Low Quality Eggs Research Articles

Protection of laying chickens against the Canadian DMV/1639 infectious bronchitis virus infection through priming with heterologous live vaccine and boosting with heterologous or homologous inactivated vaccine

The emergence of the Canadian Delmarva (DMV)/1639 infectious bronchitis virus (IBV) type strains was associated with egg production disorders in Eastern Canadian layer operations. While developing vaccines for novel IBV variants is not typically a reasonable approach, the consideration of an autogenous vaccine becomes more appealing, particularly when the new variant presents significant economic challenges. The current study aimed to compare the efficacies of two vaccination programs that included heterologous live priming by Massachusetts (Mass) and Connecticut (Conn) type vaccines followed by either a commercial inactivated Mass type vaccine or a locally prepared autogenous inactivated DMV/1639 type vaccine against DMV/1639 IBV challenge. The protection parameters evaluated were egg production, viral shedding, dissemination of the virus in tissues, gross and microscopic lesions, and immunological responses. The challenge with the DMV/1639 caused severe consequences in the non-vaccinated laying hens including significant drop in egg production, production of low-quality eggs, serious damage to the reproductive organs, and yolk peritonitis. The two vaccination programs protected the layers from the poor egg-laying performance and the pathology. The vaccination program incorporating the autogenous inactivated DMV/1639 type vaccine was more effective in reducing vial loads in renal and reproductive tissues. This was associated with a higher virus neutralization titer compared to the group that received the commercial inactivated Mass type vaccine. Additionally, the autogenous vaccine boost led to a significant reduction in the viral shedding compared to the non-vaccinated laying hens. However, both vaccination programs induced significant level of protection considering all parameters examined. Overall, the findings from this study underscore the significance of IBV vaccination for protecting laying hens.

RNA binding proteins are potential novel biomarkers of egg quality in yellow catfish

BackgroundEgg quality is a major concern in fish reproduction and development. An effective evaluation of egg quality prior to fertilization is helpful in improving the fertilization rate and survival rate of the larva. In this study, we aim to identify quality instructors from the combination study of fertilization rate, hatching rate, embryo malformation rate and gene expression profile.ResultsEggs from 25 female fish were fertilized with sperm from the same fish. The egg quality was determined by the fertilization rates, hatching rate and embryo malformation rate and divided into three categories, low-quality (< 35%), medium-quality (35 to 75%), and high-quality (> 75%). Due to the distinct difference in fertilization, hatching and embryo malformation rate between low-quality eggs and high-quality eggs, these two groups were considered for the identification of quality markers. Then RNA-seq was performed for the originally preserved eggs from the low-quality group and high-quality group. We profiled the differentially expressed genes and identified a group of RNA-binding proteins (RBPs) as potential regulators. Gene function analysis indicated that most of these genes were enriched in RNA-regulated pathways including RNA processing. The RBPs were more related to egg quality from the PLS-DA analysis. Finally, gene expression was validated by qRT-PCR.ConclusionsWe found a cluster of RBP genes including igf2bp3, zar1, elavl1, rbm25b and related regulatory factors including yy1, sirt1, anp32e, btg4 as novel biomarkers of egg quality.

Carotenoids and retinoids in the gonad of brood-stock pikeperch: accumulation during vitellogenesis and influence on egg quality in farmed pikeperch Sander lucioperca.

Carotenoids are determinants of reproductive fitness and egg quality. Here we studied the accumulation of astaxanthin (AX), canthaxanthin (CA) zeaxanthin (ZX), lutein (LU), retinol (RX) and dehydroretinol (DR) during vitellogenesis comparing previtellogenic and vitellogenic pikeperch (Sander lucioperca) eggs (n = 5 each), as well as selected tissues (liver, fat and muscles) in first süawning females (1176-1450 g). Futhermore, we compared egg batches with high (88-99% hatching rate, n = 5) or low (40-67% hatching rate, n= 5) egg quality. Vitellogenic follicles revealed higher concentrations of DR, RX, ZX and LU compared to previtellogenic follicles. Neither CA nor AX was detectable. In parallel, DR and RX were mobilized in the liver. In adipose and muscle tissue, comparing previtellogenic and vitellogenic females, no significant differences in carotenoid/retinoid content were observed. In high quality egg batches, both DR and RX were increased. LU was lower in high quality than in low quality eggs. In a conclusion, the amount of retinoids seems suboptimal in low quality egg batches and increased DR and RX are desirable in pikeperch. Since hypervitaminosis of retinoids can be problematic though, supplementation of the food with carotenoids, which can serve as precursors for retinoids, has to be carried out carefully.

A residual dense comprehensively regulated convolutional neural network to identify spectral information for egg quality traceability.

In the egg market, due to the different nutritional values of eggs laid by hens under different feeding conditions, it is common for low-quality eggs to be counterfeited as high-quality eggs. This paper proposes a residual dense comprehensively regulated convolutional neural network (RDCR-Net) to identify the quality of eggs laid by hens under different feeding conditions. Firstly, a hyperspectral system is used to obtain the spectral information of eggs. Secondly, due to the complex structure of the spectral information, a comprehensively regulated convolution (CRConv) is proposed to extract features hidden in the spectral information through feature transformation in multiple spaces. Thirdly, due to the limited availability of spectral information training samples, deep networks may suffer from feature degradation. The residual dense comprehensively regulated block (RDCR-Block) is proposed to tightly connect multiple CRConv layers with residual dense connections. Finally, the RDCR-Block is taken as the central unit, and the RDCR-Net is designed to identify egg spectral information. In the comparison of multi-model results, the RDCR-Net obtains the best classification performance with 96.29% accuracy, 97.53% precision, 97.14% recall, and 96.19% kappa coefficient. In summary, the RDCR-Net effectively extracts the deep features of spectral information, achieves high accuracy in identifying eggs laid by hens under different feeding conditions, and provides a new method for egg quality traceability.

ФИЗИОЛОГО-БИОХИМИЧЕСКИЕ АСПЕКТЫ КОРРЕКЦИИ УРАТНОГО ЛИТИАЗА У КУР В РАННЕМ ОНТОГЕНЕЗЕ ПРИ ИСПОЛЬЗОВАНИИ НЕКОТОРЫХ АНТИОКСИДАНТОВ

At present, in a number of poultry farms, urate lithiasis (uric acid diathesis) is a frequent cause of the death of young animals, the development of which is stimulated by the stresses accompanying industrial incubation, as well as the use of low-quality eggs, which leads to abnormalities in the development of the embryo, including its excretory system, causing a delay of uric acid in the body. This phenomenon is often aggravated by acidosis and a violation of the formation and functionality of thermoregulatory systems, which entails the production of nonviable young animals. The paper considers a method for preventing these negative phenomena through the transovarial use of a composition of optimal concentrations of antioxidants – sodium thioctate and citric acid. According to the results of the studies, it was revealed that the presented composition has a positive effect on the prevention of uric acid diathesis, which was expressed in a decrease in the intensity of lipoperoxidation, against the background of an increase in AOA in experimental chickens by 25,4%. This phenomenon was reflected in the intensity of the central metabolic processes, namely carbohydrate-energy and protein metabolism, which was reflected in the optimization of thermoregulation of experimental chickens. The combination of the above phenomena resulted in the absence of cases of uric acid diathesis in experimental chickens, which was a consequence of the absence of acidosis in experimental chickens (reserve blood alkalinity was 10,8% higher in experimental chickens relative to the control), which is a central aspect in the absence of this pathology. Thus, it can be argued that the composition of BAS presented by us actively levels free radical processes, which expresses the stimulation of the optimization of central metabolic processes and leads to the production of more highly viable young animals.

Avian Viruses that Impact Table Egg Production.

Simple SummaryGlobal table egg production is a multibillion-dollar industry. Poultry eggs provide an important source of protein, and consumers demand high-quality products. The industry relies on efficiency and good management, including vaccinations against the major poultry diseases, in order to ensure that they can match consumer demand. However, although there are a number of effective disease prevention protocols in place on poultry farms, viruses that affect egg production continue to be a major constraint for the sustainability of the table egg industry globally. Various factors related to either the infecting virus, the host, or the management system have been demonstrated to contribute significantly to the challenge for effective egg production. Viral infections in laying flocks have been shown to target various biological mechanisms that are essential for the quantitative and qualitative traits of egg production. This review describes the mechanisms involved in viral diseases of poultry that induce undesirable effects on egg production.Eggs are a common source of protein and other nutrient components for people worldwide. Commercial egg-laying birds encounter several challenges during the long production cycle. An efficient egg production process requires a healthy bird with a competent reproductive system. Several viral pathogens that can impact the bird’s health or induce reversible or irreversible lesions in the female reproductive organs adversely interfere with the egg industry. The negative effects exerted by viral diseases create a temporary or permanent decrease in egg production, in addition to the production of low-quality eggs. Several factors including, but not limited to, the age of the bird, and the infecting viral strain and part of reproductive system involved contribute to the form of reproductive disease encountered. Advanced methodologies have successfully elucidated some of the virus–host interactions relevant to the hen’s reproductive performance, however, this branch needs further research. This review discusses the major avian viral infections that have been reported to adversely affect egg productivity and quality and aims to summarize the current understanding of the mechanisms that underlie the observed negative effects.

Low concentrations of para-aminobenzoic acid increase the viability of fish in early stages of ontogenesis under different environmental conditions

The purpose of this work is to summarize the results of multi-year studies (1995–2017) on the effects of PABA on different fish species—peled Coregonus peled (Pallas), broad whitefish Coregonus nasus (Pallas) and common carp Cyprinus carpio L.—in early stages of ontogenesis. The conducted studies revealed the influence of PABA on viability in the early ontogenesis stages of two species of fish in the genus Coregonus with different levels of chromosomal mutability, assessed the ability of PABA to improve the survival of common carp cultivated in contrasting environmental conditions, and researched the ability of PABA to reduce the frequency of spontaneous chromosomal abnormalities in peled, broad whitefish and common carp during the embryogenesis period. The studies were conducted in laboratory and industrial conditions in the north and south of the Tyumen region. After the treatment of peled and broad whitefish roe with PABA, a significant increase (in comparison with the control) in the breaking stage of prelarvae from fertilized eggs laid for incubation was observed. The range of effective dosages of PABA for broad whitefish was wider (from 0.005% to 0.00001%) than for peled (from 0.001% to 0.00001%). The following PABA concentrations were found to be most effective: for peled, 0.001% (the increase in the breaking stage of prelarvae was 1.39 times greater than that of the control) and for broad whitefish, 0.0005% (the increase in the breaking stage of prelarvae was 1.04 times greater than that of the control). Laboratory and industrial studies performed on common carp under the conditions of the Surgut warm-water farm (the north of the Tyumen region) and the Tyumen fish hatchery (the south of the Tyumen region) showed that the greatest effect of PABA in increasing the survival rates of embryos and prelarvae is indicated in low-quality eggs. During incubation of low-quality eggs using para-aminobenzoic acid (0.0008%), the number of developing common carp embryos was 1.5 times higher than the control, and the number of prelarvae was 1.8 times higher than the control. Industrial tests showed that the positive effect of using PABA persists regardless of the conditions used for fish cultivation. Under the conditions of the fish reproduction process in a warm-water farm, the use of PABA significantly increased the survival of common carp embryos. In summary, these studies showed the ability of small doses of PABA to reduce the frequency of chromosomal abnormalities for three types of fish—peled, broad whitefish and common carp—at the gastrula stage.

Transcriptome analysis of egg viability in rainbow trout, Oncorhynchus mykiss

BackgroundMaternal transcripts are accumulated in the oocyte during oogenesis to provide for protein synthesis from oocyte maturation through early embryonic development, when nuclear transcription is silenced. The maternal mRNAs have short poly(A) tails after undergoing post-transcriptional processing necessary for stabilizing them for storage. The transcripts undergo cytoplasmic polyadenylation when they are to be translated. Transcriptome analyses comparing total mRNA and elongated poly(A) mRNA content among eggs of different quality can provide insight into molecular mechanisms affecting egg developmental competence in rainbow trout. The present study used RNA-seq to compare transcriptomes of unfertilized eggs of rainbow trout females yielding different eyeing rates, following rRNA removal and poly(A) retention for construction of the libraries.ResultsThe percentage of embryos to reach the 32-cell stage at 24 h post fertilization was significantly correlated to family eyeing rate, indicating that inviable embryos were developmentally compromised before zygotic genome activation. RNA sequencing identified 2 differentially expressed transcripts (DETs) from total mRNA sequencing comparing females with low-quality (< 5% eyeing), medium-quality (30–50% eyeing), and high-quality (> 80% eyeing) eggs. In contrast, RNA sequencing from poly(A) captured transcripts identified 945 DETs between low- and high-quality eggs, 1012 between low- and medium-quality eggs, and only 2 between medium- and high-quality eggs. The transcripts of mitochondrial genes were enriched with polyadenylated transcript sequencing and they were significantly reduced in low-quality eggs. Similarly, mitochondrial DNA was reduced in low-quality eggs compared with medium- and high-quality eggs. The functional gene analysis classified the 945 DETs between low- and high-quality eggs into 31 functional modules, many of which were related to ribosomal and mitochondrial functions. Other modules involved transcription, translation, cell division, apoptosis, and immune responses.ConclusionsOur results indicate that differences in egg quality may be derived from differences in maternal nuclear transcript activation and cytoplasmic polyadenylation before ovulation, as opposed to accumulation and storage of maternal nuclear transcripts during oogenesis. Transcriptome comparisons suggest low-quality eggs suffered from impaired oxidative phosphorylation and translation. The DETs identified in this study provide insight into developmental competence in rainbow trout eggs.

Profiling of the embryonic Atlantic halibut (Hippoglossus hippoglossus L.) transcriptome reveals maternal transcripts as potential markers of embryo quality.

BackgroundCommercial Atlantic halibut (Hippoglossus hippoglossus) farming is restricted by variable oocyte quality, slow growth, and early maturation of male fish. Maternally transferred components regulate early developmental processes; therefore, they have an effect on the future viability of the embryo. Using a newly developed Agilent 10 k custom-made oligonucleotide array, we profiled components of the transcriptome involved in immune defence as well as germline and muscle development during early developmental stages: 8-cell embryos (8CS), germ ring stage (GR), 10-somite stage (10SS), and hatched embryos (HT). In addition, we identified differentially expressed transcripts in low (≤9 ± 3% hatching) and high (≥86 ± 3°% hatching) quality eggs at 8CS to identify potential maternal markers for embryo quality.ResultsOut of 2066 differentially expressed transcripts, 160 were identified as maternal transcripts being specifically expressed at 8CS only. Twenty transcripts were differentially expressed in 8-cell embryos between low and high quality egg groups. Several immune-related transcripts were identified as promising molecular markers of hatching success including interferon regulatory factor 7 and mhc class 2A chain. Differential expression was positively validated with quantitative real-time PCR.ConclusionsWe have demonstrated maternal transfer of innate and adaptive immune system transcripts into Atlantic halibut embryos and their relation with future embryo developmental potential. We identified several transcripts as potential molecular markers of embryo quality. The developed microarray represents a useful resource for improving the commercial production of Atlantic halibut.Electronic supplementary materialThe online version of this article (doi:10.1186/1471-2164-15-829) contains supplementary material, which is available to authorized users.

Variation in embryonic mortality and maternal transcript expression among Atlantic cod (Gadus morhua) broodstock: A functional genomics study

Early life stage mortality is an important issue for Atlantic cod aquaculture, yet the impact of the cod maternal (egg) transcriptome on egg quality and mortality during embryonic development is poorly understood. In the present work, we studied embryonic mortality and maternal transcript expression using eggs from 15 females. Total mortality at 7days post-fertilization (7 dpf, segmentation stage) was used as an indice of egg quality. A 20,000 probe (20K) microarray experiment compared the 7hours post-fertilization (7 hpf, ~2-cell stage) egg transcriptome of the two lowest quality females (>90% mortality at 7 dpf) to that of the highest quality female (~16% mortality at 7 dpf). Forty-three microarray probes were consistently differentially expressed in both low versus high quality egg comparisons (25 higher expressed in low quality eggs, and 18 higher expressed in high quality eggs). The microarray experiment also identified many immune-relevant genes [e.g. interferon (IFN) pathway genes ifngr1 and ifrd1)] that were highly expressed in eggs of all 3 females regardless of quality. Twelve of the 43 candidate egg quality-associated genes, and ifngr1, ifrd1 and irf7, were included in a qPCR study with 7 hpf eggs from all 15 females. Then, the genes that were confirmed by qPCR to be greater than 2-fold differentially expressed between 7 hpf eggs from the lowest and highest quality females (dcbld1, ddc, and acy3 more highly expressed in the 2 lowest quality females; kpna7 and hacd1 more highly expressed in the highest quality female), and the 3 IFN pathway genes, were included in a second qPCR study with unfertilized eggs. While some maternal transcripts included in these qPCR studies were associated with extremes in egg quality, there was little correlation between egg quality and gene expression when all females were considered. Both dcbld1 and ddc showed greater than 100-fold differences in transcript expression between females and were potentially influenced by family. The Atlantic cod ddc (dopa decarboxylase) complete cDNA was characterized, and has a 1461bp open reading frame encoding a 486 amino acid protein that contains all eight residues of the conserved pyridoxal 5’-phosphate binding site including the catalytic lysine. This study provides valuable new information and resources related to the Atlantic cod egg transcriptome. Some of these microarray-identified, qPCR-confirmed, Atlantic cod egg transcripts (e.g. ddc, kpna7) play important roles during embryonic development of other vertebrate species, and may have similar functions in Atlantic cod.

Fusion of lipid droplets in Japanese eel oocytes: Stage classification and its use as a biomarker for induction of final oocyte maturation and ovulation

The quality of eggs obtained from maturation-induced Japanese eel, Anguilla japonica, is unstable. One of the causes for low quality eggs is that females are artificially induced to ovulate at an inappropriate maturational status. Here we investigated the feasibility to use the morphological changes of lipid droplets in oocytes as a biomarker to show the optimum timing for injections of salmon pituitary extract (SPE) for priming and maturation-inducing steroid (MIS) for final oocyte maturation and ovulation. Various sizes of oocytes (700 to 1000 μm) were collected by cannulation from maturation-induced female eels during five days until ovulation. Morphology of the lipid droplets in these oocytes were classified into 10 stages mainly on the basis of their diameter, which increased with the progress of maturation as the droplets fused together. To assess the maturational status of each female, the lipid droplet stage was determined for 10 oocytes randomly chosen among the most advanced mode of developing oocytes and the median value was defined as the stage for the individual. Relationship between the lipid droplet stage at induction of ovulation and hatchability of the ovulated eggs was examined in 23 females, in which the hatching rate markedly fluctuated. Stages of females from which good quality eggs were obtained converged around stage 4 (five largest droplets, 55 to 70 μm) at priming with SPE, stage 6 (90 to 110 μm) at injection with MIS, and stage 7 (110 to 130 μm) at ovulation, while those of females from which low quality eggs were obtained showed distributions over broader ranges. Furthermore, 466 females were induced to mature and the lipid droplet stage at induction of ovulation was examined in 13 females from which high quality eggs (hatching rate >80%) were obtained. Most of the 13 females were at stage 3 (40 to 55 μm) or 4 when primed with SPE, at stage 6 when injected with MIS, and at stage 7 when they ovulated. These results suggest that the optimum stages for SPE priming, MIS injection, and ovulation are stages 3 to 4, 6, and 7, respectively, though the optimum stage for priming will be variable if the temperature and/or the time between priming and MIS injection is varied. We conclude that the lipid droplet stage can be used as a sensitive biomarker to show the optimum timing for inducing ovulation by hormonal treatments.

One-step collagenase method for zona pellucida removal in unfertilized eggs: easy and gentle method for large-scale preparation.

Zona pellucida (ZP)-free eggs are often used for studies such as evaluating the interaction of sperm-oolemma. To acquire ZP-free eggs, the most commonly used methods employ acidified Tyrode's solution, enzymatic digestion with a trypsin-like enzyme, or mechanical methods using micropipettes. However, acidified Tyrode's solution and trypsin-like enzymes often damage the oolemma, especially when many eggs are treated at once for mass sample analyses. The mechanical method requires skill, and it is time-consuming to prepare many ZP-free eggs. Therefore, in this study, to establish an easy, reliable method for preparing ZP-free eggs, we examined the ZP digestion method originally reported by Zuccotti et al. (J Reprod Fertil 93:515-520, 1991) that uses collagenase. Mouse unfertilized eggs were treated with collagenase and acidified Tyrode's solution to compare the ZP-free rates, the effect on the oolemma, and the two-cell development rates of ZP-free eggs by in vitro fertilization. The effects on the oolemma were gauged by observing the polarity of the transmembrane protein localization of enhanced green fluorescence protein tagged CD9 protein (CD9-EGFP) and using differential interference contrast microscopy. Collagenase removed the ZP and the cumulus cells from the cumulus oocyte complex. The collagenase method had no influence on the localization of CD9-EGFP, resulting in a high two-cell development rate. Additionally, the collagenase method could exclude low quality eggs with hardened ZP, since collagenase could not digest the hardened ZP. The one-step collagenase method is an easy preparation method for large numbers of high-quality ZP-free eggs.

Maternal gene expression in Atlantic halibut (Hippoglossus hippoglossus L.) and its relation to egg quality

BackgroundThe commercial production of Atlantic halibut (Hippoglossus hippoglossus L.) suffers from a major bottleneck due to the low success of producing juveniles for on-growing. Atlantic halibut females are routinely hand-stripped and incorrect timing of stripping can result in low quality eggs due to post-ovulatory aging. Post-ovulatory aging leads to compositional changes in eggs that include maternally provided proteins and RNAs. There have been few studies of the maternally provided mRNA transcripts that control early development in commercially important fish species. The present study aimed to study maternal gene expression in Atlantic halibut and its relation to egg quality parameters including blastomere symmetry and hatching success.ResultsA maternal EST library containing 2341 sequences was constructed by suppressive subtractive hybridisation. Thirty genes were selected for expression studies; 23 novel genes and 7 genes with documented roles in early development. The expressions of twenty-one selected genes were measured by qPCR from fertilization to the 10-somite stage. Three genes were identified as strictly maternal genes that were expressed until the start of gastrulation; askopos (kop), si:dkey-30j22.9 (Tudor family member), and Tudor 5 protein (Tdrd5). The expressions of 18 genes at the 8-cell stage were correlated with egg quality parameters. The majority of genes showed either no or very minor correlations with egg quality parameter. However, two genes correlated positively with hatching success (r> 0.50, HHC00353: r = 0.58, p < 0.01; HHC01517: r = 0.56, p < 0.01) and one gene (HHC00255) was negatively correlated with the percentage of normal blastomeres (r = -0.62, p < 0.05).ConclusionsDuring this study we have related maternal levels of gene expression to hatching success in fish. Poor hatching success was not correlated with a general decrease in transcript abundance but with low transcript levels of some specific genes. Thus, the molecular mechanisms leading to low Atlantic halibut egg quality cannot be entirely explained by post-ovulatory aging.

FECUNDITY AND SURVIVAL IN RELATION TO RESISTANCE TO OXIDATIVE STRESS IN A FREE‐LIVING BIRD

Major life history traits, such as fecundity and survival, have been consistently demonstrated to covary positively in nature, some individuals having more resources than others to allocate to all aspects of their life history. Yet, little is known about which resources (or state variables) may account for such covariation. Reactive oxygen species (ROS) are natural by-products of metabolism and, when ROS production exceeds antioxidant defenses, organisms are exposed to oxidative stress that can have deleterious effects on their fecundity and survival. Using a wild, long-lived bird, the Alpine Swift (Apus melba), we examined whether individual red cell resistance to oxidative stress covaried with fecundity and survival. We found that males that survived to the next breeding season tended to be more resistant to oxidative stress, and females with higher resistance to oxidative stress laid larger clutches. Furthermore, the eggs of females with low resistance to oxidative stress were less likely to hatch than those of females with high resistance to oxidative stress. By swapping entire clutches at clutch completion, we then demonstrated that hatching failure was related to the production of low-quality eggs by females with low resistance to oxidative stress, rather than to inadequate parental care during incubation. Although male and female resistance to oxidative stress covaried with age, the relationships among oxidative stress, survival, and fecundity occurred independently of chronological age. Overall, our study suggests that oxidative stress may play a significant role in shaping fecundity and survival in the wild. It further suggests that the nature of the covariation between resistance to oxidative stress and life history traits is sex specific, high resistance to oxidative stress covarying primarily with fecundity in females and with survival in males.

Distribution of lipid droplets is an indicator for egg quality in brown trout, Salmo trutta fario

In the eggs of the brown trout, Salmo trutta fario, lipid droplets are clearly visible beneath the oolemma on the surface of the protein yolk. When examining the distribution of lipid vesicles in egg batches of different quality, four categories of lipid vesicle distribution could be distinguished: In the first category, the lipid droplets were evenly distributed throughout the egg. These egg batches had a high quality (percentage of eggs developing to eyed stage embryos: 96.2±5.2). Some lipid droplets had the tendency to coalesce in one pole of the egg in the second category. This coalescing of lipid droplets was accompanied with a reduction in egg quality to about 50% of the first category (% of eyed stage embryos: 40.6±15.2). In the last two categories, the lipid droplets were mostly coalesced in one or two poles of the egg and the egg quality was very low (% of eyed stage embryos: 3.3±2.9 and zero, respectively). In conclusion, examination of lipid droplets distribution, in brown trout, can distinguish between high and low quality eggs in an easy and reliable way.

Lipid and fatty acid composition of eggs producing larvae with high survival rate in the Japanese eel

Eggs of Japanese eel Anguilla japonica were obtained by artificial induction of maturation. Fertilization and hatching rates, and survival rates of normal larvae were examined. The levels of total lipids and polar lipids (PL) of high quality (HQ; &gt;80% survival rate at 8 days post‐hatch) eggs were significantly lower than low quality (LQ; hatching and survival rates were zero) eggs. The docosahexaenoic acid (DHA) level in PL of HQ eggs, however, was significantly higher than in LQ eggs. In contrast to DHA, arachidonic acid level was negatively correlated with egg quality variables and it was lower in HQ than in LQ eggs. Most of the egg lipid and fatty acid components showed larger variation in LQ than in HQ eggs.

The effects of temperature and salinity on egg production and hatching success of Baltic Acartia tonsa (Copepoda: Calanoida): a laboratory investigation

The functional response of the aspects of reproductive success of a southwestern Baltic population of Acartia tonsa (Copepoda: Calanoida) was quantified in the laboratory using wide ranges in temperatures and salinities. Specifically, daily egg production (EP, # female−1 day−1) was determined for 4 or 5 days at 18 different temperatures between 5 and 34°C and the time course and success of hatching were evaluated at 10 different temperatures between 5 and 23°C. The effect of salinity (0 to 34 psu) on egg hatching success was also examined. The highest mean rates of EP were observed between 22 and 23°C (46.8–50.9 eggs female−1 day−1). When studied at 18 psu, hatching success of eggs increased with increasing temperature and was highest (92.2%) at 23°C. No hatching was observed for eggs incubated at low temperatures (≤12°C) that were produced by females acclimated to temperatures ≤10°C indicating a possible thermal threshold between 10.0 and 13.0°C below which only the production of diapause (or low quality) eggs exists in this population. When tested at 18°C, the hatching success of eggs incubated at 15 different salinities increased asymptotically with increasing salinity and was maximal (81.4–84.5%) between 17 and 25 psu. The high reproductive success observed over wide ranges in temperatures and salinities in this Baltic population demonstrates one of the mechanisms responsible for the cosmopolitan distribution of this species within productive, estuarine and marine habitats.

Can ovarian infertility be treated with bone marrow- or ovary-derived germ cells?

A year ago, reproductive biologists and general public were astonished with evidence reported by Johnson et al. in Nature 428:145 that mammalian ovaries possess persisting large germline stem cells, which allegedly enable follicular renewal in adult females. Recently, the same research group declared such view obscure, and reported that mammalian oocytes originate from putative germ cells in bone marrow and are distributed by peripheral blood to the ovaries (Cell 122:303). While neglecting available data on the germ cell origin from the ovarian surface epithelium (OSE) in adult mouse and human females and complexity of follicular renewal in humans, the authors widely extrapolated their observations on formation of allogeneic oocytes after bone marrow (or blood) transplantation in ovaries of adult mice treated with cytostatics to clinical implications in the public media. Yet, the resulting outcome that such allogeneic oocytes may enable the propagation of ovarian cycles is a poor alleviation for the women with ovarian infertility. Women lacking primary follicles, or carrying follicles with low quality eggs persisting in aging ovaries, are not concerned about the lack of menstrual cycles or ovarian steroids, but about virtually no chance of having genetically related children. Johnson et al. also reported that the germ cell formation in bone marrow disappears in ovariectomized mice. Such observation, however, raises solid doubts on the bone marrow origin of oocytes. Since germ cells developing from the OSE cells of adult human ovaries during periodical follicular renewal are known to enter blood vessels in order to enable formation of primary follicles at distant ovarian sites, they also contaminate peripheral blood and hence bone marrow. Better knowledge on the complexity of follicular renewal in humans and exploration of a potential of human OSE cells to produce new oocytes in vitro are essential for novel approaches to the autologous treatment of premature ovarian failure and age induced ovarian infertility.

Large scale real‐time PCR analysis of mRNA abundance in rainbow trout eggs in relationship with egg quality and post‐ovulatory ageing

The mRNA levels of 39 target genes were monitored in unfertilized eggs of 14 rainbow trout sampled the day of ovulation and again 5, 14, and 21 days later. For all 56 collected egg batches, an egg sample was fertilized to estimate egg quality by monitoring embryonic development. Remaining eggs were used for RNA extraction and subsequent real-time PCR analysis. A significant drop of egg quality was observed when eggs were held in the body cavity for 14 or 21 days post-ovulation (dpo). During the same period, eight transcripts (nucleoplasmin or Npm2, ferritin H, tubulin beta, JNK1, cyclin A1, cyclin A2, cathepsin Z, and IGF2) exhibited a differential abundance at one or several collection time(s). Interestingly, we observed higher levels of cyclins A1 and A2 mRNAs in eggs taken 5 days post-ovulation than in eggs taken, from the same females, at the time of ovulation. In addition, seven transcripts exhibited a differential abundance between low quality and high quality eggs. Low quality eggs were characterized by lower levels of Npm2, tubulin beta, and IGF1 transcripts. In contrast, keratins 8 and 18, cathepsin Z, and prostaglandin synthase 2 were more abundant in low quality eggs than in high quality eggs. In this study, we have demonstrated differences in mRNA levels in the rainbow trout egg that are reflective of developmental competence differences induced by post-ovulatory ageing. The putative role of these transcripts in post-ovulatory ageing-induced egg quality defects is discussed with special attention for corresponding cellular functions.

Breeding performance of great tits (Parus major) along a gradient of heavy metal pollution

The possible effects of heavy metal pollution on various breeding parameters of the great tit (Parus major) were studied at four study sites in a pollution gradient near a large nonferrous smelter in Belgium during three consecutive breeding seasons. Significantly more females interrupted their laying period near the pollution source than did females farther away. At the two most polluted sites, hatching success was significantly reduced compared to two lesser-polluted study sites. Overall breeding success was also significantly lower toward the factory complex. This lowered breeding success was caused not only by the decreased hatching success toward the pollution source but also by the significant difference in fledging success among sites. Although a clear relationship was observed between the exposure to heavy metal pollution and an impaired reproduction, not all reproductive parameters were affected. We found no significant differences in the onset of laying, clutch size, length of the incubation period, and the proportion of nests with low-quality eggs among sites. Although nest box occupancy was every year slightly lower toward the smelter compared to farther away, differences among study sites were never significant.