Comparative genomic analysis of the pattern of evolution of male and female reproductive proteins in seed beetles.

Male seminal fluid proteins (SFPs) often show signs of positive selection and divergent evolution, believed to reflect male - female coevolution. Yet, our understanding of the predicted concerted evolution of SFPs and female reproductive proteins (FRPs) is limited. We sequenced, assembled and annotated the genome of two species of seed beetles allowing a comparative analysis of four closely related species of these herbivorous insects. We compare the general pattern of evolution in genes encoding SFPs and FRPs with those in digestive protein genes and well-conserved reference genes. We found that FRPs showed an overall dN/dS ratio (ω) similar to that of conserved genes, while SFPs and digestive proteins exhibited higher overall ω values. Further, SFPs and digestive proteins showed a higher proportion of sites putatively under positive selection and explicit tests showed no difference in relaxed selection between protein types. Evolutionary rate covariation analyses showed that evolutionary rates among SFPs were on average more closely correlated with those in FRPs than with either digestive or conserved genes. Gene expression showed the expected negative covariation with ω values, except for male-biased genes where this negative relationship was reversed. In conclusion, SFPs showed relatively rapid evolution and signs of positive selection. In contrast, FRPs evolved at a lower rate under selective constraints, on par with genes known to be well-conserved. Although our findings provide support for concerted evolution of SFPs and FRPs, they also suggest that these two classes of proteins evolve under partly distinct selective regimes.

Glyphosate presence in human sperm: First report and positive correlation with oxidative stress in an infertile French population.

Environmental exposure to endocrine disruptors, such as pesticides, could contribute to a decline of human fertility. Glyphosate (GLY) is the main component of Glyphosate Based Herbicides (GBHs), which are the most commonly herbicides used in the world. Various animal model studies demonstrated its reprotoxicity. In Europe, GLY authorization in agriculture has been extended until 2034. Meanwhile the toxicity of GLY in humans is still in debate. The aims of our study were firstly to analyse the concentration of GLY and its main metabolite, amino-methyl-phosphonic acid (AMPA) by LC/MS-MS in the seminal and blood plasma in an infertile French men population (n=128). We secondly determined Total Antioxidant Status (TAS) and Total Oxidant Status (TOS) using commercial colorimetric kits and some oxidative stress biomarkers including malondialdehyde (MDA) and 8-hydroxy-2'-deoxyguanosine (8-OHdG) by ELISA assays. We next analysed potential correlations between GLY and oxidative stress biomarkers concentration and sperm parameters (sperm concentration, progressive speed, anormal forms). Here, we detected for the first time GLY in the human seminal plasma in significant proportions and we showed that its concentration was four times higher than those observed in blood plasma. At the opposite, AMPA was undetectable. We also observed a strong positive correlation between plasma blood GLY concentrations and plasma seminal GLY and 8-OHdG concentrations, the latter reflecting DNA impact. In addition, TOS, Oxidative Stress Index (OSI) (TOS/TAS), MDA blood and seminal plasma concentrations were significantly higher in men with glyphosate in blood and seminal plasma, respectively. Taken together, our results suggest a negative impact of GLY on the human reproductive health and possibly on his progeny. A precaution principle should be applied at the time of the actual discussion of GLY and GBHs formulants uses in Europe by the authorities.

Complementary information concerning the suspected interindividual transmission of GW1516, a substance prohibited in sport, through intimate contact: a case report.

PURPOSE: Inadvertent and/or unknowing exposure to drugs and drug residues has been frequently debated in situations of so-called adverse analytical finding (AAF) in the context of sports drug testing programs. Transfer of drug residues via unprotected intercourse is a conceivable scenario but scientific data and authentic case reports are scarce. Herein, investigations into two AAFs with the peroxisome proliferator-activated receptor delta (PPARδ) agonist GW1516 are reported and discussed. METHODS: To probe for a contamination scenario involving sexual intercourse, two assays were used to determine semenogelin in human urine, with one employing an immunochromatographic lateral flow approach and another based on liquid chromatography-tandem mass spectrometry. Further, drug-residue testing using patients' ejaculate was conducted by utilizing liquid chromatography in conjunction with a triple quadrupole mass spectrometer, followed by re-analysis of suspect samples (i.e., samples indicating the presence of relevant compounds) using high resolution/high mass accuracy mass spectrometry. RESULTS: In one case, but not the other, the possibility of intimate contact as the source of the AAF was confirmed after a thorough investigation of potential contamination scenarios. Subsequent research revealed analytical evidence for the presence of seminal fluid in one of the female athlete's doping control urine samples, and the analysis of clinical ejaculate specimens provided first data on an authentic concentration level of GW1516 and its metabolites in human seminal fluid. CONCLUSIONS: The combined facts substantiate the possibility of an AAF caused by unprotected sexual intercourse and the plausibility of the case-related arguments.

Effects of an entomopathogenic fungus on the reproductive potential of Drosophila males.

While mortality is often the primary focus of pathogen virulence, non-lethal consequences, particularly for male reproductive fitness, are less understood; however, they are essential for understanding how sexual selection contributes to promoting resistance. We investigated how the fungal pathogen Metarhizium brunneum affects mating ability, fertility, and seminal fluid protein (SFP) expression of male Drosophila melanogaster paired with highly receptive virgin females in non-competitive settings. Depending on sex and dose, there was a 3-6-day incubation period after infection, followed by an abrupt onset of mortality. Meanwhile, the immune response was strongly induced already 38 h after infection and continued to increase as infection progressed. Latency to mate somewhat increased during the incubation period compared to sham-treated males, but even on Day 5 post infection >90% of infected males mated within 2 h. During the incubation period, M. brunneum infection reduced male reproductive potential (the number of offspring sired without mate limitation) by 11%, with no clear increase over time. Approaching the end of the incubation period, infected males had lower ability to convert number of mating opportunities into number of offspring. After repeated mating, infected males had lower SFP expression than sham controls, more so in males that mated with few mates 24 h earlier. Overall, despite strong activation of the immune response, males' mating ability and fertility remained surprisingly little affected by the fungal infection, even shortly before the onset of mortality. This suggests that the selection for resistance acts mainly through mortality, and the scope for fertility selection to enhance resistance in non-competing settings is rather limited.

The evolution of seminal fluid gene expression and postmating reproductive isolation in Drosophila.

Seminal fluid protein (Sfp) genes show, in general, a higher rate of sequence divergence than genes from other categories, which is often attributed to forms of postcopulatory sexual selection or sexual conflict. Recently, the relaxation of selective constraints has been proposed as an alternative explanation for the rapid sequence evolution of Sfps and other genes with sex-limited expression. The expression of Sfp genes is a likely target of selection, but the evolution of differences in their expression levels is less understood. Here, we explore both polymorphism and divergence in Sfp gene expression between Drosophila melanogaster and Drosophila simulans, how selection might have influenced their expression, and whether changes in expression might trigger the evolution of reproductive isolating barriers. In our analysis, Sfp genes showed higher divergence, but not higher polymorphism, in expression than the average male reproductive glands gene. Sfp genes with reproductive-tissue-specific expression were enriched for both directional and stabilizing selection, while relaxed selection was the predominant mode of evolution among Sfp genes with any other nonreproductive tissue-specific or nontissue-specific expression. The knockdown of single genes known to affect intraspecific sperm competition, and with patterns of expression divergence and polymorphism suggestive of directional selection, was not enough to break down postmating reproductive isolation barriers between species. Our results identify the expression of male-specific Sfp genes as an enriched target of selection and suggest a complex molecular relationship between postcopulatory sexual selection on a single gene's expression and its effect on the onset of speciation.

Impact of semen microbiota on the composition of seminal plasma.

Several studies have found associations between specific bacterial genera and semen parameters. Bacteria are known to influence the composition of their niche and, consequently, could affect the composition of the seminal plasma. This study integrated microbiota profiling and metabolomics to explore the influence of seminal bacteria on semen metabolite composition in infertile couples, revealing associations between specific bacterial genera and metabolite profiles. Amino acids and acylcarnitines were the predominant metabolite groups identified in seminal plasma. Different microbiota profiles did not result in globally diverse metabolite compositions in seminal plasma. Nevertheless, levels of specific metabolites increased in the presence of a dysbiotic microbiota. Urocanate was significantly increased in abnormal semen samples (adjusted P-value < 0.001) and enriched in samples dominated by Prevotella spp. (P-value < 0.05), which was previously linked to a negative impact on semen. Therefore, varying microbiota profiles can influence the abundance of certain metabolites, potentially having an immunomodulatory effect, as seen with urocanate.IMPORTANCEMale infertility is often considered idiopathic since the specific cause of infertility often remains unidentified. Recently, variations in the seminal microbiota composition have been associated with normal and abnormal semen parameters and may, therefore, influence male infertility. Bacteria are known to alter the metabolite composition of their ecological niches, and thus, seminal bacteria might affect the composition of the seminal fluid, crucial in the fertilization process. Our research indicates that distinct seminal microbiota profiles are not associated with widespread changes in the metabolite composition of the seminal fluid. Instead, the presence of particular metabolites with immunomodulatory functions, such as urocanate, could shed light on the interplay between seminal microbiota and variations in semen parameters.

Sast1-mediated manifold effects inhibit Plutella xylostella fertility.

BACKGROUND: Plutella xylostella (Linnaeus) is a destructive pest of cruciferous crops due to its strong reproductive capacity and extensive resistance to pesticides. Seminal fluid proteins (SFPs) are the main effective factors that determine the reproductive physiology and behaviour of both sexes. Although an increasing number of SFPs have been identified, the effects of astacins in SFPs on agricultural pests have not yet been reported. Here, we elucidated the mechanisms by which Sast1 (seminal astacin 1) regulates the fertility of Plutella xylostella (L.). RESULTS: PxSast1 was specifically expressed in the testis and accesssory gland. CRISPR/Cas9-induced PxSast1 knockout successfully constructed two homozygous mutant strains. Sast1 impaired the fertility of P. xylostella by separately regulating the reproductive capacity of males and females. Loss of PxSast1, on the one hand, significantly decreased the ability of males to mate and fertilize, mainly manifested as shortened mating duration, reduced mating competitiveness and decreased eupyrene sperm production; on the other hand, it significantly inhibited the expression of chorion genes in females, resulting in oogenesis deficits. Simultaneously, for mated females, the differentially expressed genes in signalling pathways related to oogenesis and chorion formation were significantly enriched after PxSast1 knockout. CONCLUSION: These analyses of the functions of PxSast1 as the regulator of spermatogenesis and oogenesis establish its importance in the fertility process of P. xylostella, as well as its potential as a promising target for genetic regulation-based pest control. © 2024 Society of Chemical Industry.

Proteomic differences in seminal fluid of social insects whose sperm differ in heat tolerance.

In the coming years, climate change is likely to increase the frequency and intensity of heatwaves. In many organisms, heat stress provokes physiological perturbations and can lead to decreased male fertility. Bumblebees are endo-heterothermic but display interspecific differences in thermotolerance that could have conservation implications. For the species of concern Bombus magnus, exposure to high temperatures can severely reduce sperm quality and, consequently, reproductive success. Such is not the case for B. terrestris, a ubiquitous species. To decipher the mechanisms at play, we characterized the seminal fluid proteomes of the two species. We quantified 1121 proteins, of which 522 were differentially expressed between B. terrestris and B. magnus. Several proteins with protective functions, such as proteases, antioxidant proteins and various heat-shock proteins, were present at higher levels in B. terrestris than in B. magnus under both control and heat-stress conditions. The same was true for proteins involved in cellular homeostasis, immunity, lipid/sugar metabolism and thermotolerance. Furthermore, proteins involved in the capture and elimination of reactive oxygen species also occurred at much high levels in B. terrestris. Overall, these results clearly indicate differences in the seminal proteome of the more thermotolerant B. terrestris versus B. magnus. The differences may contribute to explaining interspecific differences in sperm survival.

Post-Mating Responses in Insects Induced by Seminal Fluid Proteins and Octopamine.

Following insect mating, females often exhibit a series of physiological, behavioral, and gene expression changes. These post-mating responses (PMRs) are induced by seminal fluid components other than sperm, which not only form network proteins to assist sperm localization, supplement female-specific protein requirements, and facilitate the formation of specialized functional structures, but also activate neuronal signaling pathways in insects. This review primarily discusses the roles of seminal fluid proteins (SFPs) and octopamine (OA) in various PMRs in insects. It explores the regulatory mechanisms and mediation conditions by which they trigger PMRs, along with the series of gene expression differences they induce. Insect PMRs involve a transition from protein signaling to neuronal signaling, ultimately manifested through neural regulation and gene expression. The intricate signaling network formed as a result significantly influences female behavior and organ function, contributing to both successful reproduction and the outcomes of sexual conflict.

Females translate male mRNA transferred during mating.

Although RNA is found in the seminal fluid of diverse organisms, it is unknown whether this RNA is functional within females. Here, we develop an experimental proteomic method called VESPA (Variant Enabled SILAC Proteomic Analysis) to test the hypothesis that Drosophila male seminal fluid RNA is translated by females. We find strong evidence for 67 male-derived, female-translated proteins (mdFTPs) in female lower reproductive tracts at six hours postmating, many with predicted functions relevant to reproduction. Gene knockout experiments indicate that genes coding for mdFTPs play diverse roles in postmating interactions, with effects on fertilization efficiency, and the formation and persistence of the insemination reaction mass, a trait hypothesized to be involved in sexual conflict. These findings advance our understanding of reproduction by revealing a novel mechanism of postmating molecular interactions between the sexes that strengthens and extends male influences on reproductive outcomes in previously unrecognized ways. Given the diverse species known to carry RNA in seminal fluid, this discovery has broad significance for understanding molecular mechanisms of cooperation and conflict during reproduction.

The effect of seminal fluid gene expression on paternity.

When females mate with more than one male, competition between rival ejaculates is expected to favor adaptations that promote fertilization success. There is now compelling evidence that sperm competition selects for increased production and allocation of sperm. However, sperm comes packaged in ejaculates that also contain protein-rich seminal fluids. Predicting how males should allocate individual seminal fluid proteins in response to sperm competition is hampered by our limited knowledge of their precise function. We use gene expression studies and interference RNA to ask how seminal fluid proteins in the ejaculate of a cricket, Teleogryllus oceanicus, affect a male's paternity share when in competition for fertilizations. We find that the relative expression of one seminal fluid gene, gagein, positively affects the paternity share of competing males and that knockdown of this and two other seminal fluid protein genes renders males mating in the offensive role of sperm competition incapable of fathering living offspring. Despite having a negative effect on offspring viability these seminal fluid genes have been found to be up regulated in response to rival males, consistent with a role in promoting competitive fertilization success. Our data contribute to a growing body of evidence that, like sperm, seminal fluid gene expression is subject to post-mating sexual selection via sperm competition.

A predominant role of genotypic variation in both expression of sperm competition genes and paternity success in Drosophila melanogaster.

Sperm competition is a crucial aspect of male reproductive success in many species, including Drosophila melanogaster, and seminal fluid proteins (Sfps) can influence sperm competitiveness. However, the combined effect of environmental and genotypic variation on sperm competition gene expression remains poorly understood. Here, we used Drosophila Genetic Reference Panel (DGRP) inbred lines and manipulated developmental population density (i.e. larval density) to test the effects of genotype, environment and genotype-by-environment interactions (GEI) on the expression of the known sperm competition genes Sex Peptide, Acp36DE and CG9997. High larval density resulted in reduced adult body size, but expression of sperm competition genes remained unaffected. Furthermore, we found no significant GEI but genotypic effects in the expression of SP and Acp36DE. Our results also revealed GEI for relative competitive paternity success (second male paternity; P2), with genes' expression positively correlated with P2. Given the effect of genotype on the expression of genes, we conducted a genome-wide association study (GWAS) and identified polymorphisms in putative cis-regulatory elements as predominant factors regulating the expression of SP and Acp36DE. The association of genotypic variation with sperm competition outcomes, and the resilience of sperm competition genes' expression against environmental challenges, demonstrates the importance of genome variation background in reproductive fitness.

Quality of testicular spermatozoa improves with changes in composition of culture medium.

BACKGROUND: Spermatozoa retrieved from the testis and epididymis are deprived of the beneficial effects of seminal fluid. Thus applying an artificial medium with normal seminal fluid characteristics, known as artificial seminal fluid (ASF), may provide an appropriate condition for improving some sperm parameters in azoospermia. The objective was to investigate the impact of in vitro exposure of testicular and epididymal spermatozoa to ASF on sperm quality. The study was conducted on testicular (n = 20) and epididymal (n = 20) sperm specimens obtained from azoospermic men. Each sample was divided into two equal parts: Part I) for processing and incubation with Ham's F10 medium; Part II) for processing and incubation with ASF. RESULTS: After 2 h incubation, testicular sperm motility was significantly higher in ASF than in Ham's F10 medium. In comparison to 0 h, mitochondrial membrane potential levels of testicular spermatozoa were significantly higher after 2 h and 24 h in ASF and after 24 h in Ham's F10 medium. Furthermore, the data indicated significantly lower rates of epididymal spermatozoa with high MMP in both media after 24 h. There were no significant differences in the DNA fragmentation index of testicular and epididymal spermatozoa between ASF and Ham's F10 medium at different time points. CONCLUSION: The results demonstrated that in vitro incubation of testicular spermatozoa improved their motility more effectively than Ham's F10 medium in the short term (2 h), but had no effect on epididymal spermatozoa. Since the physiology of testicular spermatozoa is different from that of ejaculated spermatozoa, it seems that a special environment should be designed and used for each of them.

RéSUMé: CONTEXTE: Les spermatozoïdes prélevés dans les testicules et les épididymes sont privés des effets bénéfiques du liquide séminal. Ainsi, l'utilisation d'un milieu artificiel avec des caractéristiques normales du liquide séminal, connu sous le nom de liquide séminal artificiel (ASF), peut constituer une condition appropriée pour améliorer certains paramètres des spermatozoïdes obtenus dans l'azoospermie. L'objectif était d'étudier l'impact de l'exposition in vitro de spermatozoïdes testiculaires et épididymaires à l'ASF sur la qualité du sperme. L'étude a été menée sur des échantillons de spermatozoïdes testiculaires (n = 20) et épididymaires (n = 20) obtenus chez des hommes azoospermiques. Chaque échantillon a été divisé en deux parties égales: Partie I) pour le traitement et l'incubation avec le milieu F10 de Ham; Partie II) pour la transformation et l'incubation avec l'ASF. RéSULTATS: Après 2 h d'incubation, la mobilité des spermatozoïdes testiculaires était significativement plus élevée dans l'ASF que dans le milieu F10 de Ham. Par rapport à 0 h, les niveaux du potentiel de membrane mitochondriale (PMM) des spermatozoïdes testiculaires étaient significativement plus élevés après 2 h et 24 h dans l'ASF, et après 24 h dans le milieu F10 de Ham. En outre, les données ont indiqué des taux significativement plus faibles de spermatozoïdes épididymaires avec un PMM élevé dans les deux milieux après 24 heures. Il n'y avait pas de différences significatives dans l'indice de fragmentation de l'ADN des spermatozoïdes testiculaires et épididymaires entre l'ASF et le milieu F10 de Ham aux différents temps. CONCLUSION: Les résultats ont montré que l'incubation in vitro de spermatozoïdes testiculaires dans l'ASF améliorait leur mobilité plus efficacement que le milieu F10 de Ham à court terme (2 h), mais n'avait aucun effet sur les spermatozoïdes épididymaires. Étant donné que la physiologie des spermatozoïdes testiculaires est différente de celle des spermatozoïdes éjaculés, il semble qu'un environnement spécial devrait être conçu et utilisé pour chacun d'eux.

A Multicenter Retrospective Cohort Study of the Profile of Seminal Fluid Analyses of Men Seeking Fertility Care at Different Hospitals.

BACKGROUND: Male infertility contributes 40 % of couple infertility. The prevalence of abnormal semen parameters has been on the increase. Age among other factors affects the fertility potential of males. This study analysed the pattern of seminal fluid parameters of males, seeking fertility treatment in hospitals and the relationship between age, volume and liquefaction time on these other semen parameters. METHODS: This is a multicentre retrospective cohort study conducted in eight secondary and tertiary hospitals in Nigeria. The case notes of couples that sort fertility care at the Gynaecology and Urology clinics of these hospitals from January 1st 2022 to December 31st 2022 were retrieved after receiving ethical approval. A purposeddesigned proforma based on the WHO manual for the examination of human semen was used for data collection. Outcome measures were time of semen collection and examination, volume of semen, sperm number, sperm concentration, PH, agglutination, liquefaction, motility,morphology, vitality, and white blood cell count. Data was analysed using SPSS version 23. Data were presented as means and proportions. P-value of < 0.05 was used as the level of significance. RESULTS: Overall, 1063 couples attended gynaecology and urology clinics with fertility-related concerns within the study period with a retrieval rate of 98.3%. The mean age of participants was 38.24 ± 8 years, while the mean semen volume and sperm concentrations were 2.62 ± 1.6 mls and 34.32 ± 7.4 million respectively. The age of participants significantly affected motility, volume and morphology (p-values of 0.001, 0.001 and 0.004 respectively). The total motility and sperm concentration have an inverse relationship with the age of the participants. CONCLUSION: This study shows that sperm motility decreases with the age of participants. It was also observed that the most common combined abnormality was oligoasthenozoospermia.

CONTEXTE: L'infertilité masculine représente 40 % de l'infertilité des couples. La prévalence des paramètres anormaux du sperme est en augmentation. L'âge, entre autres facteurs, affecte le potentiel de fertilité des hommes. Cette étude a analysé le profil des paramètres du liquide séminal des hommes cherchant un traitement de fertilité dans les hôpitaux et la relation entre l'âge, le volume et le temps de liquéfaction sur ces autres paramètres du sperme. MÉTHODES: Il s'agit d'une étude de cohorte rétrospective multicentrique menée dans huit hôpitaux secondaires et tertiaires au Nigeria. Les notes de cas des couples qui ont eu recours à des soins de fertilité dans les cliniques de gynécologie et d'urologie de ces hôpitaux entre le 1er janvier et le 31 décembre 2022 ont été récupérées après avoir reçu une approbation éthique. Un proforma conçu à dessein et basé sur le manuel de l'OMS pour l'examen du sperme humain a été utilisé pour la collecte des données. Les mesures des résultats étaient le temps de collecte et d'examen du sperme, le volume de sperme, le nombre de spermatozoïdes, la concentration en spermatozoïdes, le PH, l'agglutination, la liquéfaction, la motilité, la morphologie, la vitalité et la numération des globules blancs. Les données ont été analysées à l'aide de SPSS version 23. Les données ont été présentées sous forme de moyennes et de proportions. Une valeur P < 0,05 a été utilisée comme niveau de signification. RÉSULTATS: Dans l'ensemble, 1 063 couples ont fréquenté les cliniques de gynécologie et d'urologie pour des problèmes de fertilité au cours de la période d'étude, avec un taux de récupération de 98,3 %. L'âge moyen des participants était de 38,24 ± 8 ans, tandis que le volume moyen de sperme et les concentrations de spermatozoïdes étaient respectivement de 2,62 ± 1,6 ml et 34,32 ±7,4 millions. L'âge des participants a affecté de manière significative la motilité, le volume et la morphologie (valeurs p de 0,001, 0,001 et 0,004 respectivement). La motilité totale et la concentration en spermatozoïdes ont une relation inverse avec l'âge des participants. CONCLUSION: Cette étude montre que la mobilité des spermatozoïdes diminue avec l'âge des participants. Il a également été observé que l'anomalie combinée la plus fréquente était l'oligoasthénozoospermie. Mots-clés: Infertilité Masculine, Anomalies du Liquide séminal, Nigeria.

Suspected Buprenorphine-Precipitated Opioid Withdrawal following Intercourse: A Case Report.

Buprenorphine, a partial mu-opioid receptor agonist, is a commonly prescribed medication for opioid use disorder (OUD). There is evidence that drugs may enter the male genitourinary tract by an ion-trapping process, based on the lipid solubility and degree of ionization (1). While little is known about the pharmacokinetics of drugs in seminal fluid, pH is thought to play an integral role. Limited evidence exists surrounding cervical absorption of drugs via seminal fluid transmission. This also prompts survey of the frequency of this event and the influence on treatment within this population.

The relevance of false positive acid phosphatase reactions indicative of the presence of seminal fluid from oral and vaginal samples.

The Body Fluid Forum of the Association of Forensic Science Providers recognised concerns raised by forensic practitioners regarding the lack of data to inform on the incidence of significant AP (Acid Phosphatase) reactions from vaginal and oral samples, i.e. those which might be misinterpreted as indicating the presence of semen. This is particularly relevant in the light of appeal court rulings regarding the need for data to support evaluations. This paper presents collaborative data on the nature and incidence of AP reactions from vaginal and oral swabs provided by donors. The results demonstrate that caution is required with mid to strong purple AP reactions from direct testing of oral swabs and with mid purple reactions from vaginal swabs. The use of a Bayesian approach to assist with the provision of opinions regarding the presence of seminal fluid is highlighted.

Low-polarity untargeted metabolomic profiling as a tool to gain insight into seminal fluid.

INTRODUCTION: A decrease in sperm cell count has been observed along the last several decades, especially in the most developed regions of the world. The use of metabolomics to study the composition of the seminal fluid is a promising approach to gain access to the molecular mechanisms underlying this fact. OBJECTIVES: In the present work, we aimed at relating metabolomic profiles of young healthy men to their semen quality parameters obtained from conventional microscopic analysis. METHODS: An untargeted metabolomics approach focusing on low- to mid-polarity compounds was used to analyze a subset of seminal fluid samples from a cohort of over 2700 young healthy men. RESULTS: Our results show that a broad metabolic profiling comprising several families of compounds (including acyl-carnitines, steroids, and other lipids) can contribute to effectively distinguish samples provided by individuals exhibiting low or high absolute sperm counts. CONCLUSION: A number of metabolites involved in sexual development and function, signaling, and energy metabolism were highlighted as being distinctive of samples coming from either group, proving untargeted metabolomics as a promising tool to better understand the pathophysiological processes responsible for male fertility impairment.

The carboxypeptidase B and carbonic anhydrase genes play a reproductive regulatory role during multiple matings in Ophraella communa.

Seminal fluid proteins (SFPs) are key factors in sexual reproduction and are transferred to females during mating with sperm. SFPs have a nutritional value because they protect and activate sperm storage and release to optimize fecundity. Multiple matings promote ovipositioning in several insect species. Therefore, insects may obtain more SFP through multiple matings to maximize reproduction, but this process has not yet been clearly confirmed. Here, the relationship between multiple matings and the SFPs in Ophraella communa (Coleoptera: Chrysomelidae), a biological control agent of the common ragweed Ambrosia artemisiifolia (Asterales: Asteraceae), was studied. Multiple matings significantly increased female fecundity and ovary egg deposition. Carboxypeptidase B (OcCpb) and carbonic anhydrase (OcCa) genes were identified as putative SFP genes in O. communa and they showed strong male-biased expression. Additionally, OcCpb and OcCa expression was upregulated in the bursa copulatrix of mating females compared to that in virgin females, but their expression gradually declined after copulation. Furthermore, OcCpb and OcCa knockdown in males led to a decrease in insect fecundity compared to that in the control. The reproductive tract of females mated with dsRNA-treated males was dissected and observed and, notably, the ovaries produced significantly fewer eggs. These data suggest that OcCpb and OcCa play regulatory roles during multiple matings in O. communa.

Evolutionary Quantitative Proteomics of Reproductive Protein Divergence in Drosophila.

Reproductive traits often evolve rapidly between species. Understanding the causes and consequences of this rapid divergence requires characterization of female and male reproductive proteins and their effect on fertilization success. Species in the Drosophila virilis clade exhibit rampant interspecific reproductive incompatibilities, making them ideal for studies on diversification of reproductive proteins and their role in speciation. Importantly, the role of intraejaculate protein abundance and allocation in interspecific divergence is poorly understood. Here, we identify and quantify the transferred male ejaculate proteome using multiplexed isobaric labeling of the lower female reproductive tract before and immediately after mating using three species of the virilis group. We identified over 200 putative male ejaculate proteins, many of which show differential abundance between species, suggesting that males transfer a species-specific allocation of seminal fluid proteins during copulation. We also identified over 2000 female reproductive proteins, which contain female-specific serine-type endopeptidases that showed differential abundance between species and elevated rates of molecular evolution, similar to that of some male seminal fluid proteins. Our findings suggest that reproductive protein divergence can also manifest in terms of species-specific protein abundance patterns.

Post-orgasmic illness syndrome with positive intradermal test for autologous semen.

Post-orgasmic illness syndrome (POIS) is a rare disease characterized by flu-like symptoms persisting for 2-7 days after ejaculation. POIS has been chiefly attributed to allergic reactions to autologous seminal plasma. However, the exact pathophysiology remains unclear, and there is no effective treatment. We present the case of a 38-year-old man with a 10-year history of recurrent episodes of flu-like symptoms of 1-week duration after ejaculation. The patient was diagnosed with irritating bowel syndrome because of fatigue, myalgia, and lateral abdominal pain. After starting infertility treatment and increasing the frequency of intercourse with his wife, the patient noticed these symptoms after ejaculation. Based on these episodes and symptoms, POIS was suspected. To diagnose POIS, a skin prick test and an intradermal test were performed using his seminal fluid, with the latter yielding a positive result. The patient was diagnosed with POIS, and treatment with antihistamines was continued. Due to its rarity, POIS is often underdiagnosed and underreported; however, the skin test can be a valid diagnostic tool. In this case, the intradermal test result was positive according to the broadly accepted criteria for POIS. Although quality of life is often severely affected in patients with POIS, a lack of a clear understanding of the pathogenesis of POIS prevents early diagnosis. To make diagnoses earlier, it is undoubtedly important to take a detailed medical history and perform skin allergy tests, although the latter requires further validation.