The effect of dietary amino acid composition on egg production in blue tits

Most studies on the interaction between food supply and reproduction in animals have assumed that energy is likely to be the factor limiting egg number and/or size. In this paper, we investigate whether dietary protein proximately constrains egg production in birds. We provisioned breeding blue tits with two food supplements that differed only in the concentration of five essential amino acids. Birds receiving a supplementary diet containing an amino acid balance close to that required for egg protein formation laid significantly larger clutches (18% greater) than control birds, whereas birds receiving an otherwise identical supplementary diet but without a favourable amino acid balance did not increase egg production. To our knowledge, this is the first demonstration that dietary amino acid composition may limit egg production in free-living birds.     

Why are incubation periods longer in the tropics? A common-garden experiment with house wrens reveals it is all in the egg

Incubation periods of Neotropical birds are often longer than those of related species at temperate latitudes. We conducted a common-garden experiment to test the hypothesis that longer tropical incubation periods result from longer embryo development times rather than from different patterns of parental incubation behavior. House wrens, one of few species whose geographic range includes tropical equatorial and temperate high latitudes, have incubation periods averaging 1.2 days longer at tropical latitudes. We incubated eggs of house wrens in Illinois and Panama under identical conditions in mechanical incubators. Even after factoring out differences in egg size, tropical house wrens still required 1.33 days longer, on average, to hatch. We conclude that parental attendance patterns do not account for latitudinal differences in incubation period but that some other as yet unmeasured factor intrinsic to the egg or embryo, or both, extends development time in the tropics.     

Correlated evolutionary divergence of egg size and a mitochondrial protein across the Isthmus of Panama

An explicit assumption of studies that employ a mitochondrial DNA (mtDNA) molecular clock is that mtDNA evolves independently of morphology. Here we report a very strong correlation between egg size divergence and cytochrome c oxidase-1 (CO1) amino acid sequence divergence among sister species of bivalve molluscs separated by the Central American Isthmus (i.e., "geminate" species). Analyses of the molecular data reveal that CO1 sequences likely did not diverge as a function of time or evolve in response to positive natural selection. Given that an excess of CO1 amino acid polymorphism exists within species (as expected if most mutations are only slightly deleterious), a third hypothesis is that reductions in effective population size could simultaneously increase the fixation rate of nearly neutral mtDNA polymorphisms and in some way also facilitate egg size evolution. The remarkable strength of the relationship between egg size and CO1 amino acid sequence demonstrates that, even in the absence of an obvious functional relationship or clock-like evolution, the amounts of molecular and morphological change can be tightly correlated, and therefore may reflect common processes. Accordingly, the assumption that the evolutionary divergence of molecules and morphology are independent must always be carefully examined.     

Physiological and Ecological Correlates of Prolonged Incubation in Sea Birds

Sea birds with long incubation periods are identified, togetherwith the features of their incubation physiology which distinguishthem from birds in general. Most sea birds with prolonged incubationare members of the order Procellariformes. The majority of Pelecaniformesand Charadniformes with long incubation periods are tropicalspecies. The total amount of water lost from the egg duringincubation is a similar fraction of the initial egg weight insea birds with prolonged incubation as in other birds. The oxygenconsumption of the newly hatched chick is similarly relatedto the chick weight, regardless of the duration of incubation.Within the constraints imposed by these similarities, sea birdswith prolonged incubation display a number of adaptations. Thedaily rate of water loss from the egg, the water vapor conductanceof the egg shell, and the total functional pore area of theegg are all relatively low in sea birds with prolonged incubation.The eggs of sea birds with long incubation times are large inrelation to the size of the adult bird in the two species thathave been studied, the high energy content of the egg is paralleledby the greater total amount of oxygen consumed during incubation.However, the growth of the embryo is relatively slow in at leastone sea bird with a long incubation time so that prolonged incubationprobably entails a comparatively high allocation of energy resourcesto maintenance requirements. In some species with prolongedincubation, the interval between pipping and hatching is alsolong and it appears to be a period of great physiological importance.Ecologically prolonged incubation is associated with eitherpelagic feeding habits or a tropical environment, both factorsmay be related to food supply.     

Cloning and characterization of vitellogenin cDNA from the common Japanese conger (Conger myriaster) and vitellogenin gene expression during ovarian development

The major yolk protein precursor, vitellogenin (VTG) was detected in plasma from vitellogenic females and estradiol-17β (E₂)-treated immature females, but not in males and immature females by Western blotting in common Japanese conger Conger myriaster. Its molecular mass was approximately 180 kDa under denaturing and reducing conditions. The common Japanese conger VTG cDNA was cloned from the liver of vitellogenic female. It contains 5110 nucleotides including an open reading frame that encodes 1663 amino acids. The deduced amino acid sequence of the common Japanese conger VTG shares 80% identity with that of eel Anguilla japonica VTG-1, and 45–55%, 32–34% and 27–29% identity with the deduced amino acid sequences of other fish, amphibian and avian VTG with polyserin domain, respectively. In female common Japanese conger, VTG gene was highly expressed in the liver of this species similar with other oviparous vertebrates. The expression levels of VTG gene in the liver increased from the oil droplet stage to the tertiary yolk globule stage and were maintained until the migratory nucleus stage.     

Spinal seizures evoked by sudden cooling of amphibian isolated spinal cords: involvement of excitatory amino acids.

Sudden cooling of the isolated spinal cord of frogs results in characteristic seizure-like activity in the hind legs. In the present investigation, these spinal seizures induced by sudden cooling (SSSC) were studied to determine whether excitatory amino acids (EAAs) are involved in the mediation of this activity. The nonspecific EAA antagonist, L-glutamic acid diethyl ester and cis-2,3-piperidine dicarboxylic acid inhibited the clonic and tonic phase of SSSC after intralymphatic or intrathecal administration. The antagonist gamma-D-glutamylaminomethylsulfonic acid and gamma-D-glutamyltaurine also suppressed both phases after intrathecal injections. The NMDA receptor antagonist DL-2-amino-5-phosphonovaleric acid, DL-2-amino-7-phosphonoheptanoic acid, and 3-(2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid were effective inhibitors of the tonic phase and actually prolonged the duration of the clonic phase, an effect similar to that observed after low doses of gamma-D-glutamylglycine. SSSC were resistant to spinal perfusion to tetrodotoxin (1 microM). The concentrations of glutamate, aspartate, and glycine were increased in the Ringer's solution surrounding rapidly cooled spinal cord slices, but only in cords from species that elicited some magnitude of SSSC, not in cords from species resistant to induction of SSSC. Our data support the hypothesis that EAAs play a role in SSSC via activation of quisqualate receptors.     

Egg yolk triglycerides during development of the domestic chicken

The triglycerides isolated from egg yolk lipids of eggs at various stages of incubation were fractionated according to the degree of unsaturation by argentation chromatography, and individual fractions were analyzed for their fatty acids by gas liquid chromatography. The proportions of the various fractions were constant during development. Fatty acid composition of the fractions were constant also. Fractions with one saturated fatty acid and two monoenoic fatty acids (SM₂) constituted over 40% of the total triglyceride. Palmitic acid constituted over 30%, and oleic acid over 45% of the fatty acid of total triglycerides. It is suggested that during development of thick embryo there is no selective utilization of the egg yolk triglycerides.     

AMPA receptor expression in mouse testis and spermatogonial GC-1 cells: A study on its regulation by excitatory amino acids

Excitatory amino acids (EAAs) are found present in the nervous and reproductive systems of animals. Numerous studies have demonstrated a regulatory role for Glutamate (Glu), d -aspartate ( d -Asp) and N-methyl- d -aspartate (NMDA) in the control of spermatogenesis. EAAs are able to stimulate the Glutamate receptors, including the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR). Here in, we assess expression of the main AMPAR subunits, GluA1 and GluA2/3, in the mouse testis and in spermatogonial GC-1 cells. The results showed that both GluA1 and GluA2/3 were localized in mouse testis prevalently in spermatogonia. The subunit GluA2/3 was more highly expressed compared with GluA1 in both the testis and the GC-1 cells. Subsequently, GC-1 cells were incubated with medium containing l -Glu, d -Glu, d -Asp or NMDA to determine GluA1 and GluA2/3 expressions. At 30 minutes and 2 hours of incubation, EAA-treated GC-1 cells showed significantly higher expression levels of both GluA1 and GluA2/3. Furthermore, p-extracellular signal-regulated kinase (ERK), p-Akt, proliferating cell nuclear antigen (PCNA), and Aurora B expressions were assayed in l -Glu-, d -Glu-, and NMDA-treated GC-1 cells. At 30 minutes and 2 hours of incubation, treated GC-1 cells showed significantly higher expression levels of p-ERK and p-Akt. A consequent increase of PCNA and Aurora B expressions was induced by l -Glu and NMDA, but not by d -Glu. Our study demonstrates a direct effect of the EAAs on spermatogonial activity. In addition, the increased protein expression levels of GluA1 and GluA2/3 in EAA-treated GC-1 cells suggest that EAAs could activate ERK and Akt pathways through the AMPAR. Finally, the increased PCNA and Aurora B levels may imply an enhanced proliferative activity.     

The evolution of egg shape in birds: selection during the incubation period

A recent broad comparative study suggested that factors during egg formation – in particular ‘flight efficiency’, which explained only 4% of the interspecific variation – are the main forces of selection on the evolution of egg shape in birds. As an alternative, we tested whether selection during the incubation period might also influence egg shape in two taxa with a wide range of egg shapes, the alcids (Alcidae) and the penguins (Spheniscidae). To do this, we analysed data from 30 species of these two distantly related but ecologically similar bird families with egg shapes ranging from nearly spherical to the most pyriform eggs found in birds. The shape of pyriform eggs, in particular, has previously proven difficult to quantify. Using three egg‐shape indices – pointedness, polar‐asymmetry and elongation – that accurately describe the shapes of all birds’ eggs, we examined the effects of egg size, chick developmental mode, clutch size and incubation site on egg shape. Linear models that include only these factors explained 70–85% of the variation in these egg‐shape indices, with incubation site consistently explaining > 60% of the variation in shape. The five species of alcids and penguins that produce the most pyriform eggs all incubate in an upright posture on flat or sloping substrates, whereas species that incubate in a cup nest have more spherical eggs. We suggest that breeding sites and incubation posture influence the ability of parents to manipulate egg position, and thus selection acting during incubation may influence egg‐shape variation across birds as a whole.     

Rapid modulation of the n-3 docosahexaenoic acid levels in the brain and retina of the newly hatched chick

The newly hatched chick obtains its fatty acids almost completely from the lipids of the egg yolk as these are transferred to the developing embryo during its 21-day period of incubation. Since the diet of the laying hen greatly influences the fatty acid composition of the egg lipids, and presumably also the fatty acid composition of the resulting chick, we tested how quickly and to what extent varying the amount of n-3 fatty acids in the diet of the hen would modulate the level of n-3 fatty acids in the brain and retina of the newly hatched chick. White Leghorn hens were fed commercial or semi-purified diets supplemented with 10% fish oil, linseed oil, soy oil, or safflower oil. Eggs, together with the brain, retina, and serum of newly hatched chicks, were then analyzed for fatty acid composition. The fatty acids of egg yolk responded quickly to the hen's diet with most of the change occurring by 4 weeks. There was a linear relationship between the linolenic acid content of the diets and levels of this fatty acid in egg yolk and chick serum. In chicks from hens fed the fish oil diet, the total n-3 fatty acids, including 22:6(n-3), were elevated twofold in the brain and retina and sevenfold in serum relative to commercial diet controls. The safflower oil diet led to a very low n-3 fatty acid content in egg yolks and only 25% of the control n-3 fatty acid content in the brain and retina of chicks.(ABSTRACT TRUNCATED AT 250 WORDS)     

Altitudinal variation in egg retention and rates of embryonic development in oviparous Zootoca vivipara fits predictions from the cold-climate model on the evolution of viviparity

The evolution of reptilian viviparity is favoured, according to the cold‐climate hypothesis, at high latitudes or altitudes, where egg retention would entail thermal benefits for embryogenesis because of maternal thermoregulation. According to this hypothesis, and considering that viviparity would have evolved through a gradual increase in the extent of intrauterine egg retention, highland oviparous populations are expected to exhibit more advanced embryo development at oviposition than lowland populations. We tested for possible differences in the level of egg retention, embryo development time and thermal biology of oviparous Zootoca vivipara near the extreme altitudinal limits of the species distribution in the north of Spain (mean altitude for lowland populations, 235 m asl.; for highland populations, 1895 m asl.). Altitude influenced neither temperature of active lizards in the field nor temperature selected by lizards in a laboratory thermal gradient, and pregnant females selected lower temperatures in the thermal gradient than did males and nonpregnant females across altitudinal levels. Eggs from highland populations contained embryos more developed at the time of oviposition (Dufaure and Hubert's stages 33–35) than eggs of highland populations (stages 30–34) and partly because of this difference incubation time was shorter for highland embryos. When analysed for clutches from both altitudinal extremes at the same embryonic stage at oviposition (stage 33), again incubation time was shorter for highland populations, indicating genuine countergradient variation in developmental rate. Our results indicate that temperature is an environmental factor affecting the geographical distribution of different levels of egg retention in Z. vivipara, as predicted by the cold‐climate hypothesis on the evolution of viviparity.     

Biochemical composition of cuttlefish (Sepia esculenta) eggs during embryonic development

Changes in egg volume, water content, proteins, carbohydrates, lipids, amino acids and fatty acids in cuttlefish (Sepia esculenta) were determined during embryogenesis to understand the nutritional requirements in the early life phase. The egg volume and the water content decreased significantly (P<0.05) during early embryonic development, and then increased abruptly after the beginning of organ differentiation. During embryonic development, protein was the major content in the yolk and the per cent composition varied to a large extent (55.19–78.45%) with carbohydrates (9.86–15.56%) and lipids (1.62–2.97%). Proteins and lipids decreased 23.3% and 0.4%, respectively, in dry weight, while carbohydrates increased 0.46%. Total amino acids and essential amino acids (EAAs) were stable during the early embryonic developmental stage, but decreased significantly until the eggs hatched (P<0.05). The largest utilisation of the yolk protein possibly occurred with respect to EAAs (25.7%) because of a decrease in methionine (70.3%), valine (63.0%) and phenylalanine (58.5%). The most important fatty acids were saturated fatty acid (SFA) C16:0 and polyunsaturated fatty acids (PUFAs) C22:6 and C20:5. Unsaturated fatty acids (UFAs) and SFA decreased at a similar rate during embryonic development (5.69% and 6.15%, respectively). For UFAs, monounsaturated fatty acids were consumed at a greater rate than PUFAs (29.6% and 0.05%, respectively).     

Variation in maternal effects and embryonic development rates among passerine species

Embryonic development rates are reflected by the length of incubation period in birds, and these vary substantially among species within and among geographical regions. The incubation periods are consistently shorter in North America (Arizona study site) than in tropical (Venezuela) and subtropical (Argentina) South America based on the study of 83 passerine species in 17 clades. Parents, mothers in particular, may influence incubation periods and resulting offspring quality through proximate pathways, while variation in maternal strategies among species can result from selection by adult and offspring mortality. Parents of long-lived species, as is common in the tropics and subtropics, may be under selection to minimize costs to themselves during incubation. Indeed, time spent incubating is often lower in the tropical and subtropical species than the related north temperate species, causing cooler average egg temperatures in the southern regions. Decreased egg temperatures result in longer incubation periods and reflect a cost imposed on offspring by parents because energy cost to the embryo and risk of offspring predation are both increased. Mothers may adjust egg size and constituents as a means to partially offset such costs. For example, reduced androgen concentrations in egg yolks may slow development rates, but may enhance offspring quality through physiological trade-offs that may be particularly beneficial in longer-lived species, as in the tropics and subtropics. We provide initial data to show that yolks of tropical birds contain substantially lower concentrations of growth-promoting androgens than north temperate relatives. Thus, maternal (and parental) effects on embryonic development rates may include contrasting and complementary proximate influences on offspring quality and deserve further field study among species.     

Warm temperatures lead to early onset of incubation, shorter incubation periods and greater hatching asynchrony in tree swallows Tachycineta bicolor at the extremes of their range

The onset of incubation varies in birds, with many species beginning incubation prior to clutch completion. Here we examine whether early onset is more likely to occur during high temperatures, a critical prediction of the egg-viability hypothesis, which suggest that birds begin incubation prior to clutch completion in order to maintain egg-viability. We examined onset of incubation in tree swallows Tachycineta bicolor at two locations at the extremes of their breeding range, Alaska and Tennessee. A majority of individuals (68%) began incubation prior to clutch completion. While females in Tennessee were more likely to begin incubation early, there was no difference between sites when differences in temperatures inside nestboxes were controlled in analyses. Rather, early onset of incubation was predicted by the proportion of daily temperatures above physiological zero during laying, a critical prediction of the egg viability hypothesis. Both warm weather and early onset led to shorter incubation periods and increased levels of hatching asynchrony. We found no effect of timing of nesting, female body condition index or clutch size on the probability of beginning incubation prior to clutch completion. Our results are consistent with the egg viability hypothesis, not consistent with a threshold clutch size rule, and do not support the hurry-up hypothesis, that individuals breeding later in the season would begin incubation early to reduce the time spent nesting. Overall, our results suggest that broad scale geographic differences in incubation behaviour may be explained by individual-level responses to environmental conditions.     

Importance of protein quality versus quantity in alternative host plants for a leaf-feeding insect

The nutritional value of alternative host plants for leaf-feeding insects such as caterpillars is commonly measured in terms of protein quantity. However, nutritional value might also depend on the quality of the foliar protein [i.e., the composition of essential amino acids (EAAs)]. A lack of comparative work on the EAA compositions of herbivores and their host plants has hampered the testing of this hypothesis. We tested the “protein quality hypothesis” using the tree-feeding caterpillars of Lymantria dispar (gypsy moth) and two taxonomically unrelated host plants, red oak (Quercus rubra) and sugar maple (Acer saccharum). Because L. dispar has higher fitness on oak than on maple, support for the hypothesis would be found if protein were of higher quality from oak than from maple. The whole-body EAA composition of L. dispar larvae was measured to estimate its optimum dietary protein composition, which was compared with the EAA compositions of oak and maple leaves. Contrary to the protein quality hypothesis, the EAA compositions of oak and maple were not significantly different in the spring. The growth-limiting EAAs in both tree species were histidine and methionine. Similar results were observed in the summer, with the exception that the histidine composition of oak was between 10 and 15 % greater than in maple leaves. The two main factors that affected the nutritional value of protein from the tree species were the quantities of EAAs, which were consistently higher in oak, and the efficiency of EAA utilization, which decreased from 80 % in May to <50 % in August. We conclude that the relative nutritional value of red oak and sugar maple for L. dispar is more strongly affected by protein quantity than quality. Surveys of many wild herbaceous species also suggest that leaf-feeding insects would be unlikely to specialize on plants based on protein quality.     

Schistosoma mansoni: incorporation and metabolism of protein amino acids in vitro

1. The amino acid composition of total proteins in six stages of the life cycle of Schistosoma mansoni was determined by routine autoanalysis of acid hydrolysates. Aspartate, glutamate and glycine were consistently the most abundant protein amino acids in all stages. 2. Incorporation of each of the protein amino acids into adult and egg proteins was determined using 72 hr cultures in complex media. Incorporation rates varied widely and there was no correlation between abundance in protein and the rate of incorporation. 3. Only five amino acids were interconverted to other amino acids which were themselves incorporated into worm and egg proteins (ala, arg, asp, gly, ser); of these only two (glu from ala and pro from arg) appeared to be of quantitative significance. Exogenous glucose yielded only three protein amino acids (ala, asp, glu). 4. The data are considered in the light of differences in egg and adult protein synthesis and with particular regard to potential chemotherapy at this level.     

Diet preferences for specific fatty acids and their effect on composition of fat reserves in migratory Red-eyed Vireos (Vireo olivaceous)

Fatty acid composition of body fat in birds often differs between bird species and between seasons, and changes in diet may be responsible for this variation. We tested two related hypotheses using Red-eyed Vireos, a long-distance migratory songbird: (1) birds prefer diets with certain fatty acids, and (2) fatty acid composition of the diet primarily determines the composition of lipid reserves. During paired-choice experiments, vireos preferred semi-synthetic diets with triolein (81% digestive extraction efficiency) over diets with tristearin (54% digestive extraction efficiency) and, in general, ate more when offered diets with unsaturated fats compared to saturated fats. These results demonstrate that vireos can discriminate between diets differing only in fatty acid composition and prefer diets with long-chain unsaturated fatty acids. When vireos were fed one of two diets for 1 month, the primary fatty acids in each diet also predominated in the tissues of birds fed each diet. However, some fatty acids that were absent in the diet occurred in bird tissues (e.g., 22:4, 22:5) suggesting that selective metabolism of fatty acids along with diet composition determine the fatty acid composition of lipid reserves in migratory birds.     

Vitellogenin dynamics during egg-laying: daily variation, repeatability and relationship with egg size

Daily variation in circulating levels of the avian yolk precursor, vitellogenin (VTG), throughout the laying cycle was investigated in female zebra finches Taeniopygia guttata and compared with predicted ovarian follicle demand (based on a model of follicular development for this species). In general, the pattern of variation in plasma VTG matched the predicted demand from the developing ovarian follicle hierarchy. Plasma VTG was non-detectable in non-breeders, but increased rapidly with onset of yolk development, remaining high (1.43–1.82 μg/ml, zinc) through to the 3-egg stage. Plasma levels then declined at the 5-egg stage (to 0.78±0.32 μg/ml) and were undetectable at clutch completion. This result is consistent with the hypothesis that yolk precursor production is costly and that selection has matched supply and demand. While inter-individual variation in plasma VTG was marked (e.g. 0.47–4.26 μg/ml at the 1-egg stage), it also exhibited high intra-individual repeatability (r=0.87–0.93). Finally, we examined the relationship between plasma VTG and primary reproductive effort. While individual variation in plasma VTG was independent of clutch size, laying interval and laying rate, there was a complex, diet-dependent relationship between VTG and egg size, with low plasma VTG levels being associated with both very small (<0.90 g) and very large (>1.15 g) egg sizes.     

The fatty acid composition of oophagous tadpoles (Chirixalus eiffingeri) fed conspecific or chicken egg yolk

We compared the lipid content and fatty acid composition of (1) the egg yolk of three anuran species (Chirixalus eiffingeri, Rhacophorus moltrechti and Buergeria robustus) and chicken eggs; and (2) C. eiffingeri tadpoles fed conspecific eggs or chicken egg yolk. Anuran and chicken egg yolk contained more non-polar than polar lipids but the proportions varied among species. Chicken egg yolk contained low amounts of 22:5n-3 in the polar lipid fraction, and B. robustus eggs did not contain any n-3 or n-6 non-polar lipids. The specific variation of lipid contents and fatty acid composition may relate to the maternal diet and/or breeding biology. In C. eiffingeri tadpoles that fed chicken yolk or frog egg yolk, the dominant components of polar and non-polar lipids were 16:0, 18:0, 18:1, and 18:2n-6, or 20:4n-6 fatty acids. C. eiffingeri eggs contained more n-3 fatty acids (e.g. 18:3n-3 and 20:5n-3) than chicken egg yolk, and tadpoles fed conspecific eggs contained more of these fatty acids than tadpoles fed chicken egg yolk. The compositional differences in the fatty acids between C. eiffingeri tadpoles that fed frog egg or chicken egg yolk are the reflection of the variation in the dietary sources. Our results suggest a direct incorporation of fatty acids into the body without or minimal modification, which provide an important insight into the physiological aspects of cannibalism.