-scale hatcheries is being practiced. Details are given of the milkfish hatchery system, listing also the major criteria to be taken into account for site selection. Operation of the hatchery is described, outlining the following activities: broodstock rearing; hormone implantation; broodstock maintenance; egg production and harvest; and, larval rearing.

补充营养对柑橘凤蝶卵成熟和雄虫寿命的影响

在实验室条件下,定量地研究了蔗糖、葡萄糖、果糖和蜂蜜等4种补充营养物对柑橘凤蝶卵成熟和雄虫寿命的影响.结果表明:(1)补充蔗糖、果糖和蜂蜜水的雌虫孕成熟卵量都显著高于清水对照组,其中补充蔗糖的雌虫孕成熟卵量最高;(2)不同的补充营养物对雌虫孕亚成熟卵量和未成熟卵量的影响不显著;(3)饲喂蔗糖、果糖和蜂蜜的雄虫寿命显著高于对照组,其中饲喂果糖的雄虫寿命最长;(4)饲喂添加不同浓度NaCl溶液的蜂蜜水的雄虫寿命比仅饲喂蜂蜜水的雄虫短,当NaCl溶液浓度为0.001 mol/L,0.1 mol/L和 1 mol/L 时,雄虫寿命显著低于对照,其中饲喂0.001 mol/L NaCl溶液的雄虫寿命最短.

Nutritive value of fish of Lake Tanganyika [part] I: Amino acid composition

The amino acid composition was determined of three economic important fish species of Lake Tanganyika, representing 95% of the tolal catch:Stolothrissa tanganicae Regan, Limnothrissa miodon Boulenger and Luciolates stapersii Boulenger. The analysis of the whole sun dried fish indicate that, compared with Tilapia macrochir, beef and the whole hen's egg, the fish is of a very high nutritive value. The free amino acids in the dried fish count for an average of 11 %, indicating the degree of auteolysis.

The reproductive strategies of an endemic Tibetan fish, Gymnocypris selincuoensis

The reproductive traits of Gymnocypris selincuoensis from Selincuo Lake and its tributaries were investigated in 1997 and 1998. The youngest mature male was age 7 with a standard length (SL) of 172.0 mm, and the youngest mature female was age 8 with a SL of 194.0 mm. The L(50)s Of SL and age at first maturity were respectively 250.32 mm and age 9 for males and 224.71 mm and age 8 for females. The gonadosomatic index (GSI) significantly changed with seasons for mature individuals but not for immature individuals. GSIs of mature females at stages IV and V of ovary development increased with SL and reached a maximum value at the SL range from 370 mm to 390 mm; the GSIs of mature males were negatively correlated with SL. The breeding season lasted from early April to early August. Egg size did not significantly change with SL but increased with the delay of spawning. The individual absolute fecundity varied from 1,341 to 28,002 eggs (mean 12,607+/-7,349), and the individual relative fecundity varied from 6.4 to 42.0 eggs.g(-1) (mean 25.5+/-9.7). The individual fecundity increased with total body weight; it also increased with SL for those of SL less than 370 mm. There was a rest of spawning for mature individuals.

温度影响轮虫卵割速度卵割规律和星状体移动速度之研究

<正> 一.绪言从初期胚胎发育过程和发生生理的研究,知道卵的性质有全能与限能的不同。全能卵(regulative egg)的初期发育,在某一固定的温度下,卵的每次分割是按冪次式(exponential type)而进行,每完成一有丝分裂周期(mitotic cycle)所需的时间都相同。这种情形,早经 Gray,Balinsky,和 Syngajemskaja 等,备就 Echinus miliaris,Ascidia(Ciona intestinalis)和 Campanularie,等动物卵的观察,先

The reproductive traits of brown trout (Salmo trutta fario L.) from the Yadong River, Tibet

Some key aspects of the reproductive strategy of the brown trout (Salmo trutta fario L.) in the Yadong River, Tibet, including spawning season, age at sexual maturity, fecundity and egg size, have been studied. The majority of the samples were less than 215 mm and age ranged from 1 to 4 in both sexes, indicating that the majority of the fish were younger and the pressure by overfishing was high. The spawning periodicity was determined to be between the end of October and January, mainly in November and December. The ratio of male to female brown trout population (1.29:1 with P > 0.05) suggested no sex significant differences, although males were significantly more abundant than females in October (P < 0.0001) on monthly basis. Age and size of males and females at maturity was different and males matured earlier than females. Fecundity was markedly correlated with their body weight (P < 0.001, r = 0.9255), standard length (P < 0.01, r = 0.8879), and gonad weight (P < 0.001, r = 0.9366). The mean size of mature eggs in the spawning season was: 4.0 +/- 0.45 mm and tended to increase along with the female spawners size (P < 0.001, r = 0.9641). Further researches about the brown trout population in the Yadong River should be conducted on issues such as artificial reproduction, culture, conservation, management, and restocking.

Using drift nets to capture early life stages and monitor spawning of the Yangtze River Chinese sturgeon (Acipenser sinensis)

P>A sampling system for capturing sturgeon eggs using a D-shaped bottom anchored drift net was used to capture early life stages (ELS) of Chinese sturgeon, Acipenser sinensis, and monitor annual spawning success at Yichang on the Yangtze River, 1996-2004, before and just after the Three Gorges Dam began operation. Captured were 96 875 ELS (early life stages: eggs, yolk-sac larvae = eleuthero embryos, and larvae); most were eggs and only 2477 were yolk-sac larvae. Most ELS were captured in the main river channel and inside the bend at the Yichang spawning reach. Yolk-sac larvae were captured for a maximum of 3 days after hatching began, indicating quick dispersal downstream. The back-calculated day of egg fertilization over the eight years indicated a maximum spawning window of 23 days (20 October-10 November). Spawning in all years was restricted temporally, occurred mostly at night and during one or two spawning periods, each lasting several days. The brief temporal spawning window may reduce egg predation by opportunistic predators by flooding the river bottom with millions of eggs. During 1996-2002, the percentage of fertilized eggs in an annual 20-egg sample was between 63.5 to 94.1%; however, in 2003 the percentage fertilized was only 23.8%. This sudden decline may be related to the altered environmental conditions at Yichang caused by operation of the Three Gorges Dam. Further studies are needed to monitor spawning and changes in egg fertilization in this threatened population.

Effects of adrenergic agents on the expression of zebrafish (Danio rerio) vitellogenin Ao1

Teleost vitellogenins (VTGs) are large multidomain apolipoproteins, traditionally considered to be estrogen-responsive precursors of the major egg yolk proteins, expressed and synthesized mainly in hepatic tissue. The inducibility of VTGs has made them one of the most frequently used in vivo and in vitro biomarkers of exposure to estrogen-active substances. A significant level of zebrafish vtgAo1, a major estrogen responsive form, has been unexpectedly found in heart tissue in our present studies. Our studies on zebrafish cardiomyopathy, caused by adrenergic agonist treatment, suggest a similar protective function of the cardiac expressed vtgAo1. We hypothesize that its function is to unload surplus intracellular lipids in cardiomyocytes for "reverse triglyceride transportation" similar to that found in lipid transport proteins in mammals. Our results also demonstrated that zebrafish vtgAo1 mRNA expression in heart can be suppressed by both (x-adrenergic agonist, phenylephrine (PE) and beta-adrenergic agonist, isoproterenol (ISO). Furthermore, the strong stimulation of zebrafish vtgAo1 expression in plasma induced by the beta-adrenergic antagonist, MOXIsylyl, was detected by Enzyme-Linked ImmunoSorbent Assay (ELISA). Such stimulation cannot be suppressed by taMOXIfen, an antagonist to estrogen receptors. Thus, Our present data indicate that the production of teleost VTG in vivo can be regulated not only by estrogenic agents, but by adrenergic signals as well. (c) 2009 Elsevier Inc. All rights reserved.