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Insulin‐like peptides (ILPs) act as crucial reproductive neuropeptides in insects, regulating insect reproduction through the insulin signaling pathway (ISP). Our previous studies have found that the sublethal concentrations (LC1 and LC10) of lambda‐cyhalothrin (λCy) could induce severe reproductive toxicity in the lacewing, Chrysoperla sinica (Tjeder), but the toxicological mechanism remains unclear. This study discovered that λCy could inhibit CsILP transcription, leading to a decrease in insulin content and downregulation of C. sinica insulin receptor (CsInR) and C. sinica forkhead box O (CsFOXO) expression in ISP. Interference with CsILP expression resulted in downregulation of C. sinica vitellogenin (CsVg) and decreasing fecundity, while exogenous injection of bovine insulin promoted upregulation of CsVg expression and facilitated reproduction in female adults of C. sinica. Meanwhile, interference with FOXO of ILP downstream transcription factor could lead to downregulation of CsVg, hindering ovarian development and resulting in a decrease in egg production. However, exogenous injection of bovine insulin could remedy the effects caused by FOXO interference. In addition, ILP mediates juvenile hormone and 20‐hydroxyecdysone biosynthesis by acting on their synthetic regulatory enzymes and influences the signal transduction of the 2 reproductive endocrine hormones, thereby regulating the reproductive endocrine environment in C. sinica. In conclusion, λCy inhibits CsILP expression, leading to disorder of ISP, leading to the reduced fecundity of C. sinica. [ABSTRACT FROM AUTHOR]

The use of nanoparticles in the industry carries the risk of their release into the environment. Based on the presumption that the primary graphene oxide (GO) toxicity mechanism is reactive oxygen species production in the cell, the question arises as to whether well-known antioxidants can protect the cell or significantly reduce the effects of GO. This study focused on the possible remedial effect of vitamin C in Acheta domesticus intoxicated with GO for whole lives. The reproduction potential was measured at the level of Vitellogenin (Vg) gene expression, Vg protein expression, hatching success, and share of nutrition in the developing egg. There was no simple relationship between the Vg gene's expression and the Vg protein content. Despite fewer eggs laid in the vitamin C groups, hatching success was high, and egg composition did not differ significantly. The exceptions were GO20 and GO20 + Vit. C groups, with a shift in the lipid content in the egg. Most likely, ascorbic acid impacts the level of Vg gene expression but does not affect the production of Vg protein or the quality of eggs laid. Low GO concentration in food did not cause adverse effects, but the relationship between GO toxicity and its concentration should be investigated more thoroughly. [ABSTRACT FROM AUTHOR]

Simple Summary: Honeybees are vital pollinators, essential for maintaining ecosystems and biodiversity, but there is rising concern about the health of managed honey bee colonies, especially in heavily human-influenced ecosystems. Multiple factors contribute to this decline, including environmental conditions, forage quality, and pesticide use. These elements have a complex effect on the health and nutritional status of honeybee colonies and influence their response to disease and various stressors. In this study, the authors propose a new approach to assess colony health by correlating common measures of colony strength such as honey and pollen reserves, the number of bees, and brood with hemolymph proteins, common bee pathogens (Varroa destructor and Nosema spp.), and essential trace elements (iron, zinc, and copper). Significant correlations were found between hemolymph proteins and colony performance measures, and between V. destructor and hemolymph proteins and iron content. In conclusion, this study confirms the need for a more holistic approach to honeybee health considering all the relevant aspects and critical points that may affect colony survival. Environment, forage quality, management practices, pathogens, and pesticides influence honeybee responses to stressors. This study proposes an innovative approach to assess colony health and performance using molecular diagnostic tools by correlating hemolymph proteins with common measures of colony strength, prevalent honeybee pathogens (Varroa destructor and Nosema spp.), and essential trace elements (iron, zinc and copper). Colonies were selected from four apiaries located in different environmental and foraging conditions in the province of Bologna (Italy). Hemolymph samples were taken from June to October 2019. The Varroa infestation of the colonies was estimated by assessing the natural mortality of the mites, while the bees were tested for Nosema spp. spores using a microscopic method. Hemolymph proteins were quantified and separated using SDS-PAGE, and colony performance was assessed by determining adult bees, total brood, honey, and pollen reserves. The biomarkers measured proved to be useful for monitoring changes in performance and trophic conditions during summer and early autumn. Significant correlations were found between hemolymph proteins and colony performance measures. A positive correlation between pollen reserves, vitellogenin, and hexamerin 70a highlights the importance of these proteins for successful overwintering. In October, Varroa infestation was negatively correlated with total proteins, vitellogenin, apolipophorin II, transferrin, and hexamerin 70a, with negative implications for overwintering; furthermore, Varroa infestation was also negatively correlated with iron content, potentially affecting iron homeostasis. [ABSTRACT FROM AUTHOR]

Background: Haemaphysalis flava is a notorious parasite for humans and animals worldwide. The organs of H. flava are bathed in hemolymph, which is a freely circulating fluid. Nutrients, immune factors, and waste can be transported to any part of the body via hemolymph. The main soluble components in hemolymph are proteins. However, knowledge of the H. flava proteome is limited. Methods: The hemolymph was collected from fully engorged H. flava ticks by leg amputation. Hemolymph proteins were examined by both blue native polyacrylamide gel electrophoresis (BN-PAGE) and sodium dodecyl sulfate PAGE (SDS-PAGE). Proteins extracted from the gels were further identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Results: Two bands (380 and 520 kDa) were separated from tick hemolymph by BN-PAGE and were further separated into four bands (105, 120, 130, and 360 kDa) by SDS-PAGE. LC-MS/MS revealed that seven tick proteins and 13 host proteins were present in the four bands. These tick proteins mainly belonged to the vitellogenin (Vg) family and the α-macroglobulin family members. In silico structural analysis showed that these Vg family members all had common conserved domains, including the N-terminus lipid binding domain (LPD-N), the C-terminus von Willebrand type D domain (vWD), and the domain of unknown function (DUF). Additionally, two of the Vg family proteins were determined to belong to the carrier protein (CP) by analyzing the unique N-terminal amino acid sequences and the cleaving sites. Conclusion: These findings suggest that the Vg family proteins and α-macroglobulin are the primary constituents of the hemolymph in the form of protein complexes. Our results provide a valuable resource for further functional investigations of H. flava hemolymph effectors and may be useful in tick management. [ABSTRACT FROM AUTHOR]

Introduction: Akirin as a highly conserved transcription factor, exerts a profound influence on the growth, development, immune response, and reproductive processes in animals. The brown planthopper (BPH), Nilaparvata lugens, a major pest in rice production in Asia, possesses high reproductive capacity, a critical factor contributing to reduced rice yields. The aims of this study were to demonstrate the regulatory role of Akirin in the reproduction of BPH. Methods: In this study, quantitative PCR (qPCR) was used to detect the mRNA expression of genes. RNA interference (RNAi) was used to downregulate the expression of Akirin gene, and RNA sequencing (RNA-seq) was used to screen for differentially expressed genes caused by Akirin downregulation. Hormone contents were measured with the enzyme linked immunosorbent assay (ELISA), and protein content was evaluated with the bicinchoninic acid (BCA) method. Results: Using BPH genome data, we screened for an Akirin gene (NlAkirin). An analysis of tissue-specific expressions showed that NlAkirin was expressed in all tissues tested in female BPH, but its expression level was highest in the ovary. After inhibiting the mRNA expression of NlAkirin in BPH females, the number of eggs laid, hatching rate, and number of ovarioles decreased. Transcriptome sequencing was performed, following a NlAkirin double-stranded RNA treatment. Compared with the genes of the control, which was injected with GFP double-stranded RNA, there were 438 upregulated genes and 1012 downregulated genes; the expression of vitellogenin (Vg) and vitellogenin receptor (VgR) genes as well as the mRNA expression of genes related to the target of rapamycin (TOR), juvenile hormone (JH), and insulin pathways involved in Vg synthesis was significantly downregulated. As a result of NlAkirin knockdown, the titers of JH III and Ecdysone (Ecd) were downregulated in unmated females but returned to normal levels in mated females. The ovarian protein contents in both unmated and mated females were downregulated. Discussion and conclusion: Our results suggest that NlAkirin affects female BPH reproduction by regulating the mRNA expression of genes related to the Vg, VgR, TOR, JH, and insulin signaling pathways, in addition to the titers of JH III and Ecd. The findings of this research provide novel insights into the regulatory role of Akirin in insect reproductive capacity. [ABSTRACT FROM AUTHOR]

Numerous pharmaceutical and industrial chemicals are classified as endocrine‐disrupting chemicals (EDCs) that interfere with hormonal homeostasis, leading to developmental disorders and other pathologies. The synthetic estrogen 17α‐ethynylestradiol (EE2) is used in oral contraceptives and other hormone therapies. EE2 and other estrogens are inadvertently introduced into aquatic environments through municipal wastewater and agricultural effluents. Exposure of male fish to estrogens increases expression of the egg yolk precursor protein vitellogenin (Vtg), which is used as a molecular marker of exposure to estrogenic EDCs. The mechanisms behind Vtg induction are not fully known, and we hypothesized that it is regulated via DNA methylation. Adult zebrafish were exposed to either dimethyl sulfoxide or 20 ng/L EE2 for 14 days. Messenger RNA (mRNA) expression and DNA methylation were assessed in male zebrafish livers at 0, 0.25, 0.5, 1, 4, 7, and 14 days of exposure; and those of females were assessed at 13 days (n ≥ 4/group/time point). To test the persistence of any changes, we included a recovery group that received EE2 for 7 days and did not receive any for the following 7 days, in the total 14‐day study. Methylation of DNA at the vtg1 promoter was assessed with targeted gene bisulfite sequencing in livers of adult male and female zebrafish. A significant increase in vtg1 mRNA was observed in the EE2‐exposed male fish as early as 6 h. Interestingly, DNA methylation changes were observed at 4 days. Decreases in the overall methylation of the vtg1 promoter in exposed males resulted in levels comparable to those in female controls, suggesting feminization. Importantly, DNA methylation levels in males remained significantly impacted after 7 days post–EE2 removal, unlike mRNA levels. These data identify an epigenetic mark of feminization that may serve as an indicator of not only estrogenic exposure but also previous exposure to EE2. Environ Toxicol Chem 2024;43:1547–1556. © 2024 SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA. [ABSTRACT FROM AUTHOR]

To enhance our understanding of teleost reproductive physiology, we identified six Sichuan bream (Sinibrama taeniatus) vitellogenin genes (vtg1-6) and characterized their sequence structures. We categorized them into type Ⅰ (vtg1,4,5 and 6), type Ⅱ (vtg2) and type Ⅲ (vtg3) based on differences in their subdomain structure. The promoter sequence of vtgs has multiple estrogen response elements, and their abundance appears to correlate with the responsiveness of vtg gene expression to estrogen. Gene expression analyses revealed that the vitellogenesis of Sichuan bream involves both heterosynthesis and autosynthesis pathways, with the dominant pathway originating from the liver. The drug treatment experiments revealed that 17β-estradiol (E2) tightly regulated the level of vtg mRNA in the liver. Feeding fish with a diet containing 100 μg/g E2 for three weeks significantly induced vtg gene expression and ovarian development, leading to an earlier onset of vitellogenesis. Additionally, it was observed that the initiation of vtg transcription required E2 binding to its receptor, a process primarily mediated by estrogen receptor alpha in Sichuan bream. The findings of this study provide novel insights into the molecular information of the vitellogenin gene family in teleosts, thereby contributing to the regulation of gonadal development in farmed fish. [ABSTRACT FROM AUTHOR]

Since recolonizing the seas, most teleosts evolved as pelagophils, producing highly hydrated pelagic (buoyant) eggs. Osmotic drive for oocyte hydration is mainly generated through differential proteolysis of vitellogenin-Aa-derived yolk proteins. Teleost-specific water channels are selectively retained in pelagophils and coevolved with 14-3-3ζ-like binding proteins to co-operatively regulate channel trafficking in a manner that avoids competitive membrane space occupancy and accelerates oocyte water influx. Gonadotropinergic and vasopressinergic systemic and paracrine signaling systems converge to activate meiosis resumption and oocyte hydration. Developmental downregulation of dominant-negative inhibitory aquaporin variants maximizes oocyte hydration. These innovative genetic adaptations are highly conserved in modern marine teleosts, contributing to the success of their eggs in the oceans. Genetic adaptations of organisms living in extreme environments are fundamental to our understanding of where life can evolve. Water is the single limiting parameter in this regard, yet when released in the oceans, the single-celled eggs of marine bony fishes (teleosts) have no means of acquiring it. They are strongly hyposmotic to seawater and lack osmoregulatory systems. Paradoxically, modern teleosts successfully release vast quantities of eggs in the extreme saline environment and recorded the most explosive radiation in vertebrate history. Here, we highlight key genetic adaptations that evolved to solve this paradox by filling the pre-ovulated eggs with water. The degree of water acquisition is uniquely prevalent to marine teleosts, permitting the survival and oceanic dispersal of their eggs. [ABSTRACT FROM AUTHOR]

Simple Summary: Varroa destructor is a mite that parasitises honeybee hives, weakening colonies and transmitting viruses. Beekeepers apply acaricide treatments to bee colonies to limit the spread of Varroa and the resulting negative effects on bee health. Oxalic acid treatment is the most widely applied, but little is known about its potential effects on the physiology of bees, particularly their immune system. This study investigated the short-term effects of oxalic acid treatment on the immune and antioxidant systems of house bees (i.e., glucose oxidase, phenoloxidase, glutathione S-transferase, catalase activities, and vitellogenin content). Residual concentrations of oxalic acid on the bees' external body and in the haemolymph were also measured. The results showed that the treatment did not affect the concentration of oxalic acid in the haemolymph, in which it was constitutively present. Residues of oxalic acid remained on the outer body of the bees for up to 48 h after treatment. Both glucose oxidase activity and vitellogenin concentration were affected by the treatment, increasing significantly after 48 and 24 h, respectively. No effects were observed on the other parameters investigated (phenoloxidase, glutathione S-transferase, and catalase activities). Honeybees (Apis mellifera L.) have to face many challenges, including Varroa destructor infestation, associated with viral transmission. Oxalic acid is one of the most common treatments against Varroa. Little is known about the physiological effects of oxalic acid, especially those on honeybees' immune systems. In this study, the short-term effects (0–96 h) of oxalic acid treatment on the immune system components (i.e., glucose oxidase, phenoloxidase, glutathione S-transferase, catalase activities, and vitellogenin contents) of house bees were preliminarily investigated. Oxalic acid contents of bee bodies and haemolymphs were also measured. The results confirm that oxalic acid is constitutively present in bee haemolymphs and its concentration is not affected by treatment. At 6 h after the treatment, a maximum peak of oxalic acid content was detected on bees' bodies, which gradually decreased after that until physiological levels were reached at 48 h. In the immune system, the oxalic acid treatment determined a peak in glucose oxidase activity at 48 h, indicating a potential defence response and an increase in vitellogenin content at 24 h. No significant changes were recorded in phenoloxidase, glutathione S-transferase, and catalase activities. These results suggest a time-dependent response to oxalic acid, with potential immune system activation in treated bees. [ABSTRACT FROM AUTHOR]

Identifying traits for adaptation to different management and environmental regimes is key to maintaining robust honey bee populations under global climate change. We compared mite-resistant (Pol-line and Russian) and Italian honey bee stocks in variable-temperature cage experiments (200 bees per cage) with respect to food consumption, thermoregulation, gene expression, and lifespan, in 3 experiments over 2 years. The Italian stock bees consumed more syrup and pollen on average than the mite-resistant stocks, but the mite-resistant stocks maintained higher cluster temperatures and had median lifespans 8 days longer, consistent with the increased expression of vitellogenin relative to Italian stock. Model results indicated that, to maintain the same colony size as the mite-resistant stocks, Italian stock colonies would need about 13% more sealed brood to offset reduced worker lifespans. These differences among bee stocks likely influence colony-level productivity and health, and showed the importance of experimental replication. [ABSTRACT FROM AUTHOR]

We tested the expression of the multi-functional gene Vitellogenin (Vg) in four commercially available stocks of honey bees (Apis mellifera: L) under three different treatments: caged bees fed only 50% sucrose solution, caged bees fed sucrose solution and pollen paste, and free-roaming bees in a colony setting. We found that seven-day old bees fed pollen and sucrose in the lab as well as those in the colony had higher Vg expression than those fed only sucrose. There were strong differences in how the stocks responded to the treatments. This ranged from the Italian bees where expression was different between all treatment groups to the Saskatraz where there were no effects of treatment on Vg expression. [ABSTRACT FROM AUTHOR]

Wastewater treatment plants release complex mixtures of chemicals into the aquatic environment as wastewater effluent (WWE); however, the effects of these mixtures are still poorly understood. Chinook salmon (Oncorhynchus tshawytscha) are a culturally important species in the Pacific Northwest and are a vital food resource for southern resident killer whales (Orcinus orca) that are listed as ‘critically endangered’ under the U.S. Endangered Species Act (ESA). Chinook populations have declined drastically in this region and have failed to show significant recovery despite regional-to-federal efforts, resulting in many populations being listed as threatened under the ESA. One source of stress to juvenile Chinook is chemical pollution from WWE during their outmigration along river corridors and residency in estuaries. In this study, we investigated effects of WWE on juvenile Chinook health in a ten-day exposure to dilutions of WWE from 0.1% to 20%. At the end of the exposure, we measured endpoints associated with endocrine disruption, brain function, osmoregulation, stress, and metabolism. Exposure to WWE significantly (α = 0.1 for all analyses) induced vitellogenesis, indicating endocrine system disruption. We saw significant reductions in plasma glucose, an indication of stress, and brain Na+/K+-ATPase (NKA) activity, an enzyme essential for neuronal signaling. Lastly, metabolism was affected as evidenced by altered total protein, cholesterol, and albumin in plasma, a drastic decrease in whole body lipid content, and a significant increase in visible liver anomalies. We compared contaminant concentrations in exposure water with effects concentrations from the literature for chemicals known to induce vitellogenin or inhibit brain NKA. For most contaminants, concentrations in exposure waters were several orders of magnitude below effects concentrations in the literature. The exception was estrogenic hormones, which were detected at similar concentrations in this study compared to concentrations in other studies that induced vitellogenin. Based on comparisons to the literature, contaminants measured in this study could not explain the inhibition of brain NKA; however, WWE mixtures contain many quantified and undetected compounds that are likely acting together to cause harmful effects in Chinook. This research highlights the need for improved wastewater treatment to improve aquatic health and mitigate effects to threatened species like Puget Sound Chinook salmon.

The β′-component, a major allergen found in fish roe, is synthesized in the liver of female fish as part of the precursor of the egg yolk protein vitellogenin (Vg). Vg is distributed through the bloodstream and contributes to ovary development. In the present study, β′-component, present in Vg fragments, was detected in the muscles and internal organs of mature female salmon, with concentrations exceeding the recommended labeling level under Japan's food allergy labeling system. Additionally, total protein extracted from tissues of female salmon reacted with immunoglobulin E antibody against the salmon roe β′-component. No β′-component was detected in the tissues of mature male fish in the absence of sex hormone stimulation from females nor in immature salmon without sexual differentiation, but excess stimulation of the female sex hormone induced Vg synthesis containing the β′-component in the male fish. These findings suggest that taking into account fish sex, maturity, and sexual isolation of mature male fish could improve the safety of salmon meat for individuals with salmon roe allergy. [ABSTRACT FROM AUTHOR]

The impact of heavy metals on agricultural ecosystems has consistently been a topic of social concern. This study investigated the translocation of lead (Pb) along the soil – Vicia faba L. (Fabales: Fabaceae) – Megoura crassicauda (Hemiptera: Aphidoidea) – Harmonia axyridis (Coleoptera: Coccinellidae) system. Lead in soil could be transferred to broad bean, and the accumulation amount was highest in roots, followed by stems and leaves. Aphids and ladybirds could also accumulate lead through the food chain. Interestingly, the lead content of broad bean roots in the aphid feeding group was significantly higher than that in the non-aphid feeding group. Lead stress significantly reduced the germination rate and seedling height of broad bean, and inhibited root elongation. The expression levels of trehalase (TRE), trehalose-6-phosphate synthase (TPS), and vitellogenin (Vg), TRE activity, and carbohydrate content in aphids changed under lead treatment. The number of offspring produced by the second and fifth generation aphids was significantly reduced under lead treatment. Furthermore, high concentrations of lead treatment can prolong the development time of the 2nd and 3rd instar larvae of the ladybird. Feeding on aphids contaminated with lead can affect the survival of ladybirds. Our results further confirm the biological transfer of lead in the food chain and explore the adaptive mechanisms of aphids and ladybirds. These relevant results provide a theoretical basis for further exploring the molecular mechanism of lead homeostasis in plants and insects under lead stress. [ABSTRACT FROM AUTHOR]

In order to study the change rule of nutrient substance in ovaries of female Alosa sapidissima at the late stage of development and improve the fertilization rate of eggs, the contents of amino acids, fatty acids, vitellogenin （VTG）and cathepsin D （CTSD）and their mRNA expression in the ovaries of female Alosa sapidissima at the stages Ⅳ and Ⅴ of gonadal development were compared and analyzed by biochemical analysis, enzyme-linked immunosorbent assay and fluorescence quantitative PCR. The results showed that the water content （75. 73%） in stage Ⅳ ovary was significantly lower than that （90. 64%） in stage V ovary, while the crude protein content （16. 07%）, crude fat content （2. 86%） and crude ash content （1. 28%） were significantly greater than those （5. 83%, 1. 25%, 0. 81%） in stage Ⅴ ovary. Among the 18 amino acids detected in the ovaries of stages Ⅳ and Ⅴ, the content of glutamic acid was the highest（ 8. 64% and 8. 13%）. Among the 8 essential amino acids, the contents of threonine, phenylalanine, lysine and tryptophan in stage Ⅳ were significantly higher than those in stage Ⅴ, and the first limiting amino acid of both was tryptophan. The total amino acid（ TAA）, semi-essential amino acid（ HEAA） and total non-essential amino acid（ NEAA） in stage Ⅳ were significantly higher than those in stage Ⅴ. A total of 19 fatty acids were detected in the ovaries of the two stages, and the contents of C17∶0, C20∶1n9, C20∶4n6 and C20∶5n3 in stage Ⅳ were significantly higher than those in stage Ⅴ; the contents of Σ MUFA, Σ PUFA, Σ UFA, Σ n3PUFA and EPA + DHA in stage Ⅳ were greater than those in stage Ⅴ, and Σ UFA （70. 54%） reached significant differences. The content of VTG in stage Ⅳ ovary was higher than that in stage Ⅴ ovary, but it was not significant, while the content of CTSD in stage Ⅳ ovary was significantly lower than that in stage Ⅴ ovary. The expression of VTG gene in stage Ⅴ ovary was significantly higher than that in stage Ⅳ, 13. 88 times of that in stage Ⅳ, while the expression of VTG gene in liver was significantly lower than that in stage Ⅳ, accounting for 25% higher than that in stage Ⅳ. From stage Ⅳ to stage Ⅴ, CTSD mRNA expression showed down-regulation in the ovary and up-regulation in the liver, but none of them was significant. Therefore, under the current aquaculture conditions, the accumulation of nutrients in the ovary is uneven during the late development of the Alosa sapidissima, and it is recommended to appropriately adjust the feed formula, increase the ratio of limiting amino acids tryptophan, methionine, L-Phenylalanine and lysine and threonine with lower content, increase the ratio of polyunsaturated fatty acids C20∶3n6, C20∶4n6 （AA）, C22∶6n3 （DHA） and C20∶5n3 （EPA）, and decrease the ratio of C18∶2n6c in order to ensure the balance of nutrients required for the late development of the ovary. [ABSTRACT FROM AUTHOR]

Wastewater treatment plants release complex mixtures of chemicals into the aquatic environment as wastewater effluent (WWE); however, the effects of these mixtures are still poorly understood. Chinook salmon (Oncorhynchus tshawytscha) are a culturally important species in the Pacific Northwest and are a vital food resource for southern resident killer whales (Orcinus orca) that are listed as 'critically endangered' under the U.S. Endangered Species Act (ESA). Chinook populations have declined drastically in this region and have failed to show significant recovery despite regional-to-federal efforts, resulting in many populations being listed as threatened under the ESA. One source of stress to juvenile Chinook is chemical pollution from WWE during their outmigration along river corridors and residency in estuaries. In this study, we investigated effects of WWE on juvenile Chinook health in a ten-day exposure to dilutions of WWE from 0.1% to 20%. At the end of the exposure, we measured endpoints associated with endocrine disruption, brain function, osmoregulation, stress, and metabolism. Exposure to WWE significantly (a = 0.1 for all analyses) induced vitellogenesis, indicating endocrine system disruption. We saw significant reductions in plasma glucose, an indication of stress, and brain Na+/K+-ATPase (NKA) activity, an enzyme essential for neuronal signaling. Lastly, metabolism was affected as evidenced by altered total protein, cholesterol, and albumin in plasma, a drastic decrease in whole body lipid content, and a significant increase in visible liver anomalies. We compared contaminant concentrations in exposure water with effects concentrations from the literature for chemicals known to induce vitellogenin or inhibit brain NKA. For most contaminants, concentrations in exposure waters were several orders of magnitude below effects concentrations in the literature. The exception was estrogenic hormones, which were detected at similar concentrations in this study compared to concentrations in other studies that induced vitellogenin. Based on comparisons to the literature, contaminants measured in this study could not explain the inhibition of brain NKA; however, WWE mixtures contain many quantified and undetected compounds that are likely acting together to cause harmful effects in Chinook. This research highlights the need for improved wastewater treatment to improve aquatic health and mitigate effects to threatened species like Puget Sound Chinook salmon. [ABSTRACT FROM AUTHOR]

Juvenile hormone (JH) acts in the regulation of caste differentiation between queens and workers (i.e., with or without reproductive capacity) during vitellin synthesis and oogenesis in social insects. However, the regulatory mechanisms have not yet been elucidated. Here, we identified a highly expressed microRNA (miRNA), miR‐1175‐3p, in the red imported fire ant, Solenopsis invicta. We found that miR‐1175‐3p is prominently present in the fat bodies and ovaries of workers. Furthermore, miR‐1175‐3p interacts with its target gene, broad‐complex core (Br‐C), in the fat bodies. By utilizing miR‐1175‐3p agomir, we successfully suppressed the expression of the Br‐C protein in queens, resulting in reduced vitellogenin expression, fewer eggs, and poorly developed ovaries. Conversely, decreasing miR‐1175‐3p levels led to the increased expression of Br‐C and vitellogenin in workers, triggering the "re‐development" of the ovaries. Moreover, when queens were fed with JH, the expression of miR‐1175‐3p decreased, whereas the expression of vitellogenin‐2 and vitellogenin‐3 increased. Notably, the suppression of fertility in queens caused by treatment with agomir miR‐1175‐3p was completely rescued by the increased vitellogenin expression induced by being fed with JH. These results suggest the critical role of miR‐1175‐3p in JH‐regulated reproduction, shedding light on the molecular mechanism underlying miRNA‐mediated fecundity in social insects and providing a novel strategy for managing S. invicta. [ABSTRACT FROM AUTHOR]

The rice stripe virus (RSV) is transmitted by the small brown planthopper Laodelphax striatellus in a persistent and circulative–propagative manner. During the last few decades, RSV has caused serious disease outbreaks in rice fields in China. The results of long-term coevolution have led to complex and diverse relationships between viruses and vector insects, and understanding these interactions is critical for the management of diseases and vector organisms. In this study, three groups of comparative experiments were performed to investigate the effects of RSV infection on the reproductive parameters, vitellogenin (Vg), and Vg receptor (VgR) expressions of L. striatellus. The results showed that RSV infection promoted the fecundity, hatchability, and Vg and VgR expressions of L. striatellus but had no effect on the preoviposition period and oviposition period. Furthermore, the RNA interference of the nucleocapsid protein (CP) gene and the injection of the CP antibody in RSV-viruliferous L. striatellus reduced the fecundity, hatchability, and Vg and VgR expressions, further strengthening the hypothesis that RSV may manipulate the reproductive behavior of insect vectors to promote its own transmission. The results of this study can further reveal the interaction mechanism of virus–vector insects in reproduction and virus transmission, and provide new insights for the control of insect-borne viruses. [ABSTRACT FROM AUTHOR]

The caligid ectoparasite, Caligus rogercresseyi, is one of the main concerns in the Chilean salmon industry. The molecular mechanisms displayed by the parasite during the reproductive process represent an opportunity for developing novel control strategies. Vitellogenin is a multifunctional protein recognized as a critical player in several crustaceans' biological processes, including reproduction, embryonic development, and immune response. This study aimed to characterize the C. rogercresseyi vitellogenins, including discovering novel transcripts and regulatory mechanisms associated with microRNAs. Herein, vitellogenin genes were identified by homology analysis using the reference sea louse genome, transcriptome database, and arthropods vitellogenin-protein database. The validation of expression transcripts was conducted by RNA nanopore sequencing technology. Moreover, fusion gene profiling, miRNA target analysis, and functional validation were performed using luciferase assay. Six putative vitellogenin genes were identified in the C. rogercresseyi genome with high homology with other copepods vitellogenins. Furthermore, miR-996 showed a putative role in regulating the Cr_Vitellogenin1 gene, which is highly expressed in females. Moreover, vitellogenin-fusion genes were identified in adult stages and highly regulated in males, demonstrating sex-related expression patterns. In females, the identified fusion genes merged with several non-vitellogenin genes involved in biological processes of ribosome assembly, BMP signaling pathway, and biosynthetic processes. This study reports the genome array of vitellogenins in C. rogercresseyi for the first time, revealing the putative role of fusion genes and miRNA regulation in sea lice biology. [ABSTRACT FROM AUTHOR]

Neoseiulus californicus is a key natural predator of spider mites and small arthropod pests. Although vitellogenin (Vg) and vitellogenin receptor (VgR) genes are important regulators of oocyte maturation, egg formation, and embryonic development in spider mites and ticks, their role in phytoseiid mites is less understood. Investigating the reproductive mechanism of predatory mites is crucial for understanding their reproduction and evolution. In this study, three Vg genes and one VgR gene were identified, cloned, and characterized in N. californicus. Vg1, Vg2, Vg3, and VgR mRNAs were 6066, 5728, 4515, and 5331 bp long, encoding proteins comprising 1851, 1849, 1471, and 1722 amino acids, respectively. Temporal expression analysis revealed elevated Vg1, Vg2, and VgR levels in fertilized female adult mites, whereas Vg3 exhibited peak expression during the developmental stage. RNA interference–mediated suppression of Vg1 and Vg2 resulted in 14.66% and 11.59% decreases in fecundity, respectively. Vg1 silencing notably reduced egg hatching rate and female offspring ratio. In contrast, Vg3 and VgR silencing had no significant impact on fecundity and egg hatching rate. These findings provide insights into the regulatory roles of Vg and VgR in predatory mite reproduction, offering a theoretical foundation for understanding their evolution and reproductive strategies. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The use of nanoparticles in the industry carries the risk of their release into the environment. Based on the presumption that the primary graphene oxide (GO) toxicity mechanism is reactive oxygen species production in the cell, the question arises as to whether well-known antioxidants can protect the cell or significantly reduce the effects of GO. This study focused on the possible remedial effect of vitamin C in Acheta domesticus intoxicated with GO for whole lives. The reproduction potential was measured at the level of Vitellogenin (Vg) gene expression, Vg protein expression, hatching success, and share of nutrition in the developing egg. There was no simple relationship between the Vg gene’s expression and the Vg protein content. Despite fewer eggs laid in the vitamin C groups, hatching success was high, and egg composition did not differ significantly. The exceptions were GO20 and GO20 + Vit. C groups, with a shift in the lipid content in the egg. Most likely, ascorbic acid impacts the level of Vg gene expression but does not affect the production of Vg protein or the quality of eggs laid. Low GO concentration in food did not cause adverse effects, but the relationship between GO toxicity and its concentration should be investigated more thoroughly.

To elucidate the function and reproductive regulation mechanism of the Vitellogenin(GmVg) gene in Grapholita molesta, a significant pest of fruit trees, and to provide a theoretical basis for further research on the function of the GmVg gene, in this study, GmVg was cloned and identified and a comprehensive bioinformatics analysis was performed. The expression patterns of GmVg at various developmental stages, across sexes, and in different tissues were examined using quantitative reverse transcription PCR(qRT-PCR). The results revealed that the full-length of GmVg gene spanned 5 573 base pairs(bp), with an open reading frame(ORF) of 5 364 bp that encoded 1 787 amino acids. The signal peptide consisted of 15 amino acids, and the molecular formula was C9041H14090N2544O2743S57. The percentage of the amino acids alanine(Ala), glutamine(Gln), and serine (Ser) was large and they comprised 8.2%, 7.9%, and 7.8% of the protein, respectively. The protein contained 187 acidic residues(Asp+Glu) and 204 basic residues(Arg+Lys). Bioinformatics predictions indicated that the molecular weight of the GmVg protein was 204.1 ku, with an isoelectric point of 8.79, and it featured three characteristic conserved domains: Vitellogenin-N, DUF1943, and VWD. The expression of GmVg was markedly higher in adults compared to that in eggs, larvae, and pupae, reaching its peak 24 h post-emergence. The gene displayed sex-specific expression, with negligible levels in males. Within female adults, tissue-specific patterns were noted in head, ovary, and fat body; the expression in the fat body was substantially higher than in the head and ovary-21.49-fold and 16.4-fold greater, respectively. [ABSTRACT FROM AUTHOR]

Simple Summary: Recently, there has been a serious decline in honey bee colonies due to various factors, including habitat loss, diseases, pathogen infections, chemical exposures, and climatic changes. Among these factors, malnutrition-induced stress plays a significant role in colony decline. In our study, we present a novel perspective on maintaining and developing honey bee colonies. Our research demonstrates that a specific pollen substitute diet significantly improves colony performance during early spring, as evidenced by an increase in population, a greater capped brood area, altered colony weight, honey bee weight (dried head + thorax), and vitellogenin value. This finding could contribute to enhanced colony maintenance and development during the early spring season, with the potential to prevent colony decline. The nutritional quality of a colony significantly affects its health and strength, particularly because it is required for population growth in the early spring. We investigated the impact of various artificial pollen substitute diets on colony performance in the Republic of Korea during early spring, a critical period for colony health and growth. The colonies were provided with different diets, including the commercial product Megabee (positive control), our developed diet Test A, and four upgraded versions (Diet 1, Diet 2, Diet 3, and Diet 4) of Test A. The negative control group received no supplementary feed. Over 63 days, we observed 24 experimental colonies and assessed various parameters at the colony and individual levels. The results revealed that Diet 2 had the highest consumption and had the most positive impact on population growth, the capped brood area, colony weight, honey bees' weight, and vitellogenin levels. These findings suggested that Diet 2 is most attractive to honey bees and thus holds great promise for improving colony maintenance and development during the crucial early spring period. [ABSTRACT FROM AUTHOR]

In insects, vitellogenin (Vg) is generally viewed as a female‐specific protein. Its primary function is to supply nutrition to developing embryos. Here, we reported Vg from the male adults of a natural predator, Chrysopa pallens. The male Vg was depleted by RNAi. Mating with Vg‐deficient male downregulated female Vg expression, suppressed ovarian development and decreased reproductive output. Whole‐organism transcriptome analysis after male Vg knockdown showed no differential expression of the known spermatogenesis‐related regulators and seminal fluid protein genes, but a sharp downregulation of an unknown gene, which encodes a testis‐enriched big protein (Vcsoo). Separate knockdown of male Vg and Vcsoo disturbed the assembly of spermatid cytoplasmic organelles in males and suppressed the expansion of ovary germarium in mated females. These results demonstrated that C. pallens male Vg signals through the downstream Vcsoo and regulates male and female reproduction. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Insect vitellogenins are an intriguing class of complex proteins. They primarily serve as a source of energy for the developing embryo in insect eggs. Vitellogenesis is a complex hormonally and neurally controlled process that command synthesis of vitellogenin molecules and ensures their transport from the female fat bodies or ovarial cells into eggs. The representatives of all insect hormones such as juvenile hormones, ecdysteroids, and neurohormones participate in vitellogenesis, but juvenile hormones (most insect species) and ecdysteroids (mostly Diptera) play the most important roles in the process. Strikingly, not only insect females, but also males have been reported to synthesize vitellogenins indicating their further utility in the insect body. Indeed, it has recently been found that vitellogenins perform a variety of biological functions in the insect body. They participate in defense reactions against entomopathogens such as nematodes, fungi, and bacteria, as well as against venoms such as the honeybee Apis mellifera venom. Interestingly, vitellogenins are also present in the venom of the honeybee itself, albeit their exact role is unknown; they most likely increase the efficacy of the venom in the victim's body. Within the bee's body vitellogenins contribute to the lifespan regulation as antiaging factor acting under tight social interactions and hormonal control. The current minireview covers all of these functions of vitellogenins and portrays them as biologically active substances that play a variety of significant roles in both insect females and males, and not only acting as passive energy sources for developing embryo. [ABSTRACT FROM AUTHOR]

Recently, neuropeptide F2 (NPF2) has been demonstrated to increase food consumption and body weight in the cotton bollworm, Helicoverpa armigera. This study evaluated the effect of NPF2 on the adult female reproductive system of H. armigera. Daily NPF2 injections increased oocyte size, ecdysteroid titre in ovaries and haemolymph and the expression level of ecdysone receptor (EcR), ultraspiracle (USP) and vitellogenin (Vg) genes. However, NPF2 injection had no effect on the expression level of hormone receptor 3 (HR3). Fecundity and fertility were increased after the injection of NPF2. The knockdown of the NPF2 gene using RNA interference (RNAi) had opposite effects compared to NPF2 injection. Overall, these results revealed that NPF2 has a regulatory role in the female reproduction of the cotton bollworm. [ABSTRACT FROM AUTHOR]

Inadequate gonadal maturation and poor spawning performance increasingly threaten the sustainability of shrimp aquaculture. Unraveling the mechanisms regulating ovarian development and maturation hence is critical to address industry challenges. Vitellogenin (Vtg), a precursor of yolk protein found in the hepatopancreas and ovary of shrimp, plays a key role in facilitating shrimp's oocyte maturation and embryonic development after oviposition. This study found that FpVtg was specifically expressed in F. penicillatus hepatopancreas and ovary. FpVtg was localized predominantly in the oocyte cytoplasm and distributed uniformly in the hepatopancreas tissue. Silencing FpVtg led to apoptosis in both hepatopancreas and ovary tissues. Furthermore, FpVtg depletion upregulated the expression of ovarian peritrophin 1, ovarian peritrophin 2, serine proteinase inhibitor 6, and juvenile hormone esterase-like carboxylesterase 1, while downregulated that of vitellogenin, delta-9 desaturase, and insulin-like receptor. KEGG pathway analysis implicated such as PI3K-AKT signaling, RNA transport, ECM-receptor interaction, hippo signaling, oocyte meiosis, and apoptosis were enriched and involved in ovarian development. These findings have provided insights into the FpVtg's reproductive role and the associated regulatory genes and pathways in F. penicillatus. This knowledge can contribute to establishing strategies to improve the breeding and aquaculture production of F. penicillatus by elucidating its vitellogenesis regulation in redtail prawn and other penaeid species. Further characterization of the implicated pathways and genes will clarify the intricacies underlying ovarian maturation. [ABSTRACT FROM AUTHOR]

Seawater contains a variety of mineral ions, such as Na+, Mg2+, Ca2+, and K+. Although previous studies have shown that brackish water can promote ovarian development of the Chinese mitten crab (Eriocheir sinensis), it remains unclear whether these ions directly affect the ovarian development and vitellogenesis of E. sinensis. Therefore, this study was designed, using in vitro tissue incubation mediums with different concentrations of Na+ (280, 320, 360, and 400 mmol/L), Mg2+ (5, 10, and 15 mmol/L), Ca2+ (5, 10, 15, and 20 mmol/L), and K+ (5.5, 9, and 13 mmol/L), to investigate their effects on Vg/Vn contents and the expression of ovarian development-related genes in the hepatopancreas and ovaries of E. sinensis. The results showed that: (1) Different Na+ concentrations had no significant impact on vitellogenesis and the expression of ovarian development-related genes in both tissues, except for an enhancement of the esEcR2-mRNA levels in the hepatopancreas. (2) The Mg2+ (15 mmol/L) significantly increased the Vg/Vn content in hepatopancreas and ovary. However, the 10 mmol/L Mg2+ group decreased esVgR-mRNA levels and esEcR2-mRNA levels in the ovary compared to the control group. (3) Ca2+ (10 ∼ 15 mmol/L) significantly promoted Vg/Vn content and esVg-mRNA levels in both tissues, which also upregulated the expression of ovarian development-related genes (esERR, esEcR2, and esRXR). (4) Increasing K+ content significantly decreased the Vg/Vn content in hepatopancreas and ovaries. Furthermore, the K+ significantly reduced esRXR-mRNA levels in the ovaries but significantly increased esERR-mRNA levels in both tissues. In summary, under in vitro conditions, Na+ concentration had no significant effect on the vitellogenesis, while Ca2+ (10 ∼ 15 mmol/L) and Mg2+ (15 mmol/L) significantly promoted the vitellogenesis in the hepatopancreas and ovaries of E. sinensis. Additionally, Ca2+ significantly enhanced the expression of genes related to ovarian development, while K+ had a negative impact on the vitellogenesis.

Environment, forage quality, management practices, pathogens, and pesticides influence honeybee responses to stressors. This study proposes an innovative approach to assess colony health and performance using molecular diagnostic tools by correlating hemolymph proteins with common measures of colony strength, prevalent honeybee pathogens (Varroa destructor and Nosema spp.), and essential trace elements (iron, zinc and copper). Colonies were selected from four apiaries located in different environmental and foraging conditions in the province of Bologna (Italy). Hemolymph samples were taken from June to October 2019. The Varroa infestation of the colonies was estimated by assessing the natural mortality of the mites, while the bees were tested for Nosema spp. spores using a microscopic method. Hemolymph proteins were quantified and separated using SDS-PAGE, and colony performance was assessed by determining adult bees, total brood, honey, and pollen reserves. The biomarkers measured proved to be useful for monitoring changes in performance and trophic conditions during summer and early autumn. Significant correlations were found between hemolymph proteins and colony performance measures. A positive correlation between pollen reserves, vitellogenin, and hexamerin 70a highlights the importance of these proteins for successful overwintering. In October, Varroa infestation was negatively correlated with total proteins, vitellogenin, apolipophorin II, transferrin, and hexamerin 70a, with negative implications for overwintering; furthermore, Varroa infestation was also negatively correlated with iron content, potentially affecting iron homeostasis.

To enhance our understanding of teleost reproductive physiology, we identified six Sichuan bream (Sinibrama taeniatus) vitellogenin genes (vtg1-6) and characterized their sequence structures. We categorized them into type Ⅰ (vtg1,4,5 and 6), type Ⅱ (vtg2) and type Ⅲ (vtg3) based on differences in their subdomain structure. The promoter sequence of vtgs has multiple estrogen response elements, and their abundance appears to correlate with the responsiveness of vtg gene expression to estrogen. Gene expression analyses revealed that the vitellogenesis of Sichuan bream involves both heterosynthesis and autosynthesis pathways, with the dominant pathway originating from the liver. The drug treatment experiments revealed that 17β-estradiol (E2) tightly regulated the level of vtg mRNA in the liver. Feeding fish with a diet containing 100 μg/g E2 for three weeks significantly induced vtg gene expression and ovarian development, leading to an earlier onset of vitellogenesis. Additionally, it was observed that the initiation of vtg transcription required E2 binding to its receptor, a process primarily mediated by estrogen receptor alpha in Sichuan bream. The findings of this study provide novel insights into the molecular information of the vitellogenin gene family in teleosts, thereby contributing to the regulation of gonadal development in farmed fish.

Honeybees (Apis mellifera L.) have to face many challenges, including Varroa destructor infestation, associated with viral transmission. Oxalic acid is one of the most common treatments against Varroa. Little is known about the physiological effects of oxalic acid, especially those on honeybees’ immune systems. In this study, the short-term effects (0–96 h) of oxalic acid treatment on the immune system components (i.e., glucose oxidase, phenoloxidase, glutathione S-transferase, catalase activities, and vitellogenin contents) of house bees were preliminarily investigated. Oxalic acid contents of bee bodies and haemolymphs were also measured. The results confirm that oxalic acid is constitutively present in bee haemolymphs and its concentration is not affected by treatment. At 6 h after the treatment, a maximum peak of oxalic acid content was detected on bees’ bodies, which gradually decreased after that until physiological levels were reached at 48 h. In the immune system, the oxalic acid treatment determined a peak in glucose oxidase activity at 48 h, indicating a potential defence response and an increase in vitellogenin content at 24 h. No significant changes were recorded in phenoloxidase, glutathione S-transferase, and catalase activities. These results suggest a time-dependent response to oxalic acid, with potential immune system activation in treated bees.

Abstract In phytophagous insects, adaptation to a new host is a dynamic process, in which early and later steps may be underpinned by different features of the insect genome. Here, we tested the hypothesis that early steps of this process are underpinned by a shift in gene expression patterns. We set up a short-term artificial selection experiment (10 generations) for the use of an alternative host (Cicer arietinum) on populations of the bean beetle Zabrotes subfasciatus. Using Illumina sequencing on young adult females, we show the selected populations differ in the expression of genes associated to stimuli, signalling, and developmental processes. Particularly, the “C. arietinum” population shows upregulation of histone methylation genes, which may constitute a strategy for fine-tuning the insect global gene expression network. Using qPCR on body regions, we demonstrated that the “Phaseolus vulgaris” population upregulates the genes polygalacturonase and egalitarian and that the expression of an odorant receptor transcript variant changes over generations. Moreover, in this population we detected the existence of vitellogenin (Vg) variants in both males and females, possibly harbouring canonical reproductive function in females and extracellular unknown functions in males. This study provides the basis for future genomic investigations seeking to shed light on the nature of the proximate mechanisms involved in promoting differential gene expression associated to insect development and adaptation to new hosts.

Simple Summary: Microsporidia Nosema ceranae is an obligate intracellular parasite of the honey bee and causes nosemosis, a disease with negative effects on the health, reproductive and productive capabilities of bee colonies. The aim of this study was to investigate the effects of a plant-based supplement branded as "B+" on honeybees in a laboratory experiment. Food supplement branded as "B+" showed the potential to positively influence bee survival. Supplemental feeding reduced N. ceranae infection level. Treatment significantly decreased oxidative stress in Nosema-infected bees. Vitellogenin gene expression was the highest in the supplement-fed group. One of the most important approaches in the prevention and treatment of nosemosis is the use of herbal preparations as food supplements for bees. Therefore, the aim of this study was to investigate the effects of a plant-based supplement branded as "B+" on honeybees in a laboratory experiment. Four experimental groups were established: treated group (T), N. ceranae-infected and treated group (IT), N. ceranae-infected group (I) and non-infected group (NI). Survival, N. ceranae spore load and oxidative stress parameters together with expression levels of antioxidant enzyme genes and vitellogenin gene were monitored. The mortality in the T, IT and NI groups was significantly (p < 0.001) lower than in than in the I group. Within Nosema-infected groups, the IT group had a significantly lower (p < 0.001) number of N. ceranae spores than the I group. In addition, expression levels of genes for antioxidant enzymes were lower (p < 0.001) in the IT group compared to the I group. The concentration of malondialdehyde and the activities of antioxidant enzymes (superoxide dismutase, catalase and glutathione S-transferase) were significantly lower (p < 0.001) in the IT group compared to the I group. No negative effects of the tested supplement were observed. All these findings indicate that the tested supplement exerted beneficial effects manifested in better bee survival, reduced N. ceranae spore number and reduced oxidative stress of bees (lower expression of genes for antioxidant enzymes and oxidative stress parameters). [ABSTRACT FROM AUTHOR]

Simple Summary: Bees are often fed by beekeepers with supplemented artificial diets. The formulation of an integrative diet for honey bee colonies able to prevent nutritional deficiencies is yet to be found. In this work, the effects of fat-enriched diet (coconut oil) administration in newly emerged and forager bees were tested. Fat supplementation in the honey bee diet determines an increase in newly emerged bees' survival in the short term and an increase in fat content. Further investigations to evaluate the use of such a supplement food to prevent the fat loss of weak families during winter are desirable. Searching for artificial diets positively affecting the survival, immune and antioxidant systems of honey bees is one of main challenges occurring in beekeeping. Among nutrients, lipids play a significant role in insect nutrition as structural components in cell membranes, energy sources and reserves, and are involved in many physiological processes. In this context, the aim of this work was to investigate the effect of 0.5% and 1% coconut oil-enriched diet administration on newly emerged and forager bees survival rate, feed intake, immune system, antioxidant system and both fat and vitellogenin content. In newly emerged bees, supplementation with 1% coconut oil determined a decrease in feed consumption, an increase in survival rate from the 3rd to 14th day of feeding, a short-term decrease in phenoloxidase activity, an increase in body fat and no differences in vitellogenin content. Conversely, supplementation with 0.5% coconut oil determined an increase in survival rate from the 3rd to 15th day of feeding and an increase in fat content in the long term (i.e., 20 days). Regarding the forager bee diet, enrichment with 0.5% and 1% coconut oil only determined an increase in fat content. Therefore, supplementation with coconut oil in honey bee diets at low percentages (0.5 and 1%) determines fat gain. Further investigations to evaluate the use of such supplement foods to prevent the fat loss of weak families during winter are desirable. [ABSTRACT FROM AUTHOR]

Recent research on decapod crustaceans revealed the presence of multiple vitellogenin genes within a single species. Characterizing their expression dynamics and regulatory mechanisms is helpful for the sustainability of commercially relevant decapods through a greater comprehension of vitellogenesis. However, it is unclear whether multiple vitellogenin genes exist in the kuruma prawn Marsupenaeus japonicus, an essential species in several Asian countries. This study determined the full-length cDNA sequence of the new vitellogenin (Maj-Vg2) using transcriptome analysis and cDNA cloning. The deduced Maj-Vg2 consisted of 2,554 amino acids and shared only 53.8% identity with Maj-Vg1, the known vitellogenin of this species. The Maj-Vg1 gene (Maj-Vg1) was expressed in both the hepatopancreas and ovary, whereas the Maj-Vg2 gene (Maj-Vg2) was expressed only in the hepatopancreas. Maj-Vg2 expression in the hepatopancreas was low during the previtellogenic stage and increased during the early and late yolk globule stages. When eyestalks were ablated from immature prawns, Maj-Vg2 expression in the hepatopancreas increased significantly, like Maj-Vg1 expression in the ovary, suggesting possible regulation by hormones from the eyestalk ganglia. These results suggested that Maj-Vg2 and Maj-Vg1 are involved in vitellogenesis in this species. [ABSTRACT FROM AUTHOR]

The uMgeni River is one of the most polluted freshwater ecosystems in KwaZulu-Natal. The study aims to assess the response of Clarias gariepinus to water contamination at the Nagle and Inanda dams using selected biomarkers. Samples were collected and preserved as per the analyses to be carried out. Generally, Nagle Dam showed good quality water compared to Inanda Dam. The Inanda Dam population showed a relatively higher prevalence of alterations in the gills and liver than that from Nagle Dam. The degree of alterations showed some variability within each population, however, there was no significant difference (p>0.05) between the two populations. Organ indices of 20 denoting moderate alterations were observed for both populations. Although the mean AChE activity and VtG induction showed no significant difference between the two populations (p>0.05); the lowest AChE activity and the highest VtG level, were observed in the Inanda Dam populations. Both histopathologic and biochemical biomarkers show signs of severely compromised fish health in the Inanda Dam compared to those from Nagle Dam. It is thus, evident that the tributaries feeding the uMgeni River downstream of Nagle Dam are resulting in the increase of pollution level in the Inanda Dam. [ABSTRACT FROM AUTHOR]

Polycyclic musk compounds are commonly used in personal care products to replace expensive natural fragrances. Due to their huge consumption, they have become a part of the aquatic environment. In the present study, a two-month exposure effect of tonalide on juvenile zebrafish (Danio rerio) was investigated. We determined the vitellogenin concentration to define the potential endocrine-disrupting effect of tonalide and also analysed selected indices to evaluate the induction of oxidative stress. The environmentally relevant concentration of tonalide (i.e., 500 ng/l) caused a significant decrease in the catalase activity (P

The rice stripe virus (RSV) is transmitted by the small brown planthopper Laodelphax striatellus in a persistent and circulative–propagative manner. During the last few decades, RSV has caused serious disease outbreaks in rice fields in China. The results of long-term coevolution have led to complex and diverse relationships between viruses and vector insects, and understanding these interactions is critical for the management of diseases and vector organisms. In this study, three groups of comparative experiments were performed to investigate the effects of RSV infection on the reproductive parameters, vitellogenin (Vg), and Vg receptor (VgR) expressions of L. striatellus. The results showed that RSV infection promoted the fecundity, hatchability, and Vg and VgR expressions of L. striatellus but had no effect on the preoviposition period and oviposition period. Furthermore, the RNA interference of the nucleocapsid protein (CP) gene and the injection of the CP antibody in RSV-viruliferous L. striatellus reduced the fecundity, hatchability, and Vg and VgR expressions, further strengthening the hypothesis that RSV may manipulate the reproductive behavior of insect vectors to promote its own transmission. The results of this study can further reveal the interaction mechanism of virus–vector insects in reproduction and virus transmission, and provide new insights for the control of insect-borne viruses.

Vitellogenin (VTG), a biomarker for endocrine activity, is a mechanistic component of the regulatory assessment of potential endocrine-disrupting properties of chemicals. This review of VTG data is based on changes reported for 106 substances in standard fish species. High intra-study and inter-laboratory variability in VTG concentrations was confirmed, as well as discrepancies in interpretation of results based on large differences between fish in the dilution water versus solvent control, or due to the presence of outlier measurements. VTG responses in fish were ranked against predictions for estrogen receptor agonist activity and aromatase inhibition from bioactivity model output and ToxCast in vitro assay results, respectively. These endocrine mechanisms explained most of the VTG responses in the absence of systemic toxicity, the magnitude of the VTG response being proportional to the in vitro potency. Interpretation of the VTG data was sometimes confounded by an alternative endocrine mechanism of action. There was evidence for both false positive and negative responses for VTG synthesis, but overall, it was rare for substances without endocrine activity in vitro to cause a concentration-dependent VTG response in fish in the absence of systemic toxicity. To increase confidence in the VTG results, we recommend improvements in the VTG measurement methodologies and greater transparency in reporting of VTG data (including quality control criteria for assay performance). This review supports the application of New Approach Methodologies (NAMs) by demonstrating that endocrine activity in vitro from mammalian cell lines is predictive for in vivo VTG response in fish, suggesting that in vitro mechanistic data could be used more broadly in decision-making to help reduce animal testing.

Vitellogenesis is the most important stage of ovarian maturation in fish, involving the synthesis and transport of essential yolk substances and their complex mechanisms and coordination process. The liver is the main tissue involved in the vitellogenesis of oviparous animals, but studies of vitellogenesis in fish rarely include the liver and ovary as a whole. The aim of this study was to explore the molecular mechanism and associated regulatory factors behind vitellogenesis in Sichuan bream (Sinibrama taeniatus). The different stages of oogenesis were first identified by successive histological observations. Then, ovary and liver tissues that developed to 115 days (stage II, previtellogenesis stage), 165 days (stage III, vitellogenesis stage) and 185 days (stage IV, late-vitellogenesis stage) were collected for transcriptomic and metabolomic analyses, and serum testosterone (T), 17β-estradiol (E2), vitellogenin (Vtg), triiodothyronine (T3), and thyroxine (T4) levels were measured at the corresponding stages. We found that energy redistribution during vitellogenesis is mainly regulated through glycolysis, fatty acid biosynthesis and the citrate cycle pathway. In the liver, energy metabolism was promoted by activating glucolipid metabolic pathways to provide sufficient ATP, but at the same time, the ovary tends to retain nutrients rather than decompose them to produce energy. In addition, we have identified several key factors involved in the metabolism of neutral lipids, polar lipids, amino acids and vitamins, which are involved in the assembly and transport of important yolk nutrients. The initiation of vitellogenesis was found to be associated with a surge in serum E2 levels, but the sustained increase in Vtg levels in the late stage may be due more to upregulation of the estrogen receptor. These results provide valuable information about the regulation of ovarian development in cultured fish.

Municipal effluents are well-recognized as disrupting sexual differentiation and reproduction in mussels. However, the contribution to this problem made by rainfall combined with sewer overflow (increased by rain due to climate change) is not well understood. The purpose of this study was to compare the neuroendocrine effects of municipal discharge and rainfall overflow on caged endemic mussel Elliptio complanata. To this end, mussels were experimentally caged and placed for 3 months at a municipal effluent dispersion plume site and at overflow sites. Data revealed that downstream surface water contained some pharmaceuticals (caffeine and carbamazepine) and accumulated significant levels of heterotrophic bacteria, but these effects were not observed at the overflow sites. The principal effects observed at the downstream site were increased soft tissue mass (and gonad index), inflammation, and Vtg proteins in male mussels as determined by a novel immunostaining methodology. The rainfall overflow sites had no effects on these markers, but were specifically associated with reduced Vtg proteins in females, dopamine (Dop), gonad lipids, and DNA strand breaks, with increased metallothioneins. In conclusion, the observed feminizing effects of municipal effluent were not additionally observed in mussels caged at rainfall overflow sites, although the latter exhibited a different pattern of toxicity. [ABSTRACT FROM AUTHOR]

Simple Summary: Climate change is associated with warmer and drier weather on average in central California. At the same time, honeybees are being transported from all over the United States to California for the completion of pollination services that ensure increased yields for a variety of crops, resulting in bees experiencing ambient abiotic stressors. Higher temperatures can reduce air quality, further exacerbating honeybee health challenges. We investigated the relationship between higher daily temperatures and poor air quality and demonstrated associations with incidences of pests (e.g., Varroa mites) and pathogens (e.g., Nosema ceranae). We also correlated the expression of genes linked to immune functioning, oxidative stress, and buffering against temperature and air quality stressors. High daily temperatures are associated with poorer air quality, and they are both associated with lowered immune system functions and increased oxidative stress protection against pests and diseases. Higher Varroa mite loads are correlated with the lower potential ability of honey bees to buffer against temperature stress. Our study provides insights into interactions between climate change-related abiotic stressors and their relation to biotic stressors, which underlie a decline in honeybee health. Climate change-related extreme weather events have manifested in the western United States as warmer and drier conditions with an increased risk of wildfires. Honeybees, essential for crop pollination in California, are at the center of these extreme weather events. We associated the maximum daily temperature and air quality index values with the performance of colonies placed in wildfire-prone areas and determined the impact of these abiotic stressors on gene expression and histopathology. Our results indicate that poor air quality was associated with higher maximum daily temperatures and a lower gene expression level of Prophenoloxidase (ProPO), which is tied to immune system strength; however, a higher gene expression level of Vitellogenin (Vg) is tied to oxidative stress. There was a positive relationship between Varroa mites and N. ceranae pathogen loads, and a negative correlation between Varroa mites and Heat Shock Protein 70 (HSP70) gene expression, suggesting the limited ability of mite-infested colonies to buffer against extreme temperatures. Histological analyses did not reveal overt signs of interaction between pathology and abiotic stressors, but N. ceranae infections were evident. Our study provides insights into interactions between abiotic stressors, their relation to common biotic stressors, and the expression of genes related to immunity and oxidative stress in bees. [ABSTRACT FROM AUTHOR]

Little is known about the molecular mechanisms that underlie the division of labor in stingless bees. We investigated the expression of a set of candidate genes associated with the nurse-to-forager transition in the honey bee in two stingless bee species, Melipona quadrifasciata and Frieseomelitta varia. These species differ with respect to worker reproduction. The workers of M. quadrifasciata lay trophic and reproductive eggs, but those of F. varia are completely sterile. For M. quadrifasciata nurses, we found elevated vitellogenin (Vg) and low juvenile hormone esterase (jhe) transcript levels in the abdomen, in agreement with their reproductive potential. In F. varia, Vg and methyl farnesoate epoxidase (mfe) transcript levels were high in the heads of foragers, indicating an association with foraging behavior. For malvolio (mvl) and foraging (for), both involved with food search behavior, mvl showed persistent low expression in the head, but increasing levels in the abdomen of both species, speaking against a role in behavioral development. The for transcript levels in F. varia were higher in foragers, both in the head and the abdomen, i.e., consistent with foraging. In M. quadrifasciata, however, for expression was higher in the heads of nurse bees, and, thus, is unlikely to play a role in foraging. The results for the two stingless bee species, in comparison with honey bees and bumble bees, indicate that the degree of worker sterility appears to be the primary factor that shapes the expression pattern of key genes in the life history of social bees. [ABSTRACT FROM AUTHOR]

The present study has investigated the effects of Metro Vancouver's wastewater treatment plant (WWTP) effluents on English sole (Parophrys vetulus) hepatic gene expression using novel targeted gene expression assays to complement the 2017 Burrard Inlet Ambient Monitoring Program conducted by Metro Vancouver. Seven locations of varying distance to the WWTPs were included. Twelve genes involved in xenobiotic defense (CYP1A, HSP70), thyroid function (DIO1), lipid and glucose metabolism (FABP1, FASN, GLUT2, PPAR δ , PPAR γ), protein synthesis (18S rRNA, RPS4X), and reproduction (ER α , VTG) revealed several differences between these impacted sites. A key finding of the present study was that males exhibited VTG transcript levels either equivalent or exceeding female levels of this gene at all sites investigated, indicating widespread exposure of estrogenic contaminants throughout Burrard Inlet. Furthermore, the induction of hepatic CYP1A was observed due to possible downstream sites being subjected to a larger influx of certain planar halogenated and non-halogenated hydrocarbons from multiple industrial contributors. This study also revealed significant differences between the sites examined and in genes involved in transcriptional regulation and synthesis of proteins, lipids and glucose metabolism, and thyroid hormone metabolism. Collectively, this study demonstrates the potential of molecular biomarkers of urban contaminant exposure in wild caught English sole for use in diagnosing a broader range of adverse health effects when combined with conventional whole organism health indicators. [ABSTRACT FROM AUTHOR]

Maca (Lepidium meyenii Walp.) has been used in folk medicine to treat fertility disturbances, a claim that has been evidenced in some studies. However, the clinical trials validating this use have shown paradoxical findings and then maca safety is not well known. This study investigated the effects and mechanisms by which maca affects the reproductive system using an in vivo model, the nematode Caenorhabditis elegans. Tuber maca powder, obtained from local commerce, was used to prepare the aqueous extract. Worms were acutely exposed to maca extracts (40, 120, 240, and 330 μg/μl), and 48 h after treatments, physiological and biochemical assays were conducted. Maca extract caused a significant decrease in total number of eggs and in the number of eggs per worm. These effects were associated to increased lipid peroxidation, reduced triacylglycerol levels, and also impaired vit‐2 (vitellogenin) expression, besides increase in the number of apoptotic germline cells. We have found quantifiable levels of alkaloids in this maca extract, which presence could be related to this toxicity. Collectively, our data suggest that maca extract exposure causes reproductive toxicity to worms that could be, at least in part, associated to both an increase in apoptosis of germline cells and also to a decrease in vitellogenin expression, needed for egg yolk production and, consequently, successful reproduction. Maca (Lepidium meyenii Walp.) has been used in folk medicine to treat fertility disturbances, a claim that has been evidenced in some studies. This study investigated the effects and mechanisms by which maca affects the reproductive system using an in vivo model, the nematode Caenorhabditis elegans. Tuber maca powder, obtained from local commerce, was used to prepare the aqueous extract. Worms were acutely exposed to maca extracts (40, 120, 240, and 330 μg/μl), and 48 h after treatments, physiological and biochemical assays were conducted. Maca extract caused a significant decrease in total number of eggs and in the number of eggs per worm. These effects were associated to increased lipid peroxidation, reduced triacylglycerol levels, and also impaired vit‐2 (vitellogenin) expression. [ABSTRACT FROM AUTHOR]

In human and veterinary medicine, serum proteins are considered to be useful biomarkers for assessing the health and nutritional status of the organism. Honeybee hemolymph has a unique proteome that could represent a source of valuable biomarkers. Therefore, the aims of this study were to separate and identify the most abundant proteins in the hemolymph of worker honeybees to suggest a panel of these proteins that could represent useful biomarkers for assessing the nutritional and health status of the colonies and, finally, to analyze them in different periods of the year. Four apiaries were selected in the province of Bologna, and the bees were analyzed in April, May, July, and November. Thirty specimens from three hives of each apiary were sampled and their hemolymph was collected. The most represented bands obtained after 1D sodium-dodecyl-sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) were cut from the gel, and the proteins were identified using an LC-ESI-Q-MS/MS System. A total of twelve proteins were unmistakably identified; the two most abundant proteins were apolipophorin and vitellogenin, which are known biomarkers of bee trophic and health status. The two other proteins identified were transferrin and hexamerin 70a, the first being involved in iron homeostasis and the second being a storage protein. Most of these proteins showed an increase from April to November, mirroring the physiological changes of honeybees during the productive season. The current study suggests a panel of biomarkers from honeybee hemolymph worth testing under different physiological and pathological field conditions. [ABSTRACT FROM AUTHOR]

Diaphorina citri, a vector of citrus huanglongbing (HLB) disease, frequently leads to HLB outbreaks and reduces Rutaceae crop production. Recent studies have investigated the effects of RNA interference (RNAi) targeting the Vitellogenin (Vg4) and Vitellogenin receptor (VgR) genes, which are involved in egg formation in this pest, providing a theoretical foundation for developing new strategies to manage D. citri populations. This study presents RNAi methods for Vg4 and VgR gene expression interference and reveals that dsVgR is more effective than dsVg4 against D. citri. We demonstrated that dsVg4 and dsVgR persisted for 3–6 days in Murraya odorifera shoots when delivered via the in-plant system (IPS) and effectively interfered with Vg4 and VgR gene expression. Following Vg4 and VgR gene expression interference, egg length and width in the interference group were significantly smaller than those in the negative control group during the 10–30-day development stages. Additionally, the proportion of mature ovarian eggs in the interference group was significantly lower than that in the negative control group at the 10, 15, 20, 25, and 30-day developmental stages. DsVgR notably suppresses oviposition in D. citri, with fecundity decreasing by 60–70%. These results provide a theoretical basis for controlling D. citri using RNAi to mitigate the spread of HLB disease. [ABSTRACT FROM AUTHOR]