Development of European bitterling in the gills of freshwater mussels

The development of bitterling embryos within the unique environment of a freshwater mussel's gills requires a departure from typical cyprinid embryological development. Eggs are large (2·6×1·7 mm), illustrating the low risk of predation, and elliptical; a response to unionid gill morphology and a way of increasing the transfer rates of respiratory and excretory products to and from the tissues. The yolk sac develops elongated lateral processes during early ontogeny; these secure the embryo into the host's interlamellar space. Once the larva is capable of movement (8·2 mm) the lateral processes are lost and the larva becomes less dependent of the host's gills for the provision of oxygen. Hatching (3·3 mm) and pigmentation of the blood (6·4 mm) occur relatively early; this may increase the rate of oxygen supply to the tissues. Pigmentation of the eyes and appearance of the melanophores occurs relatively late (7·4 mm and 7·9 mm, respectively); embryos are not required to detect or avoid predators. Bitterling larvae generally emerge from the host once the yolk sac has been consumed (10·5 mm); this may mark a change in respiratory and nutritional requirements.     

Early ontogeny of Galeichthys feliceps from the south east coast of South Africa

The early development of Galeichthys feliceps (Valenciennes, 1840) was investigated using a combination of incubator-reared embryos and embryos removed from the buccal cavities of wild-caught parents. The eggs were 15·7 mm in diameter and the average brood size was 49. Hatching occurred at an advanced developmental state after approximately 75–80 days, when branchial and fin differentiation were complete. Exogenous feeding began within the adult buccal cavity shortly after hatching. The young were released as juveniles after approximately 140 days, at a total length of 54 mm.     

PRIMARY CELL CULTURE FOR EMBRYONIC CARDIAC MYOCYTES OF MEXICAN AXOLOTL AND DISTRIBUTION OF DESMIN AND VIMENTIN

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The effects of environmental factors on rainbow smelt Osmerus mordax embryos and larvae

Experiments were conducted to identify environmental factors that influence the survival of rainbow smelt Osmerus mordax during their early life stages. Developing rainbow smelt embryos and yolk-sac larvae were cultured under controlled conditions with different dissolved oxygen (DO; 1·09, 2·18, 4·37 and 6·55 mg l⁻¹, pH (4·0, 4·5, 5·0, 5·5, 6·0 and 7·0), nitrate ( 0·7, 3·6, 7·3, 14·6 and 29·2 mg l⁻¹), phosphate (0·04, 0·21, 0·42, 2·08 and 4·17 mg l⁻¹) and salinity (0, 5, 10, 15, 20 and 30) levels. Rainbow smelt embryos were also incubated with simulated tidal salinity fluctuations (2–28), ultraviolet radiation (irradiances of 2·8, 6·2 and 5·1 W m⁻²) and under natural conditions in two rainbow smelt spawning rivers. In the laboratory, hatch was only impaired under the lowest DO and pH conditions (0 and 13% hatch, respectively) and at highest constant salinity levels (0% hatch). Larval survival was only affected by pH levels ≤5·0. The experiment that compared hatch under natural conditions was terminated when embryos became covered with silt and fungus. These results suggest that water acidification, sediment and fungal growth may affect rainbow smelt survival during their early life stages.     

Effects of temperature on developmental performance, survival and growth of sea lamprey embryos

This study assesses the influence of thermal regime on the development, survival rates and early growth of embryos of sea lamprey Petromyzon marinus incubated at five constant temperatures (7, 11, 15, 19 and 23° C). The time from fertilization to 50% hatching and from hatching to 50% burrowing were inversely related to incubation temperature. All the embryos incubated at 7° C died at very early stages, while those maintained at 11° C did not attain the burrowing stage. Survival from fertilization to hatching was 61, 89, 91 and 89% at 11, 15, 19 and 23° C, decreasing to 58, 70 and 70% from hatching to burrowing at 15, 19 and 23° C, respectively. Larvae reared during the first 3 months of exogenous feeding in a common environment at constant 21° C, revealed maximum survival for an incubation temperature of 15° C (43% of burrowed larvae) decreasing strongly at 19° C (16%) and 23° C (one suvivor among 240 larvae). Body length at the burrowing stage was maximum for embryos incubated at 19° C, but body mass increased in the interval 15–23° C. Mean incubation temperatures experienced by 117 broods during the embryonic development in the source river were estimated in 15·3±2·30° C and 16·7±1·76° C (mean±1 s.d .) for the periods fertilization-to-hatching and hatching-to burrowing, respectively.     

The mammary gland of the echidna, Tachyglossus aculeatus' with observations on the incubation of the egg and on the newly-hatched young

During the non-breeding season the lobules of the mammary glands consist of solid cords of cells which may occasionally be embedded in a pad of fat tissue.
During the breeding season some mammary glands can exhibit a tubular grade of organization and can even secrete milk without the intervention of pregnancy. Apparently the stimulus of pregnancy and incubation of an egg in the pouch is generally, but not invariably necessary for development of an alveolar grade of organization. The ducts leading from the lobules to the skin of the areola undergo hypertrophy during the course of lactation.
The present work shows that the gestation period is at least 16 days long but evidence in the literature shows it may be as long as 27 days. The incubation period of an egg in the pouch was found to be of the order of 10–10·5 days.
A newly-hatched echidna weighed 378 mg and was 1·47 cm long; it exhibited an egg tooth and the remains of foetal membranes attached to the abdomen. The newly-hatched had well-developed olfactory organs including sensory cells in the olfactory epithelium. The alimentary tract exhibited a degree of differentiation consistent with the notion that it could digest milk.
The fatty acid composition of the triglyceride fraction of a sample of milk taken just after hatching differed from that of mature milk in that the ratio of the concentration of oleic acid to palmitic acid in the early milk was about unity whereas it was 3·7 in mature milk. A high level of arachidonic acid may be a characteristic of the milk at hatching.
Iron was present in a concentration of 8·3 μg/ml in the early milk, and of 43·0 μ/mlg/ml in a sample of mature milk. The regression of the mammary glands from the alveolar to the solid-cord state is described.
Two corpora lutea of pregnancy, taken about 11 and 19 days after egg laying, were degenerate.     

The ontogeny of haematopoiesis in the marine teleost Leiostomus xanthurus and a comparison of the site of initial haematopoiesis with Opsanus tau

The ontogeny of haematopoiesis in the perciform fish, spot Leiostomus xanthurus , differed from that reported as the norm for fishes, as exemplified by the cypriniform zebrafish Danio rerio , and observed in the batrachoidiform oyster toadfish Opsanus tau . Erythropoiesis in spot was first evident in the head kidney of yolk‐sac larvae 3 days after hatching (DAH). No embryonic intermediate cell mass (ICM) of primitive stem cells or blood islands on the yolk were apparent within embryos. Erythrocytes were first evident in circulation near the completion of yolk absorption, c . 5 DAH, when larvae were c . 2·0 mm notochord length ( L _N). Erythrocyte abundance increased rapidly with larval development for c . 14 to 16 DAH, then became highly variable following changes in cardiac chamber morphology and volume. Erythrocytic haemoglobin (Hb) was not detected within whole larvae until they were 12 DAH or c . 3·1 mm L _N, well after yolk and oil‐globule absorption. The Hb was not quantified until larvae were >47 DAH or >7 mm standard length. The delayed appearance of erythrocytes and Hb in spot was similar to that reported for other marine fishes with small embryos and larvae. In oyster toadfish, a marine teleost that exhibits large embryos and larvae, the ICM and Hb were first evident in two bilateral slips of erythropoietic tissue in the embryos, c . 5 days after fertilization. Soon thereafter, erythrocytes were evident in the heart, and peripheral and vitelline circulation. Initial haematopoiesis in oyster toadfish conformed with that described for zebrafish. While the genes that code for the development of haematopoiesis are conserved among vertebrates, gene expression lacks phylogenetic pattern among fishes and appears to conform more closely with phenotypic expression related to physiological and ecological influences of overall body size and environmental oxygen availability.     

Calcium metabolism in relation to ovarian functions during early and late pregnancy in the viviparous blenny Zoarces viviparus

Calcium metabolism was studied in relation to ovarian functions and embryonic development during early and late pregnancy in the viviparous blenny Zoarces viviparus . The level of total calcium in the maternal serum decreased during pregnancy from 21·7 (± 2·6) to 8·7 (± 1·1) mg 100 ml⁻¹. Calcium in the ovarian fluid surrounding the embryos appeared at a level which was always lower than in the maternal blood. The calcium content of the embryos increased during their development in the ovarian cavity. During early pregnancy, oestradiol-treatment increased the level of total calcium and alkkali-labile protein P (vitellogenin) in serum of the maternal organism, but did not have any effect on the level of ultrafiltrable calcium. However, in the untreated females, the post-ovulatory follicles, believed to act as calyces nutriciae in the pregnant ovary, were able to concentrate the ultrafiltrable calcium above the level in the serum. The activity of injected⁴⁵Ca per unit plasma and follicular fluid volume was always higher in oestradiol-pretreated fish when compared with controls over a time course range. Embryos from females, which were not treated with oestradiol, accumulated high levels of labelled calcium and the accumulation was time-course dependent. Embryos from oestradiol-treated females showed low levels of tracer accumulation at all samplings and their appearance indicated an overall negative effect of oestradiol. During late pregnancy, labelled calcium showed a rapid turnover within 24 h post-injection in the maternal blood and ovarian fluid and a large accumulation in the embryos. Calcium influx in embryos, which were incubated with labelled calcium during 72 h in in vitro systems was largest during the initial 4·5–24h time interval.     

Acute effects of the sodium salt of 2,4-D on the early developmental stages of bleak, Alburnm alburnus

The acute effects of a herbicide based on 2,4-D on embryos and larvae of bleak, Alburnus alburnus L. exposed to concentrations of 25–3200 mg 1⁻¹ for 12–48 hours were investigated. The 2,4-D delayed or stopped the development of embryos in their early stages, caused behavioural changes and morphological alterations. The L.C.₅₀ values for embryos varied between 12–9 and 15·9–4mg 1⁻¹ and for larvae from 51·6 to 111·2mg 1⁻¹ according to the time of exposure. The level of 2,4-D in Lake Balaton, or similar shallow lakes, without substantial injury of the littoral fish fauna should not surpass 0·5–1·0 mg 1⁻¹.     

Photoperiodism and control of summer diapause in the Mediterranean tiger moth Cymbalophora pudica

Photoperiodic responses to both constant and changing photoperiods were studied in the Mediterranean tiger moth Cymbalophora pudica. Embryos, larval instars and prepupae were all sensitive to photoperiod, and the responses of larvae and prepupae to changing photoperiods were similar. At 23+/-2 degrees C, constant 24-h photoperiods with short photophases (11, 12h) induced a long diapause (mean 88days) whereas long photophases (14, 16h) induced a short diapause (mean 52days). A change to a longer or shorter photophase during larval development or during diapause caused a significant extension (up to a maximum of 138days) or shortening (down to a minimum of 10days) of diapause, respectively, but only when at least one of the photophases was longer than 14h. Thus, shortening and prolongation of photophase had an opposite effect than constant short and long photophases, respectively. Changes within the range of photophases of 10-14h did not cause a significant change in diapause duration.Experimental results enabled us to outline the mechanisms regulating voltinism and the duration of summer diapause. For the monovoltine cycle, cold autumn/winter temperatures slow down larval development, and prepupal aestivation starts relatively late (March, April). Prepupae are then kept in diapause by the increasing daylength (>14h after late April). Pupation is synchronized by decreasing daylength after summer solstice, and imagoes emerge in September/October. For the bivoltine cycle, when the autumn/winter temperatures are relatively warm, a certain portion of the population (depending on the individual rate of growth) may be diverted to a bivoltine life-cycle. In such a case, larval development is fast and short enough to allow an early start of diapause (prior to or during February). The duration of such early diapause is not influenced by changes in daylength (<14h); pupation occurs very early (April/May), and spring generation imagoes fly and oviposit in May/June. Summer larvae and prepupae live under decreasing daylength (>14h), which shortens their diapause to 20-30days. Imagoes of the autumnal generation thus occur in September/October, together with the univoltine portion of the population.     

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.     

Maternal transfer of cadmium tolerance in larval Oreochromis mossambicus

Exposing female tilapia to Cd did not affect reproduction and egg quality, nor the survival of their offspring. The LT ₅₀ of Cd-exposed larvae (within 24 h post hatching) from Cd-treated female Oreochromis mossambicus (mean ±S.E.=10·9±0·5 days, n =4) was greater than that of Cd-exposed larvae from control females (6·9± 1·2 days, n =4). A substantial amount of metallothionein mRNA (MT mRNA) was found in both oocytes and newly hatched larvae from Cd-treated females, but MT mRNA levels were low in newly fertilized eggs. The amount of MT mRNA was very low in all these stages in the control group. Cadmium could not be detected at any stage for either control or Cd-treated group, and metallothionein protein levels were also very low in both groups. The early appearance of MT mRNA in oocytes and newly hatched larvae from Cd-treated females may result in early translation after hatching and explain their tolerance to Cd.     

Effect of delayed first feeding on development and feeding ability of Paralabrax maculatofasciatus larvae

The impact of delayed feeding early in development on late feeding ability and development of spotted sand bass Paralabrax maculatofasciatus larvae was examined. Larvae were sampled from hatching until day 19 after delayed feeding for zero (control), 1, 2, or 3 days. Feeding incidence was evaluated as the percentage of larvae with food in the gut and feeding intensity was measured by direct counting of prey after dissection of the gut. Delayed feeding effects due to starvation were observed early in development. By day 5, significant differences ( P  < 0·05) were observed between controls and larvae submitted to degrees of delayed feeding, including total length ( L _T), eye diameter, and the size of the head, liver, gut, muscles and body. Differences were still apparent in L _T, and body, liver and muscle heights at the end of the studied period. Larvae under total starvation did not survive beyond day 5. Initial feeding incidence was 35, 60, 90 and 10% for larvae first fed on day 2, 3, 4 or 5, respectively. Mean ±  s . d . feeding intensity was 3·6 ± 0·8, 2·8 ± 1·3, 5·2 ± 0·3, and 10·2 ± 1·5 rotifers per larva depending on whether larvae were initially fed on day 2, 3, 4 or 5, respectively. The point of no return occurred between 4 and 5 days after hatching.     

Survival and sublethal responses of early life stages of pike exposed to low pH in artificial post-mining lake water

The threshold pH for survival of early life stages of pike Esox lucius in post-mining lakes was determined in a laboratory experiment using artificial water characterized by high concentrations of dissolved Ca, Al, Fe, Mn and SO2/4⁻. At pH 3·50–4·00, high mortality was observed before hatching. At pH 4·00 and 4·25, hatching rates were reduced compared with the controls (pH 7·40), and many embryos died in a partly hatched state. Hatching success of embryos exposed to pH levels of 4·50 or higher was not affected. However, at pH 5·00 many newly hatched embryos were deformed. Furthermore, pike exposed to pH 4·00–5·00 did not start feeding. At pH 4·75 and below, mortality increased to 100%, and at pH 5·00, only few eleutheroembryos which were in extremely poor condition survived to the end of the experiment. At pH 5·50, survival was in the same range as in the control group, but growth was reduced. Therefore, early life stages of pike are expected to survive in Lustian post-mining lakes when a pH of 5·50 and above is reached and maintained.     

Dynamic changes in microtubules and early development of reconstructed embryos after somatic cell nuclear transfer in a non-human primate

In order to improve somatic cell nuclear transfer (SCNT) efficiency and to understand cellular changes in SCNT, the dynamic changes in microtubules/DNA and early development of SCNT embryos with single or multiple pronuclei were investigated, along with activation timing on efficiency of SCNT, were studied in the Cynomolgus monkey. The confocal images showed that microtubules assembled around condensed DNA at 1h after cell injection; normal or abnormal reconstructed spindle formed at 2 h after cell injection; and reconstructed spindle separated at 2 h after activation. The results of nuclear formation showed that 61.3% of the reconstructed embryos did not form pronuclei; 19.3% formed a single nucleus, and 11.9% and 7.5% formed two and more than two reconstructed pronuclei, respectively. The cleavage and 8-cell development rates of SCNT embryos with pronuclei were significantly higher than those without pronuclei, but there was no difference in development rates among NT embryos with single, two and more then two pronuclei. Activation at 2 h after cell injection yielded more embryos with pronuclei and yielded 8-cell NT embryos more reliably than did activation at 3-4 h. In conclusion, microtubules assembled around condensed DNA at 1-2 h after cell injection, and formed a spindle at 2 h after SCNT, which separated at 2 h after activation; early development was affected by activation time, but no different between single and multiple pronuclei.     

The relationship between plasma progesterone and the timing of ovulation and early embryonic development in the marmoset monkey (Callithrix jacchus)

Levels of progesterone in peripheral plasma samples showed a mean (± S.E.M.) ovarian cycle length of 28 ·63 ± 1·01 days ( n = 19).
The preovulatory, or follicular phase (mean ± S.E.M. length: 8·25 ± 0·30 days, n = 56) was defined as that period of the cycle during which progesterone levels remained below 10 ng/ml. The postovulatory, or luteal phase (mean ± S.E.M. length: 19·22 ± 0·63 days, n = 48) was defined as the remaining period of the cycle during which levels remained between 10 ng/ml and 150 ng/ml.
The day of ovulation (day 0) was defined as that preceding the day on which progesterone levels first exceeded 10 ng/ml (day l), at the onset of the luteal phase. Oocytes and preim-plantation embryos recovered from the reproductive tract provided supporting evidence for the timing of ovulation.
The short follicular phase indicated that follicular growth may be initiated during the previous luteal phase. The long luteal phase may be related to the extended period of preim-plantation development.     

High mortality of embryos of a distinct autochthonous Austrian coregonid population during Zuger jar incubation

For a distinct autochthonous Austrian coregonid population called Reinanke Coregonus sp., embryo viability was very low in Zuger jars (mean ± s.d. 36·1 ± 16·1% eyed stage embryos in experiment 1, 15·1 ± 17·0% in experiment 2) but high in vertical flow incubators (63·3 ± 29·0% eyed stage embryos in experiment 1, 57·8 ± 18·9% in experiment 2). Mechanical shock sensitivity is a possible explanation for high embryo mortality.     

Morphological development and allometric growth patterns in the juvenile seahorse Hippocampus kuda Bleeker

The morphological development and allometric growth patterns in the juvenile spotted seahorse Hippocampus kuda were studied under hatchery rearing conditions. Newborn spotted seahorses [mean ± s.d . standard length ( L _S) 9·33 ± 0·79 mm] were raised till the age of 124 days (119·35 ± 6·04 mm). Growth was characterized by three stages with two inflexion points occurring at day 21 and 76. The mean growth rates in the first, second and third stages were 0·68, 1·16 and 0·71 mm day⁻¹, respectively. The growth rate was most rapid in the second stage and was probably influenced by a behavioural shift from pelagic to benthic form. The mass ( M ) and L _S relationship was exponential ( M = 7·14 × 10⁻⁶ L _S^2·76), but the slope, b = 2·76, reflected negative allometric growth. Sexes could be distinguished at c. 110 days, and the sex ratio was unbiased. The L _S in males and females did not differ significantly. Morphological stageing series is proposed, which divides H. kuda juvenile development into eight stages based on the development of coronet, cheek and eye spines, keel and pigmentation. The morphometric ratios for all the body parts, except trunk length, showed considerable changes at a transition point occurring at c. 25 mm L _S. The high proportional growth in head length, head depth, pectoral fin base length, dorsal fin base length, snout length, snout depth and eye diameter at the initial stages, and the abrupt increase in tail length only after the first 2 weeks, possibly reflect development priorities during early development where important organs are being developed first for the enhancement of juvenile survival.