VPS38/UVRAG and ATG14, the variant regulatory subunits of the ATG6/Beclin1-PI3K complexes, are crucial for the biogenesis of the yolk organelles and are transcriptionally regulated in the oocytes of the vector Rhodnius prolixus.

In insects the reserve proteins are stored in the oocytes into endocytic-originated vesicles named yolk organelles. VPS38/UVRAG and ATG14 are the variant regulatory subunits of two class-III ATG6/Beclin1 PI3K complexes that regulate the recruitment of the endocytic (complex II) and autophagic (complex I) machineries. In a previous work from our group, we found that the silencing of ATG6/Beclin1 resulted in the formation of yolk-deficient oocytes due to defects in the endocytosis of the yolk proteins. Because ATG6/Beclin1 is present in the two above-described PI3K complexes, we could not identify the contributions of each complex to the yolk defective phenotypes. To address this, here we investigated the role of the variant subunits VPS38/UVRAG (complex II, endocytosis) and ATG14 (complex I, autophagy) in the biogenesis of the yolk organelles in the insect vector of Chagas Disease Rhodnius prolixus. Interestingly, the silencing of both genes phenocopied the silencing of ATG6/Beclin1, generating 1) accumulation of yolk proteins in the hemolymph; 2) white, smaller, and yolk-deficient oocytes; 3) abnormal yolk organelles in the oocyte cortex; and 4) unviable F1 embryos. However, we found that the similar phenotypes were the result of a specific cross-silencing effect among the PI3K subunits where the silencing of VPS38/UVRAG and ATG6/Beclin1 resulted in the specific silencing of each other, whereas the silencing of ATG14 triggered the silencing of all three PI3K components. Because the silencing of VPS38/UVRAG and ATG6/Beclin1 reproduced the yolk-deficiency phenotypes without the cross silencing of ATG14, we concluded that the VPS38/UVRAG PI3K complex II was the major contributor to the previously observed phenotypes in silenced insects. Altogether, we found that class-III ATG6/Beclin1 PI3K complex II (VPS38/UVRAG) is essential for the yolk endocytosis and that the subunits of both complexes are under an unknown transcriptional regulatory system.

Mathematical modeling of the interaction between yolk utilization and fish growth in zebrafish, Danio rerio.

Optimal embryonic development plays a major role in the health of an individual beyond the developmental stage. Nutritional perturbation during development is associated with cardiovascular and metabolic disease later in life. With both nutritional uptake and overall growth being risk factors for eventual health, it is necessary to understand not only the behavior of the processes during development but also their interactions. In this study, we used differential equations, image analyses, curve fittings, parameter estimation and laboratory experiments to quantify the rate of yolk absorption and its effect on early development of a vertebrate model (Danio rerio). Findings from this study establish a nonlinear functional relationship between nutrient absorption and early fish growth. We found that the rate of change in fish length and yolk utilization is logistic, that is the yolk decays rapidly for a period of time before leveling out. An interesting finding from this study is that yolk utilization reaches its maximum at 84 h post-fertilization. We validated our mathematical models against experimental observations, making them powerful tools for replication and future simulations.

Egg quality depending on the diet with different sources of protein and age of the hens.

Due to the planned limitations in the use of genetically modified soybean meal, the search for alternative sources of protein in animal nutrition is ongoing, which also supports the consumers' expectations of good quality products, such as eggs. The aim of the study was to assess and compare morphological traits of eggs, fatty acid composition in yolk lipids, and the content and activity of lysozyme in thick and thin albumen in eggs from hens fed a diet based on legume seeds as a substitute for soybean meal depending on the hens age. Analyses were carried out for 300 eggs on dates I-V (19-39 weeks age of hens), obtained from Rosa 1 hens managed in the semi-intensive system. Quality analysis was performed for 30 eggs from each group at a time. The control group of hens (A) was fed a diet based on soybean meal (SBM) and the treatment group (B) was fed a diet based on seeds from narrow-leaved lupin (Boruta), yellow lupin (Mister), and pea (Muza). Eggs were analysed for morphological traits (egg weight, the weight and density of egg components, egg shape index and egg surface area), parameters of albumen (height, Haugh units), yolk colour (La Roche, colorimetry in the CIE Lab system), lysozyme content and activity in albumen, and fatty acids composition in yolk lipids. In group B the egg shape index was higher (p = 0.002), and the eggshell colour index was lower (p = 0.007), as well as the height of thick albumen, Haugh score, and yolk colour (La Roche) were higher in group B (p < 0.05), while redness of yolk was significantly lower (p < 0.001) than in group A. Lower content of C15:0 and C18:1n9 and higher content of C18:2n6 and C18:3n3, (p < 0.05), as well as lower content of MUFA and OMEGA 9, but higher content of PUFA, OMEGA 3, OMEGA 6, PUFA/SFA, OMEGA 9/6 and OMEGA 9/3 were found in group B compared to group A (p < 0.05). There were significant differences in traits between age of hens (p < 0.05). Egg weight, yolk weight and its proportion in egg, as well as shell weight, its strength and thickness increased with the age of hens (p < 0.05). The content of lysozyme and its activity were also higher in eggs from older hens (p < 0.05). Fatty acids composition was beneficial at the beginning and end of the analysed egg production period. The proposed feed mixture based on legume seeds had no negative effect on the quality of eggs, and had a positive effect on yolk colour, the quality of albumen. There was no detrimental effect of diet and age of hens on fatty acid composition of eggs from both groups throughout the study period. To sum up, diet with legume seeds could be accepted as a substitute for soybean meal, due to the beneficial effects for some mentioned traits of eggs. Use of legumes could give a partial self-sufficient for producers from small farms, where is not possible to produce soybean meal. At the same time, the level of anti-nutrients in legumes should be considered.

Comparison of different carrier-compounds and varying concentrations of oleic acid on freezing tolerance of ram spermatozoa in tris-citric acid-egg yolk plasma semen diluent.

The current study was conducted to determine the optimal concentration carrier-compound for oleic acid (OA) among dimethyl sulfoxide (DMSO), liposome and ß-cyclodextrin on ram spermatozoa cryosurvival. The preliminary experiment was designed to ascertain the optimal concentration of egg yolk plasma. In Experiment 1, semen was placed in a diluent containing different concentrations of OA dissolved in DMSO (0.125, 0.25, 0.50, 1, 2, 4 and 8 mM). In Experiments 2 and 3, effects of liposome loaded-OA and ß-cyclodextrin-OA complexes (0.25, 0.50, 1 and 2 mM) on semen cryopreservation were evaluated. In Experiment 4, optimal concentrations of OA were determined, based on results from previous experiments. Spermatozoa viability, kinematics, plasma membrane integrity, malondialdehyde, superoxide dismutase activity and total antioxidant status of samples were evaluated. Results indicated varying concentrations of OA had different effects on sperm kinematics, viability and membrane functionality after freezing/thawing (P < 0.05). In addition, inclusion of OA in liposomes or combinations with ß-cyclodextrin resulted in greater values for spermatozoa motion variables compared with DMSO dissolved-OA (P < 0.05). Inclusions of OA at 0.25 and 0.50 mM led to a reduction in amounts of lipid peroxidation when there was inclusion of liposome and ß-cyclodextrin as carrier-compounds (P < 0.05). Activity of SOD was similar with inclusion of different concentrations of OA or carrier-compounds, but total antioxidant capacity was affected by OA concentration and carrier-compound type (P < 0.05). The results highlight the importance of carrier-compound type and concentrations of OA on ram spermatozoa during cryopreservation.

Effects of feeding broiler breeder hens a coextruded full-fat flaxseed and pulses mixture without or with multienzyme supplement.

The effects of coextruded full-fat flaxseed and pulses (FFF; 1:1 wt/wt) mixture on n-3 polyunsaturated fatty acids (PUFA) enrichment in egg yolk, hepatic attributes, apparent retention (AR) of components, and ceca metabolites were evaluated in broiler breeder hens. The diets were as follows: 1) corn-soybean control, 2) control diet plus 18% FFF (FFF-), and 3) FFF plus enzyme supplement (FFF+) containing galactanase, protease, mannanase, glucanase, xylanase, amylase, and cellulase activities. Twenty-six-week-old Cobb 500 broiler breeder hens were allocated to 30 identical cages (2 hens/cage) and given 1-week adaptation period. The 3 diets were assigned to 10 replicate cages based on postadaptation BW and fed based on breeder curve for 30 D. Excreta samples were collected from day 24 to 27 for determination of AR of components, and eggs were collected from day 28 to 30 for yolk polyunsaturated fatty acids analyses. On day 30, birds were weighed, killed via cervical dislocation, liver weighed, and stored for fat analyses. Ceca digesta samples were taken for concentration of short-chain fatty acids. Liver and yolk weights as well as total yolk FA were not influenced by diets (P > 0.05). Control birds had lower yolk concentration of α-linolenic acid than birds fed either FFF- or FFF+ (P < 0.01) corresponding to 7.5, 36.8, and 37.3 mg/g for the control, FFF-, and FFF+, respectively. Control birds also exhibited lower yolk concentration of docosahexaenoic acid (P < 0.01). Control birds had higher hepatic concentration of crude fat and apparent retention of dry matter and crude protein compared with either the FFF- or FFF+ birds (P < 0.05). Birds fed FFF- diet had lower ceca digesta concentration of lactic acid than control and FFF+ (P < 0.05) birds. Results showed broiler breeder hens enriched egg yolk with n-3 polyunsaturated fatty acids without effects on the liver while the supplemental enzyme did not improve the utilization of FFF.

A review on yolk sac utilization in poultry.

During incubation, embryonic growth and development are dependent on nutrients deposited in the egg. The content of the yolk can be transferred to the embryo in 2 ways: directly into the intestine via the yolk stalk or through the highly vascularized yolk sac membrane. It has been suggested that, as a result of genetic selection and improved management, the increase in posthatch growth rate and concurrently the increase in metabolic rate of broiler chickens during the last 50 yr has also increased embryonic metabolism. A higher metabolic rate during incubation would imply a lower residual yolk weight and possibly lower energy reserve for the hatchling. This might affect posthatch development and performance. This review examined scientific publications published between 1930 and 2018 to compare residual yolk weight at hatch, metabolic heat production, and yolk utilization throughout incubation. This review aimed to investigate 1) whether or not residual yolk weight and composition has been changed during the 88-yr period considered and 2) which abiotic and biotic factors affect yolk utilization in poultry during incubation and the early posthatch period. It can be concluded that 1) residual yolk weight and the total solid amount of the residual yolk at hatch seem to be decreased in the recent decades. It cannot be concluded whether the (lack of) differences between old and modern strains are due to genetic selection, changed management and incubation conditions, or moment of sampling (immediately after hatch or at pulling). It is remarkable that with the genetic progress and improved management and incubation conditions over the last 88 yr, effects on yolk utilization efficiency and embryonic metabolic heat production are limited; 2) factors specially affecting residual yolk weight at hatch include egg size and incubation temperature, whereas breeder age has more influence on nutrient composition of the residual yolk.

The effect of capsulated and noncapsulated egg-yolk-specific antibody to reduce colonization in the intestine of Salmonella enterica ssp. enterica serovar Infantis-challenged broiler chickens.

The antibacterial properties of egg yolk antibodies have been known for many years. Enhanced antibiotic resistance has resulted in increased need for using these antibodies as an alternative. In the present study, generation, capsulation, and inhibition growth properties of IgY directed against Salmonella enterica subsp. enterica serovar Infantis (SI) were evaluated. White Leghorn layer hens were immunized using whole cell of inactivated SI. Salmonella Infantis-specific antibody activities in sera and egg yolk were determined by ELISA. A total of 480 one-day-old male "Cobb 500" chicks were randomly divided into 8 groups, with 6 replications of 10 birds kept for 21 D. All birds from 7 challenged groups were orally inoculated with 1 mL of SI suspension (1 × 107 CFU/mL) at 3 and 4 D of age. Two groups were dietary supplemented with 5 g/kg immune powdered yolk or nonimmune powdered yolk. One group was dietary supplemented with 12.8 g/kg capsulated immune yolk (CIY). Two groups were given 8.3 mL/L of immune water-soluble yolk or nonimmune water-soluble yolk fraction in drinking water. In the antibiotic group, 1 mL/L Enrofloxacin 10% was added to drinking water. All supplements except for the antibiotic (on Day 4 for 10 D) were added on day one and continued during the experiment. Negative and positive control groups received no supplements. During the experiment, among the challenged groups, the minimum SI cecal colonization and the lowest isolation of SI from the liver (P < 0.01) was observed in the antibiotic group. Following antibiotic group, in the group receiving CIY, colonization of bacteria in ceca and liver was significantly reduced during the second and third weeks of the experiment (P < 0.01). According to the results, capsulated specific IgY has a beneficial effect in reducing the colonization of Salmonella under the conditions of this study in comparison with other forms of IgY antibody.

Effects of high-intensity ultrasonic (HIU) treatment on the functional properties and assemblage structure of egg yolk.

The effects of high-intensity ultrasonic (HIU) treatment on the functional properties of egg yolk were studied in the present work. After HIU treatment, the emulsifying, foaming and gel properties of the egg yolk solution significantly increased, but the foam stability decreased. SDS-PAGE results showed that there was no obvious change in the protein bands of egg yolk, indicating that the yolk proteins did not undergo covalent crosslinking or degradation. HIU treatment enhanced the zeta potential of egg yolk components in solution and increased the free sulfhydryl content of egg yolk proteins. Moreover, the particle size distribution of egg yolk components in solution changed markedly, and these changes demonstrated that HIU treatment caused the aggregation of yolk low-density lipoprotein and the partial dissociation of yolk granules. These results revealed that HIU treatment could change the aggregation of yolk components, which in turn could influence the solution characteristics of egg yolk, finally resulting in changes to the functional properties of egg yolk.

In ovo metabolism of estradiol to estrone sulfate in chicken eggs: Implications for how yolk estradiol influences embryonic development.

The steroid 17ß-estradiol (herein "estradiol") is a potent regulator of sexual differentiation that exerts wide-ranging effects on the developing brain and other tissues. The developing gonads are an important source of estradiol but most, if not all, vertebrate embryos are also exposed to maternally derived estradiol during development. In birds, this maternally derived estradiol is present in the egg at the time of oviposition but very little is known about how this source of estradiol influences development. A critical aspect of understanding yolk estradiol effects is deciphering how steroid metabolism may regulate embryonic exposure to yolk estradiol. In this study, we examine the metabolic fate of estradiol during the first five days of incubation in chicken (Gallus gallus) eggs. Using tritiated estradiol to trace the movement and metabolism of estradiol, we demonstrate that estradiol is metabolized to estrone, which is subsequently conjugated to estrone sulfate as the primary metabolite. Estrone sulfate then accumulates in the albumen by day five of incubation. Overall, these findings have important implications for how yolk estradiol may influence development and alter offspring phenotype. Mechanisms through which estradiol, as well as estrone sulfate, might elicit effects are discussed.

Sodium alginate potentiates antioxidants, cryoprotection and antibacterial activities of egg yolk extender during semen cryopreservation in buffalo.

The study was conducted to determine effects of sodium alginate on sperm during cryopreservation. Each ejaculate (n = 20) of five buffalo bulls (3-5 years) were divided into six equal fractions and diluted using egg yolk based extender supplemented with different concentrations of sodium alginate and cryopreserved. Frozen-thawed semen samples were evaluated using the CASA, hypo-osmotic swelling test, cervical mucus penetration capacity test, and chlortetracycline fluorescence assay (CTC). Phosphorylation of tyrosine containing proteins and malondialdehyde concentration of sperm membrane were evaluated using immunoblotting and thiobarbituric acid reactive substance assay respectively. The semen extender's anioxidative capacities were estimated by conducting 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) assays, metal chelating capacity by assessing ferrozine and antibacterial capacity using agar plate methods. Supplementation of sodium alginate in extender improved sperm longevity, plasma membrane integrity as well as capacity to transit through the cervical mucus. Supplementation of extender with sodium alginate minimises the phase transition of sperm membranes and phosphorylation of tyrosine containing proteins during cryopreservation. Malondialdehyde concentration of sperm was less in sodium alginate-treated sperm as compared with control samples. The results indicated that sodium alginate increased antioxidant capacity of semen extender. Supplementation with sodium alginate also improved the metal chelating capacity and antibacterial properties of the extender. In conclusion, supplementation of extender with sodium alginate enhances free radical scavenging, metal reduction and chelating capacities to protect sperm during cryopreservation.

Basic fibroblast growth factor promotes prehierarchical follicle growth and yolk deposition in the chicken.

Basic fibroblast growth factor (bFGF) plays a pivotal role in prompting ovarian follicular development and angiogenesis as well as inhibiting atresia. In the chicken, high laying performance depends largely on efficient healthy development of ovarian follicles. Moreover, rapid growth of oocytes resulted from abundant yolk deposition via blood circulation and intra-ovarian interactions among somatic and germ cells. The major components of yolk mass consist of very low density lipoprotein (VLDL) and vitellogenin that are taken up by maturing oocytes via VLDL receptor (VLDLR)-mediated endocytosis from blood capillaries in the theca layer and gaps between granulosa cells. Here we used immunofluorescence, BrdU, TUNEL, Western bolt and RT-qPCR methods to investigate effects of bFGF on growth and yolk deposition of chicken prehierarchical follicles. Results showed that VLDLR was mainly expressed in the granulosa cells of the prehierarchical and preovulatory follicles, and its expression declined with follicle growth. Moreover, bFGF caused a dose-dependent promoting effect on growth of small white follicles and this effect was inhibited by SU5402 (an FGFR1 antagonist). Proliferation of follicular theca externa cells was accelerated by bFGF via FGFR1-AKT signaling, coupled with augmented angiogenesis and up-regulated p-ERK expression in granulosa cells. After combined inhibition of FGFR1 and PPARγ, we found that PPARγ could also suppress VLDLR expression in granulosa cells. These results indicate that bFGF facilitated growth and yolk deposition in chicken prehierarchical follicles through promoting proliferation and angiogenesis in theca layers, and also through down-regulating VLDLR expression in granulosa cells.

Liquid storage of dromedary camel semen in different extenders.

This study was conducted to assess the effects of commercial extenders and storage temperature on dromedary camel sperm quality during liquid preservation. In Experiment 1, ejaculates (n = five males; replicated seven times) were split and diluted with synthetic (OPTIXcell, EquiPlus, INRA96, Bioxcell or AndroMed; Experiment 1a) or egg-yolk based (Biladyl, Green buffer or Triladyl; Experiment 1b) extenders and stored for 48 h at 4 °C. In Experiment 2, split ejaculates (n = five males; replicated six times) were used to directly compare Green buffer, OPTIXcell and Triladyl extenders over 48 h of storage at 4 °C. Ejaculates collected in Experiment 3 (n = five males; replicated five times) were diluted with Green buffer or Triladyl before chilled storage for 48 h at 4 or 15 °C. Sperm kinematics, viability and acrosome integrity were assessed during liquid storage. In Experiment 1a, there was the greatest total sperm motility (TM) in the OPTIXcell group following 24 and 48 h of storage, while in Experiment 1b, there was the greatest TM after 48 h of storage with Triladyl and Green buffer. In Experiment 2, there were greater TM and viable acrosome intact spermatozoa in the Triladyl and Green buffer than with OPTIXcell group. In Experiment 3, there was a greater TM in the Triladyl than Green buffer group at 24 and 48 h of storage regardless of storage temperature (which had no effect on sperm quality). In conclusion, camel sperm have greater viability when preserved in liquid form for 48 h following dilution with Triladyl and storage at either 4 or 15 °C.

Genetic parameters of the thick-to-thin albumen ratio and egg compositional traits in layer-type chickens.

1. Generating a robust egg albumen is one of the major factors contributing to interior egg quality owing to its nutritive value and superior appearance. However, the genetic factors regulating the proportion of thick albumen are poorly understood. 2. In this study, 1330 eggs were collected from 450 Rhode Island White layers, aged 40 weeks, to measure egg compositional traits for three successive days. The restricted maximum likelihood method was applied to estimate genetic parameters for the thick-to-thin albumen ratio and other egg compositional traits. A univariate animal model was fitted to calculate heritability for each trait. 3. The heritabilities of egg weight, yolk weight, albumen height, Haugh units, percentages of yolk, thick albumen, thin albumen and the thick-to-thin albumen ratio were 0.32, 0.34, 0.28, 0.47, 0.61, 0.39, 0.31, and 0.45, respectively. The percentage of thick albumen was negatively correlated genetically with all traits, and percentage of thin albumen was negatively correlated genetically with all traits except for Haugh units. The thick-to-thin albumen ratio was positively correlated genetically with egg weight, albumen height and Haugh units, with correlations ranging from 0.21 to 0.54. 4. The results indicated that the percentage of thick albumen and the thick-to-thin albumen ratio were found to be moderately to highly heritable, and selection for the thick-to-thin albumen ratio could be conducive to the improvement of egg albumen quality.

Bulk Actin Dynamics Drive Phase Segregation in Zebrafish Oocytes.

Segregation of maternal determinants within the oocyte constitutes the first step in embryo patterning. In zebrafish oocytes, extensive ooplasmic streaming leads to the segregation of ooplasm from yolk granules along the animal-vegetal axis of the oocyte. Here, we show that this process does not rely on cortical actin reorganization, as previously thought, but instead on a cell-cycle-dependent bulk actin polymerization wave traveling from the animal to the vegetal pole of the oocyte. This wave functions in segregation by both pulling ooplasm animally and pushing yolk granules vegetally. Using biophysical experimentation and theory, we show that ooplasm pulling is mediated by bulk actin network flows exerting friction forces on the ooplasm, while yolk granule pushing is achieved by a mechanism closely resembling actin comet formation on yolk granules. Our study defines a novel role of cell-cycle-controlled bulk actin polymerization waves in oocyte polarization via ooplasmic segregation.

Boat noise affects the early life history of two damselfishes.

Anthropogenic noise can have a negative effect on the physiology and survival of marine fishes. Most research has focused on later life-stages, and few studies have investigated the effects of human-induced noise on embryogenesis. The current study investigated whether playback of motorboat noise affected the embryogenesis of the coral reef damselfishes, Amphiprion melanopus and Acanthochromis polyacanthus. Embryos reared under the playback of boat noise had faster heart rates compared to the ambient reef controls. The effects of noise on morphological development differed between species and the fundamental interrelationships between early life history characteristics changed dramatically under boat noise for Ac. polyacanthus. Noise treatments did not alter the survival rates of embryos under laboratory conditions. Although species specific, our findings suggest that anthropogenic noise causes physiological responses in fishes during embryogenesis and these changes have direct impacts on their development and these alterations may have carry-over effects to later life stages.

Are there synergistic or antagonistic effects of multiple maternally derived egg components on offspring phenotype?

Eggs are 'multivariate' in that they contain multiple maternally derived egg components (e.g. hormones, antibodies, mRNA, antioxidants) which are thought to influence offspring phenotype. However, most studies have focused on single egg components and on short-term effects. Here, we simultaneously manipulated two egg components, maternally derived antibodies (MAb) and yolk testosterone, to assess potential synergistic or antagonistic effects on zebra finch offspring phenotype from hatching to sexual maturity. We found no evidence for short- or long-term effects of either MAb or yolk testosterone alone, or their interaction, on hatching mass, size at fledging (tarsus length), body mass at sexual maturity (day 82), chick survival, humoral immune function or any measured female reproductive trait at sexual maturity. There was a positive effect of yolk testosterone, but not MAb, on offspring phytohaemagglutinin (PHA) response at 26 days of age but at 82 days of age, MAb, but not yolk testosterone, had a positive effect on PHA response. There was also a MAb×sex interaction on 30 day chick mass, and a positive effect of yolk testosterone on male courtship behaviour at sexual maturity. However, we found no evidence for synergy, i.e. where offspring treated with both MAb and yolk testosterone had higher trait values than offspring treated with either MAb or yolk testosterone alone for any measured trait. Similarly, evidence for antagonistic (compensatory) effects, where offspring treated with both MAb and yolk testosterone had intermediate trait values compared with offspring treated with either MAb or yolk testosterone alone, was equivocal.

Effect of reduced glutathione (GSH) supplementation to Tris-egg yolk extender on chilled semen variables of dogs.

One reason for reduced longevity of chilled dog semen is oxidative stress. The antioxidant glutathione (GSH) improves viability of frozen-thawed dog sperm, but its effect on chilled dog semen has not been investigated. An experiment consisting of two parts was performed: Sperm rich fractions, SRF, were split, diluted with a Tris-egg yolk (TEY) extender containing 0, 5 or 10 mM GSH and stored at 4 °C for 10 days (Part 1; n = 19) or 4 days (Part 2; n = 11), respectively. For Part 1 of the study, percentage (%) of motile, viable, morphologically abnormal spermatozoa and % acrosomal deviations were assessed on days 0, 1, 2, 4 and 10 after dilution. For % sperm motility, samples from all three aliquots of each SRF (0/5/10 mM GSH) were pipetted simultaneously and analysed in a randomised order (time point of analysis, TPA). In Part 2 of the study, motility analysis was performed during 4 days storage and samples were analysed immediately after pipetting (part 2). Most investigated parameters were affected by storage time. For motility variables, there was an effect of GSH identified for circular, CM (ANOVA, Part 1: P = 0.05, Part 2: P < 0.0001) and local motility, LM (ANOVA, Part 2: P = 0.004). Furthermore, there was a trend for an interaction between time and sperm treatment for CM (Part 2: P = 0.077). In conclusion, in the present study there was not an overall positive effect of GSH addition (5/10 mM) on sperm motility in chilled dog semen samples that were characterised to be of good quality during 4- and 10-days of storage.

Dialysis of the goat semen and its effect on the quality of frozen/thawed spermatozoa processed in the presence of egg yolk.

The aim of the present study was to analyze the effect of dialysis on the quality of frozen/thawed buck semen. Ejaculates (n = 15) from three Saanen bucks were divided into three experimental groups. Semen in Group Ce (centrifugation) was processed by standard method and washed two times at 1.085 × g for 20 min. During this time, the diluted control semen (Co) was stored at room temperature. Semen in Group D was dialyzed using 300 kDa cut-off semi-permeable cellulose tubing. The semen from all groups was diluted with extender containing 20% egg yolk and frozen in liquid nitrogen vapor. After thawing, semen samples were evaluated by microscopic and biochemical analyses. Phospholipase A2 was in amounts that was 72.0 ± 11.7% less after dialysis and 21.3 ± 10.0% less after washing with centrifugation compared to the control semen (P < 0.05). Spermatozoa from Group Co had a lesser motility and viability and greater percentage of morphological abnormal spermatozoa in comparison to Groups D and Ce at 3 h after thawing and incubation on 37 °C. At the same time motility and percentage of HOST positive spermatozoa were greater in Group Ce compared with D (P < 0.05). There, however, was no difference in morphology and viability (CFDA/PI analysis) of spermatozoa between Ce and D group. Results from the present study suggest the dialysis is the promising alternative method for reducing phospholipase A2 in the buck semen before cryopreservation.

Roles of the sire and dam quail in egg, yolk, albumen, and shell weight alterations due to the parthenogenetic trait.

Infertile eggs from Chinese Painted quail exhibit parthenogenesis, and the 1st egg in a clutch sequence is more likely to develop a parthenogenetic embryo. Also, hens that exhibit parthenogenesis have shorter clutches and heavier egg weights. These larger eggs may be a result of the egg residing in the oviduct longer, allowing more time for the parthenote to develop. However, it is unknown which components of the egg are altered due to the parthenogenetic trait or the role of dams and sires from the parthenogenetic line of quails in these alterations. Therefore, our objective was to determine if the parthenogenetic trait in dams, sires, or both contributes to alterations in egg components, such as yolk, albumen, and shell weights. Two selected lines of quail, 1 line selected for parthenogenesis and 1 line that was unselected for the parthenogenetic trait (control), were utilized in a 2 x 2 factorial arrangement of dam and sire lines to create 4 breeding treatments: control dams + control sires (CC), control dams + parthenogenetic sires (CP), parthenogenetic dams + control sires (PC), parthenogenetic dams + parthenogenetic sires (PP). Daily, fresh eggs were collected, labeled, weighed, and the germinal disc was classified as fertile, unfertilized without development, or parthenote. Yolk, albumen, and shell weights were obtained, and their weights were also calculated as percentage of total egg weight. A dam main effect revealed heavier weights for total egg, yolk, albumen, and shell as well as a larger percentage of albumen and ratio of albumen to yolk in parthenogenetic line dams versus control line dams. However, the percentage of yolk was higher for control than parthenogenetic line dams. The increase in total egg and egg component weights due to the parthenogenetic trait suggests that the transit time of the egg through the oviduct is altered. Perhaps transit through the magnum and uterus is delayed the most yielding greater albumen and shell weights, respectively.

LC-MS/MS-Based Metabolome Analysis of Biochemical Pathways Altered by Food Limitation in Larvae of Ivory Shell, Babylonia areolata.

Ivory shell, Babylonia areolata, is one of the commercially important mariculture species in China and South East Asia. Survival varies in the artificial hatching and larval rearing of B. areolata. Food deprivation may be involved in rearing mortality, and so, a better understanding of how larvae respond and adjust to starvation is needed. In this study, the metabolite profiles of newly hatched larvae with yolk (I), larvae with yolk exhaustion (II), larvae suffering 24 h starvation after yolk exhaustion (III), and larvae fed with exogenous nutrients after yolk exhaustion (IV) were analyzed by LC-MS/MS. Principal component and cluster analyses revealed differential abundance of metabolite profiles across groups. When compared to metabolite levels of the I group, significantly up-regulated metabolites included polyunsaturated fatty acids, phospholipids, nucleotide, amino acids, and their derivatives were found in the II group, indicating that organisms relied predominantly on glycerophospolipid metabolism and protein-based catabolism for energy production during this stage. During starvation after yolk exhaustion, the levels of all energy related metabolites were significantly reduced, but an increase in products of purine and pyrimidine metabolism indicated an insufficient energy supply and an increase in cellular disintegration. Larvae fed exogenous nutrients can have significantly improved metabolism compared to starved larvae. These findings suggest that metabolomics, using LC-MS/MS, can be used to assess the physiological status and food-affected metabolic changes affecting B. areolata larvae.