Yolk Protein Research Articles

Effect of high-hydrostatic pressure on the digestibility of egg yolk and granule.

The impact of high hydrostatic pressure (HHP) on protein digestibility of egg yolk and egg yolk granule was evaluated by static in vitro digestion using the standardized INFOGEST 2.0 method. The degree of hydrolysis (DH) and the phospholipid content were determined during digestion, and the protein and peptide profiles were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and reverse phase-high pressure liquid chromatography (RP-HPLC). The results showed that HHP induced protein aggregation in egg yolk and granule, mainly by disulfide bridges, which were not disrupted in the oral phase. Proteolysis during the gastric phase improved egg yolk and granule protein solubility, regardless of whether HHP was applied. However, the extent of the samples' digestibility was not affected, with DH values ranging from 15% to 20%. During the intestinal phase, the DH of egg yolk protein (∼40%) was higher than that of the granule (∼25%), probably due to the denser structure of the granule reducing the accessibility of intestinal enzymes. The DH, peptide, and protein profiles of control and HHP-treated egg yolk showed similar protein digestion behaviors for both gastric and intestinal phases. Among the different proteins, only the digestibility of β-phosvitin in HHP-treated granule was enhanced. Consequently, applying HHP to granules represents an interesting process that improves the digestibility of phosvitin with the potential to generate bioactive phosvitin-derived phosphopeptides. PRACTICAL APPLICATION: High hydrostatic pressure, mainly used as a preservation process, did not impair the nutritional quality of the egg yolk and granule proteins but improved the susceptibility of phosvitin (protein contained in egg yolk) proteolysis to produce bioactive phosphopeptides. Consequently, applying HHP to granules represents an interesting process that improves the digestibility of phosvitin.

The effects of sucrose/NaCl combined pickling on the textural characteristics, moisture distribution, and protein aggregation behavior of egg yolk.

Salted egg yolks have a tender, loose, gritty, and oily texture and are commonly employed as fillings in baked goods. This study investigated the formation mechanism of egg yolk gels using three different pickling methods: NaCl, sucrose, and mixed groups. The results revealed that of these pickling methods, egg yolks pickled with the mixture had the lowest moisture content (11.59% at 25°C and 10.21% at 45°C), almost no free water content, and the highest hardness (19.11 N at 25°C and 31.01 N at 45°C). Intermolecular force measurements indicated that pickling with the mixture mitigated the surface hardening effect of sucrose and facilitated protein cross-linking. Moreover, confocal laser scanning microscopy of the egg yolk gels pickled with the mixture displayed macromolecular aggregates and oil exudation, suggesting that this method partially disrupted the lipoprotein structure and notably promoted yolk protein aggregation and lipid release. Overall, egg yolks formed a dense gel via the mixed pickling method owing to the ionic concentration and dehydration effects. These findings show the impact of NaCl and sucrose in pickling egg yolks, providing a crucial foundation for developing innovative and desirable egg yolk products. PRACTICAL APPLICATION: This study introduces a novel pickling strategy that combines sucrose and NaCl for egg yolk processing. The egg yolk pickled using this method exhibited improved quality according to the evaluated textural characteristics, moisture distribution, and protein aggregation behavior. The findings may broaden the use of sucrose as a pickling agent for egg yolk processing and provide new ideas for developing and producing pickled eggs and other food products.

Effect of salting and dehydration treatments on the physicochemical and gel properties of hen and duck egg yolks, plasma and granules.

Salted hen egg yolks are less oily and less flavorful than salted duck egg yolks. However, hen eggs have a more adequate market supply and have a broader application prospect than duck eggs. In the present study, egg yolks, plasma, and granules were dehydrated by adding 1% NaCl to simulate traditional curing process of salted egg yolk. The changes in the pickling process of hen egg yolks (HEY) and duck egg yolks (DEY) plasma and granules were compared to reveal the gelation mechanism and the underlying causes of quality differences in salted HEY and DEY. Salted HEY can be compared with the changes in DEY during the pickling process to provide a theoretical basis for the quality improvement of salted HEY to salted DEY. The results showed that both plasma and granules were involved in gel formation, but exhibited different aggregation behaviors. Based on the intermolecular forces, the HEY proteins achieved aggregation mainly through hydrophobic interactions and DEY proteins mainly through covalent binding. According to spin-spin relaxation time, HEY gels immobilized a large amount of lipid and interacted strongly with lipids. DEY gels showed much free lipid and had weak interaction with lipid. The microstructure showed that HEY proteins were easily unfolded to form a homogeneous three-dimensional gel network structure after salting, whereas heterogeneous aggregates were formed to hinder the gel development in DEY. Changes in protein secondary structure content showed that pickling can promote the transformation of the α-helices to β-sheets structure in HEY gels, whereas more α-helices structure was formed in DEY gels. The present study has demonstrated that different gelation behaviors of hen and duck egg yolk proteins (especially in plasma) through salting treatment led to the difference in the quality of salted HEY and DEY. © 2024 Society of Chemical Industry.

Co-enzymatic hydrolysis enhanced thermal stability of mayonnaise

With the accelerated pace of life, frozen bakery products and ready-to-eat hot food are gradually emerging. Mayonnaise, a commonly used flavor sauce, is also applied to these products. However, with the large amount of oil and fat precipitation of mayonnaise in the reheating process of the product, which affects its application in the food industry, the thermal stability of mayonnaise has been gradually paid attention to. In this study, the thermal stability of mayonnaise was effectively improved by the combined enzymatic hydrolysis of liquid egg yolk (LEY) with phospholipase A2 (PLA2) and flavor protease. It was found that the moderate addition of flavor protease could effectively reduce the surface hydrophobicity and interfacial tension of egg yolk protein and improve its emulsification stability. Moreover, the hydrolysis reduced the molecular weight of yolk protein, thus reducing the thermal aggregation effect of yolk protein due to heating, which in turn reduced the loss of protein at the O/W interface and improved the thermal stability of mayonnaise.

Methionine Supplementation Alleviates the Germ Cell Apoptosis Increased by Maternal Caffeine Intake in a C. elegans Model.

Caffeine (1,3,7-trimethylxanthine) is a widely consumed bioactive substance worldwide. Our recent study showed that a reduction in both reproduction and yolk protein production (vitellogenesis) caused by caffeine intake were improved by vitamin B12 supplementation, which is an essential co-factor in methionine metabolism. In the current study, we investigated the role of methionine in the reproduction of caffeine-ingested animals (CIAs). We assessed the effect of methionine metabolism on CIAs and found that caffeine intake decreased both methionine levels and essential enzymes related to the methionine cycle. Furthermore, we found that the caffeine-induced impairment of methionine metabolism decreased vitellogenesis and increased germ cell apoptosis in an LIN-35/RB-dependent manner. Interestingly, the increased germ cell apoptosis was restored to normal levels by methionine supplementation in CIAs. These results indicate that methionine supplementation plays a beneficial role in germ cell health and offspring development by regulating vitellogenesis.

Advances in Understanding the Antioxidant and Antigenic Properties of Egg-Derived Peptides.

Pepsin, trypsin and proteinase K were used in the present study to hydrolyse the proteins from whole eggs, yolks or whites, and the resulting hydrolysates were characterised in terms of antioxidant and IgE-binding properties, using a combination of in vitro and in silico methods. Based on the degree of hydrolysis (DH) results, the egg yolk proteins are better substrates for all the tested enzymes (DH of 6.2-20.1%) compared to those from egg whites (DH of 2.0-4.4%). The SDS-PAGE analysis indicated that pepsin and proteinase K were more efficient compared to trypsin in breaking the intramolecular peptide bonds of the high molecular weight egg proteins. For all the tested substrates, enzyme-assisted hydrolysis resulted in a significant increase in antioxidant activity, suggesting that many bioactive peptides are encrypted in inactive forms in the parent proteins. The hydrolysates obtained with proteinase K exhibited the highest DPPH radical scavenging activity (124-311 µM Trolox/g protein) and the lowest residual IgE-binding capacity. The bioinformatics tools revealed that proteinase K is able to break the integrity of the main linear IgE-binding epitopes from ovalbumin and ovomucoid. It can be concluded that proteinase K is a promising tool for modulating the intrinsic properties of egg proteins.

Proteomic Analysis of Egg Yolk Proteins During Embryonic Development in Wanxi White Goose.

To investigate the alterations of yolk protein during embryonic development in Wanxi white goose, the egg yolk protein composition at days 0, 4, 7, 14, 18, and 25 of incubation (D0, D4, D7, D14, D18, and D25) was analyzed by two-dimensional gel electrophoresis combined with mass spectrometry. A total of 65 spots representing 11 proteins with significant abundance changes were detected. Apolipoprotein B-100, vitellogenin-1, vitellogenin-2-like, riboflavin-binding protein, and serotransferrin mainly participated in nutrient (lipid, riboflavin, and iron ion) transport, and vitellogenin-2-like showed a lower abundance after D14. Ovomucoid-like were involved in endopeptidase inhibitory activity and immunoglobulin binding and exhibited a higher expression after D18, suggesting a potential role in promoting the absorption of immunoglobulin and providing passive immune protection for goose embryos after D18. Furthermore, myosin-9 and actin (ACTB) were involved in the tight junction pathway, potentially contributing to barrier integrity. Serum albumin mainly participated in cytolysis and toxic substance binding. Therefore, the high expression of serum albumin, myosin-9, and ACTB throughout the incubation might protect the developing embryo. Apolipoprotein B-100, vitellogenin-1, vitellogenin-2-like, riboflavin-binding protein, and serotransferrin might play a crucial role in providing nutrition for embryonic development, and VTG-2-like was preferentially degraded/absorbed.

Foaming and structural properties of egg yolk components microgel particles and their adsorption characteristics at the air-water interface

In this study, microgel particles of egg yolk and its components were constructed by inducing and modifying their structure via the high-pressure homogenization (HPH). In order to compare their structural characterization and reveal their adsorption mechanism at the air-water interface, yolk components were also combined with egg white protein (EWP) to achieve their microgels for investigation. Results suggested that after HPH, foam stabilities of egg yolk protein microgel (EYM) and egg yolk plasma microgel (EPM) were significantly improved, whereas no significance occurred on egg yolk granules microgel (EGM). Interestingly, after HPH, the adsorption capacity of the microgels from combination of yolk components and EWP exhibited a significant increasing trend. However, their interfacial adsorption stability was poor. Fluorescence spectrum and surface tension revealed that more hydrophilic groups and lower surface tension would occur on EY and EP after HPH, whilst no significant effects would happen on EG, which was consistent with the results of foaming characteristics. SDS-PAGE exhibited that there was no obvious change in the protein bands of egg yolk and its components, indicating that egg yolk and component proteins did not undergo covalent crosslinking or degradation with HPH treatment. In summary, our study prepares for the first time of the microgel particles from egg yolk components for stabilizing the air-water interface. In addition, we have revealed the relationship between the physicochemical properties, microstructures and interfacial adsorption properties of the microgel particles from egg yolk components, and have explored their mechanisms in depth.

Molecular Modeling of the Adsorption of an Egg Yolk Protein on a Water-Oil Interface.

Egg yolk contains several molecular species with emulsifying properties, such as proteins and phospholipids. In particular, these molecules have both polar and nonpolar parts and thus can act as surfactants. One of the most surface-active proteins from egg yolk low-density lipoproteins is the so-called Apovitellenin-1. Experimental studies have been hindered by difficulties in isolating individual species from egg yolk lipoproteins. The purpose of this work was to assess the emulsifying properties of Apovitellenin-1 and any potential cooperative or competitive behavior in the presence of phospholipids. To do so, molecular simulations were carried out in a liquid-liquid interfacial system consisting of water and soybean oil, with varying concentrations of phospholipids and for different spatial configurations. To evaluate the conformational stability of the protein at the water-oil interface, the Gibbs free energy was computed from Metadynamics simulations as a function of the distance from the interface and of the radius of gyration. Moreover, a detailed analysis was also performed to determine which peptide residues were responsible for the protein adsorption at the oil-water interface as well as the lowering of the interfacial tension. Lastly, we combined the simulation results with a thermodynamic model to predict the interfacial tension behavior at increasing protein bulk concentration, which cannot be measured experimentally.

Investigation into the impact of enzyme treatment on the structural characteristics of egg yolk protein and its correlation with flavor constituents

The effects of three different enzyme treatments on the structure and volatile compounds of egg yolk protein were investigated. The secondary and tertiary structures, as well as volatiles, were characterized using ultraviolet spectroscopy (UV), fourier transform infrared (FTIR) spectroscopy, free sulfhydryl group analysis, and gas chromatography-mass spectrometry (GC-MS). FTIR analysis revealed a decrease in the contents of intermolecular, intramolecular, and intermolecular antiparallel β-sheets while an increase in random coils was observed. The contents of α-helices and β-turns did not show significant changes, indicating a reduction in protein compactness. Enzymatic treatment led to an increase in hydrophobic interaction, electrostatic interaction, and hydrogen bond formation within the egg yolk hot gel (EYG). Structural characterization demonstrated that EYG formed large uniform air chamber cavities after lipase hydrolysis (LEYG), while EYG hydrolyzed by papain (PEYG) and by flavor protease (FEYG) exhibited more compact structures. In addition, the binding of proteins to flavor substances inhibited the release of volatile flavor compounds. These findings provide insights into enhancing the flavor and texture of egg yolk by modulating its protein structure.

Diets comprising hen egg yolk and milk proteins as potential alternatives to shark egg-based diets for larvae of the Japanese eel Anguilla japonica

Diets comprising hen egg yolk and milk proteins as potential alternatives to shark egg-based diets for larvae of the Japanese eel Anguilla japonica

RNAi-Mediated Knockdown of Acidic Ribosomal Stalk Protein P1 Arrests Egg Development in Adult Female Yellow Fever Mosquitoes, Aedes aegypti.

After taking a blood meal, the fat body of the adult female yellow fever mosquito, Aedes aegypti, switches from a previtellogenic state of arrest to an active state of synthesizing large quantities of yolk protein precursors (YPPs) that are crucial for egg development. The synthesis of YPPs is regulated at both the transcriptional and translational levels. Previously, we identified the cytoplasmic protein general control nonderepressible 1 (GCN1) as a part of the translational regulatory pathway for YPP synthesis. In the current study, we used the C-terminal end of GCN1 to screen for protein-protein interactions and identified 60S acidic ribosomal protein P1 (P1). An expression analysis and RNAi-mediated knockdown of P1 was performed to further investigate the role of P1 in mosquito reproduction. We showed that in unfed (absence of a blood meal) adult A. aegypti mosquitoes, P1 was expressed ubiquitously in the mosquito organs and tissues tested. We also showed that the RNAi-mediated knockdown of P1 in unfed adult female mosquitoes resulted in a strong, transient knockdown with observable phenotypic changes in ovary length and egg deposition. Our results suggest that 60S acidic ribosomal protein P1 is necessary for mosquito reproduction and is a promising target for mosquito population control.

Yolk proteins of the schistosomiasis vector snail Biomphalaria glabrata revealed by multi-omics analysis

Vitellogenesis is the most important process in animal reproduction, in which yolk proteins play a vital role. Among multiple yolk protein precursors, vitellogenin (Vtg) is a well-known major yolk protein (MYP) in most oviparous animals. However, the nature of MYP in the freshwater gastropod snail Biomphalaria glabrata remains elusive. In the current study, we applied bioinformatics, tissue-specific transcriptomics, ovotestis-targeted proteomics, and phylogenetics to investigate the large lipid transfer protein (LLTP) superfamily and ferritin-like family in B. glabrata. Four members of LLTP superfamily (BgVtg1, BgVtg2, BgApo1, and BgApo2), one yolk ferritin (Bg yolk ferritin), and four soma ferritins (Bg ferritin 1, 2, 3, and 4) were identified in B. glabrata genome. The proteomic analysis demonstrated that, among the putative yolk proteins, BgVtg1 was the yolk protein appearing in the highest amount in the ovotestis, followed by Bg yolk ferritin. RNAseq profile showed that the leading synthesis sites of BgVtg1 and Bg yolk ferritin are in the ovotestis (presumably follicle cells) and digestive gland, respectively. Phylogenetic analysis indicated that BgVtg1 is well clustered with Vtgs of other vertebrates and invertebrates. We conclude that, vitellogenin (BgVtg1), not yolk ferritin (Bg yolk ferritin), is the major yolk protein precursor in the schistosomiasis vector snail B. glabrata.

Maternal High-Fat Diet Affects the Contents of Eggs and Causes Abnormal Development in the Medaka Fish.

Maternal nutritional status can affect development and metabolic phenotypes of progeny in animals. The effects of maternal diet are thought to be mediated mainly by changes inside oocytes such as organelles, maternal RNAs, and metabolites. However, to what extent each factor contributes to offspring phenotypes remains uncertain, especially in viviparous mammalian systems, where factors other than oocytes, such as placenta and milk, need to be considered. Here, using the medaka fish as an oviparous vertebrate model, we examined whether maternal high-fat diet (mHFD) feeding affects offspring development and what kind of changes occur in the contents of mature eggs. We found that mHFD caused the high frequency of embryonic deformities of offspring, accompanied by downregulation of transcription- and translation-related genes and zygotic transcripts at the blastula stage. Transcriptomic and metabolomic analyses of mature eggs suggested decreased catabolism of amino acids and glycogen, moderate upregulation of endoplasmic reticulum stress-related genes, and elevated lipid levels in mHFD eggs. Furthermore, high-fat diet females showed a higher incidence of oocyte atresia and downregulation of egg protein genes in the liver. These data suggest that attenuated amino acid catabolism triggered by decreased yolk protein load/processing, as well as elevated lipid levels inside eggs, are the prime candidates that account for the higher incidence of embryonic deformities in mHFD offspring. Our study presents a comprehensive data on the changes inside eggs in a mHFD model of nonmammalian vertebrates and provides insights into the mechanisms of parental nutritional effects on offspring.

Genome-Wide Identification of Vitellogenin Gene Family and Comparative Analysis of Their Involvement in Ovarian Maturation in Exopalaemon carinicauda.

Vitellogenin (Vtg) is a precursor of yolk proteins in egg-laying vertebrates and invertebrates and plays an important role in vitellogenesis and embryonic development. However, the Vtg family remains poorly characterized in Exopalaemon carinicauda, a major commercial mariculture species found along the coasts of the Yellow and Bohai Seas. In this study, 10 Vtg genes from the genomes of E. carinicauda were identified and characterized. Phylogenetic analyses showed that the Vtg genes in crustaceans could be classified into four groups: Astacidea, Brachyra, Penaeidae, and Palaemonidae. EcVtg genes were unevenly distributed on the chromosomes of E. carinicauda, and a molecular evolutionary analysis showed that the EcVtg genes were primarily constrained by purifying selection during evolution. All putative EcVtg proteins were characterized by the presence of three conserved functional domains: a lipoprotein N-terminal domain (LPD_N), a domain of unknown function (DUF1943), and a von Willebrand factor type D domain (vWD). All EcVtg genes exhibited higher expression in the female hepatopancreas than in other tissues, and EcVtg gene expression during ovarian development suggested that the hepatopancreas is the main synthesis site in E. carinicauda. EcVtg1a, EcVtg2, and EcVtg3 play major roles in exogenous vitellogenesis, and EcVtg3 also plays a major role in endogenous vitellogenesis. Bilateral ablation of the eyestalk significantly upregulates EcVtg mRNA expression in the female hepatopancreas, indicating that the X-organ/sinus gland complex plays an important role in ovarian development, mostly by inducing Vtg synthesis. These results could improve our understanding of the function of multiple Vtg genes in crustaceans and aid future studies on the function of EcVtg genes during ovarian development in E. carinicauda.

Vitamin B12 Supplementation Improves Oocyte Development by Modulating Mitochondria and Yolk Protein in a Caffeine-Ingested Caenorhabditis elegans Model.

Vitamin B12 is an essential cofactor involved in the function of two enzymes: cytosolic methionine synthase and mitochondrial methylmalonic-CoA mutase. In our previous studies, caffeine (1,3,7-trimethylxanthine), the most popular bioactivator, was shown to reduce yolk protein (vitellogenin) and fertility in a Caenorhabditis elegans model. Based on the previous finding that methionine supplementation increases vitellogenesis in C. elegans, we investigated the role of vitamin B12 in methionine-mediated vitellogenesis during oogenesis in caffeine-ingested animals (CIA). Vitamin B12 supplementation improved vitellogenesis and reduced oxidative stress by decreasing mitochondrial function in CIA. Furthermore, the decreased number of developing oocytes and high levels of reactive oxygen species in oocytes from CIA were recovered with vitamin B12 supplementation through a reduction in mitochondrial stress, which increased vitellogenesis. Taken together, vitamin B12 supplementation can reverse the negative effects of caffeine intake by enhancing methionine-mediated vitellogenesis and oocyte development by reducing mitochondrial stress.

Study on the mechanism of improving the quality of salted egg yolks by ultrasonic synergistic NaCl dry-curing

The dry separate curing of duck egg yolks was carried out by ultrasonic synergize NaCl (sodium chloride) and NaCl alone. The mechanism of the amelioration of salted egg yolk quality by ultrasonic synergistic NaCl dry-curing was studied. The quality variations of the salted egg yolks were analyzed for the same curing time and NaCl content achieved by ultrasonic synergistic NaCl curing and NaCl curing alone. The results showed that under the same salting time, the NaCl content, oil exudation and chewiness of U48-SEY (ultrasonic for 48 h-salted egg yolk) were higher than those in SEY (salted egg yolk). At the same NaCl content, the oil exudation and chewiness of U44-SEY (ultrasonic for 44 h-salted egg yolk) were still significantly increased. Compared to SEY, the soluble protein content and H0 of U44-SEY and U48-SEY were augmented. Scanning electron microscopy (SEM) indicated that the polyhedral particles in the salted egg yolks prepared by ultrasonic synergistic NaCl dry-curing were closely aligned and evenly distributed, and the salted egg yolks were sandier. Structural analysis revealed that the secondary and tertiary structures of egg yolk protein were changed although the ultrasonic synergistic NaCl dry-curing did not cause the fragmentation or aggregation of the peptide chain structure. The above results suggested that ultrasonic not only perfected the quality of salted egg yolk by promoting NaCl penetration, but also modified the structures of egg yolk protein by the action of ultrasonic itself, which prominently improved the quality of salted egg yolks.

Development of high internal phase Pickering emulsions stabilized by egg yolk and carboxymethylcellulose complexes to improve β-carotene bioaccessibility for the elderly

The work aimed to develop the multi-protein mixture of egg yolk as natural particles to stabilize high internal phase Pickering emulsions (HIPPEs) to improve the bioaccessibility of β-carotene in the elderly. The results showed that the depletion attraction drove the adsorption of egg yolk protein particles at the oil–water interface and the formation of osmotic droplet clusters due to the attachment of particle-coated droplets in the dispersed phase, leading to kinetic blocking and stable gelation of HIPPEs. Rheological measurements showed that HIPPEs had shear thinning, low shear stress, viscoelastic properties, and structural recovery properties, which facilitated easy consumption for the elderly. The stability of HIPPEs was verified by ionic and centrifugal stability tests, demonstrating their potential for application to complex gastric environments. HIPPEs have been applied to the International Dysphagia Dietary Standardization Initiative (IDDSI) test and simulated in vitro digestion in older adults, demonstrating their safe swallowability and high β-carotene bioaccessibility. Our findings suggest solutions for food practitioners facing the aging problem and provide new insights for preparing age-friendly foods.

The timings of vitellogenesis and choriogenesis in the Henosepilachna vigintioctopunctata oocytes

In order to provide a biological basis for the development of population management strategies, a critical need is to better understand the reproduction of agricultural pests. In the current paper, we timed the previtellogenesis, vitellogenesis and choriogenesis processes in Henosepilachna vigintioctopunctata (Fabricius), (Coleoptera, Coccinellidae), a serious polyphagous defoliator damaging a wide range of crops in Solanaceae and Cucurbitaceae. The female reproductive system was comprised of two ovaries, two lateral oviducts, a median oviduct, a spermathecal sac and a genital chamber. Each ovary had an average of 28 (±0.4) telotrophic meroistic ovarioles. In the ovarioles of 4-day-old females, only previtellogenesis process was observed. The vitellogenesis occurred at 5 to 7 days after female emergence, while the choriogenesis took place at 8 and 9 days post eclosion. Copulation and oviposition occurred before and at 8 days post emergence. In order to confirm the observation, HvVgR and HvVg were identified and their expression was examined. HvVgR was abundantly transcribed in the ovary, whereas HvVg was richly expressed in the fat body. The temporal expression profiles showed that the high levels of HvVgR and HvVg were accompanied with the active accumulation of yolk proteins and the occurrence of intercellular spaces among follicle cells. Moreover, only one oocyte was observed per ovariole in active vitellogenesis. The observation demonstrates that oocytes are continuously matured and the oviposition persists in females from 8 days to at least 13 days post eclosion.

Survival, Growth, and Reproduction Responses in a Three-Generation Exposure of the Zebrafish (Danio rerio) to Perfluorooctane Sulfonate.

A prior multigenerational perfluorooctane sulfonic acid (PFOS) exposure investigation in zebrafish reported adverse effects at 0.734 µg/L, among the lowest aquatic effect levels for PFOS reported to date. The present three-generation PFOS exposure quantified survival, growth, reproduction, and vitellogenin (VTG; egg yolk protein) responses in zebrafish, incorporating experimental design and procedural improvements relative to the earlier study. Exposures targeting 0.1, 0.6, 3.2, 20, and 100 µg/L in parental (P) and first filial (F1) generations lasted for 180 days post fertilization (dpf) and the second filial generation (F2) through 16 dpf. Survival decreased significantly in P and F2 generation exposures, but not in F1, at the highest PFOS treatment (100 µg/L nominal, 94-205 µg/L, measured). Significant adverse effects on body weight and length were infrequent, of low magnitude, and occurred predominantly at the highest exposure treatment. Finally, PFOS had no significant effects on P or F1 egg production and survival or whole-body VTG levels in P or F1 male fish. Overall, the predominance and magnitude of adverse PFOS effects at <1 µg/L reported in prior research were largely nonrepeatable in the present study. In contrast, the present study indicated a threshold for ecologically relevant adverse effects in zebrafish at 117 µg/L (SE 8 µg/L, n = 10) for survival and 47 µg/L (SE 11 µg/L, n = 19) for all statistically significant negative effects observed. Environ Toxicol Chem 2024;43:115-131. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.