Unravelling the complex reproductive tactics of male humpback whales : an integrative analysis of paternity, age, testosterone, and genetic diversity

How the underlying forces of sexual selection impact reproductive tactics including elaborate acoustic displays in cetaceans remains poorly understood. Here, I combined 26 years (1995-2020) of photo-identification, behavioural, (epi)genetic, and endocrine data from an endangered population of humpback whales (New Caledonia), to explore male reproductive success, age, physiology, and population dynamics over almost a third of the lifespan of a humpback whale. First, I conducted a paternity analysis on 177 known mother-offspring pairs and confirmed previous findings of low variation in reproductive success in male humpback whales. Second, epigenetic age estimates of 485 males revealed a left-skewed population age structure in the first half of the study period that became more balanced in the second half. Further, older males (> 23 years) more often engaged in certain reproductive tactics (singing and escorting) and were more successful in siring offspring once the population age structure stabilised, suggesting reproductive tactics and reproductive success in male humpback whales may be age-dependent. Third, using enzyme immunoassays on 457 blubber samples, I observed a seasonal decline in male testosterone in the population over the breeding season. Testosterone levels appeared highest during puberty, then decreased and levelled off at the onset of maturity, yet were highly variable at any point during the breeding season and across males of all ages. Lastly, I investigated the influence of genetic diversity at the major histocompatibility complex (MHC) class I and class IIa (DQB and DRB-a) on patterns of male reproductive success in humpback whales. Mating pairs shared fewer alleles than expected under random mating at MHC class I and IIa, thus, providing evidence of an MHC-mediated female mate choice in humpback whales. This thesis provides novel, critical insights into the evolutionary consequences of commercial whaling on the demography, patterns of reproduction and sexual selection of exploited populations of baleen whales."This work was supported by a University of St Andrews School of Biology Ph.D. Scholarship and the Louis M. Herman Research Scholarship 2022 to Franca Eichenberger. Sample collection and analyses from 2018-2020 were supported by grants to Ellen C. Garland (Royal Society University Research Fellowship (UF160081 & URF\R\221020), Royal Society Research Fellows Enhancement Award (RGF\EA\180213), Royal Society Research Grants for Research Fellows 2018 (RGF\R1\181014), National Geographic Grant (#NGS-50654R-18), Carnegie Trust Research Incentive Grant (RIG007772), British Ecological Society Small Research Grant (SR18/1288) and School of Biology Research Committee funding)."--Fundin

Reproductive tactics in baleen whales

Funding: FE is supported by a University of St Andrews School of Biology Ph.D. scholarship and a Royal Society Research Fellows Enhancement Award (RGF\EA \180213 to ECG), ECG is funded by a Royal Society University Research Fellowship (UF160081 & URF\R\221020), and ELC is funded by a Rutherford Discovery Fellowship from the Royal Society of New Zealand Te Apārangi.While a variety of reproductive tactics are readily witnessed in the odontocetes, such behaviors can be far more elusive, and in some cases, are yet to be observed, in baleen whales. This leads researchers to employ a variety of research methods, some of which have improved greatly in recent decades, to study reproductive behaviors in mysticetes. Genetics and genomics tools can provide invaluable information on maternity, paternity, age, diversity, and kinship, while acoustic tools can provide new insights into the function of sexual displays such as song. In this chapter, we explore what is known about the reproductive strategies and tactics of baleen whales, with a particular focus on the comparatively well-studied right whales (Eubalaena spp.) and humpback whales (Megaptera novaeangliae). Finally, we showcase that by integrating multiple data types, we can explore the interactions between anatomy, physiology, reproductive success, age, population dynamics, and acoustic displays to better understand the mating systems of baleen whales.Publisher PD

A pan-cetacean MHC amplicon sequencing panel developed and evaluated in combination with genome assemblies

This study was funded by a Royal Society Research Grants for Research Fellows (RGF\R1\181014) to E.C.G. E.C.G. is funded by a Royal Society University Research Fellowship (UF160081 & URF\R\221020). F.E. is supported by a University of St Andrews School of Biology Ph.D. scholarship and a Royal Society Research Fellows Enhancement Award (RGF\EA\180213 to E.C.G). E.L.C. is funded by a Rutherford Discovery Fellowship from the Royal Society of New Zealand Te Apārangi.The major histocompatibility complex (MHC) is a highly polymorphic gene family that is crucial in immunity, and its diversity can be effectively used as a fitness marker for populations. Despite this, MHC remains poorly characterised in non-model species (e.g., cetaceans: whales, dolphins and porpoises) as high gene copy number variation, especially in the fast-evolving class I region, makes analyses of genomic sequences difficult. To date, only small sections of class I and IIa genes have been used to assess functional diversity in cetacean populations. Here, we undertook a systematic characterisation of the MHC class I and IIa regions in available cetacean genomes. We extracted full-length gene sequences to design pan-cetacean primers that amplified the complete exon2 from MHC class I and IIa genes in one combined sequencing panel. We validated this panel in 19 cetacean species and described 354 alleles for both classes. Furthermore, we identified likely assembly artefacts for many MHC class I assemblies based on the presence of class I genes in the amplicon data compared to missing genes from genomes. Finally, we investigated MHC diversity using the panel in 25 humpback and 30 southern right whales, including four paternity trios for humpback whales. This revealed copy-number variable class I haplotypes in humpback whales, which is likely a common phenomenon across cetaceans. These MHC alleles will form the basis for a cetacean branch of the Immuno-Polymorphism Database (IPD-MHC), a curated resource intended to aid in the systematic compilation of MHC alleles across several species, to support conservation initiatives.Peer reviewe

Reproductive tactics in baleen whales

While a variety of reproductive tactics are readily witnessed in the odontocetes, such behaviors can be far more elusive, and in some cases, are yet to be observed, in baleen whales. This leads researchers to employ a variety of research methods, some of which have improved greatly in recent decades, to study reproductive behaviors in mysticetes. Genetics and genomics tools can provide invaluable information on maternity, paternity, age, diversity, and kinship, while acoustic tools can provide new insights into the function of sexual displays such as song. In this chapter, we explore what is known about the reproductive strategies and tactics of baleen whales, with a particular focus on the comparatively well-studied right whales (Eubalaena spp.) and humpback whales (Megaptera novaeangliae). Finally, we showcase that by integrating multiple data types, we can explore the interactions between anatomy, physiology, reproductive success, age, population dynamics, and acoustic displays to better understand the mating systems of baleen whales

A pan-cetacean MHC amplicon sequencing panel developed and evaluated in combination with genome assemblies

<p>The major histocompatibility complex (MHC) is a highly polymorphic gene family that is crucial in immunity, and its diversity can be effectively used as a fitness marker for populations. Despite this, MHC remains poorly characterised in non-model species (e.g., cetaceans: whales, dolphins and porpoises) as high gene copy number variation, especially in the fast-evolving class I region, makes analyses of genomic sequences difficult. To date, only small sections of class I and IIa genes have been used to assess functional diversity in cetacean populations. Here, we undertook a systematic characterisation of the MHC class I and IIa regions in available cetacean genomes. We extracted full-length gene sequences to design pan-cetacean primers that amplified the complete exon2 from MHC class I and IIa genes in one combined sequencing panel. We validated this panel in 19 cetacean species and described 354 alleles for both classes.  Furthermore, we identified likely assembly artefacts for many MHC class I assemblies based on the presence of class I genes in the amplicon data compared to missing genes from genomes. Finally, we investigated MHC diversity using the panel in 25 humpback and 30 southern right whales, including four paternity trios for humpback whales. This revealed copy-number variable class I haplotypes in humpback whales, which is likely a common phenomenon across cetaceans. These MHC alleles will form the basis for a cetacean branch of the Immuno-Polymorphism Database (IPD-MHC), a curated resource intended to aid in the systematic compilation of MHC alleles across several species, to support conservation initiatives.</p><p>Funding provided by: Royal Society<br>Crossref Funder Registry ID: https://ror.org/03wnrjx87<br>Award Number: RGF\R1\181014</p><p>A total of 85 tissue samples were taken from individual animals across several cetacean species. The type of tissue were either from strandings or biopsies. Stranding samples in New Zealand were taken by the Department of Conservation New Zealand and sent to the New Zealand Cetacean Tissue Archive (NZCeTA) housed at the University of Auckland Waipapa Taumata Rau with approval from mana whenua (Māori indigenous groups). Biopsy samples from New Zealand cetaceans include two Hector's dolphin (<em>Chephalorhyncus hectori</em>) (Hamner et al., 2017) and two bottlenose dolphins (<em>T. truncatus</em>) (Tezanos-Pinto et al., 2009). Further biopsies include two rough-toothed dolphins (<em>Steno bredanensis</em>) and two Blainville beaked whales (<em>Mesoplodon densirostris</em>) from French-Polynesia (Albertson et al., 2017; Oremus et al., 2012). Details on samples and associated permit numbers can be found in the published manuscript.</p> <p>DNA was extracted from tissue samples and genomic DNA underwent PCR for five Major Histocompatibility Complex loci. PCR products were pooled for each individual, indexed with Nextera indexes supplied by IDT, and sequenced on Illumina NanoSeq and MiSeq. Each individual amplicon-pool was sequenced multiple times on different sequencing runs. The reads provided here are the paired and merged reads from several sequencing runs combined for each individual in a fastq file.</p&gt

A pan-cetacean MHC amplicon sequencing panel developed and evaluated in combination with genome assemblies

The major histocompatibility complex (MHC) is a highly polymorphic gene family that is crucial in immunity, and its diversity can be effectively used as a fitness marker for populations. Despite this, MHC remains poorly characterised in non-model species (e.g., cetaceans: whales, dolphins and porpoises) as high gene copy number variation, especially in the fast-evolving class I region, makes analyses of genomic sequences difficult. To date, only small sections of class I and IIa genes have been used to assess functional diversity in cetacean populations. Here, we undertook a systematic characterisation of the MHC class I and IIa regions in available cetacean genomes. We extracted full-length gene sequences to design pan-cetacean primers that amplified the complete exon2 from MHC class I and IIa genes in one combined sequencing panel. We validated this panel in 19 cetacean species and described 354 alleles for both classes. Furthermore, we identified likely assembly artefacts for many MHC class I assemblies based on the presence of class I genes in the amplicon data compared to missing genes from genomes. Finally, we investigated MHC diversity using the panel in 25 humpback and 30 southern right whales, including four paternity trios for humpback whales. This revealed copy-number variable class I haplotypes in humpback whales, which is likely a common phenomenon across cetaceans. These MHC alleles will form the basis for a cetacean branch of the Immuno-Polymorphism Database (IPD-MHC), a curated resource intended to aid in the systematic compilation of MHC alleles across several species, to support conservation initiatives

Data for: Competing adaptations maintain non-adaptive variation in a wild cricket population

Methods used to collect data are described in the associated publication: Rayner et al. (Accepted, 2024) Competing adaptations maintain non-adaptive variation in a wild cricket population. PNAS.Funding from the Natural Environment Research Council (NE/T0006191/1

Competing adaptations maintain nonadaptive variation in a wild cricket population

Funding: Natural Environment Research Council (NE/T0006191/1) for funding to N.W.B., O.E.G., and J.G.R. that supported this study. We acknowledge computational resources from the James Hutton Institute Bioinformatics HPC (BBSRC grant BB/S019669/1).How emerging adaptive variants interact is an important factor in the evolution of wild populations, but the opportunity to empirically study this interaction is rare. We recently documented the emergence of an adaptive phenotype “curly-wing” in Hawaiian populations of field crickets (Teleogryllus oceanicus). Curly-wing inhibits males’ ability to sing, protecting them from eavesdropping parasitoid flies (Ormia ochracea). Surprisingly, curly-wing co-occurs with similarly protective silent “flatwing” phenotypes in multiple populations, in which neither phenotype has spread to fixation. These two phenotypes are frequently coexpressed, but since either sufficiently reduces song amplitude to evade the fly, their coexpression confers no additional fitness benefit. Numerous “off-target” phenotypic changes are known to accompany flatwing, and we find that curly-wing, too, negatively impacts male courtship ability and affects mass and survival of females under lab conditions. We show through crosses and genomic and mRNA sequencing that curly-wing expression is associated with variation on a single autosome. In parallel analyses of flatwing, our results reinforce previous findings of X-linked single-locus inheritance. By combining insights into the genetic architecture of these alternative phenotypes with simulations and field observations, we show that the co-occurrence of these two adaptations impedes either from fixing, despite extreme fitness benefits, due to fitness epistasis. This co-occurrence of similar adaptive forms in the same populations might be more common than is generally considered and could be an important force inhibiting adaptive evolution in wild populations of sexually reproducing organisms.Peer reviewe

A pan-cetacean MHC amplicon sequencing panel developed and evaluated in combination with genome assemblies

The major histocompatibility complex (MHC) is a highly polymorphic gene family that is crucial in immunity, and its diversity can be effectively used as a fitness marker for populations. Despite this, MHC remains poorly characterised in non-model species (e.g., cetaceans: whales, dolphins and porpoises) as high gene copy number variation, especially in the fast-evolving class I region, makes analyses of genomic sequences difficult. To date, only small sections of class I and IIa genes have been used to assess functional diversity in cetacean populations. Here, we undertook a systematic characterisation of the MHC class I and IIa regions in available cetacean genomes. We extracted full-length gene sequences to design pan-cetacean primers that amplified the complete exon2 from MHC class I and IIa genes in one combined sequencing panel. We validated this panel in 19 cetacean species and described 354 alleles for both classes. Furthermore, we identified likely assembly artefacts for many MHC class I assemblies based on the presence of class I genes in the amplicon data compared to missing genes from genomes. Finally, we investigated MHC diversity using the panel in 25 humpback and 30 southern right whales, including four paternity trios for humpback whales. This revealed copy-number variable class I haplotypes in humpback whales, which is likely a common phenomenon across cetaceans. These MHC alleles will form the basis for a cetacean branch of the Immuno-Polymorphism Database (IPD-MHC), a curated resource intended to aid in the systematic compilation of MHC alleles across several species, to support conservation initiatives