Artificial steps mitigate the effect of fine sediment on the survival of brown trout embryos in a heavily modified river

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Transfer of blood containing primordial germ cells between chicken eggs development of embryonic reproductive tract

The present study was carried out to investigate development of recipient chicken embryonic reproductive tracts which are transferred chicken primordial germ cells (PGCs). It is thought that differentiation of PGCs is affected by the gonadal somatic cells. When female PGCs are transferred to male embryos, it is possible that they differentiate to W-spermatogonia. However, the relationship development between PGCs and gonads has not been investigated. At stage 12–15 of incubation of fertilized eggs, donor PGCs, which were taken from the blood vessels of donor embryos, were injected into the blood vessels of recipient embryos. The gonads were removed from embryos that died after 16 days of incubation and from newly hatched chickens and organs were examined for morphological and histological features. The survival rate of the treated embryos was 13.6% for homo-sexual transfer of PGCs (male PGCs to male embryo or female PGCs to female embryo) and 28.9% for hetero-sexual transfer PGCs (male PGCs to female embryo or female PGCs to male embryo) when determined at 15 days of incubation. The gonads of embryos arising from homo-sexual transfer appeared to develop normally. In contrast, embryos derived from hetero-sexual transfer of PGCs had abnormal gonads as assessed by histological observation. These results suggest that hetero-sexual transfer of PGCs may influence gonadal development early-stage embryos.     

Open-pulled straw vitrification differentiates cryotolerance of in vitro cultured rabbit embryos at the eight-cell stage

The objective was to determine cryotolerance of in vitro cultured rabbit embryos to the open-pulled straw (OPS) method. Overall, 844 rabbit embryos at pronuclear, 2- to 4-cell, 8-cell, and morula/blastocyst stages were vitrified, and ≥ 1 mo later, were sequentially warmed, rehydrated, and subjected to continuous culture (n = 691) or embryo transfer (ET, n = 153). Embryos vitrified at the 8-cell stage or beyond had greater survival, expanded blastocyst and hatched blastocyst rates in vitro, and better term development than those vitrified at earlier stages. The 8-cell group had 70.1% expanded blastocysts, 63.7% hatched blastocysts, and 25.7% term development, as compared to 1.5-17.7%, 1.5-4.3% and 2.8-3.7% in the pronuclear, 2-cell and 4-cell embryos, respectively (P < 0.05). The expanded and hatched blastocyst rates in vitrified morula/blastocyst post-warming were higher than that in the 8-cell group; however, their term development after ET was similar (8-cell vs morula/blastocyst: 25.7 vs 19.4%, P > 0.05). Development after ET was comparable between vitrified-warmed embryos and fresh controls at 8-cell and morula/blastocyst stages (19.4-25.7 vs 13.7-26.6%, P > 0.05). For embryos at pronuclear or 2- to 4-cell stages, however, term rates were lower in the vitrified-warmed (2.8-3.7%) than in fresh controls (28.6-35.6%, P < 0.05). Therefore, cultured rabbit embryos at various developmental stages had differential crytolerance. Under the present experimental conditions, the 8-cell stage appeared to be the critical point for acquiring cryotolerance. We inferred that for this OPS cryopreservation protocol, rabbit embryos should be vitrified no earlier than the 8-cell stage, and stage-specific protocols may be needed to maximize embryo survival after vitrification and re-warming.     

Thermotolerance of brown trout,Salmo trutta,gametes and embryos to increased water temperatures

The present study investigated the effect of increased water temperature on sperm motility when activated in water, on egg water hardening, on the fertilization process and on embryonic development in brown trout, Salmo trutta. Brown trout gametes had a broad temperature optimum ranging from 3 to 15°C. In this temperature range the processes of egg water hardening and fertilization were unaffected. Motility duration was also not affected. The percentages of initial sperm motility and swimming velocity were lower at 9–15°C than at 3–7°C, while the percentage of locally motile spermatozoa was higher. Exposure of brown trout embryos to ≥11°C resulted in reduced percentages of hatched larvae and in increased percentages of malformed larvae. More advanced ontogenetic stages (stages ≥9 according to Ballard [1973]) showed higher tolerance to increased temperature than did less advanced stages (stages <9) when the percentage of eyed stage embryos was used as the evaluation end point. However, when the percentages of hatched larvae and normal shaped larvae were used as evaluation end points, no differences in thermotolerance could be found between the different otogenetic stages.     

Effects of reduced oxygen level on the mortality and hatching rate of Chondrostoma nasus embryos

Exposure of embryos of Chondrostoma nasus to a strongly reduced oxygen content (about 10% of air saturation) enhanced mortality and depressed hatching success. Viable larvae hatched only from the controls and from the group exposed to reduced oxygen during the earliest embryogenesis (from fertilization to gastrula). In the groups exposed from gastrula to eyed stage and from eyed stage to hatching, a high percentage of fish hatched, but all were deformed and almost all died during the following 1–5 days. Hatching period was prolonged in the groups exposed to reduced oxygen content. At a spawning area C. nasus had deposited a large portion of eggs in areas of reduced oxygen content. On the other hand, eggs placed at the substratum surface were exposed to high predation pressure from Barbus barbus, Lota lota and ducks ( Anas platyrhynchos ). The trade-off between predation pressure at the substratum surface and reduced oxygen conditions in the interstices resulted in a narrow space for successful development of embryos of this endangered rheophilic fish species under natural conditions.     

The importance of measuring biotic and abiotic factors in the lower egg pocket to predict coho salmon egg survival

Based on results from simulated redds of coho salmon Oncorhynchus kisutch , the amount of fine sediment <0·5 mm in the lower half of the egg pocket, rather than the entire egg pocket of the redd, was a strong predictor of egg survival to hatching (  r ²  = 0·62). The relationship was much stronger than observed in other studies, which typically ignore egg pocket structure. Abundance of a fish egg-eating worm, Haplotaxis ichthyophagous , an oligochaete that may have been attracted to fine sediment and dead eggs in the egg pocket, was also associated with a decrease in egg survival. The worm, however, accounted for little of the variance in survival compared to fine sediment. Only 10% fine sediment (<0·5 mm) in the lower pocket was required to decrease survival from 100 to 5%. Other abiotic factors had weaker (gravel permeability) or non-existent (dissolved oxygen) correlations with survival.     

Uptake and utilization of14C-glycine by embryos ofSebastes melanops

Synopsis The ability of embryos of the viviparous scorpaenidSebastes melanops to take up nutrients from an exogenous substrate was demonstrated by incubating embryos at various stages of development (18–30 days after fertilization) in¹⁴C-labeled glycine for 24 h. Uptake was highest for embryos at the latest stages (28–30 days) and increased at a linear rate during the incubation period. Nutrient uptake was not time dependent in embryos at the early stages (18–22 days). Nutrient utilization byS. melanops embryos was measured by the oxidation of¹⁴C-labeled glycine to¹⁴CO₂. The amount of respired¹⁴CO₂ by the oldest embryos increased significantly at a linear rate over the 24 h incubation period. There was no evidence of nutrient utilization by the youngest embryos. The developmental changes we observed in the uptake and utilization of exogenous glycine are supported by our previous findings that the oldest embryos have fully developed mouths and guts, and require additional nutrition from intraovarian sources at this stage of development.     

Hatching and viability of rotifer diapausing eggs collected from pond sediments

1. Planktonic rotifers inhabiting variable environments produce diapausing eggs that accumulate in the sediment of lakes and ponds, forming egg banks that may withstand adverse periods. A common assumption in zooplankton diapausing egg bank studies is to count as viable all eggs in the sediment that look healthy. This assumption should be challenged by asking how effectively ‘healthy‐looking’ eggs represent viable eggs. 2. In this study, viability of more than 1100 ‘healthy‐looking’ diapausing eggs belonging to the Brachionus plicatilis species complex was assessed in a laboratory hatching experiment. Eggs were collected at different depths from sediment cores obtained from 15 ponds located in coastal and inland areas of Eastern Spain. 3. Only approximately one half of the ‘healthy‐looking’ diapausing eggs hatched after incubation in experimental conditions. Almost all the hatchlings (99.4%) survived to maturity. The proportion of ‘healthy‐looking’ diapausing eggs that hatched varied among areas and among ponds within area, and substantially declined with sediment depth. Most of the hatchlings (88%) were obtained from the uppermost 2 cm of sediment. ‘Healthy‐looking’ eggs from upper sediment layers hatched after significantly shorter incubation times than eggs recovered from deeper layers. 4. Both decreased hatching success and increased incubation time for hatching with sediment depth suggest that older ‘healthy‐looking’ eggs are less responsive to hatching stimuli and could become unviable. However, the strong correlation found between the number of ‘healthy‐looking’ eggs and the number of hatchlings indicates that the abundance of ‘healthy‐looking’ eggs is a good index of egg bank viability.     

The effects of increased constant incubation temperature and cumulative acute heat shock exposures on morphology and survival of Lake Whitefish (Coregonus clupeaformis) embryos

Increasing incubation temperatures, caused by global climate change or thermal effluent from industrial processes, may influence embryonic development of fish. This study investigates the cumulative effects of increased incubation temperature and repeated heat shocks on developing Lake Whitefish (Coregonus clupeaformis) embryos. We studied the effects of three constant incubation temperatures (2 °C, 5 °C or 8 °C water) and weekly, 1-h heat shocks (+3 °C) on hatching time, survival and morphology of embryos, as these endpoints may be particularly susceptible to temperature changes. The constant temperatures represent the predicted magnitude of elevated water temperatures from climate change and industrial thermal plumes. Time to the pre-hatch stage decreased as constant incubation temperature increased (148 d at 2 °C, 92 d at 5 °C, 50 d at 8 °C), but weekly heat shocks did not affect time to hatch. Mean survival rates and embryo morphometrics were compared at specific developmental time-points (blastopore, eyed, fin flutter and pre-hatch) across all treatments. Constant incubation temperatures or +3 °C heat-shock exposures did not significantly alter cumulative survival percentage (~50% cumulative survival to pre-hatch stage). Constant warm incubation temperatures did result in differences in morphology in pre-hatch stage embryos. 8 °C and 5 °C embryos were significantly smaller and had larger yolks than 2 °C embryos, but heat-shocked embryos did not differ from their respective constant temperature treatment groups. Elevated incubation temperatures may adversely alter Lake Whitefish embryo size at hatch, but weekly 1-h heat shocks did not affect size or survival at hatch. These results suggest that intermittent bouts of warm water effluent (e.g., variable industrial emissions) are less likely to negatively affect Lake Whitefish embryonic development than warmer constant incubation temperatures that may occur due to climate change.     

Influence de quelques paramètres du milieu d'incubation sur la survie d'alevins de saumon atlantique,Salmo salar L., en ruisseau expérimental. Influence of some Environmental Parameters upon Survival during Embryonic Development of Atlantic salmon,Salmo solar L., in an Experimental Stream Channel

The effect of some environmental parameters of the redd (substrate composition, interstitial dissolved oxygen concentration) upon survival of Atlantic salmon embryos was studied in a large artificial stream channel. Batches of 3000 eyed eggs were buried 0.25 m deep in the gravel substrate of 8 different channel sections, where natural or artificial sediment deposition took place. Upon emergence, fry were captured in drift nets or by electrofishing. Survival varied from 32 to 94 % in the different sections. It was strongly affected by volume of sediment less than 10 mm in particle size and more particularly by the fraction less than 0.2 mm, the main component of natural sediment deposition. Survival was less strongly determined by oxygen concentration variations except for values less than 2 mg/l obtained in the section with highest sediment deposition. Available volume of interstitial void space appeared to be crucial for good survival during larval development.     

Vitrification of Mouse Embryos at Various Stages by Open-Pulled Straw (OPS) Method

This study was performed to pursue the optimal condition for the cryopreservation of mouse morulae by a two-step OPS method and to investigate the feasibility of the optimal condition for vitrification of embryos at other developmental stages. First, the mouse morulae were vitrified in OPS using one-step procedure—that is, embryos were vitrified after direct exposure to EDFS30 (15% ethylene glycol (EG), 15% dimethyl sulfoxide (DMSO), Ficoll and sucrose), or two-step method—that is, embryos were first pretreated in 10%E+10%D (10% EG and 10% DMSO in mPBS) for 30 sec, then exposed to EDFS30 for 15 to 60 sec, respectively. After vitrification and warming, the embryos were morphologically evaluated and assessed by their development to blastocysts, expanded/hatched blastocysts, or to term after transfer. The result showed that all the vitrified-warmed morulae had similar blastocyst rate compared to that of control (91.7% vs. 100%), and the highest developmental rate to expanded blastocysts (100%) or hatched blastocysts (62.3%) was observed when the morulae were pretreated with 10%E+10%D for 0.5 min, exposed to EDFS30 for 25 sec before vitrification and warming in 0.5 M sucrose for 5 min. After transfer, the survival rate (33.1%) in vivo of the vitrified morulae was higher (P > 0.05) than that of the fresh embryos (24.6%). Secondly, embryos at different stages were cryopreserved and thawed following the above program. Most (93.4 to 100%) of the embryos recovered after vitrification were morphologically normal at all the developmental stages. The blastocyst rates of the vitrified one-cell (52.5 to 66.7%) and the two-cell (63.3 to 68.9%) embryos were lower (P < 0.05) than those of the vitrified four-cell embryos (81.7 to 86.4%), the eight-cell embryos (90.0 to 93.3%), morulae (96.7 to 100%), and the expanded blastocysts rate (98.3 to 100.0%) of the vitrified early blastocysts. The highest survival rate in vivo of vitrified embryos were from the early blastocysts (40.4%), which was similar to that of fresh embryos (48.6%). The data demonstrate that the optimal protocol for the cryopreservation of morulae was suitable for the four-cell embryos to early blastocyst stages and that the early blastocyst stage is the most feasible stage for mouse embryo cryopreservation under our experimental conditions.     

Interaction between maternal effects and temperature affects diapause occurrence in the cricket Allonemobius socius

The induction of diapause can be adaptive for egg survival during unfavorable conditions, while direct development can be advantageous under favorable conditions by allowing additional generations to exploit abundant resources. Therefore, the physiological capability of a female to respond to environmental cues indicative of habitat quality by producing eggs of the appropriate developmental phenotype should be under strong selection. Additionally, developing embryos may alter the developmental trajectory initiated by the female in response to changing environmental conditions. In this study, we used a cross-fostering approach to isolate the maternal effects of parental diapause history (not previously studied) and egg-laying temperature from the influence of the incubation environment experienced by the developing embryo on the proportion of diapause eggs produced by the striped ground cricket, Allonemobius socius. We found that an interaction between egg-incubation temperature and parental diapause history strongly affected the proportion of diapause eggs produced and the proportion of eggs that hatched within a 16–18 day incubation period, while egg-laying temperature and all other interactions did not. These novel results indicate that embryos can respond directly to the environmental conditions they experience during development, but that their ability to do so is influenced by maternal effects such as parental diapause history. The results of this study not only provide evidence, for the first time, of parental diapause history affecting diapause proportions, but also raise additional questions about the mechanism by which environmental information is transmitted from parent to offspring and how offspring are able to respond to conditions experienced during their own development.     

Additional observations of endobenthic behaviour in the early life history of Melanostigma atlanticum (Zoarcidae) in muddy bottom sediments of the Laurentian Trough,eastern Canada

Synopsis While adult Melanostigma atlanticum continue to be sporadically noted living within bottom sediments of the Laurentian Trough, the observation of late embryo/early juvenile stages has not previously been reported. The occurrence of 22 live specimens between 20–25 mm total length in the subsurface anoxic sediment in early May suggests that development of eggs and embryos may take place in such environments during the winter.     

Effect of temperature on early development of white and lake sturgeon,Acipenser transmontanus and A. fulvescens

Synopsis The effect of constant incubation temperatures (between 10°C and 26°C) on the developmental rates was found to fit a similar exponential relationship in both the lake and white sturgeon embryos and larvae. Although the lake sturgeon had an overall slower rate of development than the white sturgeon, no statistically significant difference was detected in the slopes of the exponential equations describing the effect of temperature on developmental rate. The effect of these incubation temperatures on embryonic survival also did not differ between these two species. Both species exhibited optimal survival between 14–17° C and incipient mortalities occurred at 20°C. Temperatures above 20°C were lethal for white sturgeon embryos. No effect of low incubation temperature on survival was evident from this study. A comparison of these North American species with Eurasian acipenserids suggests that all the sturgeon that have been examined exhibit a similar influence of incubation temperature on developmental rate.     

Hatching ofCoregonus albula andC. lavaretus embryos at different stages of development

Synopsis Hatching in Coregoninae embryos is not closely connected with a determined stage of embryonic development. Electrical stimulation (AC) induced precocious hatching, and incubation at low temperature caused delay of hatching compared to controls. Embryos hatched precociously had shorter bodies and larger yolksacs than those whose hatching was normal or delayed. This was accompanied by respective changes in duration of the eleutheroembryonic phase (DEP) — inCoregonus lavaretus DEP decreased from 7 days in embryos hatched precociously to 0 days in embryos from delayed hatching.     

Immunohistochemical study of the appearance of somatotropic hormone in the adenohypophysis of chick embryos

The differentiation of somatotropocytes was studied in the Leghorn chick embryos during 11 to 18 days of incubation and in chickens during the first week of life using the immunohistochemical method of nonlabelled antibodies with PAP complex and antiserum against human somatotropic hormone (STH). Unlike in humans, the fixation of pituitaries in the Carnoy mixture is optimal for STH to be immunohistochemically estimated in the chickens and chick embryos. STH was found in adenohypophysis from 12-13 days of development. Besides predominant localization of somatotropocytes in the adenohypophysis caudal lobe, individual STH-positive cells are also present in the cephalic lobe of chickens and chick embryos. The results obtained suggest a relatively late appearance of STH during histotypical development of adenohypophysis in chick embryos.     

Hypoxia and temperature: Does hypoxia affect caiman embryo differentiation rate or rate of growth only?

In crocodilians, the rate of embryonic development and consequently many posthatch attributes are affected by temperature. Since temperature exhibits strong influences on fitness (embryo survivorship and phenotype) by shaping development, we manipulated oxygen concentration in order to uncouple the effects of developmental rate from the direct effects of temperature. Here we consider whether oxygen constrains either differentiation rate (progression from one stage to the next) or embryonic growth (size). Thus, we incubated Caiman latirostris eggs at various oxygen concentrations, and at two temperatures (31 °C, 100% female-producing temperature, and 33 °C, 100% male-producing temperature). We monitored the developmental stages of these embryos within the thermosensitive period (stages 20–24), and assessed several physiological and morphological hatchling traits. While embryonic size was strongly influenced by oxygen, differentiation rate did not seem to be affected. Very low oxygen concentrations and high temperatures inhibited embryo survival. In addition, oxygen availability affected incubation period and hatchling size, whereas temperature did not cause a significant variation in hatchling size. By investing energy in differentiation hypoxic embryos decreased their size.     

Sex ratios in mule duck embryos at various stages of incubation

Mule duck hatcheries have long reported varying degrees of unbalance in the sex ratio, with a preponderance of male mules at hatching. The aim of the present study was to assess the distributions of sex ratios at various stages of development in embryos originating from intra- and intergeneric crosses between parental lineages (Muscovy male x Muscovy female, Pekin male x Pekin female, Muscovy male x Pekin female or Mule, and Pekin male x Muscovy female or Hinny). In Experiment I, embryo sexing was performed on Days 1 and 5 of incubation (by multiplex PCR) and at hatching (by vent observation). The sex ratio was not significantly modified during the early stages of embryo development whatever the genetic origin (P>0.05, Days 1 and Day 5) but our results in mule and hinny ducklings confirmed the preponderance of males among normally hatched ducklings originating from the intergeneric lineage (58.9 and 55.4% males in mules and hinnies, respectively; P<0.05 in both cases). Sex ratio (vent sexing) in second grade (cull) ducklings revealed that 68% of these ducklings were females (P<0.05). In Experiment II, the distribution of sex ratio was also performed in mule duck eggs from 6 batches (400,000 eggs/batch) first examined for fertility (candling) on Day 18 of incubation. These results indicate that the percentage of males present in the population of normally hatched ducklings increases when fertility decreases. In addition, this experiment also revealed that 83.7-90.5% of viable male mule embryos develop up to hatching, compared to only 43.0-51.0% of female mule embryos. Given that a deviation in sex ratio during the first stages of incubation is unlikely (Experiment I), it is concluded that the skewed sex ratio of mule ducks at hatching is primarily due to increased late mortality in female mule embryos occurring between egg transfer and hatching. This mortality originated, at least in part, from the intergeneric origin of female mules, and was marked to a greater or lesser extent depending on the initial success of fertilization in a given batch, a possible indication that the initial quality of gametes may selectively exert its influence at the later stages of embryo development.     

Fine sediment affects on survival to emergence of robust redhorse

Robust redhorse (Moxostoma robustum) is a rare riverine sucker for which life history information is scarce. Spawning occurs over loose gravel substrate and eggs and larvae may be adversely affected by fine sediments among the gravel. A 2-year study was conducted to determine the threshold at which fine sediments are detrimental to successful egg incubation and larval emergence. Year 1 gravel treatments contained 0, 25, 50, and 75% fine sediments. Mean survival during Year 1 ranged from 63.5% in the 0% fine sediment treatment to 0% in the 75% fine sediment treatment. The results also indicated an adverse affect threshold between 0 and 25% fine sediment. Year 2 gravel treatments contained 0, 5, 10, 15, 20, and 25% fine sediments. Mean survival during Year 2 ranged from 69.8% in the 0% treatment to 9.1% in the 25% treatment. Year 2 results also identified the 15% fine sediment treatment as the threshold at which survival began to decline. Substrates at one known spawning area used by robust redhorse typically contain 25 to 50% fine sediment, but the spawning act cleans some fines from the egg pocket. Whether the “cleaning” that results from the spawning act reduces the fines sufficiently to avoid adverse effects is unknown. According to our results, survival rates of robust redhorse eggs and larvae are predicted to be about 8.0% or less when fine sediment is >25%.     

Invertebrate bioturbation can reduce the clogging of sediment: an experimental study using infiltration sediment columns

1. Invertebrate bioturbation can strongly affect water‐sediment exchanges in aquatic ecosystems. The objective of this study was to quantify the influence of invertebrates on the physical characteristics of an infiltration system clogged with fine sediment. 2. Two taxa (chironomids and tubificids) with different bioturbation activities were studied in experimental slow infiltration columns filled with sand and gravel and clogged with a 2 cm layer of fine sediment at the surface. We measured the effects of each taxon separately and combined on hydraulic head, water mobility and sediment reworking. 3. The results showed that invertebrates could reduce sediment clogging and this effect was linked to the functional mode of bioturbation of each group. Tubificid worms dug networks of galleries in the fine sediment, creating pathways for water flow, which reduced the clogging of sediment. In contrast, the U‐shaped tubes of chironomids were restricted to the superficial layer of fine sediments and did not modify the hydraulic conductivity of experimental columns. The combination of invertebrates did not show any interactive effects between tubificids and chironomids. The occurrence of 80 tubificids in the combination was enough to maintain the same hydraulic conductivity that 160 worms did in monospecific treatment. 4. The invertebrates like tubificid worms can have a great benefit on functioning of clogged interfaces by maintaining high hydraulic conductivity, which contributes to increased water‐sediment exchanges and stimulates biogeochemical and microbial processes occurring in river sediments.