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The aim of this study was to apply biochemical parameters to Onychostoma barbatulum to understand whether Wuling Farm activities have caused organic pollution from xenoestrogens to flow into surrounding streams and thereby affected area aquatic ecosystems. Individuals of the Taiwan shovel-jaw carp (O. barbatulum) were collected by fyke netting in the Cijiawan, Kaoshan, and Yusheng Rivers in the protected area of Shei-Pa National Park and in the Ikawan River outside the protected areas from 4–5 June 2015 to 20–21 September 2015. The collected male individuals of O. barbatulum were divided into two groups. In the first group monooxygenase (Mon) activity, glutathione S-transferase (GST) activity, and vitellogenin (VTG) concentration in the liver were measured right after they were captured. Those in the second group were acclimated for 14 days and then exposed to 0 (control) or 10 ng/L 17β-estradiol for 10 days and then measured for changes in Mon, GST, and VTG in their livers. The value of VTG concentrations in O. barbatulum individuals collected from the Ikawan River was 2.12 ± 1.62 μg PO42+/g protein, which was the highest among all samples, followed by those collected from the Yusheng River (0.78 ± 1.00 μg PO42+/g protein). Individuals collected from the Yusheng River had the highest liver Mon activity (4.16 ± 1.08△A650/30 min/g protein) and the highest GST activity (1.58 ± 1.13 △A340/20 min/g protein), followed by those from the Ikawan River (Mon: 3.63 ± 1.13 △A650/30 min/g protein; GST: 1.24 ± 0.73 △A340/20 min/g protein). Comprehensive analyses showed that Mon and GST activities and VTG concentrations in livers measured right after being collected were lower than individuals exposed to 10 ng 17β-estradiol/L in the laboratory. Induced VTG concentrations were 9.87 ± 0.89 μg PO42+/g protein, Mon activity 8.02 ± 1.74 △A650/30 min/g protein, and GST activity 3.24 ± 0.62 △A340/20 min/g protein, indicating that farming activities have not significantly affected these aquatic organisms. However, pollution sources are still releasing pollutants containing xenoestrogens into the Yusheng River and Ikawan River. The impact of xenoestrogens is worthy of continuous long-term follow-up monitoring. [ABSTRACT FROM AUTHOR]

Background: Vitellogenin (Vg), known as the yolk protein precursor for oocyte development in female insects, can be secreted to plant host from salivary glands of hemipterans, including rice leafhopper Recilia dorsalis. The aim of this study was to investigate the function of salivary-secreted Vg of R. dorsalis (RdVg) in rice host. We propose that RdVg possibly regulates the rice defense against insects, benefiting R. dorsalis feeding., Results: RdVg was released into rice phloem along with saliva during R. dorsalis feeding. Knocking down RdVg increased the level of H 2 O 2 and improved H 2 O 2 metabolism in rice plants, making it difficult for R. dorsalis to feed. The transient expression or overexpression of the lipoprotein N-terminal domain of RdVg (RdVg2) significantly reduced hydrogen peroxide (H 2 O 2 ) metabolism in plants. This suggests that salivary-secreted RdVg acts as an effector suppressing the H 2 O 2 burst in rice plants, and RdVg2 is the key domain. RdVg2 could interact with rice sulfite oxidase (OsSO), which catalyzes the oxidation of SO 3 2- and produces H 2 O 2 . Exposure of rice plants to R. dorsalis, overexpression of RdVg2 or knocking out OsSO reduced OsSO accumulation and SO 3 2- oxidation, benefiting R. dorsalis feeding. However overexpression of OsSO increased SO 3 2- oxidation and H 2 O 2 metabolism, inhibiting R. dorsalis feeding., Conclusion: RdVg inhibits H 2 O 2 generation via suppressing OsSO accumulation, ultimately benefiting R. dorsalis feeding. These findings identify RdVg as an effector that suppresses plant defense to insects, and provide insights into the function of salivary-secreted Vg in other Hemiptera insects. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

Panonychus citri is one of the most destructive pests in citrus orchards, exhibiting varying degrees of tolerance to numerous insecticides, such as spirodiclofen. To effectively manage pests, this study explores the response of P. citri to spirodiclofen stress from the perspectives of life history, enzymatic parameters, and reproduction. The effects of two concentrations (LC 30 and LC 50 ) of spirodiclofen on the biological parameters of P. citri were evaluated by the life table method. The results showed that the development duration, fecundity, oviposition days, and lifespan were shortened, though the pre-oviposition period of two treatments was prolonged in comparison with the control. A significant decrease was recorded in the net reproductive rate ( R 0 ) and the mean generation time ( T ) for the two treatments. Nevertheless, the intrinsic rate of increase ( r ) and the rate of increase ( λ ) were not significantly affected in the LC 30 treatment. The enzyme activity assay resulted in higher activities of catalase (CAT), superoxide dismutase (SOD), peroxidase (POD), and carboxylesterase (CarE), among the treatments than the control. In contrast, the treatments recorded lower cytochromeP450 (CYP450) and Glutathione S-transferase (GST) activities than the control. Furthermore, the study detected that relative mRNA expression of Vitellogenin (Vg) and Vitellogenin receptor (VgR) for two treatments were lower than the control. In summary, two concentrations of spirodiclofen inhibited progeny growth and fecundity of 50 treatment. The enzyme activity assay resulted in higher activities of catalase (CAT), superoxide dismutase (SOD), peroxidase (POD), and carboxylesterase (CarE), among the treatments than the control. In contrast, the treatments recorded lower cytochromeP450 (CYP450) and Glutathione S-transferase (GST) activities than the control. Furthermore, the study detected that relative mRNA expression of Vitellogenin (Vg) and Vitellogenin receptor (VgR) for two treatments were lower than the control. In summary, two concentrations of spirodiclofen inhibited progeny growth and fecundity of P. citri . Additionally, the results of this study may support further research on tolerance of P. citri in response to spirodiclofen stress.

Background: The nano-particle of zinc oxide (nZn) and selenium (nSe) has recently attracted in animal feeding because of their high absorbability, catalytic efficiency, and low toxicity than organic and non-organic forms. The present study was done to evaluate the effect of nZn and nSe supplementation in the honey bee's diet under heat stress on antioxidant status, aminotransferase activity, and the relative gene expression of vg and sod-1. Materials and methods: Fifty honey bee colonies were randomly divided into five treatments and 10 replicates. Treatments were sugar syrup without or with nanoparticle supplementation, as: i) the control group (no additives); ii) 100 µgL−1 nano-zinc oxide (nZn-1); iii) 200 µg L − 1 of nano-zinc oxide (nZn-2); iv) 100 µg L − 1 nano-selenium (nSe-1); v) 200 µg L − 1 nano-selenium (nSe-2). On days 30 and 60 of experiment, the bee's samples were taken and antioxidant status, aminotransferase activities and relative genes expression of vg and sod-1 were measured. Results: The highest (P

Honey bees exhibit an elaborate social structure based in part on an age-related division of labor. Young workers perform tasks inside the hive, while older workers forage outside the hive, tasks associated with distinct diets and metabolism. Critical to colony fitness, the work force can respond rapidly to changes in the environment or colony demography and assume emergency tasks, resulting in young foragers or old nurses. We hypothesized that both task and age affect the gut microbiota consistent with changes to host diet and physiology. We performed two experiments inducing precocious foragers and reverted nurses, then quantified tissue-specific gut microbiota and host metabolic state associated with nutrition, immunity and oxidative stress. In the precocious forager experiment, both age and ontogeny explained differences in midgut and ileum microbiota, but host gene expression was best explained by an interaction of these factors. Precocious foragers were nutritionally deficient, and incurred higher levels of oxidative damage relative to age-matched nurses. In the oldest workers, reverted nurses, the oxidative damage associated with age and past foraging was compensated by high Vitellogenin expression, which exceeded that of young nurses. Host-microbial interactions were evident throughout the dataset, highlighted by an age-based increase of Gilliamella abundance and diversity concurrent with increased carbonyl accumulation and CuZnSOD expression. The results in general contribute to an understanding of ecological succession of the worker gut microbiota, defining the species-level transition from nurse to forager.

The increasing efflux on a large scale of organic contaminants holding endocrine disrupting activity from sewage treatment plants produces detrimental biological effects to various fish species. However, the impact of small-scale sewage treatment plant-efflux in small river streams and narrow creeks is largely unknown. Extensive pesticide output especially in vineyards and orchards also causes adverse effects on the endocrine system of wildlife fish species inhabiting nearby rivers. To elaborate whether fish species and populations in the areas of interest were at risk of experiencing endocrine disruption, we identified different biomarkers related to endocrine disruption in Cottus gobio and Salmo trutta fario and applied this approach to selected Austrian freshwater streams pre and post sewage treatment plants and permanent cultures with extensive pesticide output in South Tyrol. Overall, mRNA expression levels of vitellogenin, estrogen receptor α and zona pellucida genes in wildlife fish, compared to a control population reared under constant conditions in the laboratory were significantly increased. Sewage-treatment plant efflux did not significantly affect the mRNA expression levels while extensive use of pesticides altered mRNA expression significantly in C. gobio. C. gobio and S. trutta fario display different levels of mRNA expression. Cadmium and copper concentrations in liver tissues varied but did not indicate significant levels of contamination. Our results demonstrate the presence of endocrine disrupting chemicals in the tested freshwater streams. We anticipate our study to be a starting point for further studies focusing on the effects of endocrine disrupting chemicals on individuals and populations. Especially the fact that the two selected species reveal highly different levels of mRNA expression levels is of interest when applying biomarker approaches which can be a useful tool for monitoring projects and risk-assessment associated studies.

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.

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, PPARδ, PPARγ), protein synthesis (18S rRNA, RPS4X), and reproduction (ERα, 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.

Simple Summary: The tobacco cutworm, Spodoptera litura (F.), exemplifies strong reproductive capacities and damages many agricultural crops. The insulin signaling pathway is known as a key determinant of female reproduction in insects. However, the detailed molecular mechanisms in these processes are poorly studied. Here, we injected bovine insulin into the newly emerged moth, resulting in gene expression changes in the insulin pathway, while knockdown of SlInR caused an inverse gene expression change involved in the insulin pathway. Further studies indicated that the content of JH-III, Vg, total proteins and triacylgycerol could be suppressed by SlInR dsRNA injection. Furthermore, stunted ovaries and lower fecundity were observed by RNAi. Our studies indicated that SlInR plays a key role in JH-III synthesis and the ovarian development in S. litura. Insulin signaling can regulate various physiological functions, such as energy metabolism and reproduction and so on, in many insects, including mosquito and locust. However, the molecular mechanism of this physiological process remains elusive. The tobacco cutworm, Spodoptera litura, is one of the most important pests of agricultural crops around the world. In this study, phosphoinositide 3-kinase (SlPI3K), protein kinase B (SlAKT), target of rapamycin (SlTOR), ribosomal protein S6 kinase (SlS6K) and transcription factor cAMP-response element binding protein (SlCREB) genes, except transcription factor forkhead box class O (SlFoxO), can be activated by bovine insulin injection. Then, we studied the influence of the insulin receptor gene (SlInR) on the reproduction of S. litura using RNA interference technology. qRT-PCR analysis revealed that SlInR was most abundant in the head. The SlPI3K, SlAKT, SlTOR, SlS6K and SlCREB genes were decreased, except SlFoxO, after the SlInR gene knockdown. Further studies revealed that the expression of vitellogenin mRNA and protein, Methoprene-tolerant gene (SlMet), could be down-regulated by the injection of dsRNA of SlInR significantly. Furthermore, a depletion in the insulin receptor by RNAi significantly decreased the content of juvenile hormone III (JH-III), total proteins and triacylgycerol. These changes indicated that a lack of SlInR could impair ovarian development and decrease fecundity in S. litura. Our studies contribute to a comprehensive insight into reproduction, regulated by insulin and the juvenile hormone signaling pathway through nutrition, and a provide theoretical basis for the reproduction process in pest insects. [ABSTRACT FROM AUTHOR]

During their breeding season, estrogen induces vitellogenin (VTG) production in the liver of teleost fish through estrogen receptors (ERs) that support oocyte vitellogenesis. There are at least three ER subtypes in teleost fish, but their roles in mediating E2-induced VTG expression have yet to be ascertained. In this study, we investigated the expression of vtgs and ers in the liver of orange-spotted grouper (Epinephelus coioides). Their expression levels were significantly increased in the breeding season and were upregulated by an estradiol (E2) injection in female fish, except for the expression of erβ1. The upregulation of vtgs, erα and erβ2 by E2 was also observed in primary hepatocytes, but these stimulatory effects could be abolished by ER antagonist ICI182780 treatment. Subsequent studies showed that ERβ antagonist Cyclofenil downregulated the E2-induced expression of vtg, erα, and erβ2, while the ERβ agonist DPN simulated their expression. Knockdown of erβ2 by siRNA further confirmed that ERβ2 mediated the E2-induced expression of vtgs and erα. To reveal the mechanism of ERβ2 in the regulation of erα expression, the erα promoter was cloned, and its activity was examined in cells. E2 treatment simulated the activity of the erα promoter in the presence of ERβ2. Deletions and site-directed mutations showed that the E2 up-regulated transcriptional activity of erα occurs through a classical half-estrogen response element- (ERE) dependent pathway. This study reveals the roles of ER subtypes in VTG expression in orange-spotted grouper and provides a possible explanation for the rapid and efficient VTG production in this species during the breeding season. [ABSTRACT FROM AUTHOR]

We investigated the effects of fine particulate matter from hydrothermal ore on the physiological and molecular responses of the marine copepod Tigriopus japonicus. We evaluated its life cycle parameters including survival, development, and fecundity and the transcriptional profiles of its genes regulating detoxification (cytochrome P450 [CYP]), antioxidation (glutathione S-transferase [GST]), and development (vitellogenin [Vtg]) in response to hydrothermal ore and beneficiation leachates. There were preponderances of copper, iron, and zinc in the hydrothermal ore and the leachates derived from it. There were no significant changes in copepod developmental time in response to leachate exposure. However, copepod survival and fecundity significantly changed in response to the leachates. Moreover, the transcriptional profiles of the detoxification, antioxidation, and development-related genes were significantly modulated following leachate treatment. Hence, hydrothermal ore and beneficiation leachates promoted the transcriptional expression of detoxification, antioxidation, and development-related genes and altered survival and fecundity in T. japonicus. The results of the present study may help to assess the potential impacts of ore mining, mineral extraction, and beneficiation on the physiological and molecular responses of marine organisms. [ABSTRACT FROM AUTHOR]

Proteins are under selection to maintain central functions and to accommodate needs that arise in ever‐changing environments. The positive selection and neutral drift that preserve functions result in a diversity of protein variants. The amount of diversity differs between proteins: multifunctional or disease‐related proteins tend to have fewer variants than proteins involved in some aspects of immunity. Our work focuses on the extensively studied protein Vitellogenin (Vg), which in honey bees (Apis mellifera) is multifunctional and highly expressed and plays roles in immunity. Yet, almost nothing is known about the natural variation in the coding sequences of this protein or how amino acid‐altering variants might impact structure–function relationships. Here, we map out allelic variation in honey bee Vg using biological samples from 15 countries. The successful barcoded amplicon Nanopore sequencing of 543 bees revealed 121 protein variants, indicating a high level of diversity in Vg. We find that the distribution of non‐synonymous single nucleotide polymorphisms (nsSNPs) differs between protein regions with different functions; domains involved in DNA and protein–protein interactions contain fewer nsSNPs than the protein's lipid binding cavities. We outline how the central functions of the protein can be maintained in different variants and how the variation pattern may inform about selection from pathogens and nutrition. [ABSTRACT FROM AUTHOR]

β‐Amyloid toxicity (Aβ) is an important pathological factor of Alzheimer's disease (AD). Studies have shown that genistein can reduce the toxicity of Aβ to a certain extent; however, the specific mechanism is still uncertain. In the study, we applied Caenorhabditis elegans strains expressing Aβ peptides to evaluate the role of genistein inhibiting Aβ toxicity and the undying mechanism. Genistein influencing the sterol metabolism pathway, the HSP‐16.2 pathway, and lipofuscin in different strains of C. elegans were studied using reverse transcription‐polymerase chain reaction, fluorescence labeling, RNA interference (RNAi), and so forth. Our results showed that genistein alleviated the paralysis of transgenic C. elegans strains. Furthermore, in AD C. elegans, genistein reduced the fluorescence of lipofuscin, downregulated the messenger RNA (mRNA) level of vit‐3 and vit‐6 which were related to the sterol metabolism pathway, significantly increased the mRNA level and protein level of HSP‐16.2, increased the nuclear translocation of the DAF‐16 transcription factor and increased the survival rate after heat stress, which was closely associated with HSP‐16.2 levels. However, the paralysis‐alleviating effect of genistein was greatly reduced because of RNAi‐mediated inhibition of hsp‐16.2, indicating that the anti‐Aβ toxicity effect of genistein was greatly dependent on HSP‐16.2. The above results suggest that genistein inhibiting the toxicity of Aβ in C. elegans, is involved in the modulation of the sterol metabolism pathway by promoting transcription factor DAF‐16 translocation into the nucleus, increasing the expression level of HSP‐16.2, and reducing the levels of lipofuscin through its antioxidant activity. [ABSTRACT FROM AUTHOR]

Abstract Vitellogenin (Vg), a storage protein, has been significantly studied for its egg yolk precursor role in oviparous animals. Recent studies found that vitellogenin and its Vg‐like homologs were fundamentally involved in many other biological processes in social insects such as female caste differences and oxidative stress resilience. In this study, we conducted the first large‐scale molecular evolutionary analyses of vitellogenin coding genes (Vg) and Vg‐like genes of bumble bees, a primitively eusocial insect belonging to the genus Bombus. We obtained sequences for each of the four genes (Vg, Vg‐like‐A, Vg‐like‐B, and Vg‐like‐C) from 27 bumble bee genomes (nine were newly sequenced in this study), and sequences from the two closest clades of Bombus, including five Apis species and five Tetragonula species. Our molecular evolutionary analyses show that in bumble bee, the conventional Vg experienced strong positive selection, while the Vg‐like genes showed overall relaxation of purifying selection. In Apis and Tetragonula; however, all four genes were found under purifying selection. Furthermore, the conventional Vg showed signs of strong positive selection in most subgenera in Bombus, apart from the obligate parasitic subgenus Psithyrus which has no caste differentiation. Together, these results indicate that the conventional Vg, a key pleiotropic gene in social insects, is the most rapidly evolving copy, potentially due to its multiple known social functions for both worker and queen castes. This study shows that concerted evolution and purifying selection shaped the evolution of the Vg gene family following their ancient gene duplication and may be the leading forces behind the evolution of new potential protein function enabling functional social pleiotropy.

[Display omitted] Vitellogenin (Vg), a yolk protein precursor, plays an important role in the oocyte development of insects and is an important target of genetic pest management. Vg is synthesized in the fat body, transported through haemolymph and accumulates in developing oocytes. In this regard, the eggplant shoot and fruit borer, Leucinodes orbonalis (Lepidoptera: Crambidae) is the major pest in South and South East Asia and a serious concern for farmers. Therefore, in the present study, we have cloned and characterized V g from L. orbonalis (LoVg) for further applications. The cloned Vg consisted of 5,370 base pairs encoding 1,790 amino acid residues long protein. Further, sequence alignment revealed that LoVg has three conserved domains: a Vitellogenin N domain (LPD-N), a domain of unknown function protein families (DUF1943), and a von Willebrand factor type D domain (VWD). Using phylogenetic analysis, it was found that LoVg evolved alongside homologous proteins from different insects. The real-time expression levels of LoVg were significantly greater in female adults followed by the pupal stage. This suggests that Vg production and absorption in L. orbonalis occurs in the later pupal stage. Our studies showed that editing LoVg using CRISPR/Cas9 did not affect the total number of eggs laid but affected egg hatchability. These studies help us to design newer approaches in insect pest management through genetic suppression for sustainable pest management. [ABSTRACT FROM AUTHOR]

Insects of different orders produce elaborate structures to protect their eggs from the many threats they may face from the environment and natural enemies. In the weevil genus Gonipterus, their dark, hardened egg capsule is possibly generated by a mixture of the insects' excrement and glandular substances. To test this hypothesis, this study focused on the elucidation of protein components present in the egg capsule cover and interrogated them through comparative analysis and gene expression to help infer potential functions. First, female Gonipterus sp. n. 2 reproductive and alimentary tissues were isolated to establish a reference transcriptome-derived protein database. Then, proteins from weevil frass (excrement) and egg capsule cover were identified through mass spectrometry proteomics. We found that certain egg capsule cover proteins were both exclusive and shared between frass and egg capsule cover, including those of plant origin (e.g. photosystem II protein) and others secreted by the weevil, primarily from reproductive tissue. Among them, a mucin/spidroin-like protein and novel proteins with repetitive units that likely play a structural role were identified. We have confirmed the dual origin of the egg capsule cover substance as a blend of the insects’ frass and secretions. Novel proteins secreted by the weevils are key candidates for holding the egg case cover together.

Nutritional stress is one of the major factors affecting the health of honey bees. Supplementing the pollen patty with microalgae enhances the protein content of the patty and therefore is supposed to improve bee’s health. The objective of the present study was to investigate the effect of Chlorella as a dietary supplement on the health and physiology of the honey bee (Apis mellifera). We formulated the honey bee feed by supplementing Chlorella sorokiniana, obtained commercially, with commercially available rapeseed pollen patty in different amounts—0.5, 2, 5, and 10%, and the treatment groups were named P0.5, P2, P5, and P10, respectively. Pollen patty was set as a positive control and only 50% sucrose solution (no protein) was set as a negative control. Diets were supplied ad libitum to newly emerged workers in cages; food consumption patterns; longevity; and physiology including the development of the brain in terms of protein (i.e., total amino acids), thorax muscle, fat body mass, and glands (hypopharyngeal and venom); and gene expression of nutrition-related gene vitellogenin (Vg) of honey bee at different points of time of their age were observed. The addition of Chlorella significantly increased the food consumption pattern, longevity, gland development, muscle formation, and Vg gene expression significantly in comparison to only a pollen or sugar diet. However, the response varied depending on the level of Chlorella supplementation. As depicted in most of the cases, P2, that is, the pollen diet with 2% of Chlorella supplement exhibited the best outcome in terms of all the tested parameters. Therefore, based on the results obtained in the present study, we concluded that 2% Chlorella supplementation to pollen patty could enhance the health of honey bees, which in turn improves their performance.

ABSTRACT Candidatus Liberibacter solanacearum (CLso) haplotype D, transmitted by the carrot psyllid Bactericera trigonica, is a major constraint for carrot production in Israel. Unveiling the molecular interactions between the psyllid vector and CLso can facilitate the development of nonchemical approaches for controlling the disease caused by CLso. Bacterial surface proteins are often known to be involved in adhesion and virulence; however, interactions of CLso with carrot psyllid proteins that have a role in the transmission process has remained unexplored. In this study, we used CLso outer membrane protein (OmpA) and flagellin as baits to screen for psyllid interacting proteins in a yeast two-hybrid system assay. We identified psyllid vitellogenin (Vg) to interact with both OmpA and flagellin of CLso. As Vg and autophagy are often tightly linked, we also studied the expression of autophagy-related genes to further elucidate this interaction. We used the juvenile hormone (JH-III) to induce the expression of Vg, thapsigargin for suppressing autophagy, and rapamycin for inducing autophagy. The results revealed that Vg negatively regulates autophagy. Induced Vg expression significantly suppressed autophagy-related gene expression and the levels of CLso significantly increased, resulting in a significant mortality of the insect. Although the specific role of Vg remains obscure, the findings presented here identify Vg as an important component in the insect immune responses against CLso and may help in understanding the initial molecular response in the vector against Liberibacter. IMPORTANCE Pathogen transmission by vectors involves multiple levels of interactions, and for the transmission of liberibacter species by psyllid vectors, much of these interactions are yet to be explored. Candidatus Liberibacter solanacearum (CLso) haplotype D inflicts severe economic losses to the carrot industry. Understanding the specific interactions at different stages of infection is hence fundamental and could lead to the development of better management strategies to disrupt the transmission of the bacteria to new host plants. Here, we show that two liberibacter membrane proteins interact with psyllid vitellogenin and also induce autophagy. Altering vitellogenin expression directly influences autophagy and CLso abundance in the psyllid vector. Although the exact mechanism underlying this interaction remains unclear, this study highlights the importance of immune responses in the transmission of this disease agent.

ABSTRACT The highly social honey bee has dense populations but a significantly reduced repertoire of immune genes relative to solitary species, suggesting a greater reliance on social immunity. Here we investigate immune gene expression and gut microbial succession in queens during colony introduction. Recently mated queens were placed into an active colony or a storage hive for multiple queens: a queen-bank. Feeding intensity, social context, and metabolic demand differ greatly between the two environments. After 3 weeks, we examined gene expression associated with oxidative stress and immunity and performed high-throughput sequencing of the queen gut microbiome across four alimentary tract niches. Microbiota and gene expression in the queen hindgut differed by time, queen breeder source, and metabolic environment. In the ileum, upregulation of most immune and oxidative stress genes occurred regardless of treatment conditions, suggesting postmating effects on gut gene expression. Counterintuitively, queens exposed to the more social colony environment contained significantly less bacterial diversity indicative of social immune factors shaping the queens microbiome. Queen bank queens resembled much older queens with decreased Alpha 2.1, greater abundance of Lactobacillus firm5 and Bifidobacterium in the hindgut, and significantly larger ileum microbiotas, dominated by blooms of Snodgrassella alvi. Combined with earlier findings, we conclude that the queen gut microbiota experiences an extended period of microbial succession associated with queen breeder source, postmating development, and colony assimilation. IMPORTANCE In modern agriculture, honey bee queen failure is repeatedly cited as one of the major reasons for yearly colony loss. Here we discovered that the honey bee queen gut microbiota alters according to early social environment and is strongly tied to the identity of the queen breeder. Like human examples, this early life variation appears to set the trajectory for ecological succession associated with social assimilation and queen productivity. The high metabolic demand of natural colony assimilation is associated with less bacterial diversity, a smaller hindgut microbiome, and a downregulation of genes that control pathogens and oxidative stress. Queens placed in less social environments with low metabolic demand (queen banks) developed a gut microbiota that resembled much older queens that produce fewer eggs. The queens key reproductive role in the colony may rely in part on a gut microbiome shaped by social immunity and the early queen rearing environment.

The ridgetail white shrimp Exopalaemon carinicauda has the potential to be used as a useful experimental organism in the field of crustacean research due to its transparent body, large egg, and short reproductive cycle. However, little is known about the regulatory mechanisms of ovarian maturation in E. carinicauda. In this study, we applied PacBio single-molecule long-read sequencing technology to unveil the whole transcriptome landscape of Exopalaemon carinicauda to better understand the molecular mechanisms of ovarian development. A total of 49.05 G polymerase read bases were generated, finally, 15, 151 unigenes were successfully identified with a mean length of 4, 257 bp and an N50 of 4, 884 bp from ovary tissues (five different ovarian developmental stages). A total of 14, 007 unigenes were successfully annotated in at least one public database. In addition, 8, 861 coding sequences, 4, 594 lncRNAs and 895 transcription factors were identified. Furthermore, we performed RNA-Seq analysis combined with Iso-Seq results to investigate ovarian maturation regulation mechanism and validated the results by quantitative real-time PCR. We annotated five vitellogenin isoforms in the full-length transcriptome, meanwhile these five genes and vitellogenin receptor suggested an important role during previtellogenesis of E. carinicauda. Genes were involved in PI3K-Akt signaling pathway, retinol metabolism, cell cycle and hedgehog signaling pathway, which showed obvious associations with ovarian maturation of E. carinicauda. We identified 40 DEGs, such as forkhead box protein L2, which may be related to ovarian development in the comparisons. The expression profiles of genes such as ecdysone receptor, ecdysone-induced protein 74EF, ecdysone inducible gene E75 and post-molt protein suggest that they were significantly associated with reproductive molting in the ovarian mature stage. In conclusion, the present study identified important genes and pathways involved in ovarian maturation, which might be useful for studying the reproductive regulation and mechanisms of ovarian maturation in E. carinicauda.

Neuroparsins (NP) are small-size cysteine-rich neuropeptides first discovered in insects. They are known to be involved in insect reproduction. In this study, we have cloned two neuroparsin cDNAs (i.e., MrNP1 and MrNP2) from the freshwater shrimp Macrobrachium rosenbergii. The two neuroparsins consist of 12 cysteines, which is characteristic of the neuroparsin family. These cysteines are arranged in identical relative positions that form 6-disulfide bonds. MrNP1 and MrNP2 are most similar to the corresponding neuroparsin counterparts of the shrimp Macrobrachium nipponense. Phylogenetic study results suggested that MrNP1 and MrNP2 are closely related to MnNP1 and MnNP3, respectively. Also, an additional MrNP gene similar to MnNP2 is expected to exist in M. rosenbergii. The MrNP1 expression level is the highest in the ovary, and MrNP2 expression is higher in the brain and heart of the females. In addition, during the ovary maturation cycle, MrNP1 expression in the hepatopancreas is highest in stage V; in the ovary it is variable. MrNP2 expression in the hepatopancreas and ovary is the highest in stage II and stage I, respectively. In vivo and in vitro bioassay experiment results indicate that MrNP1 and MrNP2 recombinant proteins can stimulate the expression of the MrVg gene. In contrast, silencing of MrNP1 and MrNP2 genes would suppress MrVg, VgR, and CyclinB gene expressions. The results indicate that the products of both genes can stimulate vitellogenesis by up-regulating the MrVg gene expression. Results from their difference in expression patterns indicate that they might have different regulatory roles in vitellogenin synthesis. Since gene silencing of either MrNP1 or MrNP2 affected the expression of the other NP, we have hypothesized that coordinated regulatory action between MrNP1 and MrNP2 may be necessary for the normal vitellogenesis in M. rosenbergii.

Objective To amplify and analyze the full-length cDNA of Anopheles dirus vitellogenin (AdVg) gene, and express and purify its functional domain. Methods Based on the cDNA fragments of AdVg gene cloned earlier, the full-length sequence was obtained through rapid amplification of cDNA ends (RACE). The sequences of AdVg cDNA fragment, 5' end and 3' end were corrected and assembled to obtain a complete AdVg cDNA full-length sequence by Vector NTI software. Then the full-length sequence of AdVg was used to predict the functional domain. Finally, recombinant plasmid pet28A (+) was used to express and purify the AdVg functional domain. Results The full-length cDNA sequence of the AdVg gene was successfully obtained, including a 117 bp 5' UTR sequence, a 192 bp 3'UTR sequence, and a 6 186 bp open reading frame (ORF) sequence. Meanwhile, the analysis showed that the AdVg protein mainly contained 3 structural domains, including Vit N, DUF1943 and vWD. Then we successfully expressed and purified the functional domain AdVg Vit N by constructing a recombinant plasmid. Conclusion The full-length cDNA of AdVg is successfully cloned, and the recombinant protein of its functional domain Vit N is obtained in this study.

Effluents from municipal wastewater treatment plants (MWTPs) are complex mixtures of chemicals including endocrine-disrupting compounds (EDCs) and 17α-ethynylestradiol (EE2). The objective of this study was to evaluate selected responses of two fish species, in two different years, exposed in situ to MWTP effluent. Biological markers of exposure (plasma vitellogenin (VTG) and antioxidant enzymes) were measured in two species of male fish, rainbow trout (Oncorhynchus mykiss) and fathead minnow (Pimephales promelas), caged at sites associated with wastewater outfall. The estrogenicity of the final effluent in 2010 was determined to be 17.0 + 0.4 ng/L estrogen equivalents (EEQ) and reduced to 7.5 + 2.9 ng/L EEQ after infrastructure upgrades. Pharmaceuticals and personal care products in the effluent and surface water in both years confirmed the exposures at each downstream site. Despite the presence of estrogenic compounds in the MWTP effluent, no effluent-caged male fish demonstrated plasma VTG induction. Minnows and trout that received an intraperitoneal injection of 5 mg/g EE2 showed VTG induction at both field sites. In 2012, the liver somatic index (LSI) of both species increased with exposure, as did changes in antioxidant enzymes, and reactive oxygen species (ROS) activity. Multiple biological mechanisms are modified by effluent exposure, and multiple endpoints are needed to assess risk. HIGHLIGHTS Effluent exposure resulted in induction of antioxidant enzymes and ROS activity in caged fish.; Despite measurable estrogen equivalents in the effluent, downstream caged male fathead minnow did not demonstrate VTG induction.; EE2 injected, control male fish exposed to wastewater or upstream reference conditions responded with elevated VTG concentrations.; Several factors may have reduced the VTG response in caged fish.;

Abstract Background We previously reported the results of CRISPR/Cas9 knock-out (KO) of type-I and type-III vitellogenins (Vtgs) in zebrafish, which provided the first experimental evidence on essentiality and disparate functioning of Vtgs at different stages during early development. However, the specific contributions of different types of Vtg to major cellular processes remained to be investigated. The present study employed liquid chromatography and tandem mass spectrometry (LC-MS/MS) to meet this deficit. Proteomic profiles of zebrafish eggs lacking three type-I Vtgs simultaneously (vtg1-KO), or lacking only type III Vtg (vtg3-KO) were compared to those of wild type (Wt) eggs. Obtained spectra were searched against a zebrafish proteome database and identified proteins were quantified based on normalized spectral counts. Results The vtg-KO caused severe changes in the proteome of 1-cell stage zebrafish eggs. These changes were disclosed by molecular signatures that highly resembled the proteomic phenotype of poor quality zebrafish eggs reported in our prior studies. Proteomic profiles of vtg-KO eggs and perturbations in abundances of hundreds of proteins revealed unique, noncompensable contributions of multiple Vtgs to protein and in energy homeostasis. The lack of this contribution appears to have a significant impact on endoplasmic reticulum and mitochondrial functions, and thus embryonic development, even after zygotic genome activation. Increased endoplasmic reticulum stress, Redox/Detox activities, glycolysis/gluconeogenesis, enrichment in cellular proliferation and in human neurodegenerative disease related activities in both vtg1- and vtg3-KO eggs were found to be indicators of the aforementioned conditions. Distinctive increase in apoptosis and Parkinson disease pathways, as well as the decrease in lipid metabolism related activities in vtg3-KO eggs implies compelling roles of Vtg3, the least abundant form of Vtgs in vertebrate eggs, in mitochondrial activities. Several differentially abundant proteins representing the altered molecular mechanisms have been identified as strong candidate markers for studying the details of these mechanisms during early embryonic development in zebrafish and possibly other vertebrates. Conclusions These findings indicate that the global egg proteome is subject to extensive modification depending on the presence or absence of specific Vtgs and that these modifications can have a major impact on developmental competence.

• Java medaka were exposed to environmentally relevant concentrations of E2 and BPA with fragmented PET MPs. • Exposure to the PET MPs induced no significant effect on the growth parameters of Java medaka. • Exposure to the PET MPs induced an additive toxic effect on egg production in Java medaka exposed to BPA. • An additive effect of the PET MPs on the GSI value was observed in Java medaka exposed to BPA. • Additive effects of the PET MPs were observed on VTG levels in males exposed to E2 and BPA. To investigate whether environmental concentrations of fragmented polyethylene terephthalate (PET) microplastics (MPs) have additional or combined effects on endocrine-disrupting activity, Java medaka (Oryzias javanicus) were exposed to 17β-estradiol (E2; 5, 10, 50, and 100 ng L−1), bisphenol A (BPA; 5, 10, 50, and 100 µg L −1), and E2 and BPA combined with PET MPs (1 and 100 particles L −1) for 200 days. The growth parameters, such as body length and weight, were significantly decreased by the highest concentrations of E2 and BPA. A significant reduction in egg production was observed in female fish exposed to BPA, with an additive toxic effect of PET MPs. A female-biased sex ratio was observed in fish exposed to both chemicals. Exposure to E2 significantly increased the hepatosomatic index (HSI) in both sexes, while no significant effect was observed in the gonadosomatic index (GSI). Exposure to BPA significantly increased the HSI in female fish and decreased the GSI in both sexes of fish. An additive effect of PET MPs was observed on the GSI value of female exposed to BPA. Significant elevations in vitellogenin (VTG) levels were observed in both sexes due to exposure to E2 and BPA. Additive effects of PET MPs were observed on VTG levels in males exposed to E2 and BPA. Taken together, even long-term treatment with PET MPs induced only a negligible additive effect on the endocrine-disrupting activity in Java medaka at environmentally relevant concentrations. [ABSTRACT FROM AUTHOR]

• Oreochromis niloticus hepatocytes were exposed to a mixture of PAHs. • Exposure affected the expression of genes related to the estrogen pathway. • Vtg expression increased 120% in response to PAH exposure. • Antioxidant defense system responded to PAH exposure. • Multixenobiotic resistance-associated proteins activity increased. This study aimed to investigate the toxicity and endocrine disrupting potential of a complex mixture of polycyclic aromatic hydrocarbons (PAHs) in the estrogen pathway using hepatocytes of Nile tilapia Oreochromis niloticus , the hepatocytes were exposed to various concentrations of the PAH mixture, and multiple endpoints were evaluated to assess their effects on cell viability, gene expression, oxidative stress markers, and efflux activity. The results revealed that the PAH mixture had limited effects on hepatocyte metabolism and cell adhesion, as indicated by the non-significant changes observed in MTT metabolism, neutral red retention, and crystal violet staining. However, significant alterations were observed in the expression of genes related to the estrogen pathway. Specifically, vitellogenin (vtg) exhibited a substantial increase of approximately 120% compared to the control group. Similarly, estrogen receptor 2 (esr2) showed a significant upregulation of approximately 90%. In contrast, no significant differences were observed in the expression of estrogen receptor 1 (esr1) and the G protein-coupled estrogen receptor 1 (gper1). Furthermore, the PAH mixture elicited complex responses in oxidative stress markers. While reactive oxygen species (ROS) and reactive nitrogen species (RNS) levels remained unchanged, the activity of catalase (Cat) was significantly reduced, whereas superoxide dismutase (Sod) activity, glutathione S-transferase (Gst) activity, and non-protein thiols levels were significantly elevated. In addition, the PAH mixture significantly influenced efflux activity, as evidenced by the increased efflux of rhodamine and calcein, indicating alterations in multixenobiotic resistance (MXR)-associated proteins. Overall, these findings, associated with bioinformatic analysis, highlight the potential of the PAH mixture to modulate the estrogen pathway and induce oxidative stress in O. niloticus hepatocytes. Understanding the mechanisms underlying these effects is crucial for assessing the ecological risks of PAH exposure and developing appropriate strategies to mitigate their adverse impacts on aquatic organisms. [ABSTRACT FROM AUTHOR]

Tick infestations transmit various infectious agents and result in significant socioeconomic consequences. Currently, the primary focus of tick control efforts is identifying potential targets for immune intervention. In a previous study, we identified a highly conserved protein abundant in tick haemolymph extracellular vesicles (EVs) known as translationally controlled tumour protein (TCTP). We have found that native TCTP is present in various tissues of the Rhipicephalus haemaphysaloides tick, including salivary glands, midgut, ovary, and fat body. Notably, TCTP is particularly abundant in the tick ovary and its levels increase progressively from the blood-feeding stage to engorgement. When the TCTP gene was knocked down by RNAi, there was a noticeable delay in ovarian development, and the reproductive performance, in terms of egg quantity and survival, was also hindered. Our investigations have revealed that the observed effects in ovary and eggs in dsRNA-treated ticks are not attributable to cell death mechanisms like apoptosis and autophagy but rather to the reduction in the expression of vitellogenin (Vg1, Vg2, and Vg3) and ferritin (ferritin 1 and ferritin 2) proteins crucial for ovarian development and embryo survival in ticks. Additionally, phylogenetic analysis and structural comparisons of Rh TCTP and its orthologues across various tick species, vertebrate hosts, and humans have shown that TCTP is conserved in ticks but differs significantly between ticks and their hosts, particularly in the TCTP_1 and TCTP_2 domains. Overall, TCTP plays a vital role in tick reproductive development and presents itself as a potential target for tick control in both humans and animals. • TCTP is involved in tick ovary development and oviposition. • RNA interference of tctp affects vitellogenin and ferritin protein expression. • TCTP is highly conserved in ticks but different from their host animals. [ABSTRACT FROM AUTHOR]

Background: Honeybees are one of the most important pollinators in the world, and their products are nowadays included in most anticancer, antiallergic, antimicrobial drugs and are included in cosmetic treatments. In the present study, honeybee venom and Egyptian ethanolic propolis extract (EP) were focused to test their effect on health and some genes for honeybee workers (defensin2, abaecin, hymenoptaecin, vitellogenin, and juvenile hormone esterase). Results: Honeybee venom and Egyptian propolis extract (EP) were used as supplements in the nutrition with different concentrations in Varroa mites-infected colonies to measure the colonies' activities after treatment. The immune-related genes and antimicrobial peptides (AMPs) were evaluated by using qRT-PCR. Treated colonies with HBV and EP showed up-regulation of immune and immune-related genes' expressions and increased the life span, activities and their density of bee workers. The data illustrated that the highest gene expression fold of juvenile hormone esterase was detected in the treated colonies with Egyptian ethanolic propolis extract (EP), while the highest vitellogenin expression fold in treated colonies was with honeybee venom. The up-regulation of antimicrobial peptides occurred in colonies with both treatments. Conclusion: The findings suggest that honeybee venom and Egyptian ethanolic propolis extract (EP) could be used as potential supplements, even at the lowest concentration to develop the immunity of worker bees to increase their efficiency and prevent loss of honeybee colonies due to several diseases closely associated with Varroa mites' infections that cause sudden death. [ABSTRACT FROM AUTHOR]

The protozoan parasite Babesia spp. invades into tick oocytes and remains in the offspring. The transovarial transmission phenomenon of Babesia in ticks has been demonstrated experimentally, but the molecular mechanisms remain unclear. Babesia invasion into oocytes occurs along with the progression of oogenesis. In the present study, to find the key tick factor(s) for Babesia transmission, we focused on molecules involved in yolk protein precursor (vitellogenin, Vg) synthesis and Vg uptake, which are crucial events in tick oogenesis. With a Haemaphysalis longicornis tick– Babesia ovata experimental model, the expression profiles of Akt , target of rapamycin , S6K , GATA , and Vg , Vg synthesis-related genes, and Vg receptor (VgR) and autophagy-related gene 6 (ATG6), Vg uptake-related genes, were analyzed using real-time PCR using tissues collected during the preovipositional period in Babesia -infected ticks. The expression levels of H. longicornis Vg-2 (HlVg-2) and HlVg-3 decreased in the fat body of Babesia -infected ticks 1 day after engorgement. In the ovary, HlVg-2 mRNA expression was significantly higher in Babesia -infected ticks than in uninfected ticks 1 and 2 days after engorgement and decreased 3 days after engorgement. HlVgR expression was significantly lower in Babesia -infected ticks than in uninfected ticks 2 and 4 days after engorgement. HlATG6 had a lower gene expression in Babesia -infected ticks compared to uninfected ticks 2 days after engorgement. Additionally, western blot analysis using protein extracts from each collected tissue revealed that H. longicornis Vg-2 (HlVg-2) accumulate in the fat body and hemolymph of Babesia -infected ticks. These results suggest that Vg uptake from the hemolymph to the ovary was suppressed in the presence of B. ovata. Moreover, HlVg-2 knockdown ticks had a lower detection rate of B. ovata DNA in the ovary and a significant reduction of B. ovata DNA in the hemolymph compared with control ticks. Taken together, our results suggest that accumulated HlVg-2 is associated with Babesia infection or transmission in the tick body. These findings, besides previous reports on VgR, provide important information to elucidate the transovarial transmission mechanisms of pathogens in tick vectors. [ABSTRACT FROM AUTHOR]

Atrazine, one of themost intensively applied herbicides worldwide, is commonly found in several water bodies, affecting the associated fauna. Autochthon crustacean species have been relatively less studied, compared to vertebrate species, particularly concerning reproductive success. In this mini-review, we summarize the relevant information about the effects of atrazine exposure on themain reproductive aspects of crustaceans. One of these effects is related to the inhibition of ovarian growth. In this respect, a diminished vitellogenin content was found in the ovary of crabs exposed to atrazine during the entire period of ovarian growth, in correlation with a reduced oocyte size and a delay of ovarianmaturation. Similar results were observed in crayfish. Atrazine was also able to affect the reproductive process, acting as an endocrine disruptor. In this sense, this herbicide was suspected to affect the secretion of some neurohormones involved in the gonadal growth, as well as to alter the circulating levels of steroid hormones which promote the synthesis of vitellogenin for ovarian growth. Moreover, atrazine induced sexual differentiation in juvenile crayfish toward a higher proportion of females,while it produced an increment ofmales in daphnids. Another aspect affected by this herbicidewas the reduction of offspring production, as well as several embryonic abnormalities; genotoxic effects have been also reported in crayfish. Finally, some metabolic imbalances, such as reduction in energy reserves, have been observed in some species, together with oxidative stress and histopathological effects. [ABSTRACT FROM AUTHOR]

Heterotis niloticus is an African species of Osteoglossiformes that presents biological peculiarities and zootechnical performances favorable for fish farming. However, the absence of a sexual dimorphism hinders the optimization of its reproduction in captivity and limits the understanding of its reproductive behavior. This study is aimed at developing a minimally invasive and reliable sexing method to detect vitellogenin (Vtg) in female plasma. A commercial sexing kit (Acobium, Montpellier, France) for Arapaima gigas—a phylogenetically sister species of H. niloticus—successfully identified only 20% of mature H. niloticus females. Enzyme-linked immunosorbent assays (ELISA) were carried out using three Vtg antibodies. The A. gigas Vtg1 antibody cross-reacted significantly with plasma dilutions of female H. niloticus ranging from 1:1000 to 1:10,000, but with relatively low intensity. The Vtg antibody from Osteoglossum bicirrhosum, another species of Osteoglossiformes, showed non-specific binding with the Vtg of H. niloticus female plasma. Finally, an antibody for H. niloticus Vtg developed in this study allowed us to differentiate the two sexes with plasma coating dilutions ranging from 1:1000 to 1:10,000. The results of the assay were validated by a proteomic approach showing that Vtg-targeted mass spectrometry analysis of H. niloticus blood protein extracts could be used to accurately determine the presence of Vtg in the plasma of mature females. The final validation of the ELISA technique using the H. niloticus Vtg antibody was confirmed by visual sexing of a significant number of blood-sampled fish gonads; 100% of the fish were correctly sexed by the ELISA method. [ABSTRACT FROM AUTHOR]

Glucose is vital to embryogenesis, as are glucose transporters. Glucose transporter 4 (Glut4) is one of the glucose transporters, which is involved in rapid uptake of glucose by various cells and promotes glucose homeostasis. Although energy metabolism in insect reproduction is well known, the molecular mechanism of Glut4 in insect reproduction is poorly understood. We suspect that Glut4 is involved in maintaining glucose concentrations in the ovaries and affecting vitellogenesis, which is critical for subsequent oocyte maturation and insect fertility. Harmonia axyridis (Pallas) is a model organism for genetic research and a natural enemy of insect pests. We studied the influence of the Glut4 gene on the reproduction and development of H. axyridis using RNA interference technology. Reverse transcription quantitative polymerase chain reaction analysis revealed that HaGlut4 was most highly expressed in adults. Knockdown of the HaGlut4 gene reduced the transcript levels of HaGlut4, and the weight and number of eggs produced significantly decreased. In addition, the transcript levels of vitellogenin receptor and vitellogenin in the fat bodies and the ovaries of H. axyridis decreased after the interference of Glut4, and decreased the triglyceride, fatty acid, total amino acid and adenosine triphosphate content of H. axyridis. This resulted in severe blockage of ovary development and reduction of yolk formation; there was no development of ovarioles in the developing oocytes. These changes indicate that a lack of HaGlut4 can impair ovarian development and oocyte maturation and result in decreased fecundity. [ABSTRACT FROM AUTHOR]

Background: Tick hemolymph bathes internal organs, acts as an exchange medium for nutrients and cellular metabolites, and offers protection against pathogens. Hemolymph is abundant in proteins. However, there has been limited integrated protein analysis in tick hemolymph thus far. Moreover, there are difficulties in differentiating tick-derived proteins from the host source. The aim of this study was to profile the tick/host protein components in the hemolymph of Haemaphysalis flava. Methods: Hemolymph from adult engorged H. flava females was collected by leg amputation from the Erinaceus europaeus host. Hemolymph proteins were extracted by a filter-aided sample preparation protocol, digested by trypsin, and assayed by liquid chromatography–tandem mass spectrometry (LC–MS/MS). MS raw data were searched against the UniProt Erinaceidae database and H. flava protein database for host- and tick-derived protein identification. Protein abundance was further quantified by intensity-based absolute quantification (iBAQ). Results: Proteins extracted from hemolymph unevenly varied in size with intense bands between 100 and 130 kDa. In total, 312 proteins were identified in the present study. Therein 40 proteins were identified to be host-derived proteins, of which 18 were high-confidence proteins. Top 10 abundant host-derived proteins included hemoglobin subunit-α and subunit-β, albumin, serotransferrin-like, ubiquitin-like, haptoglobin, α-1-antitrypsin-like protein, histone H2B, apolipoprotein A-I, and C3-β. In contrast, 169 were high-confidence tick-derived proteins. These proteins were classified into six categories based on reported functions in ticks, i.e., enzymes, enzyme inhibitors, transporters, immune-related proteins, muscle proteins, and heat shock proteins. The abundance of Vg, microplusin and α-2-macroglobulin was the highest among tick-derived proteins as indicated by iBAQ. Conclusions: Numerous tick- and host-derived proteins were identified in hemolymph. The protein profile of H. flava hemolymph revealed a sophisticated protein system in the physiological processes of anticoagulation, digestion of blood meal, and innate immunity. More investigations are needed to characterize tick-derived proteins in hemolymph. [ABSTRACT FROM AUTHOR]

Zeugodacus cucurbitae (Coquillett), Bactrocera dorsalis (Hendel), and Ceratitis capitata (Wiedemann) are important pests of fruit and vegetable crops and are difficult to control because of their rapid reproduction rate and egg production. To investigate the key genes regulating reproduction in three fruit fly species, we selected genomic information of three fruit fly species, screened specific genes and single-copy homolog genes, and performed KEGG and GO enrichment analysis on specific genes and single-copy homolog genes of the strong positive select (SP); the results showed that Z. cucurbitae (Coquillett), B. dorsalis (Hendel), and C. capitata (Wiedemann) had seven, 11, and one Vitellogenin -related genes, respectively; Z. cucurbitae (Coquillett) had 84 specific genes enriched in immune system-related pathways; B. dorsalis (Hendel) had 1,121 specific genes enriched in signaling pathways related to cell growth and differentiation; C. capitata (Wiedemann) had 42 specific genes enriched in the degradation and metabolism pathways of exogenous organisms; Z. cucurbitae (Coquillett) may have a stronger immune system; B. dorsalis (Hendel) has a faster developmental and reproductive rate; and C. capitata (Wiedemann) has a higher detoxification capacity. Only one SP single-copy homolog gene (gene name: very long-chain specific acyl-CoA dehydrogenase, mitochondrial) is enriched in the fatty acid metabolic pathway in both Z. cucurbitae (Coquillett) and B. dorsalis (Hendel) as well as in Z. cucurbitae (Coquillett) and C. capitata (Wiedemann). This study provides a molecular basis for studying the reproductive mechanisms of three fruit fly species and provides a scientific basis for developing effective control strategies for fruit flies. [ABSTRACT FROM AUTHOR]

Simple Summary: Vitellogenin (Vg) is the precursor of the yolk protein gene, which is crucial for insect reproduction. In this study, we identified four Vg genes in Zeugodacus cucurbitae (Coquillett). Their molecular characteristics and expression patterns were analyzed. The four genes are mainly expressed in the fat body tissue of adult female melon flies, and their expression is regulated by the juvenile hormone and ecdysone. Nutritional stress significantly down-regulated their expression, indicating that nutrition-dependent vitellogenic development occurs during ovarian development. RNAi-mediated inhibition of the expression of the four genes resulted in significantly delayed ovarian development in Z. cucurbitae. The results indicate that the four genes play an important role in the development of ovaries in Z. cucurbitae. Vitellogenin (Vg) genes encode the major egg yolk protein precursor in arthropods. In this study, four Vgs were identified in Zeugodacus cucurbitae (Coquillett). Sequence analysis showed that four ZcVgs had the conserved Vg domain. Phylogenetic analysis indicated that four ZcVgs were homologous to the Vgs of Tephritidae insects. The temporal and spatial expression patterns of ZcVgs were analyzed by quantitative real-time polymerase chain reaction (RT-qPCR), and the four ZcVgs showed high expression levels in female adults, especially in the fat body. The expression of ZcVg1 and ZcVg3 was down-regulated by a low dosage (0.5 μg) of 20-hydroxyecdysone (20E), and ZcVg2, ZcVg3, and ZcVg4 were up-regulated by a high dosage (1.0 and 2.0 μg) of 20E. The expression of ZcVg1 and ZcVg2 was up-regulated by 5 μg of juvenile hormone (JH), while all of the ZcVgs were down-regulated by a low and high dosage of JH. Expression of ZcVgs was down-regulated after 24 h of starvation and recovered to normal after nutritional supplementation. After micro-injection of the gene-specific double-stranded RNA, the ZcVgs' expression was significantly suppressed, and ovarian development was delayed in Z. cucurbitae females. The results indicate that RNA interference of reproduction-related genes is a potential pest control method that works by manipulating female fertility. [ABSTRACT FROM AUTHOR]

Induction of vitellogenin (VTG) is widely used as a biomarker of exposure of male or immature fish to chemicals that are agonists of the estrogen receptor (i.e., xenoestrogens). Analysis of VTG in samples of epidermal mucosa collected from fish is a non-invasive method for evaluating whether wild fish are exposed to xenoestrogens. In this study, the mean levels of VTG in the mucus of immature brook trout (Salvelinus fontinalis) collected from the Credit River in Ontario, Canada downstream of aging residential septic systems and in an agricultural watershed were 0.67 ng per mg protein, which was significantly elevated relative to the mean VTG levels of 0.22 ng per mg protein in the mucus of immature brook trout collected from a less impacted site. To validate the mucus assay, immature brook trout were exposed in the laboratory to 17α-ethinylestradiol (EE2) at nominal concentrations of 10, 50 and 100 ng/L and VTG levels in mucus from these fish showed a concentration-dependent increase relative to fish from the control treatment. This study illustrates the utility of this non-lethal method for assessing whether wild fish have been exposed in situ to xenoestrogens. Exposures to xenoestrogens from non-point sources may be impacting brook trout populations in urban watersheds in southern Ontario. [ABSTRACT FROM AUTHOR]

We evaluated the impact of a combined vitamin E (vit E), a mixture of nucleotide (mixed-NT), Haematococcus pluvialis (H. pluvialis), arachidonic acid (AA), and yeast extract powder (YEP) supplementations on the relative mRNA expression of reproduction and antioxidant-related genes in red claw crayfish Cherax quadricarinatus using an orthogonal design (L16 45) plus control diet for 70 days feeding trial. Five factors with four levels: A, vit E 40, 160, 240, and 320 mg/kg diet; B, mixed-NT 2, 4, 6, and 8 g/kg diet; C, H. pluvialis2, 4, 6, and 8 g/kg diet; D, AA 5, 10, 15, and 20 g/kg diet; E, YEP 2.5, 5, 10, and 15 g/kg diet were studied. The relative mRNA expression of reproduction-related genes (vitellogenin (vtg) and fatty acid-binding protein (fabp)), and antioxidant-related genes (heat shock protein 70 (hsp70), glutathione peroxidase (gpx), glutathione S-transferase (gst), mitochondrial manganese superoxide dismutase (mmnsod), and ferritin) in hemocyte, hepatopancreas, and ovary tissues were measured. There was a major depression in gene expression levels throughout the experimental diet at high vit E 320 mg/kg level with other factors. Expression of antioxidant enzyme genes significantly increased in treatments of T9, T10, T11, and T12, while significantly decreasing in treatment of T13 and other treatment groups with high vit E 320 mg/kg levels with the other factors. Gene expression of vtg and fabp demonstrated similar patterns to those of antioxidant genes. These results provide evidence that combined supplementations of vit E, mixed-NT, H. pluvialis, AA, and YEP with vit E 240 mg/kg, mixed-NT 8 g/kg, H. pluvialis4 g/kg, AA 5 g/kg, and YEP 10 g/kg may present a novel strategy for improving reproduction performance and antioxidant activities in RC-crayfish, C. quadricarinatus by the positive impact on the molecular level of reproduction and antioxidant genes. [ABSTRACT FROM AUTHOR]

Simple Summary: Transgenerational immune priming (TGIP) to elicit social immunity in the honeybee Apis mellifera has two axes: the first is the ingested pathogen fragments–vitellogenin (Vg)–queen's ovary axis for the developing embryo, and the second is the ingested pathogen fragments–Vg–nurse's hypopharyngeal gland axis for the queen and young larvae through royal jelly. However, the dynamics of the expression of the major royal jelly proteins (MRJPs) in the hypopharyngeal glands of A. mellifera nurse bees after bacterial ingestion must be determined to improve our understanding of the second axis of TGIP. In this study, we investigated the expression patterns of MRJPs 1–7 and defensin-1 in the hypopharyngeal glands and Vg in the fat body of nurse bees fed with live or heat-killed Paenibacillus larvae over 12 h or 24 h by using northern blot analysis. We found that the expression of MRJPs and defensin-1 in the hypopharyngeal glands and Vg in the fat body was significantly induced in nurse bees upon bacterial ingestion, indicating that the differential expression patterns of MRJPs, defensin-1, and Vg were dependent on the bacterial status and timing of bacterial ingestion. We also found that antimicrobial peptide (AMP) genes showed induced expression in young larvae upon bacterial ingestion. In summary, our findings indicate that MRJPs in the hypopharyngeal glands are upregulated along with Vg in the fat body of nurse bees upon bacterial ingestion, providing novel insights into the ingested pathogen fragments–Vg–nurse's hypopharyngeal gland axis for TGIP. Honeybee vitellogenin (Vg) transports pathogen fragments from the gut to the hypopharyngeal glands and is also used by nurse bees to synthesize royal jelly (RJ), which serves as a vehicle for transferring pathogen fragments to the queen and young larvae. The proteomic profile of RJ from bacterial-challenged and control colonies was compared using mass spectrometry; however, the expression changes of major royal jelly proteins (MRJPs) in hypopharyngeal glands of the honeybee Apis mellifera in response to bacterial ingestion is not well-characterized. In this study, we investigated the expression patterns of Vg in the fat body and MRJPs 1–7 in the hypopharyngeal glands of nurse bees after feeding them live or heat-killed Paenibacillus larvae. The expression levels of MRJPs and defensin-1 in the hypopharyngeal glands were upregulated along with Vg in the fat body of nurse bees fed with live or heat-killed P. larvae over 12 h or 24 h. We observed that the expression patterns of MRJPs and defensin-1 in the hypopharyngeal glands and Vg in the fat body of nurse bees upon bacterial ingestion were differentially expressed depending on the bacterial status and the time since bacterial ingestion. In addition, the AMP genes had increased expression in young larvae fed heat-killed P. larvae. Thus, our findings indicate that bacterial ingestion upregulates the transcriptional expression of MRJPs in the hypopharyngeal glands as well as Vg in the fat body of A. mellifera nurse bees. [ABSTRACT FROM AUTHOR]

Metformin is a glucose-lowering drug taken for diabetes. It is excreted by humans in urine and detected in municipal wastewater effluents and rivers. Fathead minnows (Pimephales promelas) were exposed over a life cycle to measured concentrations of metformin: 3.0, 31, and 322 μg/L. No significant changes were observed in survival, maturation, growth, condition factor, or liver size. Relative ovary size of females exposed to 322 μg/L metformin was significantly larger than controls. There was no induction of vitellogenin in plasma of minnows, and gonad maturation was not statistically different from controls. The start of breeding was delayed by 9–10 d in the mid- and high metformin treatments (statistically significant only in the mid-concentration), but numbers and quality of eggs were not statistically different from controls. There were no effects of metformin on survival or growth of offspring. Exposure to metformin at environmentally relevant concentrations (i.e., 3.0 and 31 μg/L metformin) caused no adverse effects in fathead minnows exposed for a life cycle, with the exception of a delay in time to first breeding (that did not impact overall egg production). The results of the study are important to help understand whether metformin concentrations in rivers and lakes can harm fishes.

The protozoan parasite Babesia spp. invades into tick oocytes and remains in the offspring. The transovarial transmission phenomenon of Babesia in ticks has been demonstrated experimentally, but the molecular mechanisms remain unclear. Babesia invasion into oocytes occurs along with the progression of oogenesis. In the present study, to find the key tick factor(s) for Babesia transmission, we focused on molecules involved in yolk protein precursor (vitellogenin, Vg) synthesis and Vg uptake, which are crucial events in tick oogenesis. With a Haemaphysalis longicornis tick–Babesia ovata experimental model, the expression profiles of Akt, target of rapamycin, S6K, GATA, and Vg, Vg synthesis-related genes, and Vg receptor (VgR) and autophagy-related gene 6 (ATG6), Vg uptake-related genes, were analyzed using real-time PCR using tissues collected during the preovipositional period in Babesia-infected ticks. The expression levels of H. longicornis Vg-2 (HlVg-2) and HlVg-3 decreased in the fat body of Babesia-infected ticks 1 day after engorgement. In the ovary, HlVg-2 mRNA expression was significantly higher in Babesia-infected ticks than in uninfected ticks 1 and 2 days after engorgement and decreased 3 days after engorgement. HlVgR expression was significantly lower in Babesia-infected ticks than in uninfected ticks 2 and 4 days after engorgement. HlATG6 had a lower gene expression in Babesia-infected ticks compared to uninfected ticks 2 days after engorgement. Additionally, western blot analysis using protein extracts from each collected tissue revealed that H. longicornis Vg-2 (HlVg-2) accumulate in the fat body and hemolymph of Babesia-infected ticks. These results suggest that Vg uptake from the hemolymph to the ovary was suppressed in the presence of B. ovata. Moreover, HlVg-2 knockdown ticks had a lower detection rate of B. ovata DNA in the ovary and a significant reduction of B. ovata DNA in the hemolymph compared with control ticks. Taken together, our results suggest that accumulated HlVg-2 is associated with Babesia infection or transmission in the tick body. These findings, besides previous reports on VgR, provide important information to elucidate the transovarial transmission mechanisms of pathogens in tick vectors.

Atrazine, one of the most intensively applied herbicides worldwide, is commonly found in several water bodies, affecting the associated fauna. Autochthon crustacean species have been relatively less studied, compared to vertebrate species, particularly concerning reproductive success. In this mini-review, we summarize the relevant information about the effects of atrazine exposure on the main reproductive aspects of crustaceans. One of these effects is related to the inhibition of ovarian growth. In this respect, a diminished vitellogenin content was found in the ovary of crabs exposed to atrazine during the entire period of ovarian growth, in correlation with a reduced oocyte size and a delay of ovarian maturation. Similar results were observed in crayfish. Atrazine was also able to affect the reproductive process, acting as an endocrine disruptor. In this sense, this herbicide was suspected to affect the secretion of some neurohormones involved in the gonadal growth, as well as to alter the circulating levels of steroid hormones which promote the synthesis of vitellogenin for ovarian growth. Moreover, atrazine induced sexual differentiation in juvenile crayfish toward a higher proportion of females, while it produced an increment of males in daphnids. Another aspect affected by this herbicide was the reduction of offspring production, as well as several embryonic abnormalities; genotoxic effects have been also reported in crayfish. Finally, some metabolic imbalances, such as reduction in energy reserves, have been observed in some species, together with oxidative stress and histopathological effects.

Vitellogenin is an essential protein involved in ovary maturation in many animals. Detection of this protein correlated with reproductive capacity may be important if carried out on marine organisms such as the red spiny lobster Palinurus elephas, a crustacean that is an economically important crop from wild fish catches. Moreover, in recent years, vitellogenin has assumed an important role as a possible biomarker of marine environmental pollution, as its expression levels can be influenced by the presence of similar estrogen pollutants and can affect the reproductive sphere of marine organisms such as crustaceans. The P. elephas vitellogenin protein and its coding gene have never been isolated, so there is little information about its presence in this lobster. The aim of the present study was to develop a molecular strategy to create, for the first time, an antibody for the detection and quantization of vitellogenin in P. elephas.

Zeugodacus cucurbitae (Coquillett), Bactrocera dorsalis (Hendel), and Ceratitis capitata (Wiedemann) are important pests of fruit and vegetable crops and are difficult to control because of their rapid reproduction rate and egg production. To investigate the key genes regulating reproduction in three fruit fly species, we selected genomic information of three fruit fly species, screened specific genes and single-copy homolog genes, and performed KEGG and GO enrichment analysis on specific genes and single-copy homolog genes of the strong positive select (SP); the results showed that Z. cucurbitae (Coquillett), B. dorsalis (Hendel), and C. capitata (Wiedemann) had seven, 11, and one Vitellogenin-related genes, respectively; Z. cucurbitae (Coquillett) had 84 specific genes enriched in immune system-related pathways; B. dorsalis (Hendel) had 1,121 specific genes enriched in signaling pathways related to cell growth and differentiation; C. capitata (Wiedemann) had 42 specific genes enriched in the degradation and metabolism pathways of exogenous organisms; Z. cucurbitae (Coquillett) may have a stronger immune system; B. dorsalis (Hendel) has a faster developmental and reproductive rate; and C. capitata (Wiedemann) has a higher detoxification capacity. Only one SP single-copy homolog gene (gene name: very long-chain specific acyl-CoA dehydrogenase, mitochondrial) is enriched in the fatty acid metabolic pathway in both Z. cucurbitae (Coquillett) and B. dorsalis (Hendel) as well as in Z. cucurbitae (Coquillett) and C. capitata (Wiedemann). This study provides a molecular basis for studying the reproductive mechanisms of three fruit fly species and provides a scientific basis for developing effective control strategies for fruit flies.

Vitellogenin (VTG) plays a very important role in the development of oocytes. This study aims to explore the effect of swimming on the VTG synthesis of conger eel (Conger myriaster). Circulating water flow and hormone injection regimen were two factors set in the following trials: A1 (no injection, no water flow), A2 (no injection, water flow), B1 (injection, no water flow), and B2 (injection, water flow). The flow velocity of the flowing water was 0.4 m/s (12 h, 60 days). We examined VTG, estrogen receptor (Er) gene expression, and VTG content in the liver and serum on the 30th and 60th days. VTG gene expression in A1 and A2 gradually decreased and was not significantly different between the two groups. The expression of Er gene in A1 was significantly higher than that in A2. The expression of VTG gene in group B1 was significantly higher than that in group B2, whereas the expression level of Er gene in group B1 was slightly higher than that in group B1. The VTG content in the liver in group B1 reached 1,396.93 μg/L, which was significantly higher than that in group B2 (1,302.06 μg/L). Results showed that the flowing water factor can inhibit the expression of VTG and Er genes in the liver and reduce the synthesis of VTG in the liver. Stimulation of flowing water can inhibit the yolk accumulation during the ovarian development of conger eel.

Current BLASTP search analysis results suggested that the lobster (Homarus americanus) HaCHH-A and HaCHH-B may be derived from two different four-exon genes. Repeated tissue expression studies have revealed much different expression patterns of these two genes from those reported in the past. With RT-PCR, rapid amplification of complementary DNA (cDNA) ends (RACE), and genomic DNA cloning, we confirmed that the HaCHH-A and HaCHH-B transcripts were derived from two different four-exon CHH genes. By an alternative splicing mechanism, each gene can produce different but larger transcript variants (i.e., sHaCHH-A and sHaCHH-B) mainly in different non-eyestalk tissues of the females. The larger and unspliced transcripts can be detected in the hepatopancreas, gill, heart, nerve cord, brain, ovary, and thoracic ganglion of the reproductive females. The expression patterns of sHaCHH-A and sHaCHH-B in other non-eyestalk tissues suggest that these transcripts have a wide spectrum of expressions during the female reproductive cycle. An in vitro organ explant culture system was developed to investigate the reproductive function of these cDNAs. The results showed that the recombinant proteins for sHaCHH-A and sHaCHH-B inhibited the gene expression of vitellogenin, whereas the double-stranded RNA (dsRNA) for sHaCHH-A and sHaCHH-B stimulated the expression of the vitellogenin gene in vitro. The results of the study may provide insights for the development of techniques to induce gonad development without using eyestalk ablation operation. This is the first in-depth report of the characterization of two four-exon CHH genes in a crustacean.

17β-Estradiol (E2) and 17α-methyltestosterone (MT) are steroid hormones with dramatic regulatory effects on the growth performance and gonadal development of aquatic species. The aim of the present study was to investigate the regulatory effects of E2 and MT on the growth performance and gonadal development of juvenile prawns (Macrobrachium nipponense). Male and female juvenile prawns were separately maintained and fed different concentrations of E2 and MT, respectively. Treatment with both E2 and MT at 500 mg/kg dramatically promoted the growth of male and female juvenile prawns, respectively (p 0.05). The expression levels of vitellogenin and vitellogenin receptor were dramatically decreased with increased MT concentrations, and the lowest expression levels were observed in the group treated with 1000 mg/kg (p

Vitellogenin (Vg) is a conserved protein used by nearly all oviparous animals to produce eggs. It is also pleiotropic and performs functions in oxidative stress resistance, immunity, and, in honey bees, behavioral development of the worker caste. It has remained enigmatic how Vg affects multiple traits. Here, we asked whether Vg enters the nucleus and acts via DNA-binding. We used cell fractionation, immunohistology, and cell culture to show that a structural subunit of honey bee Vg translocates into cell nuclei. We then demonstrated Vg-DNA binding theoretically and empirically with prediction software and chromatin immunoprecipitation with sequencing (ChIP-seq), finding binding sites at genes influencing immunity and behavior. Finally, we investigated the immunological and enzymatic conditions affecting Vg cleavage and nuclear translocation and constructed a 3D structural model. Our data are the first to show Vg in the nucleus and suggest a new fundamental regulatory role for this ubiquitous protein. [ABSTRACT FROM AUTHOR]