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Vitellogenin (Vtg) serves as the precursor of yolk protein and exhibits widespread distribution in tissues, including in the ovary of both vertebrates and invertebrates. Vtg plays a critical role in facilitating oocyte maturation and embryonic development following oviposition. In this study, we have successfully elucidated the complete transcript sequence of TtVtg6-like from an ancient chelicerate Tachypleus tridentatus. The TtVtg6-like transcript encompassed a length of 4887 bp and encoded 1629 amino acids residues. Notably, TtVtg6-like was found to contain 25 exons. Furthermore, the molecular weight and isoelectric point of TtVtg6-like were determined to be 191.6 KDa and 6.73, respectively. Subsequent mRNA expression analysis demonstrated the specific expression of TtVtg6-like in ovary and yellow connective tissue. In addition, TtVtg6-like was located and distributed in both ovary and yellow connective tissue. Intriguingly, employing an siRNA approach to silence TtVtg6-like resulted in a decrease in TtVtg6-like transcription levels. Concomitantly, TtVtg6-like silencing led to increase production of ROS, ultimately resulting in DNA damage and cell apoptosis within the ovarian primary cell. The induction of apoptosis ovarian primary cells due to TtVtg6-like silencing was further corroborated through TUNEL assay and flow cytometry analysis. Overall, our findings underscore the significance of TtVtg6-like in ovarian cell development, revealing its potential association with ovarian cell apoptosis. Consequently, the insights gained from this study contribute to the future exploration of vitellogenesis and ovarian development in T. tridentatus. • TtVtg6-like exhibited a length of 4887 bp and encoded a protein with 1629 amino acid residues. • TtVtg6-like was found in both ovary and yellow connective tissue. • Silencing TtVtg6-like led to a reduction in its transcription levels, highlighting its role in ovarian cell development. • A potential association between TtVtg6-like and ovarian cell apoptosis, and its significance in reproductive processes. [ABSTRACT FROM AUTHOR]

In the viviparous teleost species belonging to the family Goodeidae, intraovarian embryos absorb maternal supplements while they grow during the gestation period. They take up the components via trophotaeniae, a hindgut-derived placental structure. Our previous study using a goodeid species Xenotoca eiseni revealed that intraovarian embryos absorb the yolk protein vitellogenin (Vtg) via the trophotaenia. However, another group indicated yolk components accumulate in the intestinal lumen of X. eiseni embryos. Here, we investigated whether the intestinal duct is capable of protein uptake, as is the trophotaenia. Immunohistochemical studies indicated that endogenous vitellogenin is detected in the intestinal epithelial cells of the intraovarian embryo. Tracer analysis using FITC-Vtg also indicated that intestinal tissues can take up protein. The endocytosis-related genes expressed in trophotaenia were also detected in the intestinal tissues of the embryo. Lipid transporter genes which are not expressed in the trophotaenia were detected in the embryonic intestine. This evidence suggests that the intraovarian embryo of X. eiseni possesses two distinct sites for uptake of the maternal proteins. However, the presumed functions of the embryonic intestine and trophotaenia might be not identical. The study provides a new perspective on how mother-to-embryo matrotrophic interactions have changed in the evolution of viviparous teleosts. • Intestinal ducts can take up vitellogenin in Xenotoca eiseni embryos. • Endocytosis-related genes are expressed in the embryonic intestine. • Lipid transporter genes undetectable in trophotaenia are expressed in intestine. [ABSTRACT FROM AUTHOR]

The present study evaluated the effects of water-soluble curcuminoid-rich extract in a solid dispersion form (CRE-SD) on goat sperm qualities and sperm protein CSNK2A2 expression during liquid storage. Semen was collected from five fertile goats, using an artificial vagina. Ejaculates with a motility above 70% were cooled to 4 °C using TRIS-citric acid-fructose diluent with 10% egg yolk containing various concentrations of CRE-SD (0, 0.1, 1, 10 and 100 μg/mL). Chilled sperm were evaluated for sperm characteristics, casein kinase II catalytic subunit alpha (CSNK2A2) protein level and oxidative status up to 15 days. After 12 days of preservation, sperm motility, viability, acrosomal integrity and mitochondrial activity were significantly higher in the group preserved with 10 μg/mL CRE-SD as compared with the control group. Supplementation of CRE-SD at this concentration was also able to conserve the CSNK2A2 a significantly higher than that in control group until 9 days of cold storage, possibly by reducing oxidative stress. The molecular mass of the sperm CSNK2A2 protein detected in this study was 37 kDa; it was mostly located in the post-acrosomal region, midpiece and flagellum. These results demonstrate the possibility to use the CRE-SD as a natural antioxidant during liquid semen storage in goats. • CRE-SD is easily dissolved in semen extender and has a benefit on chilled sperm quality. • CRE-SD can extend chilled buck sperm longevity to 9 days that is still sufficient for AI. • The 37 kDa-sized CSNK2A2 proteins were detected mainly in sperm flagellum. • CRE-SD can preserve sperm CSNK2A2 during cold storage via oxidative stress reduction. [ABSTRACT FROM AUTHOR]

In many species, prenatal maternal stress leads to long-lasting changes in physiological and behavioural individual characteristics. However, these prenatal effects can be modulated by the postnatal environment and especially by maternal care. The impact of maternal care on the modulation of prenatal maternal stress effects has been investigated mainly in mammalian models but, surprisingly, remains little explored in avian species. In this study, we determined the extent to which the subsequent effects of prenatal maternal stress on offspring growth and emotional responses are modulated by the presence of an adoptive mother during postnatal development using a precocial bird, the Japanese quail, Coturnix coturnix japonica , as our study species. All experimental quail chicks were the offspring of females that experienced an environmental stressor induced by a socially unstable environment during laying and egg production. Previous reports showed that social instability of laying quail increased their offspring's emotional reactivity and that this was related to an increase in yolk testosterone levels in their eggs. In this study, offspring hatched from socially unstable females were raised either with or without adoptive mothers. We found that mothered chicks were lighter during their first 3 weeks of age and that their emotional reactivity was lower (i.e. as shown by increased exploratory behaviours in a novel environment, decreased neophobic responses in the home cage and decreased responses to social separation) relative to nonmothered chicks. Thus, our results highlight that maternal care could counteract the phenotypic consequences of prenatally stress-exposed birds. Our results clearly highlight that even in precocial species the presence of the mother has a fundamental importance in modulating the effects of prenatal conditions on the ontogeny of offspring behaviour and that this might be a key mechanism driving phenotypic plasticity in wild populations. • Maternal care in Japanese quail counteracts prenatal stress effects. • Mothered chicks have a lower level of emotional reactivity than nonmothered ones. • Maternal care's influences are similar in mammals and birds. [ABSTRACT FROM AUTHOR]

"Double-yolk egg-like" Fe 3 O 4 /PBA@PPy nanoreactor is synthesized, where the confinement effect from the cavity, the protective effect from the PPy shell, and the magnetic property were successful integrated together. Therefore, it displayed the high degradation efficiency, low leaching, and rapid separation in Fenton-like reaction. [Display omitted] • k of Fe 3 O 4 /PBA@PPy-10 was 0.38 L·min·mol−1, 5.1 times than that of reference Fe 3 O 4 /PBA-10 in Fenton-like reaction. • The concentration of leached cobalt ions was reduced from 1.47 mg/L to 0.174 mg/L by the protective function from the PPy shell. • 84.5% of the degradation efficiency of the 1st cycle was still maintained after 4 cycles. • The core of Fe 3 O 4 make sure the catalyst can be rapidly separated by a magnet. • Mechanism of Fenton-like reaction is revealed in detail. Peroxymonosulfate (PMS)-based Fenton-like reaction is an effective technique for the pollutant degradation, and the Co-based metal organic frameworks displayed the excellent activity for the PMS activation. Nevertheless, how to further improve the catalytic activity, suppress the leaching of toxic cobalt ions, and realize the rapid separation were still challenges for practical application. In this work, a novel solution was proposed: encapsulating Fe 3 O 4 and Prussian blue analogue (PBA) into the polypyrrole (PPy) shell and constructing a "double-yolk egg-like" Fe 3 O 4 /PBA@PPy as a nanoreactor. In Fe 3 O 4 /PBA@PPy-10, the catalytic performance was remarkably enhanced with the help of confinement effect, and the degradation rate (0.38 L·min·mol−1) was 5.1 times than that of reference Fe 3 O 4 /PBA-10 (0.074 L·min·mol−1). In addition, the concentration of leached cobalt ions was reduced to only 0.174 mg/L by the protective function from the PPy shell. Moreover, the nanoreactor could be magnetically separated from the reaction solution due to the encapsulation of Fe 3 O 4 nanospheres, and 84.5% of activity still preserved after the 4th cycle. The main active species involved in Fe 3 O 4 /PBA@PPy-10 system was 1O 2 , while that in reference Fe 3 O 4 /PBA-10 system was OH. Electron spin resonance analysis and radical trapping experiment revealed that the different catalytic mechanisms were attributed to the confinement effect inside the hollow cavity. This work not only presents a feasible way to prepare rarely-reported double-yolk egg-like nanoreactor, but also provides a new insight to solve the bottlenecks in Fenton-like reaction. [ABSTRACT FROM AUTHOR]

Egg yolk granule is a promising ingredient in the food industry. Recent studies have shown that ultra-high pressure homogenization (UHPH) improved the techno-functional properties of the granule, but they did not evaluate its impact on digestibility. Consequently, this work aimed to evaluate the impact of UHPH-treatment (300 MPa and 4 passes) on granule digestibility by using a static in vitro digestion method. While UHPH destabilized the high molecular weight granule lipoproteins, the treatment did not affect the in vitro digestibility of egg yolk granule since similar protein/peptide profiles were obtained from gel electrophoresis and RP-HPLC analysis. Moreover, the degree of hydrolysis was similar between control and UHPH-treated granules with values ranging from 11.5 to 25.2% for the gastric and intestinal phases, respectively. Finally, apovitellin 3-4, apovitellenin IV and phosvitin were specifically digested during the gastric and intestinal phases. Consequently, modifications induced by UHPH on egg yolk granule were not correlated with in vitro digestion experiments. Nevertheless, this work represents an interesting added-value to support the development of innovative egg yolk granule-based ingredients for the food industry. • UHPH destabilized yolk granule lipoproteins. • The in vitro digestibility of yolk granule was not affected by UHPH. • Apovitellin 3-4, apovitellenin IV and phosvitin were specifically digested. [ABSTRACT FROM AUTHOR]

Egg yolk antibodies (IgY) can be prepared in large quantities and economically, and have potential value as polyvalent passive vaccines (against multiple bacteria) in aquaculture. This study prepared live and inactivated Vibrio fluvialis IgY and immunized Carassius auratus prior to infection with V. fluvialis and Aeromonas hydrophila. The results showed that the two IgY antibodies hold effective passive protective rates against V. fluvialis and A. hydrophila in C. auratus. Further, the serum of C. auratus recognized the two bacteria in vitro, with a decrease in the bacteria content of the kidney. The phagocytic activity of C. auratus plasma was enhanced, with a decrease in the expression of inflammatory and antioxidant factors. Pathological sections showed that the kidney, spleen, and intestinal tissue structures were intact, and apoptosis and DNA damage decreased in kidney cells. Moreover, the immunoprotection conferred by the live V. fluvialis IgY was higher than that of the inactivated IgY. Addition, live V. fluvialis immunity induced IgY antibodies against outer membrane proteins of V. fluvialis were more than inactivated V. fluvialis immunity. Furthermore, heterologous immune bacteria will not cause infection, so V. fluvialis can be used to immunize chickens to obtain a large amount of IgY antibody. These findings suggest that the passive immunization effect of live bacterial IgY antibody on fish is significantly better than that of inactivated bacterial antibody, and the live V. fluvialis IgY hold potential value as polyvalent passive vaccines in aquaculture. • The IgY antibodies against live and inactivated Vibrio fluvialis were prepared. • The two IgY antibodies can activate the innate immune responses in Carassius auratus. • The two IgY antibodies have the passive and passive cross-protective abilities to C. auratus. • The IgY antibody against live V. fluvialis can be considered polyvalent passive vaccine candidates in aquaculture. [ABSTRACT FROM AUTHOR]

Streptococcosis, the most common bacterial disease of fish in recent years, is highly infectious and lethal, and has become an important factor hindering the healthy and sustainable development of aquaculture. Chicken egg yolk antibody (IgY) has the advantages of high antigen specificity, inexpensive and easy to obtain, simple preparation, no toxic side effects, and in line with animal welfare, which is a green and safe alternative to antibiotics. In this study, the potential of specific IgY in the treatment of gastrointestinal pathogens was explored by observing the effects of specific IgY on intestinal flora, pathological tissue, apoptosis, oxidative stress, and inflammatory response of tilapia. We used the specific IgY prepared in the early stage to feed tilapia for 10 days, and then the tilapia was challenged with Streptococcus agalactiae. The results showed that feeding IgY before challenge had a small effect on the intestinal flora, and after challenge specific IgY decreased the proportion of Streptococcus and increased the diversity of the intestinal flora; in histopathology, specific IgY decreased tissue damage and maintained the integrity of tissue structure. Further study found that specific IgY can reduce intestinal epithelial cell apoptosis and reduce caspase activity; at the same time, the content of MDA was decreased, and the activities of SOD, CAT, GSH-Px and GR were increased. In addition, specific IgY can down-regulate the expression levels of IL-8 and TNF-α genes and up-regulate the expression levels of IL-10 and TGF-β. The results of this study showed that specific IgY could improve the intestinal flora of tilapia infected with Streptococcus agalactiae , reduce intestinal cell apoptosis, oxidative stress injury and inflammatory response, thereby reducing tissue damage and protecting the health of tilapia. Overall, specific IgY can be further explored as a potential antibiotic alternative for gastrointestinal pathogen infections. • Specific IgY decreased the proportion of Streptococcus and increased the diversity of the intestinal flora. • Specific IgY decreased tissue damage and maintained the integrity of tissue structure. • Specific IgY reduced caspase activity and reduced intestinal cell apoptosis. • IgY reduced oxidative stress injury and inflammatory response in intestinal. [ABSTRACT FROM AUTHOR]

Offspring from females breeding in competitive social environments are often exposed to more testosterone (T) during embryonic development, which can affect traits from growth to behavior in potentially adaptive ways. Despite the important role of maternally derived steroids in shaping offspring development, the molecular mechanisms driving these processes are currently unclear. Here, we use tree swallows (Tachycineta bicolor) to explore the effects of the maternal social environment on yolk T concentrations and genome-wide patterns of neural gene expression in embryos. We measured aggressive interactions among females breeding at variable densities and collected their eggs at two timepoints, including the day laid to measure yolk T concentrations and on embryonic day 11 to measure gene expression in whole brain samples. We found that females breeding in high-density sites experienced elevated rates of physical aggression and their eggs had higher yolk T concentrations. A differential gene expression and weighted gene co-expression network analysis indicated that embryos from high-density sites experienced an upregulation of genes involved in hormone, circulatory, and immune processes, and these gene expression patterns were correlated with yolk T levels and aggression. Genes implicated in neural development were additionally downregulated in embryos from high-density sites. These data highlight how early neurogenomic processes may be affected by the maternal social environment, giving rise to phenotypic plasticity in offspring. • Aggression and yolk testosterone (T) allocation is greater at high density. • We measured gene expression patterns in embryonic day 11 whole brains. • Genes in circulatory and immune processes correlated positively with density and T. • Genes in neurodevelopmental processes correlated negatively with density. • Effects were observed in both male and female embryos. [ABSTRACT FROM AUTHOR]

[Display omitted] • The absorption property of ZnS/Ni 3 S 4 @GO yolk-shell can be changed by adjusting the thickness of air layer. • The minimum reflection loss of S8 samples at 4.8 GHz is −63.0 dB, and the effective absorption bandwidth (EAB) at 1.5 mm is 4.1 GHz. • The RLC equivalent resonant circuit shows that the yolk-shell structure with different thickness of air layer has different response to different frequency electromagnetic wave. Yolk-shell structure materials with the light weight, excellent impedance matching and electromagnetic wave (EMW) loss ability were widely used in the field of absorbing materials. However, the previous researches on this kind of structure always focused on the comparison between solid structure and empty structure. Different from previous studies, in this paper, the effect of yolk-shell structure with different air layer thickness on EMW absorption was studied for the first time. Graphene oxide (GO) supported yolk-shell ZnS/Ni 3 S 4 absorbers with adjustable air layer were prepared by a simple two-step hydrothermal method. Through the equivalence of RLC resonant circuit and the elimination of the influence of polarization relaxation and conduction loss, it was found that yolk-shell structure with different air layer thickness will resonate with EMW of different frequencies, thus increasing the loss capacity of materials to EMW of this frequency. At the same time, Compared with the solid structure, the yolk-shell structure can not only make the material lighter, but also cause multiple reflections and scattering of EMW. Noteworthy, yolk-shell structure composite material exhibits the maximum reflection loss (RL) of −63.0 dB at 4.8 GHz and an effective absorption bandwidth (EAB) of 4.1 GHz at a thickness of 1.6 mm. This research provides an idea and basis for the design of absorbing materials that respond to different frequency EMW. [ABSTRACT FROM AUTHOR]

While cryopreservation of cauda epididymal sperm (SpCau) allows the preservation of post-mortem bulls' gametes, the process triggers sperm damage. Although improving post-thaw sperm quality, using egg yolk extenders (EY) raises biosafety concerns which forces the use of EY-free extenders (EYFE). Since EYFE are less efficient in preserving post-thaw sperm quality, a strategy for ejaculated sperm (SpEj) frozen with EYFE is to add an Equilibrium Time (ET) step period to the cryopreservation process. However, the ET effect on the quality of SpCau cryopreserved in EYFE remains unknown. Distinct from SpEJ, SpCau physiologically displays cytoplasmic droplets (CDs) in the flagellum that may benefit cell exchange during ET. We hypothesized that using ET in SpCau cryopreserved with EYFE impacts sperm morphofunctional features, CD area, and in vitro fertility ability. Extender nanoparticles were also assessed. Following collection from the cauda epididymis of six Nellore bulls by retrograde flow, SpCau were cryopreserved in EYFE BoviFree® (Minitube, Germany) using three ET protocols: ET0 (no-ET); ET2.5 (2.5 h-ET); and ET5 (5 h-ET). SpCau from ET2.5 and ET5 showed a higher (P ≤ 0.05) percentage of motility and integrity of plasma and acrosome membranes and a smaller (P ≤ 0.05) distal CD area. There are no differences in sperm abnormalities, oxidative stress, capacitation-like events, and in vitro fertility ability. However, a better sperm recovery was found after Percoll® selection for ET2.5 and ET5. Interestingly, the number of nanoparticles in the extender decreased in post-thawed samples. In conclusion, an ET of 2.5 or 5 h is required for an efficient SpCau cryopreservation using an EYFE. [ABSTRACT FROM AUTHOR]

l - carnitine (LC) transports fatty acids to the mitochondria for energy production, reducing lipid availability for peroxidation through β- oxidation. This research examines the effect of LC supplementation to two skimmed milk-based extenders on the cryosurvival of chilled (5°C) and frozen-thawed Peruvian Paso horse spermatozoa.An initial experiment determined the optimal LC concentration (0, 1, 5, 10, 25, and 50 mM) when added to INRA-96® and UHT (skimmed milk + 6% egg yolk) extenders, using nine ejaculates from three stallions chilled for up to 96 h. Subsequently, the effect of 25 mM LC supplementation (the optimal concentration) on chilling (INRA-96) and freezing (INRA-Freeze®) extenders was evaluated using eight pooled samples from sixteen ejaculates (2 ejaculates/pool) from four stallions. Results indicated that all LC concentrations produced significantly higher values (P<0.05) for kinematic variables (total [TM] and progressive motilities, curvilinear [VCL] and straight-line [VSL] velocity, and beat-cross frequency [BCF]), and the integrity of plasma/acrosome membranes (IPIA) compared to non-supplemented chilled sperm samples for up to 96 h with both extenders. Moreover, the use of 25 mM LC was more efficient (P<0.05) in preserving the post-chilled values of velocity, BCF, and IPIA for the long term than lower LC concentrations (1–10 mM). Post-thaw values of total motility, the amplitude of lateral head displacement (ALH), and IPIA were significantly improved (P<0.05) when INRA-Freeze extender was supplemented with 25 mM LC. In conclusion, supplementation of l -carnitine to skimmed milk-based extenders enhanced kinematic variables and protected the membrane integrity in chilled and frozen-thawed Peruvian Paso horse spermatozoa. [ABSTRACT FROM AUTHOR]

Salted egg is made by brining eggs in saturated saline for 30 days. However, producers usually increase the curing temperature of salted eggs from 20 °C to 45 °C to shorten the production cycle by nearly 50%, leading to the increase in the probability of softening salted egg yolk with a declined edible and processing quality. To elucidate the softening mechanism, this study compared the microstructure, texture properties, and composition of key proteins in non-softening salted egg yolk cured at 20 °C (NSSEY) and softening salted egg yolk cured at 45 °C (SSEY). The results showed that SSEY exhibited a higher carbonyl content, a lower free sulfhydryl content, a weaker intermolecular potential, and a smaller size of aggregates, as well as a loose network composed of numerous spheroidal nanoparticles connected in series. Salting with higher temperature also significantly decreased the water holding and protein-lipid binding capacity, with the release of more oil droplets and water. Quantitative proteomics analysis revealed an obvious decrease in the content of apolipoproteins from high-density lipoprotein (HDL) in SSEY, indicating that HDL may play a key role in the formation of SSEY. • Curing temperature significantly changed functional properties of salted duck egg yolk. • 91 different proteins were identified between softening and non-softening salted egg yolk. • Degradation of high-density lipoprotein led to the deterioration of salted duck egg yolk gel at 45 °C. [ABSTRACT FROM AUTHOR]

Hen egg yolk lipids from different regions were determined by ultra-high-performance liquid chromatography with high-resolution mass spectrometry. A total of 15 lipid classes and 346 lipid molecular species were tentatively identified. Principal component analysis showed that the egg lipid profiles varied a certain extent across different production regions. Hierarchical cluster analysis indicated that the egg lipid compositions from Jiangsu and Chungking were relatively similar, but significantly different to those from Inner Mongolia, Jiangxi, and Hainan. Differences in the content of some lipid molecules are obvious, for instance, the concentration of lysophosphatidylcholine (18:1/0:0) in Jiangsu egg and Chungking egg, and phosphatidylinositol (18:0/20:4) in Hainan egg, Changde egg and Jiangxi egg varied significantly from other origins. This study provides a foundation for comprehending the impact of diverse geographical environments on egg lipid profiles. • The lipid profiles of eggs from different origins were analyzed. • 346 molecular species of egg lipids were identified. • Differential markers of various eggs were screened out. • The phospholipid levels of eggs from different origins were significantly different. [ABSTRACT FROM AUTHOR]

During commercial production of liquid egg yolk (LEY), phospholipase A 2 (PLA 2) is used to improve its emulsification capacity and thermal stability. The enzymatic treatment may occur at elevated temperatures such as 50 °C, potentially allowing foodborne pathogens, such as Bacillus cereus, to grow. Little knowledge is available concerning growth of B. cereus in LEY during PLA 2 treatment. Therefore, the objective of this study was to investigate the growth kinetics of B. cereus during PLA 2 treatment using pathogenic B. cytotoxicus NVH391-98, the most thermotolerant member in the B. cereus group, as a surrogate. Inoculated LEY samples were placed in precision programmable incubators to observe the growth of B. cytotoxicus NVH391-98 under multiple isothermal and dynamic temperature conditions between 20 and 53 °C. The bacterial growth was described using the differential Baranyi model coupled with two different secondary models. The kinetic parameters were determined using one-step dynamic inverse analysis of multiple growth curves. The least square method was used in combination with the 4th order Runge-Kutta method to solve the differential Baranyi model using multiple growth curves to determine the cardinal kinetic parameters. The results showed that B. cytotoxicus NVH391-98 can grow prolifically at 50 °C. The estimated minimum, optimum and maximum temperatures were 16.7 or 18.5, 47.8 or 48.1, and 52.1 or 52.4 °C, respectively, depending on the secondary models, with an optimum growth rate of 2.1 log colony-forming-unit (CFU)/g per hour. The dynamic model is validated using isothermal curves with reasonable accuracy. B. cytotoxicus died off slowly at 15 °C. At 55 °C, thermal inactivation was observed, with a D value of approximately 2.7 h. Holding at 55 °C or below 15 °C can effectively prevent the growth of B. cytotoxicus in egg yolk. • B. cytotoxicus is a pathogen that may grow during liquid egg yolk (LEY) production. • The growth of B. cytotoxicus in LEY during Phospholipase A 2 treatment was studied. • B. cytotoxicus may grow rapidly at certain temperatures (30–50 °C) in LEY. • Mathematical models were developed to describe the growth of B. cytotoxicus. • A strategy for managing its growth was identified. [ABSTRACT FROM AUTHOR]

Pine nut skin is a byproduct arising from the pine nut industry, whose composition and bioactive potential are currently unexplored. For this purpose, its characterization is required for the development of sustainable food ingredients. Pine nut skin was separated into finer (<500 μm) and coarser (≥500 μm) fractions, composed of carbohydrates (35 g/100 g and 11 g/100 g, respectively), lignin (47 g/100 g and 72 g/100 g), lipids (8 g/100 g and 9 g/100 g), and protein (7 g/100 g and 3 g/100 g). The lipophilic extract of both fractions had similar amounts of β-sitosterol (30 mg/g), differing in the amount of long-chain aliphatic alcohols, namely nonacosan-10-ol (8 mg/g and 51 mg/g for finer and coarser fractions, respectively) and secondary/secondary alkanediols (18 mg/g and 185 mg/g), known as emulsifier compounds. The composition of pine nut skin fractions indicated their potential in affecting the cholesterol absorption in the intestinal lumen. The lipophilic extract from the finer fraction showed to decrease cholesterol micellar solubilization, and the polysaccharides and lignin had bile salt sequestration properties, rendering this byproduct an hypolipidemic potential. The coarser fraction showed potential as a substitute of egg yolk (3.4 g/100 g dry weight) in a mayonnaise formulation (49 g/100 g oil-in-water emulsion), an emulsifying capacity ascribed to the dietary fiber content and to its lipophilic fraction. From pine nut skin it is possible to obtain low-cost food ingredients with relevance as emulsifiers and potential hypocholesterolemic activity. • Pine nut skin granulometric separation yields 2 fractions with distinct composition. • Fine fraction, rich in β-sitosterol and fiber, is able to sequester bile salts. • Finer fraction has hypolipidemic potential by lowering cholesterol bioaccessibility. • Coarser fraction alkanediols richness explains its high emulsifying capacity. • Pine nut skin coarse fraction can substitute egg yolk in a mayonnaise formulation. [ABSTRACT FROM AUTHOR]

During metazoan early embryogenesis, the intracellular properties of proteins and organelles change dynamically through rapid cleavage. In particular, a change in the nucleus size is known to contribute to embryonic development-dependent cell cycle and gene expression regulation. Here, we compared the nuclear sizes of various blastomeres from developing Xenopus embryos and analyzed the mechanisms that control the nuclear expansion dynamics by manipulating the amount of intracellular components in a cell-free system. Nuclear expansion was slower in blastomeres from vegetal hemispheres during a longer interphase than in those from animal hemispheres. Furthermore, upon recapitulating interphase events by manipulating the concentration of yolk platelets, which are originally rich in the vegetal blastomeres, in cell-free cytoplasmic extracts, nuclear expansion and DNA replication became slower than that in normal yolk-free conditions. Under these conditions, the supplemented yolk platelets accumulated around the nucleus in a microtubule-dependent manner and impeded the organization of the endoplasmic reticulum network. Overall, we propose that yolk platelets around the nucleus reduce membrane supply from the endoplasmic reticulum to the nucleus, resulting in slower nuclear expansion and cell cycle progression in the yolk-rich vegetal blastomeres. [Display omitted] • The nucleus expands slowly in blastomeres from the vegetal side in Xenopus embryos. • The nuclear-to-cell volume ratio is not different between animal and vegetal blastomeres. • Yolk platelets, much in vegetal blastomeres, decrease the nuclear expansion speed. • Yolk platelets accumulate around the nucleus via microtubules-based mechanisms. • The yolk platelets accumulation physically impedes nuclear expansion. [ABSTRACT FROM AUTHOR]

The Toll-Dorsal signaling pathway controls dorsal-ventral (DV) patterning in early Drosophila embryos, which defines specific cell fates along the DV axis and controls morphogenetic behavior of cells during gastrulation and beyond. The extent by which DV patterning information regulates subcellular organization in pre-gastrulation embryos remains unclear. We find that during Drosophila cleavage, the late endosome marker Rab7 is increasingly recruited to the yolk granules and promotes the formation of dynamic membrane tubules. The biogenesis of yolk granule tubules is positively regulated by active Rab7 and its effector complex HOPS, but negatively regulated by the Rab7 effector retromer. The occurrence of tubules is strongly biased towards the ventral side of the embryo, which we show is controlled by the Toll-Dorsal signaling pathway. Our work provides the first evidence for the formation and regulation of yolk granule tubulation in oviparous embryos and elucidates an unexpected role of Toll-Dorsal signaling in regulating this process. • Rab7 is recruited to yolk granules and promotes granule tubulation during cellularization. • Yolk granule tubulation is specifically upregulated at the ventral region. • Toll-Dorsal signaling triggers upregulation of yolk granule tubulation. • Yolk granule tubulation requires microtubules and the Rab7 effector complex HOPS. • The Rab7 effector complex retromer represses yolk granule tubulation. [ABSTRACT FROM AUTHOR]

Hierarchical yolk-shell Co 3 O 4 -C@CoSiO x nanoreactor with multi-active component was synthesized. The degradation efficiencies of CIP reach up to ∼98.2% only in 17 min under the optimal conditions. The fundamental improvement in performance might be due to the synergistic effect of structure and components: (1) hierarchical structure with nanosheet walls and yolk-shell structure endowed the catalyst with high surface area (∼232.47 m2/g) and fully exposed active sites; (2) abundant highly active ≡Co-OH+ were formed on the surface of CoSiO x , which was the primary active species to active PMS; (3) a large amount of oxygen vacancies and nitrogen-doped carbon promoted the decomposition of PMS through a non-radical process. The results revealed both the radical (SO 4 ∙-, ∙OH and O 2 ∙-) and non-radical (1O 2 and direct charge transfer) were the dominated mechanisms of CIP degradation, in which the 1O 2 and SO 4 ∙- mainly contributed to the degradation process. Our work highlights that multi-component catalysts derived from MOFs with special structure have broad application prospects in AOPs. [Display omitted] The reasonable design of the structure and composition of catalysts was essential to improve the catalytic performance of advanced oxidation processes (AOPs). Herein, we reported a simple strategy to synthesize hierarchical Co 3 O 4 -C@CoSiO x yolk-shell nanoreactors with multiple active components by using metal–organic frameworks (MOFs). The novel nanoreactors are further used to activate peroxymonosulfate (PMS) for ciprofloxacin (CIP) degradation. The effects of reaction parameters (pH value, co-existing ions, reaction temperature, etc.) on CIP degradation were systematically investigated. Especially, ∼98.2% of CIP was degraded within 17 min under the optimal conditions, together with the low cobalt leaching and excellent reusability. The appreciable catalytic performance improvement might be due to the synergistic effect of the structure and component design: (1) the hierarchical yolk-shell structure endowed the catalyst with high surface area (∼232.47 m2/g) and fully exposed active sites; (2) abundant highly active ≡Co-OH+ were formed on the surface of CoSiO x ; (3) the presence of oxygen vacancies and nitrogen-doped carbon promoted the decomposition of PMS through a non-radical process. The results revealed both the radical (SO 4 ∙-, ∙OH and O 2 ∙-) and non-radical (1O 2 and direct charge transfer) should be responsible for the CIP degradation. Moreover, the possible degradation pathways of CIP were proposed through the identification of intermediates using LC-MS/MS techniques and density functional theory (DFT) calculation. Our work highlights that multi-component catalysts derived from MOFs with novel structure have broad application prospects in AOPs. [ABSTRACT FROM AUTHOR]

This study was aimed to assess the effectiveness of two methods for cryopreservation of dog epididymal spermatozoa, one by conventional freezing (CF) with shortening both equilibration and cooling times, and the other by ultra-rapid freezing (URF) with nonpermeable cryoprotectant. Sixty epididymides were recovered from thirty orchiectomized adult dogs and the sperm samples were retrieved by retrograde flushing using TCG-EY (tris, citric acid, glucose + 20% egg yolk) extender and then 20 pools were conformed. Each pool was divided into 2 aliquots and then cryopreserved by CF and URF methods respectively. The CF method maintained the cooled-pool samples for 2h (1h without and 1h with 5% glycerol) and then were frozen by liquid nitrogen (LN 2) vapors for 2 min. The URF method cryopreserved the cooled-pool samples using TCG-EY+250 mM sucrose, equilibrating during 30 min (5 °C) and submerging 30-μL drops directly in LN 2. The results showed that the URF method produced a lower percentage of total and progressive motilities and acrosome integrity (P < 0.05) than the CF method. However, the kinetic variables (curvilinear and straight-line velocities, straightness, linearity, wobble, amplitude of lateral head displacement, and beat-cross frequency) and plasma membrane integrity did not differ (P > 0.05) between both cryopreservation methods. Unlike the URF method, the width, area and perimeter of sperm head were reduced after the CF method (P < 0.05). In conclusion, despite the low motility achieved after the ultra-rapid freezing method, the similar values of kinetic, viability and head morphometric dimensions to those obtained after conventional freezing, suggest that ultra-rapid freezing with sucrose may be a useful alternative for the cryopreservation of canine epididymal sperm. • Conventional freezing with shortening equilibration and cooling times achieved better sperm motility. • Ultra-rapid and conventional freezing yielded similar kinetic and plasma membrane integrity. • Sperm head dimensions were maintained after the ultra-rapid freezing-thawing process. • Width, area, and perimeter of sperm head were reduced after conventional freezing. [ABSTRACT FROM AUTHOR]

In recent decades, biologist have focused on the spatiotemporal regulation and function of genes to understand embryogenesis. It is clear that maternal diet impacts fetal development but how nutrients, like lipids and vitamins, modify developmental programs is not completely understood. Fish are useful research organisms for such investigations. Most species of fish produce eggs that develop outside the mother, dependent on a finite amount of yolk to form and grow. The developing embryo is a closed system that can be readily biochemically analyzed, easily visualized, and manipulated to understand the role of nutrients in tissue specification, organogenesis, and growth. Natural variation in yolk composition observed across fish species may be related to unique developmental strategies. In this review, we discuss the reasons that teleost fishes are powerful models to understand nutritional control of development and highlight three species that are particularly valuable for future investigations: the zebrafish, Danio rerio , the African Killifish, Nothobranchius furzeri , and the Mexican tetra, Astyanax mexicanus. This review is a part of a special issue on nutritional, hormonal, and metabolic drivers of development. • Fish can be used to investigate how nutrients influence embryonic development. • Natural variation in egg yolk composition exists among fish species. • The zebrafish yolk proteome and lipidome has been characterized and can be manipulated. • African Killifish embryos can be used to study the timing of nutrient utilization. • Cave and river-adapted Mexican tetra can be used to study the evolution of yolk metabolism. [ABSTRACT FROM AUTHOR]

We compared the results of using egg yolk plasma (EYP) instead of egg yolk (EY) in a TRIS-based Equex STM Paste freezing extender system for dog semen [25]. We also tested whether the addition of lecithin and catalase to the EYP extenders would improve results. Fractionated semen collection was done in 17 stud dogs and the sperm rich fraction diluted with different extenders in 2 steps: (I) TRIS-fructose-citric acid extender (TRIS) containing 20% egg yolk (EY) and 3% glycerol [25], (II) TRIS containing 20% egg yolk plasma (EYP) and 3% glycerol, and (III) TRIS containing 20% EYP and 0.8% lecithin (EYP-L) and 3% glycerol. After equilibration the second dilution step was done: samples with (I) were diluted with TRIS-EY with 7% glycerol and 1% Equex STM paste [25]; samples with (II) and (III) were divided in 2 aliquots each, and one part diluted with TRIS-EYP or TRIS-EYP-L, both containing 7% glycerol and 1% Equex STM paste, and the other one part with the same extenders containing additionally 300 I.U./mL catalase. After freezing and thawing, samples were analyzed by CASA and sperm chromatin structure assay (SCSA); reactive oxygen species (ROS), degree of apoptosis and zona binding ability were determined. Semen samples with TRIS-EY with a final concentration of 5% glycerol and 0.5% Equex STM paste [25] showed best post thaw progressive motility (P), most intact cells, lowest percentage of ROS, acrosome damages, dead and apoptotic cells. Curvilinear velocity (VCL), DNA fragmentation, morphological abnormalities and zona binding ability did not differ between groups. Replacement of egg yolk by EYP increased the ROS and late apoptotic cells. Addition of lecithin and catalase to EYP containing extenders decreased motility and increased complete apoptosis. We conclude that egg yolk is superior to EYP in the here investigated extenders. The TRIS-based extender [25] with EYP could not be improved by addition of lecithin and catalase; however, in-vivo fertilization capacity of the here examined extenders remains to be investigated. • TRIS-EY + Gly + Equex extender was superior regarding progressive motility, intact and live cells. • TRIS-EY + Gly + Equex was superior regarding acrosome damages and ROS. • Replacement of EY by EYP increased the ROS and late apoptotic cells. • Lecithin and catalase in TRIS-EYP + Gly + Equex decreased motility and increased complete apoptosis. • There was no difference between extenders regarding VCL and zona binding. [ABSTRACT FROM AUTHOR]

In this study, the effects of polyamines, Spermine and Spermidine, on long-term preservation and post-thaw spermatological parameters were evaluated. Moreover, determination of the most suitable polyamine and its dose that can be added to standard extenders were aimed. Four adult Arabian stallions were used in the study. Five ejaculates were collected from each of four stallions via artificial vagina two days interval. Each ejaculate was divided into 13 aliquots. INRA96 (95,5%), egg yolk (2%), and glycerol (2,5%) were used as a control extender. Extenders of experimental groups were prepared with different doses of Spermine and Spermidine (0,1 mg/ml; 0,2 mg/ml; 0,4 mg/ml; 1 mg/ml; 2 mg/ml; 4 mg/ml). Stallion semen that were cryopreserved with Control and experimental extenders were evaluated in terms of Total Motility, Progressive Motility, Plasma Membrane Integrity, Capacitation Index, Acrosome Integrity and DNA Fragmentation Index. At the end of the evaluations, it was determined that 0,2 mg/ml Spermine and 0,4 mg/ml Spermidine showed better Total Motility and Progressive Motility, numerically. On the other hand, it was observed that 4 mg/ml Spermine and Spermidine had the lowest statistically significant values (p < 0,001). While statistically similar differences were obtained between groups in terms of the Plasma Membrane and Acrosome Integrity, it was determined that all experimental groups had lower and statistically significant values in terms of Capacitation and DNA Fragmentation Index (p < 0,001). As result, it was observed that the stallion semen cryopreservation success can be increased by the addition of 1–2 mg/ml Spermine that had effective protection on Capacitation and DNA Fragmentation Index without damaging other spermatological properties. [ABSTRACT FROM AUTHOR]

Addressing the establishment of biobanks for the conservation of wild hystricomorph rodents' germplasm, we verified the effects of different extenders and distinct concentrations of non-permeant cryoprotectants on the sperm parameters of Spix's yellow-toothed cavies. Nine testis-epididymis complexes were used for sperm collection by retrograde washing using Tris or a powdered coconut water extender (ACP®-116c). Spermatozoa were diluted and frozen with the same extenders supplemented with egg yolk or Aloe vera at a 10% or 20% concentration. After recovery and cryopreservation, all samples were evaluated for sperm kinetic parameters, morphology, membrane integrity, osmotic response, and sperm-binding capability using an egg yolk perivitelline membrane assay. After recovery, no differences were observed between Tris and ACP®-116c that provided 515.4 × 106 sperm/mL and 561.6 × 106 sperm/mL, presenting >65% motile sperm, respectively. After cryopreservation, most effective preservation of sperm kinetic parameters (68.1 ± 5.9% motile sperm) and membrane integrity (48.2 ± 7.4%) was provided by Tris extender supplemented with 10% egg yolk. However, both extenders supplemented with any concentration of egg yolk or Aloe vera presented similar preservation of osmotic response and sperm-binding ability after cryopreservation. In summary, we suggest the use of a Tris extender supplemented of 10% egg yolk for cryopreservation of Spix's yellow-toothed cavy epidydimal sperm. [ABSTRACT FROM AUTHOR]

Cryopreservation consist of a set of methods to preserve cells and tissues by drastically reducing the temperature. Among some undesired effects, cryopreservation might generate reactive oxygen species that lead to an increase of oxidative stress, causing damage to cells. This study aimed to test taurine, cysteine, and melatonin on the freezing of Prochilodus brevis sperm and assess its effects on post-thawed sperm quality. Sperm was collected and seven pools were formed (n = 7). They were diluted (1:9) in standard medium (5% glucose, 10% dimethyl sulfoxide and 5% egg yolk) supplemented or not (control) with taurine (0.3, 1.0, 3.16 or 10.0 mM), cysteine (0.3, 1.0, 3.16 or 10.0 mM) or melatonin (0.6, 1.12, 2.0 or 3.56 mM). Post-thawed sperm was evaluated for kinetic (total motility, velocities, and percentage of rapid cells), morphology and membrane and DNA integrity. Differences were found when melatonin was used as an antioxidant. For the variables rapid sperm and sperm velocities, 3.56 mM melatonin presented higher results than the control (melatonin 0 mM). Melatonin 2 mM was similar to 3.56 mM on rapid sperm, average path velocity (VAP) and curvilinear velocity (VCL). No difference was found between concentration 0 mM (control) and taurine treatments. As for cysteine, 0.3 mM presented the best results for rapid sperm than 10 mM, and higher VCL and VAP than 1 mM. Melatonin 3.56 mM presented higher results on kinetic parameters (rapid motility, VCL, VSL and VAP) than other tested antioxidants. Therefore, melatonin 3.56 mM is recommended to be added to the sperm freezing medium of P. brevis. • Supplementation with melatonin improves the post-thawing sperm quality of P. brevis • Taurine, at the tested concentrations, did not influence the analyzed variables • The lowest concentration of cysteine showed better results than the highest • Melatonin presented better results for some variables than other antioxidants [ABSTRACT FROM AUTHOR]

Sperm quality is preserved through the crucial involvement of antioxidants, which play a vital role in minimizing the occurrence of reactive oxygen species (ROS) during the cryopreservation process. The suitability of the type and concentration of antioxidants are species-dependent, and this study is crucial in order to improve the quality of the climbing perch sperm post-cryopreservation. Therefore, this study aimed to determine the best type and concentration of antioxidants for cryopreservation of climbing perch Anabas testudineus sperm. To achieve this, 6 types of antioxidants, namely, ascorbic acid, beta-carotene, glutathione, butylated hydroxytoluene (BHT), myo-inositol, and alpha-tocopherol, with inclusion of a control were tested in 3 replications at three concentration levels of 0 mg/L (control), 20 mg/L, 40 mg/L, and 60 mg/L. Sperm was diluted in a glucose-base extender at a ratio of 1:60 (sperm: glucose base), then 10 % DMSO and 5 % egg yolk was added before cryopreservation for two weeks. The results showed that the type and concentration of antioxidants had a significant effect on the motility and viability of cryopreserved climbing perch sperm (P < 0.05), where the best results for ascorbic acid, beta-carotene, glutathione, myo-inositol, and alpha-tocopherol were obtained at a concentration of 60 mg/L, while BHT was at a concentration of 20 mg/L. The best results for glutathione, myo-inositol, and alpha-tocopherol were significantly different from other treatments, while the best results for ascorbic acid and beta-carotene (60 mg/L) were not significantly different from the 40 mg/L concentration, while the best results for BHT were not significantly different from the control treatments. Therefore, the best concentration of glutathione, myo-inositol, and alpha-tocopherol was 60 mg/L, while for ascorbic acid and beta-carotene it was 40 mg/L, and BHT was not recommended. DNA integrity analysis indicated the absence of fragmentation in all samples, including fresh, control, and treated sperm. Based on practical and economic considerations, myo-inositol at 60 mg/L was recommended for cryopreservation of climbing perch A. testudineus sperm. [ABSTRACT FROM AUTHOR]

• Corticosterone in the yolk of chicken eggs is metabolized early in development. • Two primary metabolites of corticosterone are 5β-corticosterone and 20β-corticosterone. • 5β-reductase and 20-hydroxysteroid dehydrogenase produce these two metabolites. • Transcripts for these two enzymes are expressed in the extraembryonic membranes. • Transcript levels for both genes change over the course of early development. When females experience stress during reproduction, developing embryos can be exposed to elevated levels of glucocorticoids, which can permanently affect offspring development, physiology, and behavior. However, the embryo can regulate exposure to glucocorticoids. In placental species, the placenta regulates embryonic exposure to maternal steroids via metabolism. In a comparable way, recent evidence has shown the extraembryonic membranes of avian species also regulate embryonic exposure to a number of maternal steroids deposited in the yolk via metabolism early in development. However, despite the known effects of embryonic exposure to glucocorticoids, it is not yet understood how glucocorticoids are metabolized early in development. To address this knowledge gap, we injected corticosterone into freshly laid chicken (Gallus gallus) eggs and identified corticosterone metabolites, located metabolomic enzyme transcript expression, tracked metabolomic enzyme transcript expression during the first six days of development, and determined the effect of corticosterone and metabolites on embryonic survival. We found that yolk corticosterone was metabolized before day four of development into two metabolites: 5β-corticosterone and 20β-corticosterone. The enzymes, AKR1D1 and CBR1 respectively, were expressed in the extraembryonic membranes. Expression was dynamic during early development, peaking on day two of development. Finally, we found that corticosterone exposure is lethal to the embryos, yet exposure to the metabolites is not, suggesting that metabolism protects the embryo. Ultimately, we show that the extraembryonic membranes of avian species actively regulate their endocrine environment very early in development. [ABSTRACT FROM AUTHOR]

• Female' incubation behavior may regulate embryo exposure to maternal hormones. • Incubation temperature can affect testosterone metabolization into 5α-dihydrotestosterone or estradiol. • Incubation temperature at 36° C increased concentration of 5α-dihydrotestosterone, but did not affect estradiol titers. • Incubation temperature may constitute an additional pathway for hormone-mediated maternal effects. In birds, maternal hormones deposited into eggs in response to environmental stimuli can impact offspring phenotype. Although less studied, environmental conditions can also influence females' incubation behavior, which might play a role in regulating embryo exposure to maternal hormones through changes in incubation temperature that affect the activity of the enzymes responsible for converting testosterone (T) to 5α-dihydrotestosterone (DHT) or estradiol. Here, we tested the hypothesis that the initial T content of the yolk and incubation temperature determine exposure to T metabolites during early embryo development. In the Japanese quail (Coturnix japonica), we experimentally manipulated yolk T and incubation temperature (38° C versus 36° C) and analyzed DHT and estradiol titers on day four of incubation. We found that eggs with experimentally increased T and those incubated at 36° C showed higher DHT concentration in egg yolk (with no synergistic effect of the two treatments). Estradiol titers were not affected by T manipulation or incubation temperature. Our study suggests that incubation temperature influences DHT titers and may act as an understudied source of maternal influence on offspring phenotype. [ABSTRACT FROM AUTHOR]

This study investigated selenium (Se) levels in poultry feed, chicken (meat and bones), and eggs in Khyber Pakhtunkhwa Province (Pakistan) using inductively coupled plasma mass spectrometry. Selenium levels in all poultry feed, meat, bone, and egg samples were below the permissible limit of 500 µg kg−1 set by the European Food Safety Authority. Selenium level was highest in bones (362 µg kg−1), followed by eggs (150 µg kg−1), feed (96 µg kg−1), and meat (59 µg kg−1). Selenium level differed significantly (P < 0.05) in all quantified samples collected from the study area, except between eggs among the selected districts. In feed, Se levels (116 µg kg−1) were highest in Charsadda samples, while in meat (112 µg kg−1) and bone (413 µg kg−1) in Batkhela samples, and egg (170 µg kg−1) samples of Peshawar. Chicken liver, neck bone, and egg yolk stored the highest Se levels compared to other organs. Daily food intake and health risk indices indicated that chicken meat and eggs were free from Se toxicity. These findings elucidate that Se levels in poultry chicken and eggs in the study sites are safe and pose no human health risk. [Display omitted] • Selenium levels in poultry feed, meat, bone and egg samples were below permissible. • Selenium concentration was highest in bones, followed by eggs, feed, and meat. • Chicken liver, neck bone, and egg yolk stored the highest selenium levels. • All chicken meat and eggs posed no human health risks. [ABSTRACT FROM AUTHOR]

The current study intended to optimize the concentration of Oxyrase in the semen dilutor and to evaluate its effect on freezability of spermatozoa of Sahiwal bulls. Supplementation of Oxyrase at 0.125 IU/mL concentration significantly reduced dissolved oxygen (DO) in the dilutor to 4 ppm in 16–18 min at 35 °C. For supplementation studies, a total of 24 ejaculates were categorized into poor and good ejaculates categories (n = 12 each) based on their initial progressive motility. Each ejaculate was further divided into two aliquotes. The first aliquote was diluted with tris-egg yolk extender without Oxyrase (control group) whereas, in the treatment group, Oxyrase was supplemented at the concentration of 0.125 IU/mL of extender. The parameters evaluated include cholesterol and plasma membrane phospholipids (PMP) at fresh, while IPM, acrosomal and plasma membrane integrity, cholesterol, PMP and oxidative stress parameters like lipid peroxidation (LPO), total antioxidant capacity (TAC) and reactive oxygen species (ROS) were evaluated at pre-freeze and post-thaw stages. The IPM and acrosomal intactness were higher (p < 0.05) in treatment group at post-thaw stage in good ejaculates. Oxyrase supplementation resulted in lower (p < 0.05) cholesterol leakage in both categories and lower (p < 0.05) LPO in good ejaculates at post-thaw stage. No statistical difference in ROS was observed between control and treatment groups at all stages whereas, level of TAC was higher (p < 0.05) in the treatment group compared to control group at post-thaw stage of both categories. Therefore, Oxyrase as an oxygen scavenging agent could preserve the post-thaw quality of Sahiwal bull spermatozoa. [ABSTRACT FROM AUTHOR]

Sustainability of channel catfish, Ictalurus punctatus ♀ × blue catfish, Ictalurus furcatus ♂ hybrid aquaculture relies on new innovative technologies to maximize fry output. Transplanting spermatogonial stem cells (SSCs) from blue catfish into channel catfish hosts has the potential to greatly increase gamete availability and improve hybrid catfish fry outputs. Cryopreservation would make these cells readily accessible for xenogenesis, but a freezing protocol for blue catfish testicular tissues has not yet been fully developed. Therefore, the objectives of this experiment were to identify the best permeating [dimethyl sulfoxide (DMSO), ethylene glycol (EG), glycerol, methanol] and non-permeating (lactose or trehalose with egg yolk or BSA) cryoprotectants, their optimal concentrations, and the best freezing rates (−0.5, −1.0, −5.0, −10 ° C/min until −80 ° C) that yield the highest number of viable type A spermatogonia cells. Results showed that all of these factors had significant impacts on post-thaw cell production and viability. DMSO was the most efficient permeating cryoprotectant at a concentration of 1.0 M. The optimal concentration of each cryoprotectant depended on the specific cryoprotectant due to interactions between the two factors. Of the non-permeating cryoprotectants, 0.2 M lactose with egg yolk consistently improved type A spermatogonia production and viability beyond that of the 1.0 M DMSO control. The overall best freezing rate was consistent at −1 °C/min, but similar results were obtained using −0.5 °C/min. Overall, we recommend cryopreserving blue catfish testicular tissues in 1.0 M DMSO with 0.2 M lactose and egg yolk at a rate of either -0.5 or −1 °C/min to achieve the best cryopreservation outcomes. Continued development of cryopreservation protocols for blue catfish and other species will make spermatogonia available for xenogenic applications and genetic improvement programs. • Effects of cryoprotectants and freezing rates were evaluated. • Optimal concentration varied by cryoprotectant. • 1.0 DMSO with 0.2 M lactose and egg yolk was the best treatment combination. • Freezing rate of −1 °C/min was more effective than −5 or −10 °C/min. • Cryopreservation protocol established for blue catfish Type A spermatogonia cells. [ABSTRACT FROM AUTHOR]

The genetic resources of Indonesia's indigenous Kacang goat require preservation. Artificial insemination is expected to accelerate population increases and preserve genetic resources simultaneously. The present study was the maiden attempt for cryopreservation of Kacang buck sperm. The objectives of this study were to determine whether the supplementation of superior Simmental bull seminal plasma protein in egg yolk-citrate extender could improve the quality of post-thawed Kacang buck sperm, increase conceptions rates, and improve kidding rates. Buck semen was diluted without supplementation (T0) and with supplementation of 2.5 mg (T1) and 5 mg (T2) of Simmental bull seminal plasma protein per mL egg yolk-citrate extender. Extended semen was packed in 0.25 mL straw as cryopreserved frozen semen. Post-thawed semen samples were evaluated for viability, motility, intact plasma membranes, malondialdehyde level, and DNA fragmentation. Estrus was synchronized for sixty Kacang does, which were divided randomly into three groups and inseminated using post-thawed semen. The progesterone serum concentration of the does was measured 7 and 22 days post-insemination to detect ovulation and conception. Pregnancy was confirmed using abdominal palpation at 43 days post-insemination and by observing birth. The T1 group showed the highest (P < 0.05) post-thawed viability, motility, and intact plasma membrane. Conception, pregnancy and kidding rates were also higher in T1 than other treatment groups. In conclusion, the 2.5 mg Simmental bull seminal plasma protein supplementation per mL egg yolk-citrate extender provided the best seminal quality and fertility of post-thawed Kacang buck semen. • Effect of Simmental bull seminal plasma protein in egg yolk-citrate extender on Kacang buck semen fertility. • Present study is maiden attempt for the cryopreservation of Kacang buck sperm. • Artificial insemination is efficient to faster propagation and germplasm conservation in Indonesia's indigenous Kacang goats. • Supplementation of Simmental bull seminal plasma proteins enhanced quality and fertility of post thawed Kacang goat sperm. [ABSTRACT FROM AUTHOR]

Edwardsiellosis, an extremely harmful disease can be caused by Edwardsiella tarda , severely restricts the development of turbot (Scophthalmus maximus) farming worldwide, especially in China. This study aimed to establish an effective and feasible prophylaxis by feeding chitosan-alginate coated egg yolk immunoglobulin (IgY) against E. tarda 2CDM001 infections in the process of turbot farming. Enzyme-linked immunosorbent assays proved that the obtained specific IgY could specifically target E. tarda 2CDM001 and five other E. tarda isolates (1a5p, Hz-s, 1a1s, fs-a1 and 58p8). In-vitro, the bacteriostatic effects of specific IgY showed dose dependencies at concentrations ranging from 1 to 10 mg/mL. Moreover, E. tarda 2CDM001 incubated with 10 mg/mL specific IgY could induce the destruction of cell wall structures and significantly decrease the bacterial surface hydrophobicity (p < 0.05). In this study, turbots were challenged with 107 CFU E. tarda 2CDM001 after seven days of continuous feeding with basal diets containing microencapsulated IgYs. Survival rates of the 5%, 3% and 1% microencapsulated specific IgY groups were 63.3%, 56.7% and 20% on the tenth day post infection, respectively, while the turbots in the positive control and non-specific IgY groups all died within ten days. Oral administration of basal diets containing 5% microencapsulated specific IgY significantly reduced IL-1β, IL-8, TNF-α and C3 transcript levels in the head kidney and spleen of turbots compared with the positive and non-specific IgY groups at 24 h after E. tarda 2CDM001 challenging (p < 0.05). Pathological increase of leukocytes in the specific IgY group was significantly lower than that in the positive control and non-specific IgY groups (p < 0.05), decreasing slowly after 24 h of infection and showing a recovery trend. Erythrocyte counts and hemoglobin concentrations of turbots in positive and non-specific IgY groups showed a marked decrease compared with the negative and specific groups at 96 h after E. tarda 2CDM001 infection (p < 0.05). These results suggest that passive immunity via feeding microencapsulated specific IgY could be used as a valuable preventative in turbot against E. tarda 2CDM001 infections. • Specific IgY could effectively inhibit the growth of E. tarda 2CDM001 in vitro. • Pre-feeding chitosan-alginate coated specific IgY could improve the survival rate of E. tarda 2CDM001-infected turbots. • Specific IgY could significantly reduce the inflammatory responses of turbots after E. tarda 2CDM001 infection. • CRediT authorship contribution statement. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the effects of different concentrations of soybean lecithin (SL; 0.5%, 1%, and 1.5%) and egg yolk (EY) in Tris-based extenders on the semen quality parameters of post-thawed goat semen. Sixteen ejaculates were collected from eight healthy, mature Chongming White goats (3–5 years of age). Each ejaculate was divided into five equal aliquots, and then each pellet was diluted with one of the five Tris-based extenders containing 20% EY, 0.5% SL, 1% SL, 2% SL, or 3% SL. The cooled diluted semen was loaded into 0.5 mL polyvinyl French straws and cryopreserved in liquid nitrogen. Frozen semen samples were thawed at 37 °C and assessed for sperm motility, viability, plasma acrosome integrity, membrane integrity, and mitochondria integrity, and the spermatozoa were assessed for reactive oxygen species (ROS), superoxide dismutase (SOD), and malondialdehyde (MDA). The semen extended in the 2.0% SL extract tended to have a higher sperm viability (57.44%), motility (52.14%), membrane integrity (45.31%), acrosome integrity (52.96%), and mitochondrial activity (50.21%) than the other SL-based extender concentrations (P < 0.05). The 2.0% SL treatment group was equivalent to the semen extended in 20% EY (P > 0.05). The extenders supplemented 20% EY or 2.0% SL significantly increased the SOD activity and decreased the ROS and MDA activities compared to the other groups (P < 0.05). In conclusion, the extenders supplemented with 20% EY and 2.0% SL had similar effects on spermatozoa preservation. These results indicate that a soybean lecithin-based diluent may be used as an alternative extender to egg yolk for the cryopreservation of goat semen. • A tris-based extender containing SL for goat semen cryopreservation is advantageous. • The performance of 2% SL supplemented extender was superior to any of the other concentrations of SL. • 2% SL supplemented extender was similar and even better than the 20% egg yolk extender. [ABSTRACT FROM AUTHOR]

Avian leukosis virus (ALV) is one of the main causative agent of tumor development, which brings enormous economic losses to the poultry industry worldwide. ALV can be transmitted horizontally and vertically, and the latter often give rise to more adverse pathogenicity. However, the propagation and evolution of ALV underlying vertical transmission remain not-well understood. Herein, an animal model for the evolution of variants of ALV subgroup J (ALV-J) in the vertical transmission was built and different organs from infected hens and plasma from their ALV-positive progenies were collected, and then three segments in the hypervariable regions of ALV (gp85-A, gp85-B, LTR-C) were amplified and sequenced using conventional Sanger sequencing and MiSeq high-throughput sequencing, respectively. The results showed that the genomic diversity of ALV-J occurred in different organs from ALV-J infected hen, and that the dominant variants in different organs of parental hens, especially in follicle, changed significantly compared with original inoculum strain. Notably, the dominant variants in progenies exhibited higher homologies with variants in parental hens' follicle (88.9%–98.9%) than other organs (85.6%–91.1%), and most consistent mutations in the variants were observed between the progenies and parental hen's follicle. Furthermore, HyPhy analysis indicated that the global selection pressure value (ω) in the follicle is significantly higher than those in other organs. In summary, an animal model for vertical transmission was built and our findings revealed the evolution of variants of ALV in the process of vertical transmission, moreover, the variants were most likely to be taken to the next generation via follicle, which may be related to the higher selection pressure follicle underwent. • An animal model for vertical transmission of ALV-J was built. • Significant differences of the dominant variants of gp85-B were observed between follicle and other organs of parental hens. • The dominant variants in the progeny had higher homology with that in the follicle of the parental hens. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate the effects of different concentrations of catalase in a TRIS-egg yolk extender on sperm quality and embryonic development after in vitro fertilization of frozen-thawed bull sperm. For this purpose, from each of 7 bulls 2 ejaculates were collected and diluted with a TRIS-egg yolk extender containing 0, 5, 10, 15 or 20 IU catalase/mL. Sperm quality was evaluated 0, 3, 6, 12 and 24 h after thawing by using computer assisted analysis of motility and by flow cytometric assays. Embryonic development was determined after in vitro fertilization of bovine oocytes. Semen diluted with TRIS-egg yolk extender containing different concentrations of catalase showed more motile sperm, more sperm with intact plasma membranes, acrosomes and DNA, a high mitochondrial membrane potential, a high esterase activity, a low calcium level, a lower amount of synthesis of reactive oxygen species and lower degree of lipid peroxidation of sperm compared to semen frozen without catalase (P < 0.05), but not before 3 h after thawing. There was a dose-response relationship with the most prominent effect of 20 IU catalase/mL. However, the improvement of sperm quality had no effect (P ≥ 0.05) on embryonic development after in vitro fertilization with 20 IU catalase/mL. In conclusion, the addition of catalase to the sperm extender improved sperm quality with no obvious effect on in vitro fertility. [ABSTRACT FROM AUTHOR]

The synergy obtained by the combination of cryoprotectants is a successful strategy that can be beneficial on the optimization of zebrafish sperm cryopreservation. Recently, a protocol was established for this species using an electric ultrafreezer (−150 °C) performing cooling rate (−66 °C/min) and storage within one step. The ultimate objective of sperm cryopreservation is to generate healthy offspring. Therefore, the objective of this study was to select the most adequate cryoprotectant combination, for the previously established protocol, that generate high quality offspring with normal skeletogenesis. Among the permeating cryoprotectant concentrations studied 12.5% and 15% of N , N -dimethylformamide (DMF) yielded high post-thaw sperm quality and hatching rates. For these two concentrations, the presence of bovine serum albumin (10 mg/mL), egg yolk (10%), glycine (30 mM) and bicine (50 mM) was evaluated for post-thaw sperm motility, viability, in vitro fertilization success and offspring skeletal development (30 days post fertilization). Higher concentration of permeating cryoprotectant (15%) decreased the incidence of deformed arches and severe skeletal malformations, which suggests higher capacity to protect the cell against cold stress and DNA damage. Extender containing 15% DMF with Ctrl, Bicine and egg yolk were the non-permeating cryoprotectants with higher post-thaw quality. The use of these compounds results in a reduction in vertebral fusions, compressions and severity of skeletal malformations in the offspring. Therefore, these extender compositions are beneficial for the quality of zebrafish offspring sired by cryopreserved sperm with −66 °C/min freezing rate. To the best of our knowledge, this is the first report on skeletal development of the offspring sired by cryopreserved sperm performed with different freezing media compositions in zebrafish. Image 1 • Cryoprotectant composition of extender affect skeletal development of the offspring. • 15% dimethylformamide in extender decrease offspring severe skeletal malformations. • Cryopreservation with 15% of DMF with bicine or egg yolk improve zebrafish post-thaw sperm quality. • Bicine or egg yolk in extender reduce offspring vertebral fusions and compressions. [ABSTRACT FROM AUTHOR]

Polyvalent antibodies can resist multiple bacterial species, and immunoglobulin Y (IgY) antibody can be economically prepared in large quantities from egg yolk; further, IgY polyvalent antibodies have application value in aquaculture. The outer membrane proteins (OMPs) PF1380 and ExbB of Pseudomonas fluorescens were expressed and purified, and the corresponding IgY antibodies were prepared. PF1380, ExbB, and the corresponding IgY antibodies could activate the innate immune responses of chicken and Carassius auratus. The passive immunization to C. auratus showed that the IgY antibodies of PF1380 and ExbB had an immune protection rate, down-regulated the expression of antioxidant-related factors (MDA, SOD, GSH-Px, and CAT) to reduce the antioxidant reaction, down-regulated the expression of inflammation-related genes (IL-6, IL-8, TNF-α, and IL-1β) to reduce the inflammatory reaction, maintained the integrity of visceral tissue structure, and reduced apoptosis and damage of tissue cells in relation to P. fluorescens and Aeromonas hydrophila infections. Thus, the IgY antibodies of PF1380 and ExbB could be considered as passive polyvalent vaccine candidates in aquaculture. • The PF1380 and ExbB of Pseudomonas fluorescens were expressed and purified, and the corresponding IgY antibodies were prepared. • The two OMPs and their IgY antibodies can activate the innate immune responses in chicken and Carassius auratus. • The IgY antibodies of PF1380 and ExbB have passive protective abilities against P. fluorescens and Aeromonas hydrophila infections in C. auratus. • The IgY antibodies of PF1380 and ExbB could be considered as passive polyvalent vaccine candidates in aquaculture. [ABSTRACT FROM AUTHOR]

This study investigated whether the origin of sperm (epididymal vs. ejaculate) affects the cryopreservation efficiency in agouti (Dasyprocta leporina). Five sexually mature agoutis underwent electroejaculation, resulting in obtaining four semen samples. After 15 days, the same animals were euthanized, and through retrograde flushing, sperm samples were obtained from the epididymis tails. In both collection methods, samples were evaluated for sperm parameters (sperm concentration, motility, vigor, membrane integrity, osmotic response, and morphology). Then, samples were diluted in ACP 109c, added with 20% egg yolk, and a final concentration of 6% glycerol. Finally, the samples were packaged in 0.25 mL straws and frozen in liquid nitrogen. After one week, samples were thawed and evaluated in the same way as fresh samples, with the addition of membrane integrity analysis using fluorescent probes (C-FDA/PI) and computerized analysis (CASA). Immediately after obtaining the sperm, samples obtained directly from the epididymis presented higher values (P ≤ 0.05) than those obtained by electroejaculation concerning the parameters of volume, sperm concentration, and total number of sperm (1,398.25 ± 206.0 x106 and 184.5 ± 78.0 x106 sperm). On the other hand, in the classical evaluation of the other sperm parameters and the computerized analysis (CASA) after thawing, such as total motility, no statistical differences were observed between sperm from both origins (ejaculate: 16.7 ± 8.2% and epididymal: 24.8 ± 12.0%, P > 0.05). This demonstrates the possibility of direct application of the cryopreservation protocol for agouti (D. leporina) sperm obtained via the epididymis or ejaculate. • Both methods, eletroejaculation and epididymal flushing, can be used to obtain agouti sperm. • A new step in cryopreservation of agouti sperm was established by using sperm via ejaculate. • Epidydimal retrograde flushing provides larger amount of sperm than electroejaculation in agouti. • Epididymal sperm cryopreservation protocol is applicable to ejaculate samples from agoutis. [ABSTRACT FROM AUTHOR]

Penthiopyrad (PO), a succinate dehydrogenase inhibitor (SDHI) fungicide, poses a potential risk to fish. Here, we investigated the adverse effects of PO on endocrine regulation and reproductive capacity in zebrafish during a 21-d sublethal exposure to PO concentrations ranging from 0.02 to 2.00 mg/L. Following exposure to PO (0.20 and 2.00 mg/L), female-specific effects including follicle necrosis, structural disturbance of the yolk follicle, fusion of cortical follicles appeared in ovarian tissue of adult females, which led to a significant reduction in fertility. Correspondingly, 0.20 and 2.00 mg/L PO led to a marked reduction in the GSI values of females, and 2.00 mg/L PO caused a 31% decline in the proportion of perinucleolar oocytes (PCO) in oocytes. In addition, testosterone (T) level was obviously suppressed and 17β-estradiol (E2) level was increased in females after exposure to 2.00 mg/L PO. Male zebrafish treated with 0.20 and 2.00 mg/L of PO exhibited significant interstitial enlargement, edema in the testes, and reduced diameter of seminiferous tubules, along with a thinner basement membrane. The effects of PO on males were associated with significant increase in E2 level, suggesting that PO has an estrogenic effect on male fish. Greater E2 levels in serum were further supported by increased transcription levels of genes linked to the hypothalamic-pituitary-gonad-liver (HPGL) axis. Notably, transcription levels of cyp19a , er2b , era , and cyp19b was remarkably increased, exhibiting a clear link with variations in E2 levels. Overall, the present study demonstrates that PO induces reproductive impairment in zebrafish by promoting steroidogenesis. [Display omitted] • PO significantly suppressed the synthesis of T and enhanced the production of E2 in female zebrafish. • PO at 0.2 and 2.0 mg/L led to a noteworthy decline in female fertility and GSI value. • PO has estrogenic effects on male zebrafish, leading to increased synthesis of E2 and VTG in males. • PO exposure caused significant changes in transcription levels associated with HPGL axis in zebrafish. [ABSTRACT FROM AUTHOR]

In order to clarify the quality formation process of low-NaCl salted eggs (LSEs) under the dual action of vacuum and salt, changes of physicochemical properties, molecular structure, and microstructures of egg white as well as interior and exterior yolks were systematically studied. Results showed that the salt content of egg whites and egg yolks increased to around 5% and 1% respectively, which were only about half of their traditional counterparts. Exterior yolk exudes more oil than the interior yolk, which was consistent with the observation of a more compact microstructure by scanning electron microscopy (SEM). Rheological testing demonstrated a significant increase in the viscosity of yolks but a decrease of egg whites. At the molecular level, proteins of egg white and yolk had gone through the process of denaturation and aggregation, but their change tendencies of free sulfhydryl group, surface hydrophobicity and secondary structure were slightly different. In summary, vacuum assisted two-stage pickling could effectively shorten the ripening time of salted eggs and reduce salt content without compromising their good quality. [Display omitted] • Vacuum technology can accelerate dehydration and reduce salt content. • The product can be pickled successfully in 15 days with excellent quality. • Salt contents of final egg product were 5% of whites and 1% of yolks. • The quality of salted eggs is formed by osmotic dehydration of salt, protein and lipid. [ABSTRACT FROM AUTHOR]

Frozen egg yolk has the advantages of high nutritional value, convenient transportation, and long-term storage; however, freezing causes gelation of egg yolk, resulting in the deterioration of its functional properties. To minimize freezing-induced changes in the function and structure of egg yolks, the inhibitory effects of d -galactose, trypsin, and a combination of d -galactose and trypsin on freezing-induced gelation of egg yolk were evaluated. The results revealed that d -galactose enhanced the solubility, reduced hardness and viscosity, increased fluidity, and inhibited freeze-induced gelation of egg yolks. Trypsin reduced the turbidity and particle size of egg yolks, further enhanced the polarity and solubility, and partially inhibited freeze-induced gelation. The combination of trypsin and d -galactose exhibited a synergistic effect, with trypsin playing an important role. Overall, this combination exerted a remarkable inhibitory effect on freeze-induced gelation of egg yolk. • d -galactose, trypsin, and their combination improved the properties of frozen yolk. • d -galactose reduced the hardness and increased the fluidity of frozen yolk. • Trypsin reduced the turbidity and particle size of frozen yolk. • The combination (d -galactose and trypsin pretreatment) exhibited synergistic effects. [ABSTRACT FROM AUTHOR]

OptiXcell (OP) was tested as an animal protein-free alternative to an egg yolk-based extender for rhinoceros semen cryopreservation and shorter chilling/equilibration periods were evaluated. Semen was collected from three rhinoceros species: black (Diceros bicornis ; n = 2), white (Ceratotherium simum ; n = 2), and greater one-horned (GOH; Rhinoceros unicornis ; n = 3). Controls were diluted with equine extender (EQ) or OP and equilibrated for 1 h. Treatments were diluted with extender and cooled for 15 min (fast: FEQ; FOP) or not cooled (immediate: IEQ; IOP), prior to cryopreservation. Motility decreased post-thaw (EQ: 50.7 ± 5.2%; OP: 52.9 ± 3.4%) from fresh (82.9 ± 2.9%), was higher in OP than IOP (38.6 ± 4.9%; P ≤ 0.05) and decreased over time (P ≤ 0.05). Post-thaw acrosomal integrity was lower in EQ, FEQ, and IEQ (56.9 ± 0.7; 56.6 ± 4.5; 54.9 ± 2.9%) than OP, FOP, IOP (71.8 ± 4.7; 71.9 ± 3.8; 69.9 ± 4.5%) and fresh (72.6 ± 1.4%; P ≤ 0.05). Progression and viability were lower in EQ (2.8 ± 0.2; 61.9 ± 7.4%) and OP (3.1 ± 0.2; 53.4 ± 6.9%) than fresh (3.7 ± 0.2; 87.2 ± 1.3%), decreased over time (P ≤ 0.05) but not different among treatments (P > 0.05). Morphology did not differ between fresh (75.0 ± 4.9% normal) and any treatment group (70.0–77.8%) or over time (P > 0.05). OptiXcell is comparable to egg yolk-based EQ when used for rhinoceros semen cryopreservation. Furthermore, chilling/equilibration can be reduced with little impact on sperm characteristics. [ABSTRACT FROM AUTHOR]

A study was conducted to establish a sustainable and effective manual freezing technique for cryopreservation of Bangladeshi ram semen. Three diluents and freezing techniques were tested, both as treatment combinations (diluent × freezing technique) and fixed effects (diluent or freezing technique) on post-thaw sperm motility (SM), viability (SV), plasma membrane integrity (SPMI) and acrosome integrity (SAI). Ten rams were selected, based on semen evaluation. Eight ejaculates were used for each treatment combination. Semen samples were diluted using a two-step protocol for home-made Tris-based egg yolk (20%, v/v) diluents: D1 (7% glycerol, v/v) and D2 (5% glycerol, v/v), and one-step for commercial diluent: D3 (Triladyl®, consists of bi-distilled water, glycerol, tris, citric acid, fructose, spectinomycin, lincomycin, tylosin and gentamycin) at 35 °C. Fraction-A (without glycerol) was added at 35 °C, and following cooling of sample to 5 °C (−0.30 °C/min), Fraction-B (with glycerol) was added. The diluted semen samples were aspirated into 0.25 ml French straws, sealed, and equilibrated at 5 °C for 2 h. The straws were frozen in liquid nitrogen (LN) vapour, in a Styrofoam box. The freezing techniques were; One-step (F1): at −15.26 °C/min from +5 °C to −140 °C; Two-step (F2): at −11.33 °C/min from +5 °C to −80 °C, and −30 °C/min from −80 °C-140 °C; and Three-step (F3): at −11.33 °C/min from +5 °C to −80 °C, at −26.66 °C/min from to −80 °C to −120 °C, and at −13.33 °C/min from −120 °C to −140 °C. Two semen straws from each batch were evaluated before and after freezing. The group F3D3 exhibited significantly higher (p < 0.05) post-thaw SM 63.1 ± 2.5%, SV 79.0 ± 2.1% and SPMI 72.9 ± 1.7%, whereas SAI 72.9 ± 1.7% was significantly higher (p < 0.05) in group F3D2. The freezing technique F2 and F3 had significantly higher (p < 0.05) post-thaw sperm values compared to F1. The post-thaw SM and SV were above 50% and 65% with the freezing technique F2 and F3 but differed non-significant. The SPMI 67.6 ± 2.0% and SAI 76.1 ± 1.4% were significantly higher (p < 0.05) with F3. Likewise, the diluent D2 and D3 had significantly higher (p < 0.05) post-thaw sperm values compared to D1. The post-thaw SM, SV and SPMI were above 50%, 65% and 55% with the diluents D2 and D3 but differed non-significant. The SAI 76.1 ± 1.1% was significantly higher (p < 0.05) with D3. We concluded that the use of a simple home-made Tris-based diluent containing 20% (v/v) egg yolk and 5% glycerol (v/v), two-step dilution and a three-step freezing technique is a sustainable and effective method for freezing ram semen. For further validation, the fertility of ewes artificially inseminated with the frozen semen will be observed. • A novel manual ram semen freezing technique was established. • Home – made Tris-based diluent (egg yolk 20% v/v and glycerol 5% v/v) is suitable for ram semen cryopreservation. • Two – step dilution and a three – step freezing technique is an effective for freezing ram semen. [ABSTRACT FROM AUTHOR]

This study assessed the effect of l -carnitine (LC) in sheep semen extenders containing or not egg yolk for cryopreservation in sheep. Two extenders (TRIS-egg yolk or the commercial optiXcell™ IMV medium) were used, totaling six groups: IMV – (0, 5 and 10 mM LC) and TRIS – (0, 5 and 10 mM LC). After the freezing-thawing process and throughout incubation at 38 °C for up to 3 h, several parameters were evaluated: sperm kinetics, hypoosmotic, plasma membrane integrity, capacitation status and lipid peroxidation level. The supplementation of either 5 or 10 mM LC randomly affected some parameters and, overall, TRIS was superior (P < 0.05) than IMV extender. In the LC-groups, IMV had greater (P < 0.05) oxidative stress than TRIS. In conclusion, although LC affected isolated parameters, its supplementation in semen extender had no consistently beneficial effect on freezing-thawing ram sperm. [ABSTRACT FROM AUTHOR]

Egg yolk powder (EYP) is widely used in food processing due to its excellent emulsifying properties, but its poor solubility limits its market application as a powdered beverage. In this study, papain, trypsin, and subtilisin were used to hydrolyze egg yolk, and the effects of enzymatic hydrolysis on the solubility of EYP was investigated. The results showed that subtilisin treatment significantly improved the water dispersibility, powder solubility and stability coefficient of EYP (P < 0.05) by 41.67%, 98.16%, and 66.21%, respectively, while its solubility did not change significantly after the treatment with papain and trypsin. The degree of hydrolysis (DH) was related to the enzyme active site and the treatment with papain and subtilisin resulted in a higher DH. Additionally, the results of sodium dodecyl sulfate-polyacrylamide gel electrophoresis, particle size distribution and scanning electron microscopy proved that the molecular weight of subtilisin-treated EYP was lower than EYP after the other treatments. Moreover, surface hydrophobicity and zeta-potential results indicated that molecular hydrophobicity and the amount of surface charge increased after enzymatic hydrolysis. The hydratability of papain-treated EYP increased to 20.91%. Low-field nuclear magnetic resonance results showed that enzymatic hydrolysis changed the content of bound water in the EYP, resulting in the changes of its ability to combine with water. • Three hydrolyze proteases enzymes can affect the yolk solubility. • Subtilisin treatment significantly improved the solubility of egg yolk powder. • Papain and trypsin treatment did not significantly change the solubility. • The different effects due to the difference in the hydrolysis site of the enzyme. • Hydrophobic groups combined with lipoproteins can reduce the surface hydrophobicity. [ABSTRACT FROM AUTHOR]

Abstract Human enterovirus 71 (EV71) is one of the major pathogens that causes hand-foot-and mouth disease, and there is an urgent need for rapid diagnosis of EV71 virus infection for early antiviral treatment. The aim of this study was to prepare chicken egg yolk immunoglobulin Y (IgY) for the diagnosis of enterovirus type 71 infection. The antibodies were raised by intramuscular immunization of laying hens with inactivated human EV71 and isolated from the egg yolk by multiple steps of polyethylene glycol 6000 extraction. The average concentration of IgY antibody was 26.60 mg/mL during the whole immunization. After the first immunization, the IgY titer gradually increased, and reached the peak on the 55th days. Meanwhile, the use of western blotting test demonstrated that specific IgY binds specifically to capsid proteins VP2 and VP3 of EV71 virus. Furthermore, a facile one-step method based on turn-on fluorescence sensing was developed by using IgY antibodies, which can detect EV71 virus at low concentrations of 104 PFU/mL and was 94.44% coincidence with RT-PCR in 30 clinical samples. These findings indicate that EV71-IgY antibodies are an easily prepared and rich source of antibodies that offers a potential alternative strategy for routine screening of EV71 infection. Highlights • The specific IgY antibody against EV71 was prepared and identified. • A facile one-step method based on turn-on fluorescence sensing was developed. • The proposed method exhibited satisfying sensitivity and selectivity for clinical samples detection of EV71 infection. [ABSTRACT FROM AUTHOR]

Abstract Sperm quality in donkeys (Equus asinus) after freezing thawing is still considered lower than that from other animals, including horses. The aim of this study was to test a new freezing extender supplemented with jenny colostrum on donkey sperm. After thawing, we evaluated sperm motility by means of computer-assisted analysis, viability by SYBR-14 and propidium iodide (PI), membrane functional integrity by HOS-test and acrosome integrity by isothiocyanate conjugated with peanut agglutinin (FITC-PNA) and PI. Ejaculates were collected from five fertile Donkeys. Sperm samples were pooled, diluted and cryopreserved into three experimental extender groups: BotuCrio®, lactose-extender supplemented with egg yolk (20%) and lactose-extender supplemented with jenny colostrum (20%). The results demonstrated that lactose-jenny colostrum samples displayed significantly higher values in almost all parameters evaluated (p < 0.05) compared with the other two extenders after thawing (BotuCrio® and lactose-egg yolk based extender, respectively) –Total Motility, Viability, HOS test, VCL, VSL and VAP. Acrosome status, LIN, STR and WOB despite showing lower values, none of them were statistically significant (p > 0.05). In conclusion, the extender containing jenny colostrum can be successfully used for donkey semen crypreservation and could effectively improve donkey sperm qualities after freezing -thawing. Graphical abstract Schema of the experimental design. Image 1 Highlights • Jenny colostrum includes high concentrations of immunoglobulins, antimicrobial and immunomodulatory factors in its composition. • No studies have described the use of colostrum in extenders for freezing semen. • The results for the Jenny Colostrum samples were better than the results for the other two extenders compared. • Jenny colostrum in extenders improves quality and could be effectively used for freezing donkey semen. [ABSTRACT FROM AUTHOR]

Abstract Hen egg is commonly used in bakery products, such as cakes, bread, and cookies, which are all abound with starch. Egg yolk lipids (EYL) with different polarity can affect the structure and properties of starch, as well as steamed bread-making performance. EYL were mainly divided into neutral lipids, cholesterol and phospholipids with increasing polarity by thin layer chromatography. Results from Rapid Visco Analyzer showed that both cholesterol and phosphatidylcholine (PC) promoted gelatinization of wheat starch, but neutral lipids did not. Texture analysis indicated that hardness of wheat starch gels significantly decreased only with PC, caused by an increasing in the spacing between double helical cross-links as characterized by differential scanning calorimetry. Furthermore, a single helix structure of amylose-PC complex was confirmed by molecular simulation and X-ray diffraction. Finally, yolk can be used to soften the firmness of starch-based food such steamed bread. Highlights • Yolk lipids were separated into neutral lipids, cholesterol and phosphatidylcholine. • Cholesterol and phosphatidylcholine increased pasting parameters of wheat starch. • By the increase of phosphatidylcholine level, the hardness of starch gels was decreased. • Phosphatidylcholine-containing gels had larger crosslink spacing between starch polymers. • Phosphatidylcholine softened the crumb of steamed bread. [ABSTRACT FROM AUTHOR]

The effects of hyperbaric storage (HS) at 200 MPa on the microbial, chemical, physical and techno-functional properties of liquid egg yolk were investigated and compared to refrigeration. Inoculated Salmonella enterica (3.35 ± 0.12 logCFU g−1) and Staphylococcus aureus (2.78 ± 0.19 logCFU g−1) resulted below the detection limit after 24 and 48 h HS, respectively. Liquid egg yolk oxidative status remained unaffected for up to 28 days under pressure, probably due to the presence of egg yolk antioxidants. The decrease in egg yolk denaturation enthalpy and the increase in aromatic amino acid exposure indicated that egg yolk proteins unfolded according to structural changes other than those observed during refrigeration. Liquid egg yolk viscosity progressively increased during HS, eventually leading to gelling. Protein modification did not affect liquid egg yolk foaming and emulsifying properties but impaired its solubility and thermally-induced gelling. HS could be used for microbial decontamination of liquid egg yolk while maintaining oxidative stability and the typical capacity to stabilize foams and emulsions. [Display omitted] • Hyperbaric storage at 200 MPa (HS) inactivated S. enterica and S. aureus within 48 h. • HS caused protein unfolding and led to egg yolk gelling. • HS decreased egg yolk solubility with no effect on foaming and emulsifying properties. • Egg yolk oxidative status was almost unaffected during HS for 28 days. [ABSTRACT FROM AUTHOR]