The study of bacteria on artificially incubated noble crayfish eggs

Data on the abundance of heterotrophic bacteria on the surface of artificially incubated noble crayfish (Astacus astacus L.) eggs are presented. The highest abundance of heterotrophic bacteria was observed on crayfish juveniles (stage 1). The abundance of bacteria in the water of the incubator’s system was found to be considerably lower than that on crayfish eggs, and bacteria counts on dead eggs were a 100 times lower than on healthy eggs. The abundance of pigmented bacteria on dead eggs was considerably higher than on healthy eggs. Aeromonas and Pseudomonas bacteria, constituting up to 70% of all bacteria identified, were found to be dominant on noble crayfish eggs.     

The stimulation of amino acid incorporation into the protein of unfertilized sea urchin eggs by gamma-irradiation

Control and γ-irradiated sea urchin (Arbacia punctulata) eggs were incubated with H³-leucine. No incorporation of the amino acid into the proteins of most unirradiated eggs was detectable by autoradiography. All irradiated eggs incorporated the amino acid and the amount of autoradiographic labelling varied greatly from cell to cell.     

Structure of shells from eggs of kinosternid turtles

Shells from eggs of five species of kinosternid turtle (Sternotherus minor, Kinosternon flavescens, K. baurii, K. Hirtipes, and K. alamosae) were examined with light and scanning electron microscopy. Except for possible differences among species in thickness of eggshells, structure of shells from all eggs was similiar. In general, kinosternid turtles lay eggs having a rigid calcareous layer composed of calcium carbonate in the form of aragonite. The calcareous layer is organized into individual shell units with needlelike crystallites radiating from a common center. Most of the thickness of the eggshell is attributable to the calcareous layer, with the fibrous shell membrane comprising only a small fraction of shell thickness. Pores are found in the calcareous layer, but they are not numereous. The outer surface of the eggshells is sculptured and may have a thick, organic layer in places. The outer surface of the shell membrane of decalcified eggshells is studded with spherical cores which presumably nucleate growth of shell units during shell formation. The shell membrane detaches from eggs incubated to hatching, carrying with it remnants of the calcareous layer. Such changes in shell structure presumably reflect withdrawal of calcium from the eggshell by developing embryos.     

Parasitism of frozen Halyomorpha halys eggs by Trissolcus japonicus: applications for rearing and experimentation

The brown marmorated stink bug, Halyomorpha halys (Stål), has become a well-known pest to growers and homeowners since its 1996 introduction to the United States. A classical biocontrol programme is under development using the egg parasitoid Trissolcus japonicus. Widespread implementation of biocontrol requires efficient mass rearing, which is constrained by the availability of fresh H. halys eggs. In this study, parasitism rate, developmental time, sex ratio and size were compared between wasps reared on fresh versus frozen, newly laid (<1 d old) versus variably aged (0–3 d old), and frozen egg masses stored ≤4 y. Frozen eggs yielded 56–65% fewer wasps, with parasitism rate decreasing 1–3% per month stored. Parasitism rate, sex ratio and developmental time were comparable between newly laid and variably aged eggs. Freezing eggs for any duration did not affect sex ratio or weight of emerged wasps, but delayed emergence 5–6 d. To simulate deployment of sentinel eggs in the field, we incubated frozen eggs at 20°C and 30°C for 1–9 d before exposing them to T. japonicus, then evaluated parasitism trends. Trissolcus japonicus parasitism rate decreased 5–8% per day incubated, unhatched wasps increased 9% per day incubated and sex ratio was not impacted. Variably aged, frozen and longer stored eggs can be used for T. japonicus rearing and experimentation without affecting emerged wasp sex ratio or size within one generation, but have lower parasitism and slower development. Frozen sentinel eggs are effective <3–5 d, especially in hot conditions.     

Artificial incubation of eggs of the green iguana (Iguana iguana)

Successful hatching of large numbers of artificially incubated eggs from the green iguana, Iguana iguana, are reported. Gravid females were captured at a nesting site in Summit Gardens, Soberania, Parque Nacional, Panama, and released into an enclosure with an artificial nesting area. Females dug their own tunnels and nest chambers or used artificial nest chambers for egg deposition. Eggs (n = 829) from 21 clutches were removed from the nests and artificially incubated. Average hatching success per clutch was 94.6% and the mean clutch incubation time was 92.1 days. A distinct odor was noticed in the incubation containers several days before the eggs began to hatch. At this time, the substrate layer was removed exposing the top surface of all eggs, and the eggs desiccated to some degree prior to hatching. High incubation temperature, hydric conditions, and egg arrangement are all implicated as contributing factors in low hatching success in previous studies using artificial incubation techniques.     

Chemical protection of Calvia quatuordecimguttata eggs against intraguild predation by the invasive ladybird Harmonia axyridis

Intraspecific and interspecific predation of eggs is a well documented phenomenon amongst aphidophagous coccinellids. The invasive species Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae) is known to be a top intraguild predator and reported to attack the eggs of many coccinellid species both in a laboratory setting and in the wild. A previous laboratory study highlighted that while many species’ eggs were highly palatable to H. axyridis, the eggs of Calvia quatuordecimguttata (Linnaeus) (Coleoptera: Coccinellidae) appeared to be extremely well protected from attack. Here we present the results of behavioural experiments testing the hypothesis that substances on the egg surface are responsible for this protection, and report preliminary results of GC-MS analysis of these compounds. When the coatings of C. quatuordecimguttata eggs were removed using hexane, they became significantly more susceptible to predation by neonate H. axyridis larvae. However, their overall palatability was not affected, in that complete consumption was never or rarely observed. This suggests that the surface compounds are a true indicator of unpalatability in this species. The effect of hexane-washing on already palatable conspecific eggs was also analysed but had no significant effect on the susceptibility of eggs to cannibalism. We conclude that the eggs of at least one European species are effectively protected by surface deterrents from intraguild predation by H. axyridis. This effect might be due to both the diversity and abundance of hydrocarbons present within the egg coating, the presence of alkenes and/or the presence of patches of a red substance on the eggs’ surface, which is thought to belong to the acid group. In conjunction with data on the susceptibility of other immature stages of C. quatuordecimguttata, this finding may indicate a decreased risk of the species falling victim to invasive H. axyridis, despite their coincident habitat ranges.     

Post-fertilization changes in the zona pellucida glycoproteins of rat eggs

The zona pellucida (ZP) is the extracellular coat surrounding the mammalian egg. Numerious evidence supports the role of ZP carbohydrate residues as the specific sperm receptors. In this study we used lectins to study different distribution patterns of carbohydrate residues in the rat ZP, and to follow changes at fertilization. ZP were collected from follicular, ovulated, and fertilized eggs, incubated with one of 11 different biotin-labeled lectins, followed by avidin-fluorescein isothiocyanate (FITC) complex, and visualized by epifluorescent microscopy. For electron microscope (EM) histochemistry, eggs were embedded in LR white and ultrathin sections were stained with the complexRicinus communis lectin (RCA-1)-colloidal gold. Some lectins (RCA-I,Glycine max) bound to the entire ZP while others were restricted to the inner or outer zones [Griffonia simplicifolia, Concanovalia ensiformis, Triticum vulgaris (WGA), succinyl-WGA]. Other lectins (Lens culinaris, Ulex europhaeus) were totally excluded. The RCA-1 binding pattern changed following sperm penetration, from homogeneous in ZP of ovulated eggs (57%) to uneven in ZP of fertilized (71%) or activated (68%) eggs. Our results demonstrate an uneven distribution of different sugar residues in the rat ZP, and a post-fertilization change in the distribution of β-galactose, which is specifically recognized by RCA-I, presumably correlated with other changes in the ZP that lead to the block to polyspermy. This work is in partial fulfillment of the requirements for the PhD degree of Tamar Raz at the Sackler School of Medicine, Tel Aviv University     

In vitro culture of macrobrachium eggs

Causes for the death of the eggs in the prawn Macrobrachium nobilii are: i) shedding of eggs by ovigerous female, and ii) infection by epibionts: a Saprolegnial fungus, bacteria (gram negative) and protozoans (Vorticellids and Paramecium). A cause for the death of freshly hatched larvae of some decapods is the reduction in reserve yolk energy in the larvae hatched in the last few batches. To circumvent these disadvantages, an artificial incubator was designed, in which 70% of the 3-day old eggs can successfully be incubated and hatched simultaneously. The isolted eggs are irrigated with filtered and aerated water over a diaphragm in the incubator; the water flushed from below through the diaphragm in the artificial incubator, sways and keeps the eggs continuously in a suspended motion, simulating the irrigation technique of the mother.Presented in the Second International Symposium on Invertebrate Reproduction held in Davis, California during August, 1979     

Artificial incubation of trumpeter swan eggs: Selected factors affecting hatchability

Eggs collected from captive trumpeter swans (Cygnus buccinator) in 1993 (n = 33) and 1994 (n = 42) were artificially incubated with careful monitoring to identify factors contributing to the low hatch success reported by the Ontario Trumpeter Swan Restoration Program. Fertility was > 80% in both years, whereas hatch success of fertile eggs was 14.3% (n = 4) of 28 eggs in 1993 and 37.1% (n = 13) of 35 eggs in 1994. Necropsy of non‐viable eggs indicated a high incidence of embryonic mortality during early and late incubation. Early embryonic mortality was associated with egg storage times exceeding 7 days (P < 0.05) and bacterial contamination of eggs (P < 0.01). Late mortality was associated with (P < 0.001) increased weight loss during incubation period and may have resulted from incubator temperature and humidity fluctuations. We established patterns of weight loss for eggs and determined that hatched eggs lost 11–15% of initial mass and that weight loss >15% resulted in embryo mortality. Results from this study indicate that collection and handling of eggs before incubation and precise control of the incubator environment are critical to hatchability of eggs. Zoo Biol 18:403–414, 1999. © 1999 Wiley‐Liss, Inc.     

Copepod eggs survive a decade in the sediments of the Baltic Sea

Many planktonic calanoid copepod species have been proved to spend a part of their life cycle as benthic resting eggs. In addition to avoiding seasonally occurring unfavourable conditions resting stages may also be used as a long-term survival strategy. The aim of this study was to find out for how long eggs of calanoid copepods retain their viability in the sediments of the Baltic Sea. The occurrence of viable copepod eggs in sea bottom sediment was studied in Pojovik Bay, SW coast of Finland. Eggs were found throughout a 25 cm deep core but deeper than 20 cm they were very scarce. Eggs were incubated at 12 °C in order to check the viability, and their age was estimated by determining the sediment accumulation rate with ¹³⁷Cs-method. Viable eggs were estimated to be 10–13 yrs old, some possibly even 18–19 yrs. Most eggs in the top 8 cm were viable, their age being up to 7–8 yrs. Nauplii that hatched from the eggs belonged to Acartia bifilosa and Eurytemora affinis, A. bifilosa dominating the hatchers of the top sedment layers and E. affinis the deeper layers. Preliminary evidence is presented that E. affinis produces true diapause eggs in the Baltic Sea.     

Surface alterations of fertilized surf clam (Spisula solidissima) eggs induced by concanavalin A

Fertilized Spisula eggs, incubated in ConA, were examined at periodic intervals to determine the effects of lectin binding on events of fertilization and cleavage. ConA was localized to specific regions of the vitelline layer and plasma membrane by reacting lectin-treated eggs with horseradish peroxidase and diaminobenzidine. In contrast to eggs, little reaction product was associated with the plasma membrane of spermatozoa. Sperm that fused with ConA-treated eggs failed to move into the cortex of the ovum and were observed as bulbous appendages at the surface of the zygote. Reorganization of sperm nuclei was inhibited, and male pronuclei failed to develop. ConA also inhibited polar body formation and cleavage. The maternally derived chromatin underwent meiosis, and the chromosomes normally taken into the first and second polar bodies were retained within the zygote. All of the maternally derived chromatin was organized within four or more female pronuclei which subsequently entered mitosis. The effects of ConA binding on events at the surface of fertilized Spisula eggs were abrogated by α-methyl-d-mannoside; succinyl-ConA only partially inhibited fertilization-related processes. The effects of ConA are discussed in terms of possible cross-linking of surface components of fertilized Spisula eggs which may inhibit deformation of the zygote cortex.     

Water exchange of flexible-shelled eggs and its effect on hatchling traits in the Chinese skink,<Emphasis Type="Italic"> Eumeces chinensis</Emphasis>

Flexible-shelled eggs of the Chinese skink (Eumeces chinensis) were used to determine the routes of water exchange with the environment and the effects of water exchange on hatchling traits. Eggs were incubated under three different hydric conditions at 28 °C. One group (ground eggs) was half-buried in incubation substrate. The second group (upper eggs) rested on the ground eggs but did not contact the substrate directly, and the third group was placed on a plastic platform above the substrate. Ground and upper eggs increased in mass and hatched successfully, while all eggs resting on the plastic platform lost mass and died within a week. Hence, E. chinensis eggs cannot sustain water balance through exchange of water vapor in air alone, but can absorb water from the substrate and other eggs and develop well. Water gained by upper eggs was only 20% of that absorbed by ground eggs, but hatchlings from the two treatments did not differ in morphology or locomotor performance. Therefore, the degree of water exchange of E. chinesis eggs has little effect on embryonic development and hatchling traits. The insensitivity of the eggs to variations in water exchange may be an adaptation against extreme hydric conditions in a species that deposits its eggs close to the soil surface.Communicated by I.D. Hume     

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.     

Nocturnal parasitism of moth eggs by Trichogramma wasps

Parasitoid wasps of the genus Trichogramma are used worldwide as biological control agents against lepidopteran pests. Trichogramma wasps develop inside eggs of a wide range of host species, most of them moths. They are generally considered as diurnal insects. Here, we investigated whether Trichogramma wasps can also successfully parasitise host eggs at night under controlled laboratory conditions. Eggs of the moth Ephestia kuehniella were offered under dark conditions (scotophase) to females of Trichogramma brassicae and Trichogramma evanescens either from 9:00 PM to 9:00 AM or from 11:00 AM to 5:00 PM at four different temperatures (5°C, 10°C, 15°C and 20°C). Both species are known to parasitise E. kuehniella eggs in the photophase during daytime. The results show that T. brassicae did not parasitise eggs in the scotophase at night and only very few in the artificially induced scotophase during daytime from 10°C to 20°C. In contrast, T. evanescens parasitised more eggs in the dark both at night and artificially induced scotophase during daytime. Parasitism in the scotophase already started at 5°C, with more eggs being parasitised and more offspring being produced at higher temperatures. T. evanescens displayed higher parasitism activity in the induced scotophase during daytime than in the scotophase at night. The present study suggests that Trichogramma are capable of successfully parasitising host eggs at night, even at low temperatures, but that nocturnal activity with respect to parasitism varies between wasp species.     

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.     

Fusarium solani invader of the eggs of the insectPanstrongylus geniculatus in a vivarium

A strain ofFusarium solani sensu Snyder & Hansen invaded the eggs of the insectPanstrongylus geniculatus in a vivarium. None of the invaded eggs hatched. To establish experimentally the pathogenicity of thisFusarium species against the eggs ofP. geniculatus, the fungus and the eggs were incubated together under different relative humidities and temperatures. At 64% relative humidity and 26 °C, the fungus grew well colonizing and penetrating all of the chorions.Three embryos died and were also colonized byF. solani. Only 4 nymphs hatched and survived to day 20. It is concluded that the isolate ofF. solani was capable of colonizing and invading the chorion of the eggs under certain humidity and temperature conditions and cause the death of the embryos.     

Effects of ultraviolet radiation on the eggs of landlocked Galaxias maculatus (Galaxiidae, Pisces) in northwestern Patagonia

• 1 Ultraviolet radiation (UVR) damages early life stages of several fish species. Galaxias maculatus is a small catadromous fish, with landlocked forms occurring in many lakes within the Nahuel Huapi National Park (Patagonia, Argentina). In this work, the vulnerability of G. maculatus eggs exposed to both natural and artificial UVR was investigated in relation to water transparency.
• 2 Field experiments were performed in two lakes differing in UVR attenuation. Galaxias maculatus eggs were exposed to in situ levels of UVR in quartz tubes incubated at various depths. For laboratory experiments, the eggs were exposed to five levels of artificial UVB radiation.
• 3 Exposure to natural UVR causes various degrees of egg mortality depending on water transparency and incubation depth. In the less transparent lake (K_d320 = 3.08 m^‐1), almost complete mortality was observed near the surface. At a depth of 43 cm the observed mortality was only 22%, but was still significantly different from the dark control. In the most transparent lake (K_d320 = 0.438 m^‐1), almost total mortality was observed in tubes incubated at 2.56 m or shallower. A gradual decline in mortality was recorded from that depth to 3.78 m where the values approached those in the dark control treatments.
• 4 A monotonic relationship between mortality and UV exposure could be observed both in field and laboratory experiments. Using the results from field incubations, a LD₅₀ of 2.5 J cm^‐2 nm^‐1 was estimated. In a few mountain lakes, this value would be exceeded even if the eggs were laid at the maximum depth of the lake. Thus UVR seems a sufficient cause to explain the absence of G. maculatus populations in some mountain lakes. For most lakes, however, UVR is probably one of several important environmental factors, which together determine the habitat suitability.

     

Effect of beetle size on predation of cabbage root fly eggs by ground beetles

Laboratory studies were done to determine the numbers of cabbage root fly (Delia radicum L.) (Diptera: Anthomyiidae) eggs eaten by sixty species of ground beetles (Coleoptera: Carabidae) collected from a range of habitats. For 35 species that were between 2.7 mm and 10 mm long, there was a linear relationship between the numbers of eggs eaten and beetle length. For each 1 mm increase in length above 2.7 mm, an additional 18 eggs were eaten/beetle/day. Eight further species within the above size range did not eat any fly eggs, as these species are known to be either phytophagous or to feed on moving prey. Large numbers of eggs were eaten by beetles in 13 of the 25 genera tested. Based on size, maximum numbers of eggs were eaten by 7 species of Agonum Bonelli, 5 species of Amara Bonelli and seven species of Bembidion Latreille. Only one of the 6 species of Pterostichus Bonelli tested, P. strenuus (Panz.), ate the expected numbers of eggs, the other species being too large to feed from such small prey items as cabbage root fly eggs. The numbers of beetles larger than 10 mm that ate eggs was highly variable and so 10 mm was considered the upper size limit of carabid predators of cabbage root fly eggs.     

Induction of cross-fertilization between sea urchin eggs and starfish sperm by polyethylene glycol treatment

Cross-fertilization between sea urchin eggs (Strongylocentrotus nudus) and starfish sperm (Asterina pectinifera) was induced by treatment with polyethylene glycol (PEG). Without treatment with PEG, the denuded egg surface (jelly coat- and vitelline coat-free) engulfed the head of acrosome-reacted sperm; however, sperm penetration did not occur [Kyozuka and Osanai, 1988]. When these eggs were exposed briefly to PEG (molecular weight 3,000) in seawater, the sperm entered the egg by membrane fusion. Cortical granules were discharged, and embryogenesis began following sperm penetration. PEG did not induce parthenogenesis in Strongylocentrotus eggs. Egg activation is thus closely linked with gamete membrane fusion.     

Dynamics of heat transfer to cold eggs in incubating bantam hens and a black grouse

Incubating birds transfer large amount of heat from the brood patch to the eggs during rewarming of cold eggs. If a vasoconstriction is present in the brood patch as in other parts of the body, it could possibly limit heat transfer to the eggs. To investigate this, heat transfer to water-circulated eggs was measured in incubating bantam hens (Gallus domesticus) and a black grouse hen (Lyrurus tetrix) during exposure to cold eggs. Egg temperature, egg surface temperature, heat production and cloacal temperature were also measured. At all levels of egg cooling, egg surface temperature and heat transfer to the eggs was stable throughout an exposure, except during resettling movements, which often changed egg surface temperature and the level of heart transfer. Egg surface temperature decreased linearly with egg temperature in both species, but was lower and more variable at low egg temperature in black grouse than in bantam hens. A higher proportion of the heat production was transferred to the eggs in the black grouse (corresponding to 109–118% of the increase above resting level) than previously reported in bantam hens. Clutch size did not affect this efficiency of heat transfer in black grouse. It is concluded that a vasoconstruction of the brood patch does not occur even under strong cold stress from the eggs. Heat transfer to the eggs is probably controlled more by behavioural adjustments than circulatory changes. An increase in brood patch blood flow probably occurs at relatively high egg temperature at the onset of egg rewarming. The efficiency of heat transfer, and thus the energetic cost of rewarming eggs, depends on the insulation of the bird and nest structure. The boreal/subarctic black grouse was able to reduce heat loss to the environment and transfer a higher proportion of its heat production to the eggs than the tropical bantam hen.Abbreviations AVAs arteriovenous anastomoses - HP heat production - HT heat transfer - T _a ambient temperature - T _b cloacal temperature - T _brp brood patch temperature - T _e egg temperature - T _es egg surface temperature