Sex, seasonal, and stress-related variations in elasmobranch corticosterone concentrations

Serum corticosterone was previously studied in numerous elasmobranch fishes (sharks, skates and rays), but the role of this steroid, widespread throughout many taxa, has yet to be defined. The goal of this study was to test whether corticosterone varied in response to acute and chronic capture stress, and across the reproductive cycle in the bonnethead shark, Sphyrna tiburo, and Atlantic stingray, Dasyatis sabina. Serum corticosterone in S. tiburo increased following capture and again 24 h post-capture, possibly caused by interference with 1α-hydroxycorticosterone, the primary stress hormone in elasmobranchs. Higher serum concentrations in males compared to females were observed in both species. Variations in corticosterone also occurred during the reproductive cycle in both species. Consistent with other taxa, elevations in male bonnethead sharks and stingrays coincided with peak testicular development and mating. Elevations in female bonnethead sharks occurred from the time of mating through sperm storage into early gestation. In contrast, corticosterone levels in female stingrays were low during their protracted mating season, but elevated through late gestation and parturition. These results indicate that corticosterone has a limited role, if any, in acute and chronic stress associated with capture in S. tiburo, but likely has physiological functions associated with its glucocorticoid properties across the reproductive cycle of both species.     

Predominance of genetic monogamy by females in a hammerhead shark, Sphyrna tiburo: implications for shark conservation

There is growing interest in the mating systems of sharks and their relatives (Class Chondrichthyes) because these ancient fishes occupy a key position in vertebrate phylogeny and are increasingly in need of conservation due to widespread overexploitation. Based on precious few genetic and field observational studies, current speculation is that polyandrous mating strategies and multiple paternity may be common in sharks as they are in most other vertebrates. Here, we test this hypothesis by examining the genetic mating system of the bonnethead shark, Sphyrna tiburo, using microsatellite DNA profiling of 22 litters (22 mothers, 188 embryos genotyped at four polymorphic loci) obtained from multiple locations along the west coast of Florida. Contrary to expectations based on the ability of female S. tiburo to store sperm, the social nature of this species and the 100% multiple paternity observed in two other coastal shark species, over 81% of sampled bonnethead females produced litters sired by a single male (i.e. genetic monogamy). When multiple paternity occurred in S. tiburo, there was an indication of increased incidence in larger mothers with bigger litters. Our data suggest that sharks may exhibit complex genetic mating systems with a high degree of interspecific variability, and as a result some species may be more susceptible to loss of genetic variation in the face of escalating fishing pressure. Based on these findings, we suggest that knowledge of elasmobranch mating systems should be an important component of conservation and management programmes for these heavily exploited species.     

Serum steroid hormones including 11-ketotestosterone, 11-ketoandrostenedione, and dihydroprogesterone in juvenile and adult bonnethead sharks, Sphyrna tiburo.

Previous studies in the placental viviparous bonnethead shark, Sphyrna tiburo, have correlated 17 beta-estradiol, progesterone, testosterone, and dihydrotestosterone with reproductive events in both males and females. However, several key reproductive events, including implantation, maintenance of pregnancy, and parturition, did not correlate with these four steroid hormones. Therefore, the present study investigated three steroid hormones, 11-ketotestosterone, 11-ketoandrostenedione, and dihydroprogesterone, which have demonstrably important roles in the reproductive cycles of teleosts. It was hypothesized that one or more of these three hormones would correlate with specific reproductive events in S. tiburo. Concurrently, developmental (growth and/or maturation) analyses of these three steroids plus 17 beta-estradiol, progesterone, testosterone, and dihydrotestosterone were investigated in juvenile bonnethead sharks. Serum dihydroprogesterone concentrations were highest in mature females and 11-ketotestosterone concentrations were highest in mature males. In mature females, 11-ketoandrostenedione levels were elevated from the time of mating, through six months of sperm storage and another four months of gestation. At parturition concentrations became significantly lower and remained lower until mating occurred again in another two to three months. Serum 11-ketotestosterone concentrations were the highest at implantation though not significant. In mature males, significantly elevated serum levels of dihydroprogesterone occurred in April and May, near the start of annual testicular development. During growth in males, testosterone and dihydrotestosterone increased progressively and in females, testosterone increased progressively. At maturity in males, significant increases occurred in testosterone and 11-ketotestosterone concentrations while, in females, dihydroprogesterone, 11-ketotestosterone, 17 beta-estradiol, progesterone, testosterone, and dihydrotestosterone concentrations increased. This study shows that although testosterone may be the primary androgen in the bonnethead shark, other derived androgens may have important functions in growth, maturation, and reproduction. J. Exp. Zool. 284:595-603, 1999.     

Assessment of acid-base derangements among bonnethead (Sphyrna tiburo), bull (Carcharhinus leucas), and lemon (Negaprion brevirostris) sharks from gillnet and longline capture and handling methods

Blood gasses of wild bonnethead, bull, and lemon sharks were measured with the i-STAT clinical analyzer with the CG4+ cartridge immediately after capture; and again immediately prior to release after tagging, handling and morphometric measurements were taken. Relative reference ranges of post-capture status were established. Among species, stress response to capture was similar for all parameters; however, pH declined and lactate concentrations rose over time, indicating continued insult from capture and/or response to additional handling stress. pCO(2) rose faster for S. tiburo than for C. leucas, and lactate concentrations rose faster for S. tiburo than for N. brevirostris. All species caught in gillnets experienced lower pH and higher lactate concentrations than on longlines. Discriminant analysis justified the use of blood gas analysis to assess physiological stress induced by different capture methods. From these results, we recommend 1) that gear be monitored closely and sharks be removed immediately, or suboptimally, that gear is deployed for the shortest soak time possible; 2) longline over gillnet gear; and 3) extra caution with sensitive species (e.g., S. tiburo), which may include the administration of blood buffers and other therapeutics if a shark is beyond the limits of relative reference ranges reported here.     

Structure and permeability of the egg capsule of the bonnethead shark, Sphyrna tiburo

In the viviparous bonnethead shark, Sphyrna tiburo, a fluid-filled, acellular egg capsule surrounds fertilized eggs and developing embryos throughout gestation. Like other placental shark species, the capsule remains intact even at the placental implantation site. Although its intervention between the uterine and embryonic tissues of the placenta has long been thought to mediate physiological exchange, little information is available concerning even its basic structure or permeability to solutes. The 1 mum thick capsule wall consists of an inner layer of gelatinous material and an outer layer consisting of at least three laminae of orthogonally arranged fibrous material. These fibers are irregular and often branched. Permeability experiments showed that solutes less than 1,355 Da diffuse across the egg capsule whereas those greater than 6,000 Da do not pass through the membrane. Solute movement across the capsule is a concentration-dependent phenomenon indicating diffusion rather than active transport. Experimental data also suggest that there is an increase in the permeability of the egg capsule to low molecular weight materials during mid- and late gestation. These observations are discussed in relation to the function of the egg capsule as a mediator of maternal-embryonic interactions in matrotrophic sharks.     

Age and growth of the bonnethead shark, Sphyrna tiburo, from northwest Florida, with comments on clinal variation

Age and growth rates of the bonnethead shark, Sphyrna tiburo, from northwest Florida were estimated from vertebrae collected between October 1992 and October 1995. The von Bertalanffy growth equation was fit to male and female vertebral age data. Initial growth was rapid (≈ 200 mm TL) for both sexes from age 0–1. At age 2 growth slowed for males but continued for females. Similar to many species of sharks, females grew slower than males (K = 0.28 and K = 0.69, respectively) but attained a larger maximum size (L_∞=1226 and L_∞=897). Maximum age was estimated in males and females to be 8+ and 12+ years, respectively. Growth of young-of-year sharks was 21 to 30 mm TL per month determined by three different methods. A comparison of age and growth estimates from populations at more southerly latitudes suggest that clinal variation in total length may be evident among bonnethead sharks in the Gulf of Mexico with females reaching larger sizes in northern areas as compared to south Florida. This revised version was published online in July 2006 with corrections to the Cover Date.     

An acute stressor alters steroid hormone levels and activity but not sexual behavior in male and female Ocoee salamanders (Desmognathus ocoee)

Stressors that are chronic have clear suppressive effects on reproductive behaviors in both males and females. Stressors that are acute have effects on reproductive behavior that are less clear. We measured the effects of an acute bout of handling in laboratory-housed male and female Ocoee salamanders (Desmognathus ocoee), a species with a prolonged mating season. Handling resulted in decreased locomotory activity and elevated plasma corticosterone, a hallmark of the vertebrate stress response. Handling also decreased plasma testosterone in males and elevated plasma estradiol in females. Despite the handling-induced changes in hormone levels, handling had minimal impact on courtship and mating. Other species in which reproduction is insensitive to acute stressors may live in extreme environments with limited reproductive opportunities, whereas Ocoee salamanders live in a relatively temperate environment with multiple reproductive opportunities. Together, these data indicate that an allostatic response to a stressor can alter locomotory activity and elevate corticosterone without suppressing nonessential behaviors like courtship and mating in a species in which reproductive opportunities can occur over a period of multiple months. The lack of reproductive suppression in Ocoee salamanders might be due to the low energetic cost of courtship and mating in this species combined with potentially elevated energetic stores, highlighting the importance of considering energy budgets when making predictions about behavioral effects of acute stressors.     

Calcitonin-like immunoreactivity in serum and tissues of the bonnethead shark,Sphyrna tiburo

Calcitonin is a 32-amino acid peptide hormone that is best known for its actions in maintaining skeletal integrity and calcium homeostasis in mammals. Calcitonin also appears to function in regulating certain aspects of animal reproduction, but the nature of this role remains unclear, particularly in nonmammalian vertebrates. The present study investigated the relationship between calcitonin and reproduction in the bonnethead shark (Sphyrna tiburo), a well-studied member of the oldest living vertebrate group (i.e. elasmobranchs) known to possess a calcitonin-producing organ. Serum calcitonin concentrations were measured in 28 reproductively mature female S. tiburo using a heterologous enzyme-linked immunosorbent assay (ELISA) system. Sites of calcitonin immunoreactivity were detected in tissues of mature female and embryonic S. tiburo using immunocytochemistry. Significant increases in serum calcitonin concentrations of mature female S. tiburo occurred during early stages of gestation, a period characterized by yolk-dependency of developing embryos. Immunoreactive calcitonin was detected in the duodenum and pancreas of embryonic S. tiburo sampled during the same period. The results from this study suggest that calcitonin obtained from endogenous and/or maternal sources may function in regulating yolk digestion in embryonic S. tiburo. Therefore, the association between calcitonin and reproduction in elasmobranchs may reflect an important role for this hormone in embryonic development.     

Baseline health parameters and species comparisons among free-ranging Atlantic sharpnose (Rhizoprionodon terraenovae), bonnethead (Sphyrna tiburo), and spiny dogfish (Squalus acanthias) sharks in Georgia, Florida, and Washington, USA

Sharks are of commercial, research, conservation, and exhibition importance but we know little regarding health parameters and population status for many species. Here we present health indicators and species comparisons for adults of three common wild-caught species: 30 Atlantic sharpnose sharks (Rhizoprionodon terraenovae) and 31 bonnethead sharks (Sphyrna tiburo) from the western Atlantic, and 30 spiny dogfish sharks (Squalus acanthias) from the eastern Pacific. All animals were captured during June-July 2009 and 2010. Median values and preliminary reference intervals were calculated for hematology, plasma biochemistry, trace nutrients, and vitamin A, E, and D concentrations. Significant differences, attributable to physiologic differences among the species, were found in the basic hematologic and plasma biochemistry variables. Significant species differences in arsenic and selenium plasma concentrations were found and appear to coincide with diet and habitat variability among these three species. Vitamin E was significantly higher in the bonnethead shark, again related to the foraging ecology and ingestion of plant material by this species. The Atlantic sharpnose had significantly higher vitamin A concentrations, supported by the higher proportion of teleosts in the diet. Vitamin D was below the limit of quantification in all three species. These preliminary reference intervals for health variables can be used to assess and monitor the population health and serve as indicators of nutritional status in these populations of wild elasmobranchs.     

Seasonal variation in hormonal responses of timber rattlesnakes (<Emphasis Type="Italic">Crotalus horridus</Emphasis>) to reproductive and environmental stressors

Data addressing adrenocortical modulation across taxonomic groups are limited, especially with regard to how female reproductive condition influences the sensitivity of the hypothalamus–pituitary–adrenal axis. We investigated seasonal and reproductive variation in basal and stress-induced hormone profiles in a population of free-ranging timber rattlesnakes (Crotalus horridus) in north-central Pennsylvania during spring (i.e., May), summer (i.e., July), and early fall (i.e., September). Baseline corticosterone concentrations varied seasonally and were significantly lower during the summer sampling period in July. We observed a significant negative relationship between baseline corticosterone and testosterone in male snakes, while baseline corticosterone and estradiol tended to be positively correlated in females. Treatment of snakes with 1 h of capture stress significantly increased corticosterone across all seasons. However, there was a significant interaction between corticosterone responses to capture stress and season, suggesting that adrenocortical function is modulated seasonally. Because elevated corticosterone may be associated with reproduction, we asked whether hormonal stress responses vary with female reproductive condition. Although sample sizes are low, reproductive snakes had significantly higher baseline and stress-induced corticosterone concentrations than non-reproductive or post-parturient females. Further, despite similar baseline corticosterone concentrations between non-reproductive and post-parturient rattlesnakes, post-parturient females responded to capture stress with a significantly higher increase in corticosterone. Collectively, these data suggest that the sensitivity of the hypothalamus–pituitary–adrenal axis varies both seasonally and with changing reproductive states.     

Reproductive biology,multiple paternity and polyandry of the bull shark Carcharhinus leucas

To improve understanding of bull shark Carcharhinus leucas reproductive biology, we analysed reproductive traits from 118 bull sharks caught along Reunion Island coasts (Western Indian Ocean), including 16 gravid females. Specific microsatellite loci were used to investigate the frequency of multiple paternity. Males and females reached maturity at c. 234 cm and 257 cm total length (L_T), respectively, and litter sizes ranged from 5 to 14 embryos. Analysis of the 16 litters collected in various months of the year indicated that parturition occurs between October and December, with a size at birth c. 60–80 cm L_T and that the gestation period is probably c. 12 months. Assuming a 1 year resting period and a period of sperm storage (4–5 months) between mating (in June–September) and fertilisation, the reproductive cycle of bull sharks at Reunion Island would be biennial. At least 56.25% of the litters investigated were polyandrous, sired by 2–5 males. Several males that each sired several litters conceived during the same or distinct mating seasons were detected, suggesting both a seasonal aggregation of sharks to mate and some male fidelity to mating site. Altogether, these findings provide valuable information for both shark risk management and conservation of the species in the Western Indian Ocean.     

Do sharks exhibit heterodonty by tooth position and over ontogeny? A comparison using elliptic Fourier analysis

Tooth morphology is often used to inform the feeding ecology of an organism as these structures are important to procure and process dietary resources. In sharks, differences in morphology may facilitate the capture and handling of prey with different physical properties. However, few studies have investigated differences in tooth morphology over ontogeny, throughout the jaws of a single species, or among species at multiple tooth positions. Bull (Carcharhinus leucas), blacktip (Carcharhinus limbatus), and bonnethead sharks (Sphyrna tiburo) are coastal predators that exhibit ontogenetic dietary shifts, but differ in their feeding ecologies. This study measured tooth morphology at six positions along the upper and lower jaws of each species using elliptic Fourier analysis to make comparisons within and among species over their ontogeny. Significant ontogenetic differences were detected at four of the six tooth positions in bull sharks, but only the posterior position on the lower jaw appeared to exhibit a functionally relevant shift in morphology. No ontogenetic changes in morphology were detected in blacktip or bonnethead sharks. Intraspecific comparisons found that most tooth positions significantly differed from one another across all species, but heterodonty was greatest in bull sharks. Additionally, interspecific comparisons found differences among all species at each tooth position except between bull and blacktip sharks at two positions. These morphological patterns within and among species may have implications for prey handling efficiency, as well as in providing insight for paleoichthyology studies and reevaluating heterodonty in sharks.     

Gill lesions associated with Erpocotyle tiburonis (Monogenea: Hexabothriidae) on wild and aquarium-held bonnethead sharks (Sphyrna tiburo).

Gill lesions associated with infections of Erpocotyle tiburonis (Brooks, 1934) (Monogenea: Hexabothriidae) on wild bonnethead sharks (Sphyrna tiburo (L., 1758) (Carcharhiniformes: Sphyrinidae)) were compared with those on aquarium-held ones using light and scanning electron microscopy. Uninfected gill filaments had slender, triangular, smooth-surfaced lamellae and interlamellar water channels that were approximately equal in size. Four wild sharks were each infected by 3-11 widely separated adult E. tiburonis, and 1 of these sharks hosted a juvenile specimen. Lamellae flanking or touching adult E. tiburonis were pushed aside or bent, but were otherwise identical to those of uninfected filaments. Two aquarium-held sharks were each infected by hundreds of juvenile and adult E. tiburonis. In these sharks, lamellae near juveniles were pushed apart or bent, but were otherwise normal, whereas a thick, ragged-surfaced layer of hyperplastic epithelium both filled interlamellar water channels and partially or completely covered lamellae near adults. Results of this study suggest that the intense infections of E. tiburonis were facilitated by captivity and caused severe hyperplastic lesions that ultimately led to the death of the sharks by reducing or blocking the respiratory water flow over lamellae and thus reducing the exchange of gases and ions across the lamellar epithelium. In contrast, the wild sharks were infected by fewer worms and exhibited relatively minor lesions.     

Selection and sex‐biased dispersal in a coastal shark: the influence of philopatry on adaptive variation

Sex‐biased dispersal is expected to homogenize nuclear genetic variation relative to variation in genetic material inherited through the philopatric sex. When site fidelity occurs across a heterogeneous environment, local selective regimes may alter this pattern. We assessed spatial patterns of variation in nuclear‐encoded, single nucleotide polymorphisms (SNPs) and sequences of the mitochondrial control region in bonnethead sharks (Sphyrna tiburo), a species thought to exhibit female philopatry, collected from summer habitats used for gestation. Geographic patterns of mtDNA haplotypes and putatively neutral SNPs confirmed female philopatry and male‐mediated gene flow along the northeastern coast of the Gulf of Mexico. A total of 30 outlier SNP loci were identified; alleles at over half of these loci exhibited signatures of latitude‐associated selection. Our results indicate that in species with sex‐biased dispersal, philopatry can facilitate sorting of locally adaptive variation, with the dispersing sex facilitating movement of potentially adaptive variation among locations and environments.     

Serum concentrations of reproductively-related circulating steroid hormones in the free-ranging lemon shark,Negaprion brevirostris

Synopsis We have examined the concentrations of reproductively-related steroid hormones in 5 species of carcharhinid sharks, marine fishes possessing the unique attribute of placental viviparity. Measurements of serum estradiol, testosterone, progesterone, dihydrotestosterone, and corticosterone have provided baseline data for these hormones in both immature and adult male and female placental sharks. Our studies include: (1) changes in hormonal levels during maturation, (2) concentrations of circulating steroid hormones during peak breeding season, and (3) hormonal levels during gestation, including the collection of serial samples through and beyond birth from free-ranging lemon sharks. Our data suggest that steroids, important in regulating reproduction in higher mammals, are also essential in these cartilaginous fishes.     

Food Supplementation Fails to Reveal a Trade-Off between Incubation and Self-Maintenance in Female House Wrens

Incubating birds must allocate their time and energy between maintaining egg temperature and obtaining enough food to meet their own metabolic demands. We tested the hypothesis that female house wrens (Troglodytes aedon) face a trade-off between incubation and self-maintenance by providing females with supplemental food during incubation. We predicted that food supplementation would increase the amount of time females devoted to incubating their eggs, lower their baseline plasma corticosterone levels (a measure of chronic stress), and increase their body mass, haematocrit (a measure of anaemia), and reproductive success relative to control females. As predicted, food-supplemented females spent a greater proportion of time incubating their eggs than control females. Contrary to expectation, however, there was no evidence that food supplementation significantly influenced female baseline plasma corticosterone levels, body mass, haematocrit, or reproductive success. However, females with high levels of corticosterone at the beginning of incubation were more likely to abandon their nesting attempt after capture than females with low levels. Corticosterone significantly increased between the early incubation and early nestling stages of the breeding cycle in all females. These results suggest that although food supplementation results in a modest increase in incubation effort, it does not lead to significantly lower levels of chronic stress as reflected in lower baseline corticosterone levels. We conclude that female house wrens that begin the incubation period with low levels of plasma corticosterone can easily meet their own nutritional needs while incubating their eggs, and that any trade-off between incubation and self-feeding does not influence female reproductive success under the conditions at the time of our study.     

Behavioural and hormonal responses to capture stress in the male red-sided garter snake, Thamnophis sirtalis parietalis

We measured the behavioural and hormonal responses to capture stress in male red-sided garter snakes. Four hours of capture stress resulted in no suppression of mating behaviour relative to control individuals. In contrast, the same stress resulted in a significant increase in plasma levels of corticosterone and a significant decrease in plasma levels of testosterone. There was a significant negative correlation between plasma levels of corticosterone and testosterone in both control and capture-stress groups, suggesting that the increase in corticosterone directly drives the decrease in testosterone. While there was no relation between body size and initial plasma levels of the two steroids, longer individuals had a significantly greater increase in corticosterone following capture stress than did shorter individuals. Snakes display indeterminate growth, suggesting that older individuals have decreased sensitivity to negative feedback in the hypothalamic-pituitary-adrenal axis and thus hypersecrete glucocorticoids. These results suggest that male red-sided garter snakes have uncoupled their behavioural stress response from their hormonal stress response to maximize reproductive opportunities. Copyright 2000 The Association for the Study of Animal Behaviour.     

Temporal and latitudinal comparisons of reproductive parameters in a heavily exploited shark,the bonnethead,Sphyrna tiburo (L. 1758), in the southern Gulf of Mexico

In the southern Gulf of Mexico, the bonnethead shark, Sphyrna tiburo, is one of the most frequently captured species in landings of small-scale fisheries. Based on the analysis of two fishery-dependent sampling periods (1993–1994 and 2007–2014), this study aimed to determine reproductive parameters and identify temporal differences between the two time periods. In the first sampling period, 776 males and 352 females with a size range of 28.0–120.0 cm total stretched length (L_T) were analysed, and in the second sampling period, 387 males and 432 females with a size range of 28.0–122.0 cm L_T were analysed. The size at 50% maturity in the second sampling period was significantly different between sexes, 82.6 cm L_T for females and 73.8 cm L_T for males (no estimation was possible for the first sampling period). The size at 50% maternity was not different between sampling periods, 97.3 cm L_T for the first sampling period and 99.0 cm L_T for the second sampling period. Litter size varied from 3 to 19 embryos and the average was not statistically different in both periods, 10.1 (S.D. = 3.8) for the first sampling period and 11.3 (S.D. = 3.5) for the second sampling period. The female reproductive cycle is asynchronous, and it seems to be annual, with a gestation period of 5–6 months, and a consecutive ovarian cycle and gestation period. Temporal (between sampling periods) and latitudinal (southern Gulf versus northern regions) variations occur in the synchronicity of the reproductive cycle, temporal variation in the relationship between maternal length and litter size, and latitudinal variation in average size of mature sharks.     

The energetic cost of mating in a promiscuous cephalopod

Costs that individuals incur through mating can play an important role in understanding the evolution of life histories and senescence, particularly in promiscuous species. Copulation costs, ranging from energy expenditure to reduced longevity, are widely studied in insects but have received substantially less attention in other taxa. One cost of mating, the energetic cost, is poorly studied across all taxa despite its potential importance for the many species where copulation is physically demanding and/or frequent. Here, we investigated the energetic cost of mating in both male and female dumpling squid (Euprymna tasmanica). In this species, copulation can last up to 3 h and requires that the male physically restrains the female. We report that the act of copulation halves the swimming endurance of both sexes, and that they take up to 30 min to recover. Such a reduction in post-copulatory performance may have important implications for predator avoidance, foraging ability and energy allocation. Therefore, quantifying this cost is essential to understand the evolution of reproductive strategies and behaviours such as female receptivity and male and female mating frequency.     

Utility of mesohabitat features for determining habitat associations of subadult sharks in Georgia’s estuaries

We examined the affects of selected water quality variables on the presence of subadult sharks in six of nine Georgia estuaries. During 231 longline sets, we captured 415 individuals representing nine species. Atlantic sharpnose shark (Rhizoprionodon terranovae), bonnethead (Sphyrna tiburo), blacktip shark (Carcharhinus limbatus) and sandbar shark (C. plumbeus) comprised 96.1% of the catch. Canonical correlation analysis (CCA) was used to assess environmental influences on the assemblage of the four common species. Results of the CCA indicated Bonnethead Shark and Sandbar Shark were correlated with each other and with a subset of environmental variables. When the species occurred singly, depth was the defining environmental variable; whereas, when the two co-occurred, dissolved oxygen and salinity were the defining variables. Discriminant analyses (DA) were used to assess environmental influences on individual species. Results of the discriminant analyses supported the general CCA findings that the presence of bonnethead and sandbar shark were the only two species that correlated with environmental variables. In addition to depth and dissolved oxygen, turbidity influenced the presence of sandbar shark. The presence of bonnethead shark was influenced primarily by salinity and turbidity. Significant relationships existed for both the CCA and DA analyses; however, environmental variables accounted for <16% of the total variation in each. Compared to the environmental variables we measured, macrohabitat features (e.g., substrate type), prey availability, and susceptibility to predation may have stronger influences on the presence and distribution of subadult shark species among sites.