Seminal fluid proteins (SFPs) are emerging as fundamental contributors to sexual selection given their role in post-mating reproductive events, particularly in polyandrous species where the ejaculates of different males compete for fertilisation. SFP identification however remains taxonomically limited and little is known about avian SFPs, despite extensive work on sexual selection in birds. We characterize the SF proteome of the polyandrous Red junglefowl, Gallus gallus, the wild species that gave rise to the domestic chicken. We identify 1,141 SFPs, including proteins involved in immunity and antimicrobial defences, sperm maturation, and fertilisation, revealing a functionally complex SF proteome. This includes a predominant contribution of blood plasma proteins that is conserved with human SF. By comparing the proteome of young and old males with fast or slow sperm velocity in a balanced design, we identify proteins associated with ageing and sperm velocity, and show that old males that retain high sperm velocity have distinct proteome characteristics. SFP comparisons with domestic chickens revealed both qualitative and quantitative differences likely associated with domestication and artificial selection. Collectively, these results shed light onto the functional complexity of avian SF, and provide a platform for molecular studies of fertility, reproductive ageing, and domestication.

Theory predicts that males will strategically invest in ejaculates according to the value of mating opportunities. While strategic sperm allocation has been studied extensively, little is known about concomitant changes in seminal fluid (SF) and its molecular composition, despite increasing evidence that SF proteins (SFPs) are fundamental in fertility and sperm competition. Here, we show that in male red junglefowl, Gallus gallus , along with changes in sperm numbers and SF investment, SF composition changed dynamically over successive matings with a first female, immediately followed by mating with a second, sexually novel female. The SF proteome exhibited a pattern of both protein depletion and enrichment over successive matings, including progressive increases in immunity and plasma proteins. Ejaculates allocated to the second female had distinct proteomic profiles, where depletion of many SFPs was compensated by increased investment in others. This response was partly modulated by male social status: when mating with the second, novel female, subdominants (but not dominants) preferentially invested in SFPs associated with sperm composition, which may reflect status-specific differences in mating rates, sperm maturation and sperm competition. Global proteomic SF analysis thus reveals that successive matings trigger rapid, dynamic SFP changes driven by a combination of depletion and strategic allocation.

Background The development of medicines which can genuinely make a difference requires the identification of clinical, economic, and patient-centric needs. Yet, recognising the needs and experiences of both patients and healthcare professionals (HCPs) is often more challenging in rare diseases. One approach to overcoming barriers in capturing the “whole rare disease picture” is to supplement academic research with analyses of social media data. We explore rare disease social media engagement among several key stakeholders, comparing this with engagement in academic publications over the same period. Results From 2011 to 2021, there were 228,233 published academic articles concerning the rare diseases under consideration, with publication volumes increasing year-on-year across all conditions. However, rare disease Tweets were on average 11.5-fold higher, with 2,633,366 individual rare disease Tweets over the same period, ranging from 2.9-fold in 2011 to 45.2-fold in 2020. While Tweets increased, so too did unique social media contributors, increasing 4.3-fold (from 150,790 in 2011 to 655,157 in 2021). Nurses (1,267%), pharmacists (9,717%), primary care physicians (15,801%), disease specialists (1,420%), patients and patient advocacy groups (660%) and the pharmaceutical industry (2,261%) all experienced significant increases in rare disease social media engagement from 2011 to 2021 (p<0.001). Additionally, each stakeholder specialised in the delivery of content concerning different topics, with disease specialists most likely to discuss research, patients to raise awareness, primary care physicians discussing symptoms, and nurses dedicating the greatest proportion of their communications to discussing the impact of rare diseases on family life. Conclusions Systematic harnessing and analysis of social media data may provide significant value in understanding patient, HCP and system-level needs concerning rare disease management. Tweet volumes exceeded publications year-on-year, with this difference increasing over time and each stakeholder specialising in the delivery of content on different themes. Such data may provide unrivalled insights, enabling researchers to appreciate the full spectrum of stakeholders’ most critical needs, experiences and preferences, thereby enabling meaningful exploration of what “good” looks like. If performed as an adjunct or precursor to published research, the use of social media data may be a valuable tool in identifying key clinical, economic, and patient-centric needs in rare diseases.