G. Joan Holt

Co-feeding microparticulate diets with algae: toward eliminating the need of zooplankton at first feeding in larval red drum (Sciaenops ocellatus)

The effect of adding algae to the culture water used to rear red drum larvae was evaluated in terms of growth, survival and digestive enzyme activity. Red drum larvae were subjected to one of the following dietary regimes from first feeding (day 3 post-hatch) to day 14: (1) zooplankton supplemented with algae (L-A), (2) zooplankton without algae (L-NA), (3) a microparticulate diet with algae (M-A) and (4) the microparticulate diet alone (M-NA). The presence of algae in the rearing tanks improved growth of red drum larvae for both types of feeds. Growth was significantly higher (P<0.05) in larvae reared in the presence of algae (L-A and M-A) than in larvae raised in the corresponding treatments without algae (L-NA and M-NA). Red drum larvae raised on the microparticulate diet and algae (M-A) grew as well as the zooplankton treatment with no algae (L-NA), and were not significantly different from the L-A treatment. The larvae fed the microparticulate diet in the absence of algae (M-NA) were significantly smaller than the other three treatments. These results were consistent for two separate feeding trials. Final survival was highly variable in all treatments; nevertheless, mean final survival values were 30% higher in treatment L-A compared to L-NA (14.1 and 10.1%, respectively) and 42% higher in M-A than for M-NA (6.2 and 4.0%, respectively). Significantly higher trypsin and aminopeptidase activity was observed in the presence of algae, which may have influenced the digestion of the diet. Our results demonstrate that red drum larvae may be raised on a microparticulate diet from first feeding without the use of zooplankton.

Postsettlement patterns of habitat use by sciaenid fishes in subtropical seagrass meadows

Spatial and temporal patterns of distribution and abundance were examined for postsettlement sciaenids collected from seagrass meadows in the Aransas Estuary, Texas. Overall, 5443 sciaenid larvae and early juveniles were identified from biweekly epibenthic sled collections taken from August 1994 to August 1995. Eight species were present in seagrass meadows, with five accounting for over 99.9% of sciaenids collected: silver perch (Bairdiella chrysoura), spotted seatrout (Cynoscion nebulosus), spot (Leiostomus xanthurus), Atlantic croaker (Micropogonias undulatus), and red drum (Sciaenops ocellatus). Settlement to seagrass meadows was partitioned temporally with little overlap among the five species. Postsettlers from inshore spawners (B. chrysoura, C. nebulosus, S. ocellatus) inhabited seagrass meadows during the spring and summer, while individuals from offshore spawners (L. xanthurus, M. undulatus) were present in the late fall and winter. Densities ofB. chrysoura, C. nebulosus, S. ocellatus were highest for small individuals (4–8 mm SL) and these taxa remained in seagrass sites through the early juvenile stage. Conversely,L. xanthurus andM. undulatus maintained longer pelagic periods and generally entered seagrass meadows at larger sizes (10–14 mm SL). Moreover, these taxa were only temporary residents of selected seagrass meadows, apparently migrating to alternative habitats shortly after arrival. During peak settlement, mean and maximum densities among species ranged from 0.1 m−2 to 0.8 m−2 and 0.7 m−2 to 23.8 m−2, respectively. Density and mean size of possettlement sciaenids differed significantly between seagrass species (Halodule wrightii, Thalassia testudinum) and among sites within the estuary.

Documenting the settlement history of individual fish larvae using stable isotope ratios: model development and validation

The δ13C and/or δ15N of newly settled fish larvae should change from a planktonic signature to one reflecting estuarine food sources following settlement from pelagic to demersal nursery habitat. An empirical model based on measurements of δ13C and δ15N was used to estimate size at settlement (Lsett) and time since settlement (Tsett) for red drum (Sciaenops ocellatus; Sciaenidae), an estuarine-dependent species whose larvae migrate from coastal areas to shallow seagrass habitat in estuaries. The model relies on (1) the isotopic composition of the larvae prior to a dietary switch and following equilibration to a new food source, (2) growth rate measurements and (3) estimates of the contribution of metabolic turnover to isotopic change. To test the model, a settlement event was simulated by transferring settlement-size larvae (6 mm standard length; SL) reared in the laboratory to cages deployed in nursery habitat in the Aransas Estuary, TX, USA. Most of the changes in δ13C and δ15N (ca. 10‰ for both) occurring over the 15-day study period were attributed to growth, although metabolic turnover significantly accelerated the rate of isotopic change. When growth and metabolic turnover were incorporated into the model, accurate estimates of Lsett (within 1 mm SL) and Tsett (within 1–2 days) were generated. The isotopic composition of wild-caught pre- and post-settlement larvae collected during a 2-week period was also characterized. Variability in the δ13C and δ15N of potential settlers as a function of size and sampling location was small (<1‰), although there were temporal differences. For part of the study period, there was a distinct difference between the δ13C of newly settled larvae (−19.3‰) and “large” individuals that had equilibrated to estuarine foods (−16.5‰). Nitrogen could not be used as a tracer of settlement due to the absence of differences in δ15N of pre- and post-settlement larvae. The carbon data was used to estimate size and time since settlement. The most abundant Lsett for wild-caught fish was 5–6 mm SL. Using published growth rates for post-settlement red drum in the study area, it was estimated that settlement events occurred over several consecutive days. The estimates of size and time since settlement were sensitive to the effect of metabolic turnover, the isotopic composition of pre-settlement red drum and growth rate. Utilization of stable isotope ratios allows for the finer temporal resolution of the settlement dynamics of marine fish species.

Distribution and habitat associations of billfish and swordfish larvae across mesoscale features in the Gulf of Mexico.

Ichthyoplankton surveys were conducted in surface waters of the northern Gulf of Mexico (NGoM) over a three-year period (2006–2008) to determine the relative value of this region as early life habitat of sailfish (Istiophorus platypterus), blue marlin (Makaira nigricans), white marlin (Kajikia albida), and swordfish (Xiphias gladius). Sailfish were the dominant billfish collected in summer surveys, and larvae were present at 37.5% of the stations sampled. Blue marlin and white marlin larvae were present at 25.0% and 4.6% of the stations sampled, respectively, while swordfish occurred at 17.2% of the stations. Areas of peak production were detected and maximum density estimates for sailfish (22.09 larvae 1000 m−2) were significantly higher than the three other species: blue marlin (9.62 larvae 1000 m−2), white marlin (5.44 larvae 1000 m−2), and swordfish (4.67 larvae 1000 m−2). The distribution and abundance of billfish and swordfish larvae varied spatially and temporally, and several environmental variables (sea surface temperature, salinity, sea surface height, distance to the Loop Current, current velocity, water depth, and Sargassum biomass) were deemed to be influential variables in generalized additive models (GAMs). Mesoscale features in the NGoM affected the distribution and abundance of billfish and swordfish larvae, with densities typically higher in frontal zones or areas proximal to the Loop Current. Habitat suitability of all four species was strongly linked to physicochemical attributes of the water masses they inhabited, and observed abundance was higher in slope waters with lower sea surface temperature and higher salinity. Our results highlight the value of the NGoM as early life habitat of billfishes and swordfish, and represent valuable baseline data for evaluating anthropogenic effects (i.e., Deepwater Horizon oil spill) on the Atlantic billfish and swordfish populations.

The distribution of an AVT V1a receptor in the brain of a sex changing fish, Epinephelus adscensionis.

The present study describes the distribution of an arginine vasotocin (AVT) V1a receptor (AVTr) throughout the brain of a sex-changing grouper, rock hind Epinephelus adscensionis. The objectives of this study were to describe the AVTr distribution in the brain of rock hind for potential linkages of the AVT hormone system with sex-specific behaviors observed in this species and to examine sex-specific differences that might exist. An antibody was designed for rock hind AVTr against the deduced amino acid sequence for the third intracellular loop. Protein expression, identified with immunohistochemistry showed high concordance with mRNA expression, identified with in situ hybridization. AVTr protein and mRNA expression was widely distributed throughout the brain, indicating that AVT may act as a neuromodulator via this V1a receptor subtype. AVTr protein and mRNA were present in regions associated with behavior, reproduction and spatial learning, as well as sensory functions such as vision, olfaction and lateral line sensory processing. We observed high AVTr expression in granular cell formations in the internal cellular layer of olfactory bulbs, torus longitudinalis, granular layer of the corpus cerebellum, valvula of the cerebellum, nuclei of the lateral and posterior recesses, and granular eminence. High protein and mRNA expression was also observed in the preoptic area, anterior hypothalamus, and habenular nucleus. No obvious sex differences were noted in any region of the rock hind brain.

Behavior, Color Change and Time for Sexual Inversion in the Protogynous Grouper (Epinephelus adscensionis)

Hermaphroditism, associated with territoriality and dominance behavior, is common in the marine environment. While male sex-specific coloration patterns have been documented in groupers, particularly during the spawning season, few data regarding social structure and the context for these color displays are available. In the present study, we define the social structure and male typical behavior of rock hind (Epinephelus adscensionis) in the wild. In addition, we detail the captive conditions and time period necessary to induce the onset of the sex-specific coloration and sexual change. At six oil production platform locations in the Gulf of Mexico, rock hind social group size and typical male rock hind social behavior were documented. We observed a rapid temporary color display in rock hind that could be turned on and off within three seconds and was used for confronting territory intruders and displays of aggression towards females. The male-specific “tuxedo” pattern consists of a bright yellow tail, a body with alternating dark brown and white patches and a dark bar extending from the upper mandible to the operculum. Identification and size ranges of male, female and intersex fish collected from oil platforms were determined in conjunction with gonadal histology. Rock hind social order is haremic with one dominant male defending a territory and a linear dominance hierarchy among individuals. In five captive experiments, the largest remaining female rock hind displayed the male specific color pattern within 32d after dominant male removal from the social group. To our knowledge, this is the first evidence in a grouper species of color patterning used to display territoriality and dominance outside of spawning aggregations. The behavioral paradigm described here is a key advance that will enable mechanistic studies of this complex sex change process.

Biological Filter for Conical Tanks

Abstract An inexpensive biological filter was developed to fit 150-L conical tanks used for larval fish feeding and salinity studies. The filter has maintained acceptable ammonia levels over long periods of time, reducing stress on the larvae caused by the water exchanges previously needed to reduce ammonia levels.

Effects of variable ration levels on direct and indirect measures of growth in juvenile red drum (Sciaenops ocellatus)

Relationships between somatic growth (length and weight) and two indirect measures of growth (otolith growth, RNA/DNA ratio) were assessed for red drum (Sciaenops ocellatus) under different feeding rations [0%, 2.5%, 5%, 10%, 15%, and 20% body weight (BW)/day] for 30 days. Representative samples from each ration level were taken in 10-day intervals between Day 0 and Day 30 for evaluation of direct and indirect growth measures. Positive correlations were observed between ration levels, somatic growth, and otolith growth. Statistical differences in weight and length of red drum were observed among ration levels by Days 10 and 20, respectively. Statistical differences for measures of otolith growth among ration levels were evident by Days 20 and 30. In addition, RNA/DNA ratios showed clear separation between fish that were starved and fish that were fed but demonstrated minimal separation among ration levels. Overall, the combination of a measure of somatic growth (weight) and a measure of otolith growth (otolith weight) resulted in the most statistical separation among ration levels. Findings from this study suggest that somatic growth, otolith growth and RNA/DNA ratios are suitable measures of relative growth of red drum; however, due to differences in sensitivity, caution must be exercised when using indirect growth (otolith growth, RNA/DNA ratios) measures to estimate recent growth.

The onset of cortisol synthesis and the stress response is independent of changes in CYP11B or CYP21 mRNA levels in larval red drum (Sciaenops ocellatus).

Although cortisol plays an important role in teleost development, the onset of cortisol production and the cortisol stress response in teleosts remain poorly understood. Here we have reported basal cortisol levels and the development of the cortisol stress response in larval red drum (Sciaenops ocellatus). We isolated partial nucleic acid sequences encoding two key corticosteroidogenic enzymes, CYP11B and CYP21 and assessed ontogenetic patterns of their mRNA levels relative to basal and stress-induced cortisol production. Basal cortisol was first detected 3 days post-hatch (DPH) and reached a maximum at 9 DPH. Cortisol did not increase in response to an acute stressor prior to 6 DPH. From 6 DPH forward, stress caused significant increases in larval cortisol content. Stress-induced cortisol levels in 6-9 DPH larvae were highest 1h post-stress. In larvae 11 DPH and older, the highest cortisol measurements occurred 0.5h post-stress. Elevated cortisol was still evident after 3h in 6 DPH larvae. From 11 DPH onward, basal cortisol levels were reestablished in larvae by 1h post-stress. CYP11B and CYP21 transcripts were detected in red drum 12h prior to hatching and in all post-hatch larvae examined. Changes in CYP11B and CYP21 mRNA levels did not occur in association with the ontogenetic appearance of cortisol, or the onset of the stress response. As larvae developed, the dynamics of the cortisol stress response matured from a low magnitude, slow recovery response, to a response similar to that observed in juvenile and adult fish.

Arginine vasotocin V1a2 receptor and GnRH-I co-localize in preoptic neurons of the sex changing grouper, Epinephelus adscensionis

The arginine vasotocin/vasopressin (AVT/AVP) and gonadotropin releasing hormone (GnRH) systems are known to control sexual behaviors and reproduction, respectively, in different vertebrate groups. However, a direct functional connection between these two neuroendocrine systems has not been demonstrated for any vertebrate species. Therefore, the objective of this research was to test the hypothesis that AVT acts on the GnRH system via an AVT V1a receptor in a sex changing grouper species, the rock hind, Epinephelus adscensionis. AVT V1a2 receptors were co-localized with GnRH-I on neurons in the preoptic anterior hypothalamus identifying a structural linkage between the AVT system and GnRH-I. Transcripts for avt, gnrh-I, and two AVT receptor subtypes (v1a1 and v1a2) were isolated and characterized for E. adscensionis and their expression was measured in males and females by q-RT-PCR. Translation of V1a-type cDNA sequences revealed two distinct forms of the AVT V1a receptor in E. adscensionis brain similar to those reported for other species. The observation of significantly higher gnrh-I mRNA in the POA+H of rock hind males as compared to females suggests differential regulation of the gnrh-I transcripts in the two sexes of this protogynous species. In male E. adscensionis, but not in females, a negative relationship was seen between plasma 11-ketotestosterone (11-KT) and the v1a1 receptor mRNA levels in the POA+H, while a positive trend was observed between 11-KT and v1a2 receptor mRNA levels, indicating that these receptor forms may be differentially regulated.