Publications by year
Winters A (In Press). Understanding colour and chemical diversity in nudibranchs.
Winters AE, Chan W, White AM, van den Berg CP, Garson MJ, Cheney KL
(2022). Weapons or deterrents? Nudibranch molluscs use distinct ecological modes of chemical defence against predators. Journal of Animal Ecology
Weapons or deterrents? Nudibranch molluscs use distinct ecological modes of chemical defence against predators
Defensive chemicals are used by plants and animals to reduce the risk of predation through different mechanisms, including toxins that cause injury and harm (weapons) and unpalatable or odiferous compounds that prevent attacks (deterrents). However, whether effective defences are both toxins and deterrents, or work in just one modality is often unclear. In this study, our primary aim was to determine whether defensive compounds stored by nudibranch molluscs acted as weapons (in terms of being toxic), deterrents (in terms of being distasteful) or both. Our secondary aim was to investigate the response of different taxa to these defensive compounds. To do this, we identified secondary metabolites in 30 species of nudibranch molluscs and investigated their deterrent properties using antifeedant assays with three taxa: rock pool shrimp, Palaemon serenus, and two fish species: triggerfish Rhinecanthus aculeatus and toadfish Tetractenos hamiltoni. We compared these results to toxicity assays using brine shrimp Artemia sp. and previously published toxicity data with a damselfish Chromis viridis. Overall, we found no clear relationship between palatability and toxicity, but instead classified defensive compounds into the following categories: Class I & II—highly unpalatable and highly toxic; Class I—weakly unpalatable and highly toxic; Class II—highly unpalatable but weakly toxic; WR (weak response)—weakly unpalatable and weakly toxic. We also found eight extracts from six species that did not display activity in any assays indicating they may have very limited chemical defensive mechanisms (NR, no response). We found that the different classes of secondary metabolites were similarly unpalatable to fish and shrimp, except extracts from Phyllidiidae nudibranchs (isonitriles) that were highly unpalatable to shrimp but weakly unpalatable to fish. Our results pave the way towards better understanding how animal chemical defences work against a variety of predators. We highlight the need to disentangle weapons and deterrents in future work on anti-predator defences to better understand the foraging decisions faced by predators, the resultant selection pressures imposed on prey and the evolution of different anti-predator strategies. Abstract
Winters AE, Lommi J, Kirvesoja J, Nokelainen O, Mappes J
(2021). Multimodal Aposematic Defenses Through the Predation Sequence. FRONTIERS IN ECOLOGY AND EVOLUTION
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Lindstedt C, Suisto K, Burdfield-Steel E, Winters AE, Mappes J
(2020). Defense against predators incurs high reproductive costs for the aposematic moth Arctia plantaginis. Behavioral Ecology
Defense against predators incurs high reproductive costs for the aposematic moth Arctia plantaginis
. To understand how variation in warning displays evolves and is maintained, we need to understand not only how perceivers of these traits select color and toxicity but also the sources of the genetic and phenotypic variation exposed to selection by them. We studied these aspects in the wood tiger moth Arctia plantaginis, which has two locally co-occurring male color morphs in Europe: yellow and white. When threatened, both morphs produce defensive secretions from their abdomen and from thoracic glands. Abdominal fluid has shown to be more important against invertebrate predators than avian predators, and the defensive secretion of the yellow morph is more effective against ants. Here, we focused on the morph-linked reproductive costs of secretion of the abdominal fluid and quantified the proportion of phenotypic and genetic variation in it. We hypothesized that, if yellow males pay higher reproductive costs for their more effective aposematic display, the subsequent higher mating success of white males could offer one explanation for the maintenance of the polymorphism. We first found that the heritable variation in the quantity of abdominal secretion was very low (h2 = 0.006) and the quantity of defensive secretion was not dependent on the male morph. Second, deploying the abdominal defensive secretion decreased the reproductive output of both color morphs equally. This suggests that potential costs of pigment production and chemical defense against invertebrates are not linked in A. plantaginis. Furthermore, our results indicate that environmentally induced variation in chemical defense can alter an individual’s fitness significantly.
Wilson N, Winters A, Cheney K (2018). Correction: Wilson, N.G. et al. Tropical Range Extension for the Temperate, Endemic South-Eastern Australian Nudibranch Goniobranchus splendidus (Angas, 1864). Diversity 2016, 8, 16. Diversity, 10(3), 57-57.
Winters AE, White AM, Dewi AS, Mudianta IW, Wilson NG, Forster LC, Garson MJ, Cheney KL (2018). Distribution of Defensive Metabolites in Nudibranch Molluscs. Journal of Chemical Ecology, 44(4), 384-396.
Winters AE, White AM, Cheney KL, Garson MJ (2018). Geographic variation in diterpene-based secondary metabolites and level of defence in an aposematic nudibranch,<i>Goniobranchus splendidus</i>. Journal of Molluscan Studies, 85(1), 133-142.
Winters AE, Wilson NG, van den Berg CP, How MJ, Endler JA, Marshall NJ, White AM, Garson MJ, Cheney KL
(2018). Toxicity and taste: unequal chemical defences in a mimicry ring. Proceedings of the Royal Society B: Biological Sciences
Toxicity and taste: unequal chemical defences in a mimicry ring
Mimicry of warning signals is common, and can be mutualistic when mimetic species harbour equal levels of defence (Müllerian), or parasitic when mimics are undefended but still gain protection from their resemblance to the model (Batesian). However, whether chemically defended mimics should be similar in terms of toxicity (i.e. causing damage to the consumer) and/or unpalatability (i.e. distasteful to consumer) is unclear and in many studies remains undifferentiated. In this study, we investigated the evolution of visual signals and chemical defences in a putative mimicry ring of nudibranch molluscs. First, we demonstrated that the appearance of a group of red spotted nudibranchs molluscs was similar from the perspective of potential fish predators using visual modelling and pattern analysis. Second, using phylogenetic reconstruction, we demonstrated that this colour pattern has evolved multiple times in distantly related individuals. Third, we showed that these nudibranchs contained different chemical profiles used for defensive purposes. Finally, we demonstrated that although levels of distastefulness towards Palaemon shrimp remained relatively constant between species, toxicity levels towards brine shrimp varied significantly. We highlight the need to disentangle toxicity and taste when considering chemical defences in aposematic and mimetic species, and discuss the implications for aposematic and mimicry signal evolution. Abstract
Forster LC, Winters AE, Cheney KL, Dewapriya P, Capon RJ, Garson MJ
(2017). Spongian-16-one Diterpenes and Their Anatomical Distribution in the Australian Nudibranch Goniobranchus collingwoodi. J Nat Prod
Spongian-16-one Diterpenes and Their Anatomical Distribution in the Australian Nudibranch Goniobranchus collingwoodi.
Six new (1-6) spongian-16-one analogues have been characterized from the Australian nudibranch species Goniobranchus collingwoodi, along with four known spongian-16-one derivatives. The structures and relative configuration were suggested by spectroscopic analyses informed by molecular modeling. Dissection of animal tissue revealed that the mantle and viscera differ in their terpene composition. Whole body extracts were not toxic to brine shrimp (Artemia sp.), but were unpalatable to palaemon shrimp (Palaemon serenus) at a concentration found within the nudibranch. Individual terpenes were not cytotoxic to human lung (NCIH-460), colorectal (SW620), and liver (HepG2) cancer cells. Abstract
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Winters AE, Green NF, Wilson NG, How MJ, Garson MJ, Marshall NJ, Cheney KL
(2017). Stabilizing selection on individual pattern elements of aposematic signals. Proceedings of the Royal Society B: Biological Sciences
Stabilizing selection on individual pattern elements of aposematic signals
Warning signal variation is ubiquitous but paradoxical: low variability should aid recognition and learning by predators. However, spatial variability in the direction and strength of selection for individual elements of the warning signal may allow phenotypic variation for some components, but not others. Variation in selection may occur if predators only learn particular colour pattern components rather than the entire signal. Here, we used a nudibranch mollusc,Goniobranchus splendidus, which exhibits a conspicuous red spot/white body/yellow rim colour pattern, to test this hypothesis. We first demonstrated that secondary metabolites stored within the nudibranch were unpalatable to a marine organism. Using pattern analysis, we demonstrated that the yellow rim remained invariable within and between populations; however, red spots varied significantly in both colour and pattern. In behavioural experiments, a potential fish predator,Rhinecanthus aculeatus, used the presence of the yellow rims to recognize and avoid warning signals. Yellow rims remained stable in the presence of high genetic divergence among populations. We therefore suggest that how predators learn warning signals may cause stabilizing selection on individual colour pattern elements, and will thus have important implications on the evolution of warning signals. Abstract
Cheney KL, White A, Mudianta IW, Winters AE, Quezada M, Capon RJ, Mollo E, Garson MJ
(2016). Choose Your Weaponry: Selective Storage of a Single Toxic Compound, Latrunculin A, by Closely Related Nudibranch Molluscs. PLoS One
Choose Your Weaponry: Selective Storage of a Single Toxic Compound, Latrunculin A, by Closely Related Nudibranch Molluscs.
Natural products play an invaluable role as a starting point in the drug discovery process, and plants and animals use many interesting biologically active natural products as a chemical defense mechanism against predators. Among marine organisms, many nudibranch gastropods are known to derive defensive metabolites from the sponges they eat. Here we investigated the putative sequestration of the toxic compound latrunculin A--a 16-membered macrolide that prevents actin polymerization within cellular processes--which has been identified from sponge sources, by five closely related nudibranch molluscs of the genus Chromodoris. Only latrunculin a was present in the rim of the mantle of these species, where storage reservoirs containing secondary metabolites are located, whilst a variety of secondary metabolites were found in their viscera. The species studied thus selectively accumulate latrunculin a in the part of the mantle that is more exposed to potential predators. This study also demonstrates that latrunculin-containing sponges are not their sole food source. Latrunculin a was found to be several times more potent than other compounds present in these species of nudibranchs when tested by in vitro and in vivo toxicity assays. Anti-feedant assays also indicated that latrunculin a was unpalatable to rock pool shrimps, in a dose-dependent manner. These findings led us to propose that this group of nudibranchs has evolved means both to protect themselves from the toxicity of latrunculin A, and to accumulate this compound in the mantle rim for defensive purposes. The precise mechanism by which the nudibranchs sequester such a potent compound from sponges without disrupting their own key physiological processes is unclear, but this work paves the way for future studies in this direction. Finally, the possible occurrence of both visual and chemosensory Müllerian mimicry in the studied species is discussed. Abstract
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Hirayama Y, Katavic PL, White AM, Pierens GK, Lambert LK, Winters AE, Kigoshi H, Kita M, Garson MJ
(2016). New Cytotoxic Norditerpenes from the Australian Nudibranchs Goniobranchus Splendidus and Goniobranchus Daphne. Australian Journal of Chemistry
New Cytotoxic Norditerpenes from the Australian Nudibranchs Goniobranchus Splendidus and Goniobranchus Daphne
This study reports the isolation and characterisation of six new metabolites with ‘gracilin’-type carbon skeletons and of aplytandiene-3 from the Australian nudibranch Goniobranchus splendidus. The structure of gracilin G is revised, and the C-6 configuration deduced by comparison of calculated 3JC/H values with values measured using the EXSIDE pulse sequence. A lactone isolated from Goniobranchus daphne contains a rearranged spongionellin-type skeleton. Screening of selected metabolites revealed significant cytotoxicity against a HeLa S3 cell line by five of the new terpenes. Abstract
White AM, Dewi AS, Cheney KL, Winters AE, Blanchfield JT, Garson MJ
(2016). Oxygenated Diterpenes from the Indo-Pacific Nudibranchs<i>Goniobranchus splendidus</i>and<i>Ardeadoris egretta</i>. Natural Product Communications
Oxygenated Diterpenes from the Indo-Pacific NudibranchsGoniobranchus splendidusandArdeadoris egretta
Five new diterpenes (1-5), each with a highly oxygenated spongian framework, were characterized from an organic extract of a specimen of the nudibranch Goniobranchus splendidus collected from Eastern Australia. The new diterpene 7α-hydroxydendrillol-3 (6) was identified from specimens of Ardeodoris egretta. The structures and relative configurations of the six new metabolites have been elucidated by analysis of their spectroscopic data. Abstract
White AM, Pierens GK, Forster LC, Winters AE, Cheney KL, Garson MJ
(2016). Rearranged Diterpenes and Norditerpenes from Three Australian Goniobranchus Mollusks. J Nat Prod
Rearranged Diterpenes and Norditerpenes from Three Australian Goniobranchus Mollusks.
Three new norditerpenes (1, 6, and 7) and four diterpenes (2-5) with extensively rearranged carbon skeletons have been characterized from Australian nudibranchs. The relative configuration of the cyclopropyl-containing verrielactone (1) from Goniobranchus verrieri was suggested by spectroscopic analysis at 500 MHz informed by a combination of molecular modeling and DFT calculations. The nudibranchs G. splendidus and G. cf. splendidus provided 2-7, for which the structures and stereochemistry were deduced by 2D NMR studies at either 500 or 700 MHz. Each of the seven terpenoids exhibited a carbon skeleton modified from one of the tetrahydroaplysulphurin, spongionellin, or gracilane series of terpenes. A biosynthetic pathway to terpenes 1-7 from spongialactone is proposed. Abstract
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Wilson N, Winters A, Cheney K (2016). Tropical Range Extension for the Temperate, Endemic South-Eastern Australian Nudibranch Goniobranchus splendidus (Angas, 1864). Diversity, 8(4), 16-16.
Mudianta IW, White AM, Katavic PL, Krishnaraj RR, Winters AE, Mollo E, Cheney KL, Garson MJ
(2014). Chemoecological studies on marine natural products: terpene chemistry from marine mollusks. Pure and Applied Chemistry
Chemoecological studies on marine natural products: terpene chemistry from marine mollusks
AbstractSome species of nudibranchs (Mollusca) protect themselves from predatory attacks by storing defensive terpene chemicals acquired from dietary sponges (Porifera) in specialized body parts called MDFs (mantle dermal formations), often advertising their unpalatability to potential predators by means of bright coloration patterns. Consequently, the survival of these trophic specialist species is closely related to the possibility of obtaining the defensive tools from sponges that live in their immediate vicinity; therefore, it is important to determine as precisely as possible the chemical composition of nudibranch extracts prior to any ecological studies addressing issues that involve their alimentary behavior and their defensive strategies, including the significance of their color patterns. Some of our recent studies on the chemical composition of terpene extracts from nudibranchs belonging to the genera Chromodoris and Hypselodoris are summarized. We also report the development of a method to assay extracts and purified metabolites for their feeding deterrent activity against co-occurring generalist predators. In a recent chemoecological study, showing that repugnant terpene chemicals are accumulated at extremely high concentrations in exposed parts of the nudibranchs’ bodies, the feeding deterrence assays were carried out on the generalist marine shrimp Palaemon elegans, very common in the Mediterranean. We have modified this assay for use with the Australian shrimp species P. serenus, and confirmed the ecological validity of the assay by analysis of extracts from species of sponges and mollusks that live in the same habitat as P. serenus. The deterrent properties of haliclonacyclamine alkaloids isolated from the sponge Haliclona sp. were demonstrated, with the alkaloid mixture demonstrating palatability deterrence at concentrations as low as 0.05 mg/mL, and complete deterrence at 0.75 mg/mL. In contrast, the diterpene thuridillin metabolites from the sacoglossan mollusk Thuridilla splendens did not deter feeding by P. serenus. Abstract
Cheney KL, Cortesi F, How MJ, Wilson NG, Blomberg SP, Winters AE, Umanzör S, Marshall NJ
(2014). Conspicuous visual signals do not coevolve with increased body size in marine sea slugs. J Evol Biol
Conspicuous visual signals do not coevolve with increased body size in marine sea slugs.
Many taxa use conspicuous colouration to attract mates, signal chemical defences (aposematism) or for thermoregulation. Conspicuousness is a key feature of aposematic signals, and experimental evidence suggests that predators avoid conspicuous prey more readily when they exhibit larger body size and/or pattern elements. Aposematic prey species may therefore evolve a larger body size due to predatory selection pressures, or alternatively, larger prey species may be more likely to evolve aposematic colouration. Therefore, a positive correlation between conspicuousness and body size should exist. Here, we investigated whether there was a phylogenetic correlation between the conspicuousness of animal patterns and body size using an intriguing, understudied model system to examine questions on the evolution of animal signals, namely nudibranchs (opisthobranch molluscs). We also used new ways to compare animal patterns quantitatively with their background habitat in terms of intensity variance and spatial frequency power spectra. In studies of aposematism, conspicuousness is usually quantified using the spectral contrast of animal colour patches against its background; however, other components of visual signals, such as pattern, luminance and spectral sensitivities of potential observers, are largely ignored. Contrary to our prediction, we found that the conspicuousness of body patterns in over 70 nudibranch species decreased as body size increased, indicating that crypsis was not limited to a smaller body size. Therefore, alternative selective pressures on body size and development of colour patterns, other than those inflicted by visual hunting predators, may act more strongly on the evolution of aposematism in nudibranch molluscs. Abstract
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Winters AE, Stevens M, Mitchell C, Blomberg SP, Blount JD (2014). Maternal effects and warning signal honesty in eggs and offspring of an aposematic ladybird beetle. Functional Ecology
Mudianta IW, Challinor VL, Winters AE, Cheney KL, De Voss JJ, Garson MJ
(2013). Synthesis and determination of the absolute configuration of (-)-(5R,6Z)-dendrolasin-5-acetate from the nudibranch Hypselodoris jacksoni. Beilstein J Org Chem
Synthesis and determination of the absolute configuration of (-)-(5R,6Z)-dendrolasin-5-acetate from the nudibranch Hypselodoris jacksoni.
A small sample of (-)-(5R,6Z)-dendrolasin-5-acetate, which was fully characterized by 2D NMR studies, was isolated from the nudibranch Hypselodoris jacksoni, along with the sesquiterpenes (+)-agassizin, (-)-furodysinin, (-)-euryfuran, (-)-dehydroherbadysidolide and (+)-pallescensone. A synthetic sample ([α]D -8.7) of the new metabolite was prepared by [1,2]-Wittig rearrangement of a geranylfuryl ether followed by acetylation of purified alcohol isomers. The absolute configuration at C-5 was established as R by the analysis of MPA ester derivatives of (Z)-5-hydroxydendrolasin obtained by preparative enantioselective HPLC. Abstract
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WINTERS AE, YEE DA (2012). Variation in performance of two co-occurring mosquito species across diverse resource environments: insights from nutrient and stable isotope analyses. Ecological Entomology, 37(1), 56-64.