Envelope surface ultrastructure and specific gravity of artificially fertilized Pacific cod Gadus macrocephalus eggs

The envelope surface ultrastructure and specific gravity of artificially fertilized eggs of the Pacific cod Gadus macrocephalus were examined. The unfertilized, demersal and slightly adhesive eggs of G. macrocephalus were almost spherical and had no oil globules. Wrinkled envelope surface with elaborated hexagonal reticulated patterns and type I micropyle were observed under a scanning electron microscope. The adhesiveness of the eggs was lost at the blastodermal‐cap stage after fertilization. The micropylar canal was sealed by secretion of the perivitelline fluid, and the entire surface became rough. Numerous bacilli were deposited at the micropyle and the outer envelope surface at the late germ‐ring stage and at the embryo five‐eighths around the yolk stage. The micropyle was completely deformed at the embryo seven‐eighths around the yolk stage. The specific gravity of the fertilized G. macrocephalus eggs ranged from c. 1·0316 to 1·0454. These values, however, sharply decreased towards the end stages of egg development to produce pelagic larvae. The ultrastructural changes in the micropyle and envelope surface of the G. macrocephalus eggs protected the embryo from microorganism infections and mechanical stress during the long incubation period. The adhesiveness and specific gravity of the eggs influenced their dispersion potential.     

Ultrastructural features of the egg envelope of silver carp, <Emphasis Type="Italic">Hypophthalmichthys molitrix</Emphasis> (Osteichthyes,Cyprinidae)

Synopsis We investigated the ultrastructure of the egg membrane surface (unfertilized egg) of Hypophthalmichthys molitrix using scanning electron microscopy. The eggs of silver carp like most other teleosts are surrounded by a relatively thick egg membrane, and have a type III micropyle at the animal pole. The micropyle is almost circular in shape and the micropyle canal is located in its center. The micropyle region is not flat. Round or oval accessory pores are also observed in the canal. The surface of zona radiata is wavy and uneven with a uniform distribution of almost round pores with lips. Microvilli like structure are found in the pore opening region.     

Sperm penetration and transformation of sperm entry site in eggs of the freshwater teleost Rhodeus ocellatus ocellatus

Eggs of bony fishes are enveloped by an egg envelope (chorion) in which a micropyle is present near the animal pole. Therefore, sperm penetration into the eggs is limited to the sperm entry site (SES), a region of plasma membrane just beneath the micropyle. In rose bitterling eggs, the SES transforms from a tuft of microvilli into a swollen mass (SM) that continues to plug the micropyle after sperm penetration. The present observations using the rose bitterling Rhodeus ocellatus ocellatus were conducted to examine: 1) whether or not sperm penetration is necessary for formation of the SM and 2) whether or not actin microfilaments are involved in the formation of the SM. Water activation without sperm transformed the SES from a tuft of microvilli into the SM, although it took a longer time for the transformation and the SMs were smaller than in the case of inseminated eggs. The SES presumably has the ability to transform into the SM upon activation of eggs in the present species. Cytochalasin B, which acts on actin microfilaments, did not prevent formation of the SM, irrespective of insemination or activation. The present observations suggest that sperm penetration is not necessary for SM formation and actin microfilaments do not participate in SM formation. © 1996 Wiley-Liss, Inc.     

The surface structure of Antarctic fish eggs and its use in identifying fish eggs from the Southern Ocean

Summary The surface structure of eggs of Notothenia neglecta, Nototheniops larseni, Nototheniops nudifrons (Nototheniidae), Champsocephalus gunnari and Chaenocephalus aceratus (Channichthyidae), collected near Elephant Island (Atlantic sector of the Southern Ocean) and of Notothenia rossii rossii (Nototheniidae) from Kerguelen waters (Indian sector of the Southern Ocean) was studied by scanning electron microscopy (SEM). Eggs of N. larseni differed from the other species by containing a micropyle with two micropyle pits. External morphological characteristics of the egg envelope such as the ultrastructure of the zona radiata, its interpore distances and the diameter of the micropyle were species-specific and could aid in the identification of fish eggs collected from the Southern Ocean.     

Factors inducing closure of the micropylar canal in the chum salmon egg

The micropylar canal of the chum salmon egg was almost completely closed following egg activation caused by incubation in a hypotonic salt solution (HSS) for I h. The closure occurred in both inseminated and parthenogenetically activated eggs. Incubation of isolated envelopes from non-activated eggs in HSS or perivitelline fluid (PVF) did not induce any modification in micropylar structure, indicating that normal organization of the egg is essential for inducing closure. To reduce the volume of the perivitelline fluid, the eggs were activated in PVF or HSS containing 8 mM Dextran, Although the envelope showed hardening, closure of the micropyle was not observed in these eggs. The wall of the micropylar canal, however, possessed a slightly rough surface. Following activation in a Ca-free hypotonic salt solution with 10 mM EDTA, hardening of the egg envelope was completely inhibited. Although such eggs possessed an apparent perivitelline space, neither closure of the micropylar canal nor roughening of the canal surface were detected. We conclude that the synergistic action of perivitelline turgor pressure and perivitelline material is responsible for the closure of the micropyle.     

Factors Controlling Sperm Entry into the Micropyles of Salmonid and Herring Eggs

When the micropyle area of salmonid (trout and salmon) eggs was observed continuously from the moment of insemination, spermatozoa were seen moving along the surface of the chorion and entering the micropyle one by one in a directed fashion. The ability of spermatozoa to enter the micropyle was reduced after the treatment of chorions with pronase; this reduction in sperm entry was observed even before the outer opening of the micropyle channel was narrowed due to gradual swelling of the chorion by pronase treatment. Herring spermatozoa, unlike spermatozoa of most other marine fishes, were motionless in seawater. However, they became vigorously motile on contact with the micropyle area of the herring egg chorion and entered the micropyle rapidly and efficiently. Motility initiation of herring spermatozoa in the micropyle area was dependent on extracellular calcium and potassium. Sodium also appears to be intricately involved in this process as demonstrated by the initiation of sperm movement in sodium-free seawater. When herring eggs were treated with acidic seawater, organic solvents, or glutaraldehyde, spermatozoa did not initiate movement in the micropyle area, and sperm entry was not observed. Herring spermatozoa did not initiate movement in the micropyle area of salmonid eggs. These and other observations suggest that the micropyle areas of salmonid and herring eggs possess some sperm guidance factors which facilitate entry of homologous spermatozoa into the micropyle.     

A single micropyle in the eggs of the most basal living actinopterygian fish, Polypterus (Actinopterygii, Polypteriformes)

Breeding of Polypterus , the most basal living actinopterygian, provided the opportunity to study the surface structure of its eggs with SEM for the first time. The investigation revealed the previously overlooked presence of a micropyle at the animal egg pole, thus rejecting recent textbook belief that the eggs of Polypterus slack a micropyle. Comparison with data for other gnathostomes demonstrates that eggs with a micropyle are a further synapomorphy for Actinopterygii and confirm membership of the Cladistia in that group.     

滇池金线鲃亲鱼培育、繁殖力以及卵径大小与胚胎存活率的关系

2007—2010年,对滇池金线鲃(Sinocyclocheilus grahami)进行了亲鱼培育、繁殖力和卵径(egg diameter,ED)大小的研究。滇池金线鲃的催产率、核偏位率和孵化率分别由2007年的25.2%、61.5%和30.4%到2010年的91.3%、85.2%和44.5%。四年间这3项指标均有不同程度提高,其中,催产率提高的幅度最大。提供适合滇池金线鲃营养全面的饵料可以保证亲鱼培育的最佳效果,获得高质量的鱼卵和鱼苗。滇池金线鲃绝对怀卵量与体长的关系是F=0.0004826SL3.166(R2=0.6424,P<0.05)。四年来滇池金线鲃的平均产卵量为(2118.4±899.1)粒,平均绝对怀卵量为(2402.9±881.9)粒,平均相对怀卵量为(70.4±20.8)粒。产卵量、绝对怀卵量和相对怀卵量都有随着体长增加而增加的趋势。卵的大小与胚胎存活相关,在胚胎发育的前两天,不同批次胚胎的死亡呈现一种稳定的或低的死亡率;而后5～7d不同大小的卵呈现不同的死亡率,即小的卵具有更高的死亡率;而卵径>2.0mm的胚胎死亡率趋于稳定;大的卵有更高的生存潜力,能保证仔鱼开口前继续完善身体器官的形成或发生所需要的能量。     

Fertilization triggers localized activation of Src-family protein kinases in the zebrafish egg

Fertilization triggers activation of Src-family kinases in eggs of various species including marine invertebrates and lower vertebrates. While immunofluorescence studies have localized Src-family kinases to the plasma membrane or cortical cytoplasm, no information is available regarding the extent to which these kinases are activated in different regions of the zygote. The objective of the present study was to detect the subcellular distribution of activated Src-family kinases in the fertilized zebrafish egg. An antibody specific for the active, non-phosphorylated form of Src-family PTKs was used to detect these activated kinases by immunofluorescence. The results demonstrate that Fyn, and possibly other Src family members are activated by dephosphorylation of the C-terminal tyrosine at fertilization. The activated Src-family kinases are asymmetrically distributed around the egg cortex with an area of higher kinase activity localized adjacent to the micropyle near the presumptive animal pole. Fertilization initially caused elevation of kinase activity in the cytoplasm underlying the micropyle, but this quickly spread to involve the entire zygote cortex. Later, during egg activation, formation of the blastodisc involved concentration of active Src-family kinase in the blastodisc cortex. As cytokinesis began, activated Src-family kinases were no longer limited to the cortex, but became more evenly distributed in the clear apical cytoplasm of the blastomeres. The results demonstrate that the cortex of the zebrafish egg is functionally differentiated and that fertilization triggers localized activation of Src-family kinases at the point of sperm entry, which subsequently progresses through the entire egg cortex.     

Micropyle of some salmonins and coregonins

Synopsis The micropyle of seven salmonins and four coregonins were examined by a scanning electron microscope. In addition, the micropyle of Coregonus pidschian is described for the first time. The micropyles and the surface pattern may be used for identification of eggs. The studied micropyle belongs to two different types: 1. micropyle with flat pit and long canal (type II) and 2. micropyle with canal but without pit (type III). The sides of the micropyle canal are often reinforced by annular or helicoid thickening.     

云南滇池濒危特有种滇池金线鲃繁殖初报

滇池金线鲃(Sinocyclocheilus grahami)是滇池湖泊生态系统的指示物种和特有种。因水质污染和外来物种入侵等因素的影响,该鱼类自1986年起就从湖体中消失,仅有湖周围的少数龙潭中尚保存有少量滇池金线鲃。至2007年3月初,有两次繁殖试验取得了成功。先后试验了5尾雌鱼和6尾雄鱼,获得鱼卵约1600粒,并实施干法受精。约有1320粒受精,平均受精率为73%。孵化出鱼苗约480尾,平均孵化率为36%。经10天饲养,约有95%以上的鱼苗存活,体长达8-12mm。滇池金线鲃人工繁殖成功的意义有三点:能有效保护该物种免于灭绝;繁殖的鱼苗放归湖泊合适水域,将有助于恢复滇池的土著生物多样性;有助于推动当地水产养殖业从非土著鱼类养殖向土著鱼类养殖的方向转变。     

The use of poly-L-lysine to facilitate examination of sperm entry into pelagic, non-adhesive fish eggs

The fish egg is surrounded by a thick envelope called the chorion. The fertilizing spermatozoon enters the egg through a canal-like structure in the chorion, the micropyle. Examination of micropyle at fertilization is difficult if eggs are large and have no distinct landmarks surrounding the micropyle, or if they are positively buoyant in water. Eggs of many commercially important fishes (e.g., flounder, sea bream and eel) are buoyant in water or only slightly adhere to solid objects (e.g., sands, rock and water plants), which makes observation of spermatozoa at fertilization difficult. Here, we report that such eggs can be firmly attached to plastic and glass dishes that have been previously coated with poly-L-lysine. These adhering eggs can be fertilized and develop normally on the dishes. Observations of micropyles of three fish species, before and after sperm entry are presented.     

革胡子鲶受精过程的扫描电镜观察

应用扫描电镜观察和描述了革胡子鲶成熟卵和精子的形态、卵壳膜的表面结构和形态、受精孔的位置和结构、精子入卵过程的程序和变化。讨论了精子入卵过程及精孔细胞在解体之后可能转变为一种能够吸引精子在精孔区聚集的“受精素”物质等问题。     

Fine structures of the micropyles of pelagic eggs of some marine fishes

The micropyles and their surrounding structures of pelagic fish eggs of nine species were examined and compared using a scanning electron microscope (SEM). The surface of the egg membrane possesses uniformly distributed pores. However,Lateolabrax japonicus possesses a number of small knobs. The region of the micropyle of fish eggs examined is characterized by a circular elevation in which the pores are larger than those on the other parts of the egg membrane. In contrast,Kareius bicoloratus egg has no elevation and possesses uniform pores to the edge of the micropylar canal. The funnel-like structure is seen in the opening of the micropyle inSardinops melanostictus, Saurida elongata, Sillago japonica, Parapristipoma trilineatum, Hypodytes rubripinnis andK. bicoloratus. These specific features of the micropyle may be used for the identification of pelagic eggs.     

The captive husbandry and reproduction of the pink-eared turtle (Emydura victoriae) at Perth Zoo

In 1997, Perth Zoo acquired six pink-eared turtles (Emydura victoriae) from the wild for display in the reptile facility. There is very little documented information on pink-eared turtles in captivity. This article looks at the reproductive biology, ecology, behavior, diet, and captive husbandry of the species. Eight clutches of eggs were documented over a 2-year period with an average clutch size of 10 eggs. Egg size was recorded with three clutches incubated to hatching. Ten hatchlings were maintained for a growth and development study. Measurements of weight, carapace length, width, height, and plastron length were recorded weekly for about 12 months, and then monthly for approximately 2 years. The data were analyzed and showed positive growth curves in all animals. Sexual dimorphism was observed after 20 weeks and sexual maturity in males observed after 2 years.     

泥鳅精子入卵的动力作用

在扫描电镜下,泥鳅成熟卵卵膜孔外围呈现完整的左涡旋状结构,受精时精子是顺着涡旋的流线进入卵膜孔。涡旋纹理接近对数螺线。本文分析了真骨鱼类的受精因素,除已知的化学因素外,还存在物理因素。也讨论了泥鳅成熟卵卵膜孔形态形成的必然性。     

Gymnodinioides pacifica, n. sp., an exuviotrophic ciliated protozoan (Ciliophora, Apostomatida) from euphausiids of the Northeastern Pacific

A new species of apostome ciliate, Gymnodinioides pacifica n. sp. from the euphausiid Euphausia pacifica is described. The ciliated protozoan encysts on the setae of the appendages, telson and antennae. It excysts and enters the exoskeleton of the host after moulting, where it feeds on exuvial fluid. The phoront and trophont stages of this ciliate are described. The ciliature of the trophont has the following characteristics that distinguish it from the other species of Gymnodinioides: (1) a short kinety 9a (ciliary row 9a or K9a), approximately half of the length of falciform field 8, that extends from the anterior end of the cell posteriorly to the level of K1, (2) a straight K5a, perpendicular to K5b, K6, and K7, and (3) two distinct bands of kineties, K9b-K5b and K4-K1. In addition to those on the host E. pacifica, phoront cysts of similar shape were observed on Thysanoessa spinifera, T. gregaria, T. inspinata, T. longipes, and Nematoscelis difficilis. High prevalence rates were recorded (83%) for these apostome cysts on E. pacifica and T. spinifera during summer collections from the Oregon and Washington coasts. Additionally, we report other apostome ciliate symbionts of euphausiids, including Phtorophrya sp., which preys on apostomes in the genus Gymnodinioides.     

Morphology of unfertilized mature and fertilized developing marine pelagic eggs in four types of multiple spawning flounders

Starry flounder Platichthys stellatus, spotted halibut Verasper variegates, turbot Scophthalmus maximus, and Japanese flounder Paralichthys olivaceus are four commercially cultivated multiple spawning flounders that spawn pelagic eggs. Through appropriate light and scanning electron microscope processing, the shape and surface structures (such as micropyle, pores, pore density, and paten) of unfertilized mature and fertilized developing eggs of the four species were observed and measured. First, individual or intraspecific comparisons of the surface structures of eggs at different developmental stages were made. Second, interspecific differences among the four species at the same developmental stage of unfertilized mature eggs were statistically computed and analyzed through one-way analysis of variance and hierarchical cluster analysis. Eggs of the same species collected at different stages of development tend to be different in morphology. Smoothing of the convoluted egg envelope surface and closure of the micropyle to serve as a final step of the polyspermy-preventing reaction are common after fertilization. Based on detailed morphology of micropyle of just-mature fertilizable eggs, turbot, starry flounder, and Japanese flounder each have a micropyle with a long canal but no distinct micropylar vestibule, type III of Riehl and Götting (Arch Hydrobiol 74:393–402, 1974). In contrast, spotted halibut has a micropyle with a distinct flat micropylar vestibule and a long canal, type II. Envelope surface microstructures, especially those in the micropyle region, are useful characters for egg identification among the four species. Cluster analysis using selected egg characters indicated the highest similarity between turbot and Japanese flounder and that starry flounder is obviously more similar to turbot and Japanese flounder than to spotted halibut.     

食蚜蝇科卵壳超微形态初探

本文应用扫描电镜研究了24种食蚜蝇(双翅目,食蚜蝇科)卵壳的超微形态.描述了所研究种类卵的形态并编列了检索表.研究结果表明卵的受精孔形态是和卵壳花纹同等重要的分类特征.文中讨论了受精孔形态和卵表花纹在食蚜蝇卵分类中的重要性以及造成卵壳背、腹面花纹差异的原因.     

Effects of incubation temperature on the energetics of embryonic development and hatchling morphology in the Brisbane river turtle Emydura signata

Incubation temperature and the amount of water taken up by eggs from the substrate during incubation affects hatchling size and morphology in many oviparous reptiles. The Brisbane river turtle Emydura signata lays hard-shelled eggs and hatchling mass was unaffected by the amount of water gained or lost during incubation. Constant temperature incubation of eggs at 24 °C, 26 °C, 28 °C and 31 °C had no effect on hatchling mass, yolk-free hatchling mass, residual yolk mass, carapace length, carapace width, plastron length or plastron width. However, hatchlings incubated at 26 °C and 28 °C had wider heads than hatchlings incubated at 24 °C and 31 °C. Incubation period varied inversely with incubation temperature, while the rate of increase in oxygen consumption during the first part of incubation and the peak rate of oxygen consumption varied directly with incubation temperature. The total amount of oxygen consumed during development and hatchling production cost was significantly greater at 24 °C than at 26 °C, 28 °C and 31 °C. Hatchling mass and dimensions and total embryonic energy expenditure was directly proportional to initial egg mass. Accepted: 18 March 1998