Ageing of juvenile coral grouper (Plectropomus maculatus) reveals year-round spawning and recruitment: implications for seasonal closures.

Temporal patterns in spawning and juvenile recruitment can have major effects on population size and the demographic structure of coral reef fishes. For harvested species, these patterns are crucial in determining stock size and optimizing management strategies such as seasonal closures. For the commercially important coral grouper (Plectropomus spp.) on the Great Barrier Reef, histological studies indicate peak spawning around the summer new moons. Here we examine the timing of spawning activity for P. maculatus in the southern Great Barrier Reef by deriving age in days for 761 juvenile fish collected between 2007 and 2022, and back-calculating settlement and spawning dates. Age-length relationships were used to estimate spawning and settlement times for a further 1002 juveniles collected over this period. Unexpectedly, our findings indicate year-round spawning activity generates distinct recruitment cohorts that span several weeks to months. Peak spawning varied between years with no clear association with environmental cues, and little to no alignment with existing seasonal fisheries closures around the new moon. Given the variability and uncertainty in peak spawning times, this fishery may benefit from additional and longer seasonal closures, or alternative fisheries management strategies, to maximize the recruitment contribution from periods of greatest reproductive success.

Characterisation and cross-amplification of 42 microsatellite markers in two Amphiprion species (Pomacentridae) and a natural hybrid anemonefish to inform genetic structure within a hybrid zone.

The hybrid anemonefish, Amphiprion leucokranos, is known to be the product of ongoing, introgressive hybridization between parent taxa Amphiprion sandaracinos and Amphiprion chrysopterus. Hybridization is an important evolutionary phenomenon contributing to biodiversity within marine systems, where hybrid zones provide ideal systems in which to study hybridization events. Here, a suite of 42 Amphiprion microsatellite markers (including development of 8 novel markers) were cross-amplified in individuals from parent taxa and hybrid populations to facilitate investigation into the relatedness of hybridizing species across the A. leucokranos hybrid zone. Analysis revealed 15, 20 and 24 highly polymorphic loci (PIC > 0.5) in the two parent species and hybrid, respectively, for use in population genetic and parentage studies, with 305 unique alleles found overall (ranging from 1 to 13 alleles per locus) and 7 alleles per locus on average. Observed and expected heterozygosities ranged from 0.000 to 1.000 and 0.000 to 0.978, respectively. Significant deviations from Hardy-Weinberg equilibrium were found in eight loci, possibly due to relatedness among samples or the presence of null alleles. Use of the suite of markers tested here will provide valuable insights into the contemporary population structure and introgression among species and hybrids within the Amphiprion leucokranos hybrid zone, as well as inform future ecological and evolutionary studies of anemonefishes more broadly.

Reproductive control via the threat of eviction in the clown anemonefish.

In social groups, high reproductive skew is predicted to arise when the reproductive output of a group is limited, and dominant individuals can suppress subordinate reproductive efforts. Reproductive suppression is often assumed to occur via overt aggression or the threat of eviction. It is unclear, however, whether the threat of eviction alone is sufficient to induce reproductive restraint by subordinates. Here, we test two assumptions of the restraint model of reproductive skew by investigating whether resource limitation generates reproductive competition and whether the threat of eviction leads to reproductive restraint in the clown anemonefish Amphiprion percula First, we use a feeding experiment to test whether reproduction is resource limited, which would create an incentive for the dominant pair to suppress subordinate reproduction. We show that the number of eggs laid increased in the population over the study period, but the per cent increase in fed groups was more than twice that in unfed groups (205% and 78%, respectively). Second, we use an eviction experiment to test whether the dominant pair evicts mature subordinates, which would create an incentive for the subordinates to forgo reproduction. We show that mature subordinates are seven times more likely to be evicted than immature subordinates of the same size. In summary, we provide experimental support for the assumptions of the restraint model by showing that resource limitation creates reproductive competition and a credible threat of eviction helps explain why subordinates forego reproduction. Transactional models of reproductive skew may apply well to this and other simple systems.

Homing is not for everyone: displaced cardinalfish find a new place to live.

It was tested whether the pajama cardinalfish Sphaeramia nematoptera (Apogonidae) could home by displacing individuals up to 250 m within and among isolated reefs. Contrary to expectations, only two of 37 (5·4%) displaced S. nematoptera returned home and another 16 (43·2%) were found to have joined other social groups and did not home after 26 months of observations; while over the same period, 94% of control S. nematoptera remained associated with home corals, demonstrating strong site attachment. Hence, while this species has the potential to return home, being able to do so may not be as critical as previously assumed.

Hierarchical behaviour, habitat use and species size differences shape evolutionary outcomes of hybridization in a coral reef fish.

Hybridization is an important evolutionary process, with ecological and behavioural factors influencing gene exchange between hybrids and parent species. Patterns of hybridization in anemonefishes may result from living in highly specialized habitats and breeding status regulated by size-based hierarchal social groups. Here, morphological, ecological and genetic analyses in Kimbe Bay, Papua New Guinea, examine the hybrid status of Amphiprion leucokranos, a nominal species and presumed hybrid between Amphiprion sandaracinos and Amphiprion chrysopterus. We test the hypothesis that habitat use and relative size differences of the parent species and hybrids determine the patterns of gene exchange. There is strong evidence that A. leucokranos is a hybrid of smaller A. sandaracinos and larger A. chrysopterus, where A. chrysopterus is exclusively the mother to each hybrid, based on mtDNA cytochrome b and multiple nDNA microsatellite loci. Overlap in habitat, depth and host anemone use was found, with hybrids intermediate to parents and cohabitation in over 25% of anemones sampled. Hybrids, intermediate in body size, colour and pattern, were classified 55% of the time as morphologically first-generation hybrids relative to parents, whereas 45% of hybrids were more A. sandaracinos-like, suggesting backcrossing. Unidirectional introgression of A. chrysopterus mtDNA into A. sandaracinos via hybrid backcrosses was found, with larger female hybrids and small male A. sandaracinos mating. Potential nDNA introgression was also evident through distinct intermediate hybrid genotypes penetrating both parent species. Findings support the hypothesis that anemonefish hierarchical behaviour, habitat use and species-specific size differences determine how hybrids form and the evolutionary consequences of hybridization.

From cooperation to combat: adverse effect of thermal stress in a symbiotic coral-crustacean community.

Although mutualisms are ubiquitous in nature, our understanding of the potential impacts of climate change on these important ecological interactions is deficient. Here, we report on a thermal stress-related shift from cooperation to antagonism between members of a mutualistic coral-dwelling community. Increased mortality of coral-defending crustacean symbionts Trapezia cymodoce (coral crab) and Alpheus lottini (snapping shrimp) was observed in response to experimentally elevated temperatures and reduced coral-host (Pocillopora damicornis) condition. However, strong differential numerical effects occurred among crustaceans as a function of species and sex, with shrimp (75%), and female crabs (55%), exhibiting the fastest and greatest declines in numbers. These declines were due to forceful eviction from the coral-host by male crabs. Furthermore, surviving female crabs were impacted by a dramatic decline (85%) in egg production, which could have deleterious consequences for population sustainability. Our results suggest that elevated temperature switches the fundamental nature of this interaction from cooperation to competition, leading to asymmetrical effects on species and/or sexes. Our study illustrates the importance of evaluating not only individual responses to climate change, but also potentially fragile interactions within and among susceptible species.

Multispecies spawning sites for fishes on a low-latitude coral reef: spatial and temporal patterns.

Spawning sites used by one or more species were located by intensively searching nearshore coral reefs of Kimbe Bay (New Britain, Papua New Guinea). Once identified, the spawning sites were surveyed repeatedly within fixed 5 m radius circular areas, for > 2000 h of observations ranging from before dawn to after dusk spanning 190 days between July 2001 and May 2004. A total of 38 spawning sites were identified on the seven study reefs distributed at an average of one site every 60 m of reef edge. Pelagic spawning was observed in 41 fish species from six families. On three intensively studied reefs, all 17 spawning sites identified were used by at least three species, with a maximum of 30 different species observed spawning at a single site. Spawning was observed during every month of the study, on all days of the lunar month, at all states of the tide and at most hours of the day studied. Nevertheless, the majority of species were observed spawning on proportionately more days from December to April, on more days around the new moon and in association with higher tides. The strongest temporal association, however, was with species-specific diel spawning times spanning < 3 h for most species. While dawn spawning, afternoon spawning and dusk spawning species were differentiated, the time of spawning for the striated surgeonfish Ctenochaetus striatus also differed significantly among sites. The large number of species spawning at the same restricted locations during predictable times suggests that these sites are extremely important on this low-latitude coral reef.

Frequency-dependent properties of the tectorial membrane facilitate energy transmission and amplification in the cochlea.

The remarkable sensitivity, frequency selectivity, and dynamic range of the mammalian cochlea relies on longitudinal transmission of minuscule amounts of energy as passive, pressure-driven, basilar membrane (BM) traveling waves. These waves are actively amplified at frequency-specific locations by a mechanism that involves interaction between the BM and another extracellular matrix, the tectorial membrane (TM). From mechanical measurements of isolated segments of the TM, we made the important new (to our knowledge) discovery that the stiffness of the TM is reduced when it is mechanically stimulated at physiologically relevant magnitudes and at frequencies below their frequency place in the cochlea. The reduction in stiffness functionally uncouples the TM from the organ of Corti, thereby minimizing energy losses during passive traveling-wave propagation. Stiffening and decreased viscosity of the TM at high stimulus frequencies can potentially facilitate active amplification, especially in the high-frequency, basal turn, where energy loss due to internal friction within the TM is less than in the apex. This prediction is confirmed by neural recordings from several frequency regions of the cochlea.

Sediment-induced turbidity impairs foraging performance and prey choice of planktivorous coral reef fishes.

Sedimentation is a substantial threat to aquatic ecosystems and a primary cause of habitat degradation on near-shore coral reefs. Although numerous studies have demonstrated major impacts of sedimentation and turbidity on corals, virtually nothing is known of the sensitivity of reef fishes. Planktivorous fishes are an important trophic group that funnels pelagic energy sources into reef ecosystems. These fishes are visual predators whose foraging is likely to be impaired by turbidity, but the threshold for such effects and their magnitude are unknown. This study examined the effect of sediment-induced turbidity on foraging in four species of planktivorous damselfishes (Pomacentridae) of the Great Barrier Reef, including inshore and offshore species that potentially differ in tolerance for turbidity. An experimental flow tunnel was used to quantify their ability to catch mobile and immobile planktonic prey under different levels of turbidity and velocity in the range encountered on natural and disturbed reefs. Turbidity of just 4 NTU (nephelometric turbidity units) reduced average attack success by up to 56%, with higher effect sizes for species with offshore distributions. Only the inshore species (Neopomacentrus bankieri), which frequently encounters this turbidity on coastal reefs, could maintain high prey capture success. At elevated turbidity similar to that found on disturbed reefs (8 NTU), attack success was reduced in all species examined by up to 69%. These reductions in attack success led to a 21-24% decrease in foraging rates for all mid to outer-shelf species, in spite of increasing attack rates at high turbidity. Although effects of turbidity varied among species, it always depended heavily on prey mobility and ambient velocity. Attack success was up to 14 times lower on mobile prey, leaving species relatively incapable of foraging on anything but immobile prey at high turbidity. Effects of turbidity were particularly prominent at higher velocities, as attack success was overall 20-fold lower and foraging rates 3.3-fold lower at flow velocities > 30 cm/s relative to < or = 10 cm/s. Given that many planktivorous reef fishes predominantly occupy exposed, high-flow habitats, these results provide a reasonable explanation for the lack of planktivores on inshore coral reefs and warn that the performance of visual predators could be impaired at turbidity levels of only 4 NTU.

Critical research needs for managing coral reef marine protected areas: perspectives of academics and managers.

Marine protected areas (MPAs) are a primary policy instrument for managing and protecting coral reefs. Successful MPAs ultimately depend on knowledge-based decision making, where scientific research is integrated into management actions. Fourteen coral reef MPA managers and sixteen academics from eleven research, state and federal government institutions each outlined at least five pertinent research needs for improving the management of MPAs situated in Australian coral reefs. From this list of 173 key questions, we asked members of each group to rank questions in order of urgency, redundancy and importance, which allowed us to explore the extent of perceptional mismatch and overlap among the two groups. Our results suggest the mismatch among MPA managers and academics is small, with no significant difference among the groups in terms of their respective research interests, or the type of questions they pose. However, managers prioritised spatial management and monitoring as research themes, whilst academics identified climate change, resilience, spatial management, fishing and connectivity as the most important topics. Ranking of the posed questions by the two groups was also similar, although managers were less confident about the achievability of the posed research questions and whether questions represented a knowledge gap. We conclude that improved collaboration and knowledge transfer among management and academic groups can be used to achieve similar objectives and enhance the knowledge-based management of MPAs.

Crucial knowledge gaps in current understanding of climate change impacts on coral reef fishes.

Expert opinion was canvassed to identify crucial knowledge gaps in current understanding of climate change impacts on coral reef fishes. Scientists that had published three or more papers on the effects of climate and environmental factors on reef fishes were invited to submit five questions that, if addressed, would improve our understanding of climate change effects on coral reef fishes. Thirty-three scientists provided 155 questions, and 32 scientists scored these questions in terms of: (i) identifying a knowledge gap, (ii) achievability, (iii) applicability to a broad spectrum of species and reef habitats, and (iv) priority. Forty-two per cent of the questions related to habitat associations and community dynamics of fish, reflecting the established effects and immediate concern relating to climate-induced coral loss and habitat degradation. However, there were also questions on fish demographics, physiology, behaviour and management, all of which could be potentially affected by climate change. Irrespective of their individual expertise and background, scientists scored questions from different topics similarly, suggesting limited bias and recognition of a need for greater interdisciplinary and collaborative research. Presented here are the 53 highest-scoring unique questions. These questions should act as a guide for future research, providing a basis for better assessment and management of climate change impacts on coral reefs and associated fish communities.

Estimating connectivity in marine populations: an empirical evaluation of assignment tests and parentage analysis under different gene flow scenarios.

The application of spatially explicit models of population dynamics to fisheries management and the design marine reserve network systems has been limited due to a lack of empirical estimates of larval dispersal. Here we compared assignment tests and parentage analysis for examining larval retention and connectivity under two different gene flow scenarios using panda clownfish (Amphiprion polymnus) in Papua New Guinea. A metapopulation of panda clownfish in Bootless Bay with little or no genetic differentiation among five spatially discrete locations separated by 2-6 km provided the high gene flow scenario. The low gene flow scenario compared the Bootless Bay metapopulation with a genetically distinct population (F(ST )= 0.1) located at Schumann Island, New Britain, 1500 km to the northeast. We used assignment tests and parentage analysis based on microsatellite DNA data to identify natal origins of 177 juveniles in Bootless Bay and 73 juveniles at Schumann Island. At low rates of gene flow, assignment tests correctly classified juveniles to their source population. On the other hand, parentage analysis led to an overestimate of self-recruitment within the two populations due to the significant deviation from panmixia when both populations were pooled. At high gene flow (within Bootless Bay), assignment tests underestimated self-recruitment and connectivity among subpopulations, and grossly overestimated self-recruitment within the overall metapopulation. However, the assignment tests did identify immigrants from distant (genetically distinct) populations. Parentage analysis clearly provided the most accurate estimates of connectivity in situations of high gene flow.

Connectivity and resilience of coral reef metapopulations in marine protected areas: matching empirical efforts to predictive needs.

Design and decision-making for marine protected areas (MPAs) on coral reefs require prediction of MPA effects with population models. Modeling of MPAs has shown how the persistence of metapopulations in systems of MPAs depends on the size and spacing of MPAs, and levels of fishing outside the MPAs. However, the pattern of demographic connectivity produced by larval dispersal is a key uncertainty in those modeling studies. The information required to assess population persistence is a dispersal matrix containing the fraction of larvae traveling to each location from each location, not just the current number of larvae exchanged among locations. Recent metapopulation modeling research with hypothetical dispersal matrices has shown how the spatial scale of dispersal, degree of advection versus diffusion, total larval output, and temporal and spatial variability in dispersal influence population persistence. Recent empirical studies using population genetics, parentage analysis, and geochemical and artificial marks in calcified structures have improved the understanding of dispersal. However, many such studies report current self-recruitment (locally produced settlement/settlement from elsewhere), which is not as directly useful as local retention (locally produced settlement/total locally released), which is a component of the dispersal matrix. Modeling of biophysical circulation with larval particle tracking can provide the required elements of dispersal matrices and assess their sensitivity to flows and larval behavior, but it requires more assumptions than direct empirical methods. To make rapid progress in understanding the scales and patterns of connectivity, greater communication between empiricists and population modelers will be needed. Empiricists need to focus more on identifying the characteristics of the dispersal matrix, while population modelers need to track and assimilate evolving empirical results.

Transgenerational marking of marine fish larvae: stable-isotope retention, physiological effects and health issues.

This study examined the toxicological and physiological responses of a commercially important coral-reef grouper, Plectropomus leopardus (Serranidae), to injection of enriched stable-isotope barium chloride (BaCl(2)) solution. Thirty adult P. leopardus were subject to one of two (138)BaCl(2) injection treatment groups (corresponding to dosage rates of 2 and 4 mg (138)Ba kg(-1) body mass), and a control group in which fish were injected with 0.9% sodium chloride (NaCl) solution. Fish from each group were sampled at post-injection intervals of 48 h and 1, 3, 5 and 8 weeks, at which time blood and tissue samples were removed from each fish. Residual concentrations of Ba and (138)Ba:(137)Ba ratios were measured in muscle, gonad, liver and bone tissues of each experimental fish. Elevated Ba concentrations were detected in all treatment fish tissue samples within 48 h post injection. Residual Ba concentrations decreased throughout the remainder of the 8 week experimental period in all tissues except bone. The BaCl(2) injection had no significant effects on measured whole blood variables or on the plasma concentrations of steroid hormones. Enriched Ba stable isotopes can therefore be used at low dosages to mark larvae of commercially important marine fishes, without adverse effects on the health of the fishes or on humans who may consume them.

Interaction of the plant hormone, indole-3-acetic acid, with phosphatidylcholine model membranes: effects of acyl chain length.

The interaction between the plant hormone, indole-3-acetic acid (IAA), and phosphatidylcholines (PC) of varying acyl chain length has been studied by monitoring the IAA-induced changes in 1H-NMR chemical shifts of lipid headgroup -+N(CH3)3 protons. For PCs in both micellar and vesicle bilayer systems these shifts increase with chain length although for the latter the magnitude of the shifts decreases with an increase in chain unsaturation. In systems composed of mixtures of pure PCs the headgroup -+N(CH3)3 resonance for each phospholipid is shifted by IAA to different extents, indicating that IAA is able to distinguish between individual PCs in mixtures. In di-C12PC and di-C14PC, but not di-C10PC vesicle systems, the -+N(CH3)3 resonance is split into two components reflecting differences in packing of the inside and outside lamellae. This splitting is altered by IAA indicating that IAA interacts differently with the inside and outside PC molecules.

Elimination of self-association as the source of the thermodynamic nonideality in aqueous proline solutions.

The effect of high concentrations of proline on the diffusion coefficient of water has been examined to assess the extent to which the resulting thermodynamic nonideality could be explained on the statistical-mechanical basis of excluded volume. In fact, such a space-filling role not only accounts for the proline concentration-dependence of the diffusion coefficient of water but it also accounts for the nonideality of proline in freezing point depression and isopiestic measurements. These findings refute the conclusion (Schobert, B. and Tschesche, H. (1978) Biochim. Biophys. Acta 541, 270-277) that the stabilization of enzyme structure by high concentrations of proline stems from self-association of the imino acid via intermolecular hydrogen bonding; and thereby support the concept that the protective effect of proline on enzyme stability must reside mainly in its action as an inert, space-filling solute.

Effect of resistant starch on fecal bulk and fermentation-dependent events in humans.

This study investigated the effect of two diets, which differed in resistant starch (RS) concentration, on fecal bulk and fermentation-dependent events in 11 humans. Amounts of RS consumed were 5.0 +/- 0.4 and 39.0 +/- 3.0 g/d (mean +/- SEM) for the low- and high-RS diets, respectively. The two diets were fed for 3 wk each in a randomized crossover design. Fecal collections were made in the third week of each study period. The high-RS diet produced an increase (P < 0.01) in total fecal output (from 138 +/- 22 to 197 +/- 37 g/d) and lowered fecal pH (6.9 +/- 0.1 to 6.3 +/- 0.1). There were significant increases (P < 0.05) in the fecal concentrations and daily excretion of butyrate (+38% and +100%, respectively) and acetate (+26% and +72%, respectively) during the high-RS period. The fecal excretion (g/d) of nonstarch polysaccharides (NSP) also rose by 50% during the high-RS diet, suggesting that the presence of starch in the colon may affect the fermentation of NSP. Subjects reported an increase in flatulence and easier defecation. These results demonstrate that RS has a significant impact on putative markers of colonic health in humans.