Gross morphology, histology, and ultrastructure of the olfactory rosette of a critically endangered indicator species, the Delta Smelt, Hypomesus transpacificus.

The Delta Smelt (Hypomesus transpacificus) is a small, semi-anadromous fish native to the San Francisco Bay-Delta Estuary and has been declared as critically endangered. Their olfactory biology, in particular, is poorly understood and a basic description of their sensory anatomy is needed to advance our understanding of the sensory ecology of species to inform conservation efforts to manage and protect them. We provide a description of the gross morphology, histological, immunohistochemical, and ultrastructural features of the olfactory rosette in this fish and discuss some of the functional implications in relation to olfactory ability. We show that Delta Smelt have a multilamellar olfactory rosette with allometric growth. Calretinin immunohistochemistry revealed a diffuse distribution of olfactory receptor neurons within the epithelium. Ciliated, microvillous and crypt neurons were clearly identified using morphological and immunohistochemical features. The olfactory neurons were supported by robust ciliated and secretory sustentacular cells. Although the sense of smell has been overlooked in Delta Smelt, we conclude that the olfactory epithelium has many characteristics of macrosmatic fish. With this study, we provide a foundation for future research into the sensory ecology of this imperiled fish.

Demographic History, Not Mating System, Explains Signatures of Inbreeding and Inbreeding Depression in a Large Outbred Population.

AbstractInbreeding depression is often found in small, inbred populations, but whether it can be detected in and have evolutionary consequences for large, wide-ranging populations is poorly known. Here, we investigate the possibility of inbreeding in a large population to determine whether mild levels of inbreeding can still have genetic and phenotypic consequences and how genomically widespread these effects can be. We apply genome-wide methods to investigate whether individual and parental heterozygosity is related to morphological, growth, or life-history traits in a pelagic seabird, Leach's storm-petrel (Oceanodroma leucorhoa). Examining 560 individuals as part of a multiyear study, we found a substantial effect of maternal heterozygosity on chick traits: chicks from less heterozygous (relatively inbred) mothers were significantly smaller than chicks from more heterozygous (noninbred) mothers. We show that these heterozygosity-fitness correlations were due to general genome-wide effects and demonstrate a correlation between heterozygosity and inbreeding, suggesting inbreeding depression. We used population genetic models to further show that the variance in inbreeding was probably due to past demographic events rather than the current mating system and ongoing mate choice. Our findings demonstrate that inbreeding depression can be observed in large populations and illustrate how the integration of genomic techniques and fieldwork can elucidate its underlying causes.

Ecology can inform genetics: Disassortative mating contributes to MHC polymorphism in Leach's storm-petrels (Oceanodroma leucorhoa).

Studies of MHC-based mate choice in wild populations often test hypotheses on species exhibiting female choice and male-male competition, which reflects the general prevalence of females as the choosy sex in natural systems. Here, we examined mutual mate-choice patterns in a small burrow-nesting seabird, the Leach's storm-petrel (Oceanodroma leucorhoa), using the major histocompatibility complex (MHC). The life history and ecology of this species are extreme: both partners work together to fledge a single chick during the breeding season, a task that requires regularly travelling hundreds of kilometres to and from foraging grounds over a 6- to 8-week provisioning period. Using a 5-year data set unprecedented for this species (n = 1078 adults and 925 chicks), we found a positive relationship between variation in the likelihood of female reproductive success and heterozygosity at Ocle-DAB2, a MHC class IIB locus. Contrary to previous reports rejecting disassortative mating as a mechanism for maintaining genetic polymorphism in this species, here we show that males make significant disassortative mate-choice decisions. Variability in female reproductive success suggests that the most common homozygous females (Ocle-DAB2*01/Ocle-DAB2*01) may be physiologically disadvantaged and, therefore, less preferred as lifelong partners for choosy males. The results from this study support the role of mate choice in maintaining high levels of MHC variability in a wild seabird species and highlight the need to incorporate a broader ecological framework and sufficient sample sizes into studies of MHC-based mating patterns in wild populations in general.

Development of the Visual System in a Burrow-Nesting Seabird: Leach's Storm Petrel.

Little is known about the development of vision in wild birds. It is unknown, for example, whether the ability to see can be predicted by the level of prenatal growth or whether the eyes are open at hatching in a particular species. In this study, we investigated the growth of eyes, the formation of retinal ganglion cell topography, and the appearance of simple, visually guided behaviours in chicks of a small procellariiform seabird, Leach's storm petrel (Oceanodroma leucorhoa). This semi-precocial species, which has a well-developed sense of smell, nests in underground burrows where adults provision chicks for 6-8 weeks in the dark before fledging. Retinal ganglion cell topographic maps revealed that fine-tuning of cell distribution does not happen early in development, but rather that the ganglion cell layer continues to mature throughout provisioning and probably even after fledging. While the olfactory bulbs reached adult size around 7 weeks after hatching, the eyes and telencephalon continued to grow. Optokinetic head response and artificial burrow finding experiments indicated that chicks in the 2nd week after hatching lack even the most basic visually guided behaviours and are probably blind. Thus, vision in Leach's storm petrel chicks starts to function sometime around the 3rd week after hatching, well after the eyes have opened and the olfactory system is functional.

Morphological and genetic factors shape the microbiome of a seabird species (Oceanodroma leucorhoa) more than environmental and social factors.

BACKGROUND: The microbiome provides multiple benefits to animal hosts that can profoundly impact health and behavior. Microbiomes are well-characterized in humans and other animals in controlled settings, yet assessments of wild bird microbial communities remain vastly understudied. This is particularly true for pelagic seabirds with unique life histories that differ from terrestrial bird species. This study was designed to examine how morphological, genetic, environmental, and social factors affect the microbiome of a burrow-nesting seabird species, Leach's storm petrel (Oceanodroma leucorhoa). These seabirds are highly olfactory and may rely on microbiome-mediated odor cues during mate selection. Composition and structure of bacterial communities associated with the uropygial gland and brood patch were assessed using 16S rRNA amplicon-based Illumina Mi-Seq analysis and compared to burrow-associated bacterial communities. This is the first study to examine microbial diversity associated with multiple body sites on a seabird species. RESULTS: Results indicate that sex and skin site contribute most to bacterial community variation in Leach's storm petrels and that major histocompatibility complex (MHC) genotype may impact the composition of bacterial assemblages in males. In contrast to terrestrial birds and other animals, environmental and social interactions do not significantly influence storm petrel-associated bacterial assemblages. Thus, individual morphological and genetic influences outweighed environmental and social factors on microbiome composition. CONCLUSIONS: Contrary to observations of terrestrial birds, microbiomes of Leach's storm petrels vary most by the sex of the bird and by the body site sampled, rather than environmental surroundings or social behavior.

Vision on the high seas: spatial resolution and optical sensitivity in two procellariiform seabirds with different foraging strategies.

Procellariiform or 'tubenosed' seabirds are challenged to find prey and orient over seemingly featureless oceans. Previous studies have found that life-history strategy (burrow versus surface nesting) was correlated to foraging strategy. Burrow nesters tended to track prey using dimethyl sulphide (DMS), a compound associated with phytoplankton, whereas surface-nesting species did not. Burrow nesters also tended to be smaller and more cryptic, whereas surface nesters were larger with contrasting plumage coloration. Together these results suggested that differences in life-history strategy might also be linked to differences in visual adaptations. Here, we used Leach's storm petrel, a DMS-responder, and northern fulmar, a non-responder, as model species to test this hypothesis on their sensory ecology. From the retinal ganglion cell density and photoreceptor dimensions, we determined that Leach's storm petrels have six times lower spatial resolution than the northern fulmars. However, the optical sensitivity of rod photoreceptors is similar between species. These results suggest that under similar atmospheric conditions, northern fulmars have six times the detection range for similarly sized objects. Both species have extended visual streaks with a central area of highest spatial resolution, but only the northern fulmar has a central fovea. The prediction that burrow-nesting DMS-responding procellariiforms should differ from non-responding species nesting in the open holds true for spatial resolution, but not for optical sensitivity. This result may reflect the fact that both species rely on olfaction for their nocturnal foraging activity, but northern fulmars might use vision more during daytime.

Marine plastic debris emits a keystone infochemical for olfactory foraging seabirds.

Plastic debris is ingested by hundreds of species of organisms, from zooplankton to baleen whales, but how such a diversity of consumers can mistake plastic for their natural prey is largely unknown. The sensory mechanisms underlying plastic detection and consumption have rarely been examined within the context of sensory signals driving marine food web dynamics. We demonstrate experimentally that marine-seasoned microplastics produce a dimethyl sulfide (DMS) signature that is also a keystone odorant for natural trophic interactions. We further demonstrate a positive relationship between DMS responsiveness and plastic ingestion frequency using procellariiform seabirds as a model taxonomic group. Together, these results suggest that plastic debris emits the scent of a marine infochemical, creating an olfactory trap for susceptible marine wildlife.

Evidence that dimethyl sulfide facilitates a tritrophic mutualism between marine primary producers and top predators.

Tritrophic mutualistic interactions have been best studied in plant-insect systems. During these interactions, plants release volatiles in response to herbivore damage, which, in turn, facilitates predation on primary consumers or benefits the primary producer by providing nutrients. Here we explore a similar interaction in the Southern Ocean food web, where soluble iron limits primary productivity. Dimethyl sulfide has been studied in the context of global climate regulation and is an established foraging cue for marine top predators. We present evidence that procellariiform seabird species that use dimethyl sulfide as a foraging cue selectively forage on phytoplankton grazers. Their contribution of beneficial iron recycled to marine phytoplankton via excretion suggests a chemically mediated link between marine top predators and oceanic primary production.

The neuroecology of dimethyl sulfide: a global-climate regulator turned marine infochemical.

Information transfer influences food-web dynamics in the marine environment, but infochemicals involved in these processes are only beginning to be understood. Dimethylsulfoniopropionate (DMSP) is produced by phytoplankton and other marine algae, and has been studied primarily in the context of sulfur cycling and regulation of global climate. My laboratory has been investigating DMSP and its breakdown product, dimethyl sulfide as infochemicals associated with trophic interactions in marine habitats, including sub-Antarctic and coral reef ecosystems. Using a neuroecological approach, our work has established that these biogenic sulfur compounds serve as critical signal molecules in marine systems and provides us with a more mechanistic understanding of how climate change may impact information transfer within marine food webs.

Rapid communication: experimental evidence that juvenile pelagic jacks (Carangidae) respond behaviorally to DMSP.

Dimethylsulfoniopropionate (DMSP) is produced by marine algae and released during foraging activity by zooplankton and fish. Pelagic fishes depend on patchily distributed foraging opportunities, and DMSP may be an important signaling molecule for these events. We have previously shown that the abundance of carangid jacks is positively associated with elevated DMSP levels over coral reefs in the Gulf of Mexico, suggesting that these fishes may use spatial and temporal variation in DMSP to locate foraging opportunities. Here, we extend this work by demonstrating that juveniles of two species of pelagic jack, crevalle jack, Caranx hippos, and bluefin trevally, C. melampygus, detect and respond to DMSP in a flow-through tank in the laboratory. Juveniles of these species showed elevated swimming activity in response to ecologically relevant concentrations of DMSP (10(-9) M). These results provide further evidence that this chemical may serve as a chemosensory cue for carangid species.

Individual odor recognition in procellariiform chicks: potential role for the major histocompatibility complex.

Since the groundbreaking work of Wenzel, Bang, and Grubb in the 1960s, enormous progress has been made toward elucidating the sense of smell in procellariiform seabirds. Although it is now well established that adult procellariiforms use olfaction in many behaviors, such as for foraging, nest relocation, and mate recognition, the olfactory abilities of petrel chicks are less well understood. Recent studies have shown that petrel chicks can recognize prey-related odors and odors associated with their nest before leaving their burrow for the first time. The recognition of burrow odors by petrel chicks is unlikely to be used for homing, and we have suggested that chicks may be learning personal odors associated with the nest's occupants for use later in life in the context of kin recognition or mate choice. The source of personal odors in petrels is unknown. However, in other vertebrates, the major histocompatibility complex influences body odors, which in turn influence mating preferences. It is not currently known whether this highly polymorphic gene region influences body odors and individual recognition in the procellariiforms, but this could be a fruitful area of future research.

The use of odors at different spatial scales: comparing birds with fish.

Salmon travel hundreds of kilometers of open ocean and meandering rivers to return to their natal stream to spawn; procellariiform seabirds soar over thousands of kilometers of the ocean's surface searching for foraging opportunities and accurately return to their nesting islands. These large-scale olfactory-guided behaviors are among the most dramatic examples of animal navigation ever described. At much closer ranges, the sense of smell can be used for behaviors as diverse as tracking prey, nest location, and mate selection. Both fish and birds face similar problems interpreting olfactory information in fluid mediums where odors are dispersed as filamentous patches. Similar to insects, which have served as model organisms for investigating olfactory related behaviors, the few fish and bird species that have been studied tend to use olfactory information in conjunction with other sensory modalities. Similar to insects, fish and birds also employ oscillatory or cross-stream movement as sampling mechanisms. This review compares and contrasts the use of odors by fish and birds over a range of spatial scales that span from thousands of kilometers to less than a meter. In so doing, we identify behavioral similarities and new questions that need to be addressed regarding the olfactory ecology of these diverse groups of organisms.

Sensory ecology on the high seas: the odor world of the procellariiform seabirds.

Procellariiform seabirds wander the world's oceans aided by olfactory abilities rivaling those of any animal on earth. Over the past 15 years, I have been privileged to study the sensory ecology of procellariiforms, focusing on how olfaction contributes to behaviors, ranging from foraging and navigation to individual odor recognition, in a broader sensory context. We have developed a number of field techniques for measuring both olfactory- and visually based behaviors in chicks and adults of various species. Our choice of test odors has been informed by long-term dietary studies and geochemical data on the production and distribution of identifiable, scented compounds found in productive waters. This multidisciplinary approach has shown us that odors provide different information over the ocean depending on the spatial scale. At large spatial scales (thousands of square kilometers), an olfactory landscape superimposed upon the ocean surface reflects oceanographic or bathymetric features where phytoplankton accumulate and an area-restricted search for prey is likely to be successful. At small spatial scales (tens to hundreds of square kilometers), birds use odors and visual cues to pinpoint and capture prey directly. We have further identified species-specific, sensory-based foraging strategies, which we have begun to explore in evolutionary and developmental contexts. With respect to chemical communication among individuals, we have shown that some species can distinguish familiar individuals by scent cues alone. We are now set to explore the mechanistic basis for these discriminatory abilities in the context of kin recognition, and whether or not the major histocompatibility complex is involved.

Evidence for olfactory search in wandering albatross, Diomedea exulans.

Wandering albatrosses (Diomedea exulans) forage over thousands of square kilometers of open ocean for patchily distributed live prey and carrion. These birds have large olfactory bulbs and respond to fishy-scented odors in at-sea trials, suggesting that olfaction plays a role in natural foraging behavior. With the advent of new, fine-scale tracking technologies, we are beginning to explore how birds track prey in the pelagic environment, and we relate these observations to models of odor transport in natural situations. These models suggest that odors emanating from prey will tend to disperse laterally and downwind of the odor source and acquire an irregular and patchy concentration distribution due to turbulent transport. For a seabird foraging over the ocean, this scenario suggests that olfactory search would be facilitated by crosswind flight to optimize the probability of encountering a plume emanating from a prey item, followed by upwind, zigzag flight to localize the prey. By contrast, birds approaching prey by sight would be expected to fly directly to a prey item, irrespective of wind direction. Using high-precision global positioning system (GPS) loggers in conjunction with stomach temperature recorders to simultaneously monitor feeding events, we confirm these predictions in freely ranging wandering albatrosses. We found that initial olfactory detection was implicated in nearly half (46.8%) of all flown approaches preceding prey-capture events, accounting for 45.5% of total prey mass captured by in-flight foraging. These results offer insights into the sensory basis for area-restricted search at the large spatial scales of the open ocean.

Dimethylsulfoniopropionate as a foraging cue for reef fishes.

Coral reefs resemble islands of productive habitats where fishes aggregate, forage, and spawn. Although it has been suggested that some reef fishes use biogenic chemicals as aggregation cues, specific chemicals have not been identified. Dimethylsulfoniopropionate (DMSP), a secondary metabolite of many marine algal species, is released during foraging by higher-order consumers. DMSP has been studied intensively for its role in oceanic sulfur cycles and global climate regulation, but its ecological importance to marine fishes is unknown. We present evidence that planktivorous reef fishes will aggregate to experimental deployments of DMSP over coral reef habitats in the wild.

Examining the development of individual recognition in a burrow-nesting procellariiform, the Leach's storm-petrel.

Burrow-nesting petrels use their well-developed sense of smell for foraging, homing to their nest, and mate recognition. The chicks of burrow-nesting petrels can apparently learn odours associated with prey while still in the nest, but the development of individual-specific odour recognition is less well understood. We used a simple two-choice test to determine whether 4- to 6-week-old chicks of a small, burrow-nesting species, the Leach's storm-petrel (Oceanodroma leucorhoa), prefer the scent of their own nest material to (1) the scent of similar organic material collected from the colony or (2) the scent of a conspecific's nest material. Results suggest that chicks clearly preferred the scent of their own nest material to that of similar organic material collected from the colony (96%; N=24; binomial test, P<0.001). Results further suggested that birds preferred the scent of their own nest material to that of a conspecific, though the preference was statistically less robust (67%; N=39; binomial test, P=0.05). Because Leach's storm-petrel chicks do not normally leave their burrow prior to fledging, an ability to recognise individual or nest-specific odours is not likely to be used for homing but instead may be linked to the development of individual recognition in different contexts.

Testing an ecophysiological mechanism of morphological plasticity in pupfish and its relevance to conservation efforts for endangered Devils Hole pupfish.

Imperiled species that have been translocated or established in captivity can show rapid alterations in morphology and behavior, but the proximate mechanisms of such phenotypic changes are rarely known. Devils Hole pupfish (Cyprinodon diabolis) are endemic to a single desert pool and are characterized by a small body, large head and eyes, and lack of pelvic fins. To lessen the risk of extinction, additional populations of C. diabolis were established in artificial refuges. Yet, pupfish in these refuges rapidly shifted to a larger body, smaller head and eyes, and greater body depth. Here we examined how food availability and temperature, which differ between these habitats, influence morphological development in closely related Amargosa River pupfish (Cyprinodon nevadensis amargosae). We were interested in knowing whether these environmental factors could developmentally shift Amargosa River pupfish toward the morphology typical of pupfish in Devil's Hole. By regulating food ration, we created groups of pupfish with low, medium and high growth rates. Pupfish with low growth showed proportionally larger head and eyes, smaller body depth, and reduction in pelvic fin development. Elevated temperature further inhibited pelvic fin development in all treatments. Pupfish in the low growth group also showed reduced levels of thyroid hormone, suggesting a possible physiological mechanism underlying these morphological changes. To test this mechanism further, pupfish were reared with goitrogens to pharmacologically inhibit endogenous thyroid hormone production. Pupfish given goitrogens developed larger heads and eyes, shallower bodies, and reduced pelvic fins. Taken together, our results suggest that changes in environmental factors affecting the growth and thyroid hormone status of juvenile pupfish may play a developmental role in generating the morphological differences between C. diabolis in Devil's Hole and the refuges. These findings illustrate the need to incorporate a mechanistic understanding of phenotypic plasticity into conservation strategies to preserve imperiled fishes.

Early rearing environment impacts cerebellar growth in juvenile salmon.

The size and structure of an animal's brain is typically assumed to result from either natural or artificial selection pressures over generations. However, because a fish's brain grows continuously throughout life, it may be particularly responsive to the environmental conditions the fish experiences during development. Salmon are an ideal model system for studying these effects because natural habitats differ significantly from the hatchery environments in which these fish are frequently reared. For example, in the wild, salmon alevins (i.e. yolk-sac fry) are buried in the gravel, while hatchery environments lack this structural component. We show that the simple manipulation of adding stones to a standard rearing tank can dramatically alter the growth of specific brain structures in steelhead salmon alevins (Oncorhynchus mykiss). We found that alevins reared with stones grew brains with significantly larger cerebella than genetically similar fish reared in conventional tanks. This shift to a larger cerebellar size was, in turn, accompanied by changes in locomotory behaviors--behaviors that correlate strongly to the function of this brain region. We next show that hatchery fish reared in a more naturalistic setting in the wild had significantly larger brains than their lab-reared counterparts. However, relative cerebellar volumes were similar between wild-reared alevins and those reared in the complex treatment in the laboratory. Together our results indicate that, within the first three weeks of life, variation in rearing environment can result in brain differences that are commonly attributed to generations of selection. These results highlight the need to consider enrichment strategies when designing captive rearing facilities for both conservation and laboratory use.