Neurotoxicity of 2,4-dichlorophenoxyacetic butyl ester in chick embryos

Fertilized hens' eggs were externally treated, before starting incubation, with a single dose of 2,4-Dichlorophenoxyacetic butyl ester (2,4-D b.e., 3.1 mg/egg). Chicks at different developmental stages were examined, extending from 10 day embryos to one day after hatching. Previously, we demonstrated that 2,4-D b.e. produces hypomyelination in chicks born from treated eggs. In search of the causes of this hypomyelination, myelin markers such as sulphatides, cerebrosides and 2 3-cyclic nucleotide 3-phosphohydrolase (CNPase) activity, as well as protein and nucleic acid contents were determined in the embryonic brains. We have shown in the present study that the chemical alterations occurred even before the period of active myelination, since myelin appears in chicken brain stem and cerebrum approximately after 17 days of incubation, and most of the chemical parameters studied are diminished before that time. The DNA content in brain of treated group is increased from the 14^th embryonic day (with a transient diminution at 12^th day) to the first hatching day, when compared to the control, suggesting a proliferation of glial cells, possibly oligodendrocytes.     

STUDIES OF THE MECHANISM OF DEMYELINATION REGIONAL DIFFERENCES IN MYELIN STABILITY IN VITRO1

—Incubation of slices of rat central nervous system in Krebs-Ringer bicarbonate buffer produced a lipoprotein fraction which floated on 10·5% sucrose after homogenization of the slices and centrifugation. This fraction was not found after homogenization and centrifugation of fresh tissue and appeared to depend upon incubation. The amount of the light fraction increased in the following order per 100-mg slice: cerebrum < thalamic area < cerebellum < brain stem < spinal cord. The lipid composition of this fraction was similar to that of myelin, but contained a lower protein content compared to myelin of the corresponding area. This fraction was termed ‘dissociated myelin’. Upon incubation of slices a portion of the basic protein was lost from myelin subsequently isolated, and the dissociated fraction was slightly enriched in basic protein. The distribution of myelin protein among the characteristic three groups (basic, proteolipid and high mol. wt.) was quite different in myelin from spinal cord compared to that from other CNS area. Spinal cord myelin contained about 17% protein compared to about 23% in cerebrum, with brain stem myelin intermediate (19%), and the difference appeared to be due to lesser amounts of proteolipid in the caudal areas. The amount of dissociation after incubation was about 3–5 per cent of the total myelin in the cerebral cortex, 10 per cent in the thalamic area, 20 per cent in cerebellum, 35 per cent in the brain stem, and around 45 per cent in spinal cord. The smaller amount of proteolipid protein in spinal cord myelin may result in a deficiency of cohesive forces holding lipids and proteins together, thus causing greater instability and dissociation. Myelin dissociation increased with time of incubation up to 3 h, was augmented by Ca²⁺, and was substantial at pH 11, reaching a peak at pH 7, then decreased in the acid range. A similar fraction has been isolated previously from fresh CNS tissue made edematous by chronic treatment of rats with triethyl tin. The possible relationship of swelling in the disease process and myelin dissociation are discussed.     

CHANGES OCCURRING IN THE CHEMICAL COMPOSITION OF THE CENTRAL NERVOUS SYSTEM DURING FOETAL AND POST-NATAL DEVELOPMENT OF THE SHEEP

(1) The chemical composition of the CNS (separated into cerebrum, cerebellum, brain stem and spinal cord) was determined in sheep during foetal and post-natal development and in adults. (2) The spinal cord differed from the remainder of the CNS in growing more after the period studied (50-day-old foetuses to 5-week-old lambs) than before it. This was largely attributable to lipid accumulation. (3) Chemical growth (accumulation of DNA, protein and lipid) proceeded linearly in spinal cord, logarithmically in cerebrum and cerebellum while in brain stem growth was described by a sigmoid function. (4) Fat-free dry matter, protein, total lipid, cholesterol and phospholipid concentrations increased progressively in all parts of the CNS but DNA concentrations changed little. In the cerebrum alone there was an increase in DNA concentration during maturation suggesting an increased cell population. Cholesterol was present predominantly in the free form but esters were detected in foetal tissues from 70 up to 120 days gestation. (5) Cerebroside, the characteristic lipid of myelin, increased in concentration soon after 85 days of gestation, up to which point very low values were recorded, the rate varying according to the region of the CNS examined. Rates of increase in total regional cerebroside content were used to identify periods of myelination and the results suggest that there are two periods of peak activity, one about 20 days before birth and the other at 10-20 days after birth. (6) The composition of lipids added during the two phases of myelination and during maturation were characteristically different. In the spinal cord, lipid analyses best reflect changes in myelin composition.     

Effects of feeding high dosages of vitamin E to laying hens on thyroid hormone concentrations of hatching chicks.

Long-term experimental feeding of 20,000 ppm alpha-tocopheryl acetate to laying hens caused a significant (P < 0.05) decrease in hatching rates as compared to the control group, which was fed a diet containing 19 ppm alpha-tocopherol. When the thyroid hormones in the developing chicks were checked on incubation days 16, 19, 21, and 22, the following results were ascertained: During the latter part of incubation, increases in plasma concentrations of thyroxine and triiodothyronine were observed. No significant differences in hormone concentrations (P > 0.05) between the control and the treatment group were observed during incubation days 16, 19, and 22. However, on the day of hatching (day 21 of incubation) significantly lower (P < 0.05) triiodothyronine concentrations in chick embryos of piped eggs were found in the treatment group. Moreover, thyroxine concentrations in non-piped eggs and in hatched chicks were found to be significantly higher as compared to the control group. Given these results, one concludes that extremely high dosages of vitamin E may affect thyroid hormone concentrations of hatching chicks, and therefore, the chicks might be inhibited in pipping the egg shell. Hypothetically, the hepatic enzyme 5'-monodeiodinase is involved in the mechanism of inhibition.     

Myelin Lipids in the Developing Cerebrum, Cerebellum, and Brain Stem of Normal and Undernourished Children

Abstract: The developmental lipid profiles in the human cerebrum, cerebellum and brain stem are presented, with special reference to galactolipids as myelin markers to trace myelination in the three main parts of the human CNS. A group of undernourished children were also studied to test the vulnerability of myelinogenesis in the different regions of the human brain. Myelination was well advanced in the brain stem with regard to the other brain regions, a fact reflected in the much higher concentration of myelin lipids in the brain stem of the human foetus of 26 weeks of gestational age. The cerebrum, on the other hand, had the lowest galactolipid concentration during the prenatal period, galactolipid levels in the cerebellum being four times higher. From just before the end of gestation the accretion of galactolipids accelerated enormously in the cerebrum, whereas it slowed down considerably in the cerebellum. Consequently, in relation to prenatal levels galactolipids increased most rapidly in the cerebrum, followed by the cerebellum and finally by the brain stem. These regional differences were in clear contrast to data from the rat, as was the finding that only the cerebrum of undernourished children had a galactolipid concentration significantly decreased with respect to normal values. A relationship between the different myelination patterns in the human and the rat and the distinct vulnerability of myelinogenesis in the two species is suggested.     

Observations on the breeding biology and behaviour of the Lesser Jacana,Microparra capensis

Two Lesser Jacana nests were found in Hwange National Park, Zimbabwe and were observed over a period of four months beginning in March 2000. Both sexes were involved in preparing the breeding platform, incubation, and caring for the chicks, which were not carried by the adults. Both nests had three eggs. The incubation period for one clutch of eggs was not less than 19 days. The chicks all hatched on the same day and remained in the vicinity ofthe nest for the first few days where they were brooded by an adult. Initially the adults brought food to the chicks but the chicks started feeding themselves when they moved away from the nest. Ten days after hatching the chicks had doubled in size and were walking confidently with the attending adult some distance from the nest. First flight was seen at 32 days old, and the chicks appeared to be independent 63 days after hatching.     

Incubation prior to clutch completion accelerates embryonic development and so hatch date for eggs laid earlier in a clutch in the great tit Parus major

The ability of parents to respond to changes in food supply within a season will have a large effect on fitness through the number and quality of chicks fledged. Great tits, Parus major, attempt to synchronise their production of chicks with a seasonal food peak, but when food supply fails, hatching asynchrony of chicks provides a mechanism by which some young can be fledged because more developed chicks outcompete their less developed siblings for the reduced parental food supply. We tested whether female great tits can potentially control the degree of hatching asynchrony by using incubation before clutch completion, so that early laid eggs develop faster and hatch sooner. The temperature of an artificial egg placed in 29 nests during the laying period was measured with data loggers, and nocturnal incubation of eggs similar to incubation post clutch completion was recorded in all nests. We then demonstrated that eggs removed from the nest for 72 hour periods prior to clutch completion hatched later than eggs remaining in the nest for the entirety of the laying period. Our results show that variable pre clutch completion incubation (which was mostly nocturnal) can lead to faster embryo development and earlier hatching, so potentially providing a mechanism for adaptive female control of degree of hatching asynchrony.     

Hosts improve the reliability of chick recognition by delaying the hatching of brood parasitic eggs

The reliability of information that animals use to make decisions has fitness consequences. Accordingly, selection should favor the evolution of strategies that enhance the reliability of information used in learning and decision making. For example, hosts of avian brood parasites should be selected to increase the reliability of the information they use to learn to recognize their own eggs and chicks. The American coot (Fulica americana), a conspecific brood parasite, uses cues learned from the first-hatched chicks of each brood to recognize and reject parasitic chicks. However, if parasitic eggs are among the first to hatch, recognition cues are confounded and parents then fail to distinguish parasitic chicks from their own chicks. Therefore, hosts could ensure correct chick recognition by delaying parasitic eggs from hatching until after the first host eggs. Here we demonstrate that discriminatory incubation, whereby coots specifically delay the hatching of parasitic eggs, improves the reliability of parasitic chick recognition. In effect, coots gain fitness benefits by enhancing the reliability of information they later use for learning. Our study shows that a positive interaction between two host adaptations in coots--egg recognition and chick recognition--increases the overall effectiveness of host defense.     

Hatching of Daphnia sexual eggs. II. The effect of age and a second stimulus

• 1 We examined the effect of age on the hatching response of Daphnia magna sexual eggs of specific families. For old eggs (>2 years), hatching characteristics were compared at two storage temperatures (4°C and 20°C). Also, the hatching response after a second dark incubation and subsequent incubation under conditions favourable for hatching was compared with that after the first stimulus.
• 2 Daphnia sexual eggs were found to remain viable for several (at least 4.5) years. The effect of age on the hatching rate was family dependent. At least in some families, hatching rate was higher for old (>2 years) than for young (<5 months) eggs. Low temperature (4°C) during dark incubation resulted in a higher hatching rate compared with incubation at 20°C.
• 3 The application of a second hatching stimulus resulted in a renewed hatching response. The overall hatching rate after the second stimulus was, however, lower than that of the first stimulus.
• 4 More than 80% of the hatchlings of young eggs appeared on Day 3 or 4, with minor between-family differences in time distribution of hatching. The timing of the response to hatching stimuli was more variable in old than in young eggs, with the average time at hatching being 6.4 instead of 4.0 days. The response to the application of hatching stimuli was also slower after the second stimulus compared with the first stimulus.

     

The breeding biology of the Dalmatian Pelican Pelecanus crispus

The breeding biology of the Dalmatian Pelican Pelecanus crispus , a vulnerable bird species in the Palaearctic, has been studied for 7 years at Lake Mikri Prespa, a continental wetland and Tsoukalio lagoon, a coastal wetland within the Amvrakikos Gulf. The two colonies showed stability, with fluctuations in the number of breeding attempts each year varying by about 30% over a period of 7 years. The date of arrival at the breeding sites and the date of laying of the first egg varied between mid-February and mid-March. The egglaying period (including renesting) varied between 18 and 114 days between years. The annual average clutch size varied between 1.60 and 1.94 eggs per nest. The incubation period averaged 31.4 ± 1.4 days. The overall hatching success varied from 36% to 70%. Egg losses were mainly due to nest abandonment, infertile eggs and eggs rolled from the nests. The Dalmatian Pelican chicks took 11–12 weeks to fledge. Nestling mortality was very low in contrast to other species of pelican. Such a low nestling mortality explains why the breeding success each year is largely explained by the hatching success.     

Preservation of rainbow trout (Oncorhynchus mykiss) eyed eggs using a perfluorochemical as an oxygen carrier

The objective of this study was to maintain the viability of chilled rainbow trout (Oncorhynchus mykiss) eyed eggs during storage using oxygenated perfluorochemical (PFC). Three trials were conducted using eggs at 161, 180 or 217 degree days (days from fertilization x incubation temperature in degrees C). A separate trial was conducted for 147 degree day eggs that were not at the eyed stage. For each trial, eggs were stored in a moisture-saturated atmosphere at 1 degrees C in PFC, water, and 1:1 combinations of PFC and PBS, PFC and 0.3 M glucose, PFC and mineral oil, or PFC and water. The PFC was oxygenated before each trial and all media were oxygenated at weekly intervals during the storage period. Eggs from each trial were also incubated without storage to provide Day 0 results. After 3 and 5 weeks of storage, eggs from each medium were incubated at 10 degrees C until hatch. Hatching percentage was expressed as a percentage of Day 0 results. The percentage of normal alevins that hatched was also determined. There were interactions (P < 0.01) between stage of development and treatment for hatching percentage after 3 and 5 weeks of storage. After 3 weeks of storage, eggs stored at 161, 180, or 217 degree days without PFC had hatching rates of 0-14.3% but eggs stored in any medium with PFC had hatching percentages from 75.1 to 106.4% of Day 0 values. After 5 weeks of storage, eggs stored at 161 degree days in PFC plus PBS or PFC plus water, and eggs stored at 217 degree days in PFC or PFC plus water, had higher (P < 0.05) hatching percentages than eggs stored in any of the other media. Eggs stored at 161 degree days for 5 weeks in PFC and water had a higher (P < 0.05) percentage of normal alevins hatching than eggs stored in PFC and PBS. Because of their early developmental stage, eggs stored at 147 degree days had low hatching percentages, except eggs stored for 3 weeks in PFC or PFC plus PBS. Chilling eyed eggs of rainbow trout to 1 degrees C and storing them in water with PFC as an oxygen carrier can preserve their viability for 5 weeks.     

Developmental stage affects eggshell-breaking strength in two ground-nesting birds: the partridge (Alectoris rufa) and the quail (Coturnix japonica)

We examined the relationship between embryo development and egg hardness in two ground nesting bird species, the red-legged partridge (Alectoris rufa; n = 165 eggs) and the quail (Coturnix japonica; n = 148 eggs). For both species, we observed a strong effect of developmental stage on egg hardness. Eggs near hatching were significantly weaker than unincubated eggs (partridge: 18 and 23 N, respectively, and the quail 7 and 10 N, respectively). We additionally explored the effect of incubation on egg hardness in a control sample of non-fertilised quail eggs (i.e., without embryo development). The control eggs maintained in the incubator for the full incubation time (17 days) were significantly harder (7-9 N) than eggs containing fully developed chicks (5-7 N). Thus, the incubation conditions of high temperature and humidity alone seem not to have a significant effect on egg hardness, and support the important effect of calcium uptake.     

CONGENITAL HYPOMYELINOGENESIS (BORDER DISEASE) OF LAMBS: POSTNATAL NEUROCHEMICAL RECOVERY IN THE CENTRAL NERVOUS SYSTEM

Abstract— In a neurochemical study of experimental Border Disease in lambs it was found that the fresh weights of four parts of the CNS (cerebrum, cerebellum, brain stem and spinal cord) from clinically affected lambs were significantly smaller than those of controls at birth but by 20 weeks of age the cerebrum, cerebellum and brain stem had reached near normal weights. The spinal cord was still considerably smaller, however. Clinical symptoms of the disease (muscular spasms and'hairy'birthcoat) had disappeared during this period, accompanied by a regression in the neurochemical abnormalities seen at birth. Thus the deficiency of myelin lipids was partially made up by the rapid deposition of cerebrosides and by 20 weeks differences in the fatty acid composition of this lipid fraction were no longer apparent. Myelin degeneration as indicated by the presence of elevated levels of esterified cholesterol was apparently absent at 20 weeks of age and this was parallelled by a fall in the level of'anti-myelin'antibodies in the sera of affected lambs. The altered distribution of copper in spinal cord myelin seen at birth had also become nearly normal at the end of the period.     

Embryo survival in heifers after transfer of an egg to the uterine horn contralateral to the corpus luteum and the effect of treatments with progesterone or hCG on pregnancy rates.

Day 7 eggs (oestrus = Day 0) were surgically transferred singly to the uterine horn contralateral to the CL and heifers were either slaughtered on Day 24-26 (Group 1) or palpated per rectum on Day 42 (Group 2) to determine the presence of a developing conceptus and a maintained corpus luteum. There was a fall in pregnancy rate of the control heifers in Groups 1a (6/10) and 2a (2/10) (P = 0.085). Pregnancy rate was unaffected by progesterone treatments from Day 13 to the day before slaughter in Group 1b (8/10), or to Day 23 in Group 2b (2/10). Two of the conceptuses recovered in Group 1b were developing in the presence of regressed corpora lutea. There was, however, an increase in pregnancy rate in Group 2d (6/10) compared with Group 2a (2/10) when heifers were treated with hCG from Days 13 to 35 (P = 0.085). The results indicated that embryonic death is occurring after Day 24 and suggest that a change in the nature of the embryonic signal may be occurring at this time.     

When should Common Cuckoos Cuculus canorus lay their eggs in host nests?

Capsule Brood parasitic Common Cuckoo Cuculus canorus chicks hatch earlier than the nestlings of their Great Reed Warbler Acrocephalus arundinaceus hosts, but hatching priority is less consistent when Cuckoo eggs are laid after the onset of host incubation.

Aim To reveal by field observations what the optimal stage is for Cuckoos to lay their eggs in relation to the host laying cycle to ensure prior hatching of the parasitic chicks.

Methods We monitored the hatching of Cuckoo chicks in relation to the hosts’ laying stage at which the Cuckoo eggs appeared and also monitored host incubation behaviour.

Results Great Reed Warblers incubated more on day 5 after the host's onset of laying relative to day 3. All Cuckoo eggs hatched earlier than hosts when they were laid prior to the onset of host incubation (day 4). Cuckoo eggs also maintained hatching priority in about 2/3 of the nests when laid on days 5–6.

Conclusions Most Cuckoo eggs are laid prior to the onset of host incubation and this, together with other adaptive mechanisms, ensures the prior hatching of Cuckoo eggs. Cuckoo eggs laid after the onset of incubation lose the advantage of prior hatching in approximately 30% of nests.     

Brood sex ratio in the Kentish plover

How and why do the mating opportunities of males and femalesdiffer in natural population of animals? Previously we showedthat females have higher mating opportunities than males inthe Kentish plover Charadrius alexandrinus. Both parents incubatethe eggs, and males provide more brood care than females; thusit is not obvious why the females find new mates sooner thanthe males. In this study we investigated whether the sex-biasedmating opportunities stem from biased offspring sex ratios.We determined the sex of newly hatched, precocial chicks usingCHD gene markers. Among fully sexed broods, 0.461 ± 0.024(SE) of chicks (454 chicks in 158 broods) were male, and thissex ratio was not significantly different from unity. The proportionof males at hatching decreased significantly over the breedingseason, which occurred consistently in all 3 years of the study.Large chicks were more likely to be males than females. Neitherparental age nor body size of male and female parents was relatedto brood sex ratio. We also sexed a number of chicks that werecaught after they left their nest (range of estimated ages 0–17days) and found that the proportion of males increased withbrood age. This relationship remained highly significant whencontrolling statistically for hatching date. As brood size decreaseddue to mortality after the chicks left their nest, these resultssuggest that the mortality of daughters was higher than thatof the sons shortly after hatching. Taken together, our resultsshow that the female-biased mating opportunities in the Kentishplover are not due to biased brood sex ratio at hatching but,at least in part, are due to female-biased chick mortality soonafter hatching.     

Corticosterone stimulates hatching of late-term tree lizard embryos

The regulation of hatching in oviparous animals is important for successful reproduction and survival, but is poorly understood. We unexpectedly found that RU-486, a progesterone and glucocorticoid antagonist, interferes with hatching of viable tree lizard (Urosaurus ornatus) embryos in a dose-dependent manner and hypothesized that embryonic glucocorticoids regulate hatching. To test this hypothesis, we treated eggs with corticosterone (CORT) or vehicle on Day 30 (85%) of incubation, left other eggs untreated, and observed relative hatch order and hatch time. In one study, the CORT egg hatched first in 9 of 11 clutches. In a second study, the CORT egg hatched first in 9 of 12 clutches, before vehicle-treated eggs in 10 of 12 clutches, and before untreated eggs in 7 of 9 clutches. On average, CORT eggs hatched 18.2 h before vehicle-treated eggs and 11.6 h before untreated eggs. Thus, CORT accelerates hatching of near-term embryos and RU-486 appears to block this effect. CORT may mobilize energy substrates that fuel hatching and/or accelerate lung development, and may provide a mechanism by which stressed embryos escape environmental stressors.     

Pre- and post-hatching effects of corticosterone treatment on behavior of the domestic chick

We investigated the effect of 60 ìg of corticosterone administered to domestic chicks either before or after hatching on the behavioral response to isolation in a novel arena and performance in a task involving the simultaneous identification of food and detection of a predator (overhead silhouette of a hawk moving overhead). Following release into a novel arena, chicks treated with corticosterone at 18 days of incubation emitted more distress vocalizations. In contrast, no difference in the number of vocalizations was found between chicks treated with corticosterone at day 1 post-hatching and controls. Behavior in the home cages was generally similar across treatments, though chicks treated with corticosterone at 18 days of incubation slept more than control chicks. While searching for grain against a background of pebbles, chicks treated with corticosterone at embryonic day 18, but not chicks treated on day 1 post-hatching, took longer to detect the overhead image of a predator than did controls. Corticosterone treatment at both ages increased the rate of pecking at grains and pebbles. Our findings support work on other birds indicating that corticosterone treatment during incubation influences stress reactivity. The impairment in predator detection in chicks treated with corticosterone on day 18 of incubation appears to be caused by the known effects of corticosterone treatment at this age in preventing the development of lateralization of the thalamofugal visual projections. This further supports the hypothesis that brain lateralization provides an advantage in performing more than one task simultaneously.     

Hand-rearing,growth, and development of the red bird of paradise (Paradisaea rubra) at the New York Zoological Park

Growth and development of six hand-reared red bird of paradise chicks was documented at the New York Zoological Park from March 1988 to May 1989. A total of 16 eggs were laid, of which 10 were fertile. Clutches consisted of two eggs and the female left the next infrequently during incubation. Two chicks left in the nest were apparently victims of parental abuse. Eggs were subsequently removed from the nest after 10–14 days, candled, and if fertile, were artificially incubated. The average incubation period was 16.6 days. Newly hatched chicks were without down and their eyes remained closed until approximately 6 days of age. Hand-reared chicks were maintained in Air Shield Infant Isolettes. The weight of newly hatched chicks was about 8 g, and the weight typically doubled during the first week. Ratios of food intake to body weight were highest between day 4 and 10. Pin feathers were visible on the wings after 4 days, and after 3 weeks, the chicks were fully feathered. Analysis of the diet revealed acceptable levels of iron, but vitamin A and E levels were higher than recommended for poultry chicks. This paper documents the first successful hand-rearing of any species of birds of paradise from hatching to fledging.