Boric Acid Inhibits Germination and Colonization of Saprolegnia Spores In Vitro and In Vivo

Saprolegnia infections cause severe economic losses among freshwater fish and their eggs. The banning of malachite green increased the demand for finding effective alternative treatments to control the disease. In the present study, we investigated the ability of boric acid to control saprolegniosis in salmon eggs and yolk sac fry. Under in vitro conditions, boric acid was able to decrease Saprolegnia spore activity and mycelial growth in all tested concentrations above 0.2 g/L, while complete inhibition of germination and growth was observed at a concentration of 0.8 g/L. In in vivo experiments using Atlantic salmon eyed eggs, saprolegniosis was controlled by boric acid at concentrations ranging from 0.2–1.4 g/L during continuous exposure, and at 1.0–4.0 g/L during intermittent exposure. The same effect was observed on salmon yolk sac fry exposed continuously to 0.5 g/L boric acid during the natural outbreak of saprolegniosis. During the experiments no negative impact with regard to hatchability and viability was observed in either eggs or fry, which indicate safety of use at all tested concentrations. The high hatchability and survival rates recorded following the in vivo testing suggest that boric acid is a candidate for prophylaxis and control of saprolegniosis.     

Effects of simulated cold fronts on the survival and behaviour of yellow perch Perca flavescens yolk‐sac fry

Acute reductions in water temperature (i.e. cold fronts) may influence larval fish survival directly via limits on physiological tolerance or indirectly by acting as a sublethal stressor. The primary objective was to quantify survivorship of yellow perch yolk‐sac fry exposed to two different temperature declines (4 and 8°C) and compare survivorship to that of perch fry under ambient temperatures representative of natural conditions. Behaviour of yolk‐sac fry following temperature declines was also qualitatively assessed. Mean survival in the control, ?4, and ?8 treatment tanks was 90, 91 and 97%, respectively, and no significant differences in percent survival were observed between the control and the ?4 treatment (t_s = ?0.10; df = 7; P = 0.93), the control and ?8 treatment (t_s = ?1.85; df = 7; P = 0.11) or the ?4 and ?8 treatments (t_s = ?1.33; df = 7; P = 0.22). Observations of yellow perch eggs and fry behaviour following temperature declines differed among treatments. Any remaining eggs in the control treatment and ?4 treatments continued to hatch during the experiment, and fry were documented swimming throughout the water column in all tanks. However, in the ?8 treatment, any eggs that had not hatched remained inactive and all fry within all ?8 treatment tanks ceased swimming activity and settled to the bottom of the tanks once the temperature reached 3.9°C. Fry remained at the bottom of the tanks for the entire 48 h simulated cold‐front. Fry resumed swimming activity once water temperatures began to increase (by approximately 6°C). Results indicated that drops in temperature (i.e. cold fronts) similar to or greater than those found in small impoundments did not cause direct mortality of yellow perch during the yolk‐sac fry (post‐hatch larvae) stage. Although an acute drop in temperature may not induce sudden high mortality, it may be a sub‐lethal stressor, leading to increased starvation or predation risk.     

Rates of growth and protein synthesis correlated with nucleic acid content in fry of rainbow trout, Oncorhynchus mykiss: effects of age and temperature

Groups of recently hatched fry of rainbow trout, Oncorhynchus mykiss were maintained in the laboratory in order to investigate the effects of age, ration level and temperature on whole body growth, nucleic acid concentrations, protein synthesis rates and enzyme activities. In fry of up to 30 days after hatching, which were feeding but still had some yolk sac, no significant change in mean RNA concentration was observed with ration level. In older fry of 50 days or more, when the yolk sac was completely absorbed and exogenous feeding fully established, the concentration of RNA was correlated with the rate of protein growth. RNA concentrations and activities of citrate synthase and lactate dehydrogenase were significantly different between fed and starved fry. As water temperature was raised (from 5 to 15° C), higher rates of protein growth were brought about by an increase in the rate of protein synthesis and also by increased efficiency of retention of synthesized protein (reduced protein turnover). In fed fry, no change in RNA concentration was found with increasing temperature, while the amount of RNA per cell (RNA: DNA) decreased, indicating that increased rates of protein synthesis were due to increased RNA efficiency.     

Distribution, reproduction and growth of the catfish, Chrysichthys walkeri (Gunther) in the Lekki Lagoon, Nigeria

An investigation was made on the biology of the catfish, Chrysichthys walkeri (Gunther) occurring in the Lekki Lagoon. The catfish was evenly distributed over the six sampled stations of the lagoon with the sites of the shallow water stations probably serving both as breeding and nursery grounds.
Fecundity estimates for this species ranged from 896 to 4168 eggs. The average number of eggs per female was 2084. Laboratory incubated eggs hatched after 3 days with the yolk sac being completely absorbed on the seventh day. The fry averaged 8.6 mm on their first day of life.
The catfish attained a maximum total length of 25.7 cm within a period of 2 years. There were no significant differences in the growth pattern of this species during the dry and rainy seasons.     

Feeding pattern and diet of first feeding brown trout fry under natural conditions

The feeding habits of brown trout Salmo trutta fry were studied during the critical first feeding period in a natural spawning and nursery stream. A low proportion of the fry initiated exogenous feeding before emergence from the gravel, and while having nearly 30% of the yolk sac remaining. This probably reflected low feeding motivation or limited feeding opportunities within the gravel environment. The majority of the fry started feeding after emergence, and after most or all yolk was absorbed. Some fry emerged with large amounts of yolk remaining, while others emerged after yolk exhaustion. The degree of stomach fullness revealed that feeding was more efficient after a territory had been acquired. The diets of the young fry were dominated by chironomid larvae, followed by zooplankton and Plecoptera larvae. Fry dispersing downstream and out of the nursery area were significantly smaller than resident fry, indicating displacement due to competition for territories. The majority of the downstream dispersing fry had initiated feeding, and there was at this point no evidence of starvation in any of the fry. It therefore appeared that the later emerging fry actively migrated out of the overpopulated nursery area to find available territories further downstream.     

Changes in fatty acid composition,cholesterol and fat‐soluble vitamins during development of eggs and larvae in shabbout (Barbus grypus,Heckel 1843)

Changes in fatty acid composition, cholesterol and fat‐soluble vitamins were studied during development (fertilized eggs, yolk‐sac larvae, and after yolk resorption of shabbout, Barbus grypus). Significant differences were found in the monounsaturated fatty acids (MUFAs), polyunsaturated fatty acids (PUFA), PUFA/saturated fatty acids (SFA), ∑n‐3 and eicosapentaenoic acid (EPA)/docosahexaenoic acid (DHA) ratios between eggs and larvae (P < 0.05). Significant differences were also observed in the C14:0, C16:1n‐7, C18:1n‐9, C18:3n‐6, C20:0, C20:4n‐6, C24:0, C24:1, C22:6n‐3 fatty acids between eggs and larvae after yolk‐sac resorption (P < 0.05). Vitamin α‐ Tocopherol and retinol content increased during embryogenesis, but changes were insignificant in retinol acetate, δ‐Tocopherol, K1, K2 and cholesterol content between eggs and larvae after yolk resorption (P > 0.05).     

Proteolytic activity in the yolk sac membrane of quail eggs.

Extraembryonal degradation of yolk protein is necessary to provide the avian embryo with required free amino acids during early embryogenesis. Screening of proteolytic activity in different compartments of quail eggs revealed an increasing activity in the yolk sac membrane during the first week of embryogenesis. In this tissue, the occurrence of cathepsin B, a lysosomal cysteine proteinase, and cathepsin D, a lysosomal aspartic proteinase, has been described recently (Gerhartz et al., Comp Biochem Physiol, 118B:159-166, 1997). Determination of cathepsin B-like and cathepsin D-like proteolytic activity in the yolk sac membrane indicated a significant correlation between growth of the yolk sac membrane and proteolytic activity, shown by an almost constant specific activity. Both proteinases could be localized in the endodermal cells, which are in direct contact to the yolk. The concentration of proteinases in the endodermal cells appears to be almost unaltered in the investigated early stage of quail development, whereas the amount of endodermal cells increases rapidly, seen by a complicated folding of the yolk sac membrane. In the same cells quail cystatin, a potent inhibitor of quail cathepsin B (Ki 0.6 nM), has been localized at day 8 of embryonic development. Approximately at this stage of development, the quail embryo stops metabolizing yolk. In conclusion, it is strongly indicated that the amount of available free amino acids, produced by proteolytic degradation and supporting embryonic growth, is regulated by the growth of the yolk sac membrane.     

The corn snake yolk sac becomes a solid tissue filled with blood vessels and yolk-rich endodermal cells

The amniote egg was a key innovation in vertebrate evolution because it supports an independent existence in terrestrial environments. The egg is provisioned with yolk, and development depends on the yolk sac for the mobilization of nutrients. We have examined the yolk sac of the corn snake Pantherophis guttatus by the dissection of living eggs. In contrast to the familiar fluid-filled sac of birds, the corn snake yolk sac invades the yolk mass to become a solid tissue. There is extensive proliferation of yolk-filled endodermal cells, which associate with a meshwork of blood vessels. These novel attributes of the yolk sac of corn snakes compared with birds suggest new pathways for the evolution of the amniote egg.     

Flow of egg white ovalbumin into the yolk sac during embryogenesis

Western blot analyses of yolk proteins of the White Leghorn hens showed that an ovalbumin-like molecule was included in day 16 eggs but not in the ovarian follicles, and very little in newly deposited eggs. Northern hybridization, as well as in vitro translation, of poly(A)+ RNAs prepared from the yolk sac membranes of developing embryos gave no signal for ovalbumin messages. These results imply that the present ovalbumin-like protein of yolk has its origin in egg white, not being a de novo synthesis product in the yolk sac membranes.     

Embryonic and larval development of Thai Pangas (Pangasius sutchi Fowler, 1937)

The embryonic and larval development of Thai pangas was investigated during peak (May-July 1995) and late spawning (August-October 1995) periods. The fertilized eggs are adhesive and spherical with a yellowish or greenish-brown egg capsule. The yolk sac is yellowish-brown in color and 1.20-1.80 mm in diameter. Nine hours post-fertilization, the first cleavage stage, embryonic shield, head, tail region, neural grooves and somites were evident. The incubation period ranges from 24-36 h at a temperature of 20-30 degrees C. The newly hatched larvae are quite transparent and light yellowish in color with a body length of 2.98-3.10 mm. Eye pigments appear and the heart starts to work within 12-14 h of hatching. In 1-day-old pro-larvae, the mouth becomes well developed; barbules are elongated, prominent and look like tiny threads. The yolk sac is fairly well absorbed and the palatine teeth are fully developed during the 3 day pro-larval stage. At the end of 12 days of larval development, the stomach becomes functional and aerial respiration starts. After 2 weeks, the young fry is well-developed, and is of an adult appearance, that is, measuring up to 13.56 mm in length.     

The effects of incubation temperature on the morphology and composition of Australian Brush-turkey (<Emphasis Type="Italic">Alectura lathami</Emphasis>) chicks

Environmental heterogeneity during embryonic development generates an important source of variation in offspring phenotypes and can influence the evolution of life histories. The effects of incubation temperature on offspring phenotypes in reptiles has been well documented but remains relatively unexplored in birds as their embryos typically develop over a narrow range of temperatures. Megapode birds (Order Galliformes; Family Megapodiidae) are unique in that their embryos tolerate and develop over a wide range of incubation temperatures, yet little is known of the effect that temperature has on hatchling morphology and composition. Australian Brush-turkey eggs collected on the day of laying were incubated in the laboratory under constant temperatures of 32, 34 and 36°C until hatching in order to determine the influence of temperature on hatchling mass, size and composition. The dry mass of the yolk-free body and residual yolk of hatchlings were temperature dependent, such that higher temperatures produced chicks of lesser yolk-free body mass and greater residual yolk mass than chicks incubated at lower temperatures. However the overall size (linear dimensions) and lipid, protein and ash content of chicks were independent of temperature.     

Isotopic Labeling of Embryo,Yolk Sac,and Intracellular Parasites of the Embryonated Hen's Egg by Yolk Replacement

A technique is described which allows the replacement of 50% of the yolk of the embryonated hen''s egg with large volumes of diverse but chemically defined solutions. By using an electrosurgical unit and a polyethylene tunnel, the procedure was performed on eggs from days 3 through 7 with greater than 90% surgical success and viability for the short term. More than 50% of the eggs replaced showed viability for 2 weeks, and a significant proportion went full term. ³²PO₄ and amino acids (³H and ¹⁴C) added to the replaced eggs were incorporated into the macromolecules of the embryo and yolk sac as well as into parasitic rickettsiae cultivated in the replaced eggs. The incorporated ³²PO₄ was shown to be assimilated into a variety of biochemical species.     

Isotopic Labeling of Embryo, Yolk Sac, and Intracellular Parasites of the Embryonated Hen''s Egg by Yolk Replacement

A technique is described which allows the replacement of 50% of the yolk of the embryonated hen's egg with large volumes of diverse but chemically defined solutions. By using an electrosurgical unit and a polyethylene tunnel, the procedure was performed on eggs from days 3 through 7 with greater than 90% surgical success and viability for the short term. More than 50% of the eggs replaced showed viability for 2 weeks, and a significant proportion went full term. ³²PO₄ and amino acids (³H and ¹⁴C) added to the replaced eggs were incorporated into the macromolecules of the embryo and yolk sac as well as into parasitic rickettsiae cultivated in the replaced eggs. The incorporated ³²PO₄ was shown to be assimilated into a variety of biochemical species.     

The rat visceral yolk sac internalizes maternal transferrin and secretes hydrolyzed products towards the fetus

The uptake of transferrin by the rat visceral yolk sac membranes, and the fate of this protein, were measured in a two-chambered system which allowed access to both surfaces of these membranes, i.e. that facing the maternal compartment and that facing the fetal compartment. 125I-labeled transferrin was internalized by the maternal surface of the visceral yolk sac but not by the fetal surface. Following internalization, this transferrin was degraded and the amino acids were secreted exclusively towards the fetal compartment. Transcytosis of intact transferrin was not detected in either direction. These results suggest that transport across the rat visceral yolk sac bound to maternally derived transferrin is not a major mechanism of iron transport in vivo. These results support a role for the visceral yolk sac in fetal metabolism, or supplying the fetus with amino acids derived from degradation of specific maternal plasma proteins, in this case, transferrin.     

Incubation of rigid-shelled turtle eggs: do hydric conditions matter?

Rigid-shelled eggs of the broad-shelled river turtle Chelodina expansa were incubated at 28 degrees C in wet (-100 kPa), intermediate (-350 kPa) and dry (-750 kPa) conditions. Incubation period was influenced by clutch of origin, but was independent of incubation water potential. Rates of water gained from the environment and pre-pipping egg mass were influenced by incubation water potential -- eggs incubating at higher (less negative) water potentials absorbing more water from their environment. Hatchlings from wet conditions had greater mass but a smaller amount of residual yolk than hatchlings from dry conditions and it is suggested that the amount of yolk converted to tissue is influenced by the amount of water absorbed by the egg during incubation. Water content of yolk-free hatchlings from the -100-kPa treatment was greater than those from the -350-kPa and -750-kPa treatments, but the water content of residual yolks was similar across all hydric conditions.     

Metabolism of cholesterol in the tissues and blood of the chick embryo

Three artificially inseminated laying White Leghorn hens were given 35-50 micro c of cholesterol-4-(14)C intravenously. Their subsequently produced eggs contained cholesterol-(14)C-labeled yolks. Some of the fertilized eggs were analyzed for cholesterol content and radioactivity. Other eggs were incubated until hatching. The specific activity of the cholesterol contained in the serum and tissues of newly hatched chicks was determined and compared with that of yolk sac, which was taken as representative of egg yolk cholesterol before its metabolic transfer into the chick embryo. The specific activities of cholesterol in intestine, liver, serum, heart, and skeletal muscle and the whole chick were 95-98% of that in yolk sac, but that of brain cholesterol was only 11% of this value. These results indicate that whereas most of the cholesterol in the chick originated from the egg yolk, cholesterol biosynthesis was active in the brain and provided about 90% of its cholestero content. Newly hatched chicks were found to be hyperlipemic compared with older chicks and had fatty livers with a high cholesterol content. Desmosterol was found in 9- and 15-day old chick embryos but not in the newly hatched chicks, in which the only sterol was cholesterol.     

Water quality during egg incubation influences yolk-sac fry sodium kinetics in Atlantic salmon, Salmo salar L.: a possible mechanism of adaptation to acid waters

Atlantic salmon ( Salmo salar L.) fry hatched from eggs transferred from high-Na to low-Na water during the eyed stage of development had a significantly higher V_max and lower K_m (P <0.01) of the sodium uptake mechanism than fry hatched from eggs incubated entirely in low-Na or high-Na water.
Fry hatched from eggs transferred to acid, high aluminium water during the eyed stage of development had a similar V_max and K_m to fry hatched from eggs incubated entirely in high- or low-Na water. Eggs incubated continuously in acid, high aluminium (low-Na) water produced fry with significantly lower K_m and V_max values than fry hatched from eggs incubated continuously in low-Na water. Eggs and fry in acid, high aluminium water continually lost sodium and mortality was 100% at 5 5 M O degree-days (2–3 weeks after hatching).
The results are discussed with respect to the influence of perivitelline fluid ion activities in eggs in acid, high aluminium water on the kinetic characteristics of sodium uptake in yolk-sac fry. A possible mechanism for the long-term adaptation of teleosts in acidified natural waters is also proposed.     

Survival and emergence pattern of sea trout fry in substrata of different compositions

The emergence of the sea trout Salmo trutta fry of Gotland, Sweden, was studied in the laboratory. The main limiting factor for survival up to hatching was the interstitial oxygen concentration, which depends on the gravel permeability, therefore on the substratum granulometry. Fry emergence was possible in substrata in which the geometric mean particle diameter ( d _g) was >15 mm and in substrata in which d _g was <6 mm. In the substrata of low d _g, emergence took place earlier than in the other ones. As a consequence the fry were smaller and their yolk sac only partially resorbed. In an unfavourable environment, fry started to emerge as soon as they reacted positively to light, about 2 weeks after hatching.