Myxospore density of Kudoa inornata varies significantly within symmetrical white muscle tissue replicates of its fish host, the spotted seatrout, Cynoscion nebulosus.

The spotted seatrout, Cynoscion nebulosus, is a popular game fish in the southeastern USA. It is estimated that nearly 90% of the adult population in South Carolina estuaries are infected in their skeletal muscle by the myxosporean, Kudoa inornata. However, little is known about this parasite's biology, including the distribution and densities of myxospores within tissues of infected fish, which we expect affect the physiology of their hosts. In order to correlate densities with physiological parameters in future studies, we quantified the myxospores density in muscle and characterized the variation among individual fish. Naïve juvenile seatrout was experimentally infected via presumed K. inornata actinospores exposure to raw seawater. A plug of muscle was extracted from two bilaterally symmetrical regions in the epaxial fillet from fresh and frozen carcasses. Variation in density data was calculated both within and among individuals. Within individuals, density counts were compared between left- and right-side biopsies. There was no significant difference between fresh and frozen plugs, and variation among individuals accounted for the greatest proportion of variation at 68.8%, while variation within individuals was substantial at 25.6%. Simulation and correlation tests confirmed that bilaterally symmetrical replicates varied significantly within individuals. When sampled from areas surrounding the initial biopsies, myxospore density estimates were more similar than between sides. Our findings have important implications for sampling design, particularly for studies investigating physiological parameters at the cellular or molecular level in association with parasite infection.

Limited accrual of myxospores of Kudoa inornata (Cnidaria: Myxosporea) in their wild fish hosts, Cynoscion nebulosus (Teleostei: Sciaenidae).

Kudoa inornata is a myxosporean that infects the seatrout Cynoscion nebulosus. Increased prevalence of infection as fish age and absence of inflammation against plasmodia led to the hypothesis that seatrout retain and accumulate myxospores throughout their lives. However, opportunistic observations that wild-caught seatrout cleared infection when maintained in aquaculture conditions and evidence of encapsulated infected necrotic myofibers suggested that fish develop an immunity against this parasite, or that myxospores have a limited life span. To evaluate myxospore clearance and to test putative resistance to re-infection, we examined 44 wild-caught seatrout broodstock maintained in parasite-free water for 2-6 yr. Twenty-five fish served as negative controls (time zero of experiment), and 19 were exposed to water-borne K. inornata infective stages for 18 wk. Over 73% of the exposed fish became infected, compared to ~12% of control fish, indicating that fish were susceptible to re-infection by K. inornata. Whether plasmodia degenerate because K. inornata myxospores have a limited life span or seatrout develop an adaptive immunity against these life stages remains unknown. To test for accumulation of myxospores over time, we compared myxospore densities and intensities between sexes and across ages and sizes of wild seatrout. There was no significant difference in myxospore densities with size, age, or sex. However, intensities increased significantly with increasing fish age and size, indicating accrual of myxospores over time. These results combined with evidence of infection clearance suggest that K. inornata myxospores do not persist but nevertheless accrue in wild seatrout due to continuous contact with infective stages.

Reproductive resilience of an estuarine fish in the eye of a hurricane.

Spatial and temporal patterns of spawning activity are important measures of resilience in fishes that directly link environmental disturbances with reproductive success. We acoustically monitored spawning in spotted seatrout (Cynoscion nebulosus) from April through September 2017 at 15 sites near Port Aransas, Texas, which coincided with the landfall of a category 4 hurricane (Harvey) on 25 August. Spawning sounds were recorded every day of the study across all sites and were also confirmed during the hurricane at two sites located within the eye of the storm. Daily spawning continued after the hurricane, but the onset of spawning shifted 2.12 h earlier for 5 days, after which it returned to the pre-storm schedule. These results illustrate the resilience of seatrout to intense, episodic disturbances and offer insights on the phenotypic plasticity of estuarine fishes to cope with projected increases in environmental variability.

Development of a modified cortisol extraction procedure for intermediately sized fish not amenable to whole-body or plasma extraction methods.

The corticosteroid hormone cortisol is the central mediator of the teleost stress response. Therefore, the accurate quantification of cortisol in teleost fishes is a vital tool for addressing fundamental questions about an animal's physiological response to environmental stressors. Conventional steroid extraction methods using plasma or whole-body homogenates, however, are inefficient within an intermediate size range of fish that are too small for phlebotomy and too large for whole-body steroid extractions. To assess the potential effects of hatchery-induced stress on survival of fingerling hatchery-reared Spotted Seatrout (Cynoscion nebulosus), we developed a novel extraction procedure for measuring cortisol in intermediately sized fish (50-100 mm in length) that are not amenable to standard cortisol extraction methods. By excising a standardized portion of the caudal peduncle, this tissue extraction procedure allows for a small portion of a larger fish to be sampled for cortisol, while minimizing the potential interference from lipids that may be extracted using whole-body homogenization procedures. Assay precision was comparable to published plasma and whole-body extraction procedures, and cortisol quantification over a wide range of sample dilutions displayed parallelism versus assay standards. Intra-assay %CV was 8.54%, and average recovery of spiked samples was 102%. Also, tissue cortisol levels quantified using this method increase 30 min after handling stress and are significantly correlated with blood values. We conclude that this modified cortisol extraction procedure provides an excellent alternative to plasma and whole-body extraction procedures for intermediately sized fish, and will facilitate the efficient assessment of cortisol in a variety of situations ranging from basic laboratory research to industrial and field-based environmental health applications.

Boat noise in an estuarine soundscape - A potential risk on the acoustic communication and reproduction of soniferous fish in the May River, South Carolina.

The impact of boat related noise on marine life is a subject of concern, particularly for fish species that utilize acoustic communication for spawning purposes. The goal of this study was to quantify and examine the risk of boat noise on fish acoustic communication by performing acoustic monitoring of the May River, South Carolina (USA) from February to November 2013 using DSG-Ocean recorders. The number of boats detected increased from the source to the mouth with the highest detections near the Intracoastal Waterway (ICW). Boat noise frequency ranges overlapped with courtship sounds of silver perch (Bairdiella chrysoura), black drum (Pogonias cromis), oyster toadfish (Opsanus tau), spotted seatrout (Cynoscion nebulosus), and red drum (Sciaenops ocellatus). In the May River estuary, red drum may experience the greatest risk of auditory masking because of late afternoon choruses (21% time overlap with boat noise) and only one spawning location near the noisy ICW.

Co-exposure to sunlight enhances the toxicity of naturally weathered Deepwater Horizon oil to early lifestage red drum (Sciaenops ocellatus) and speckled seatrout (Cynoscion nebulosus).

The 2010 Deepwater Horizon oil spill resulted in the accidental release of millions of barrels of crude oil into the Gulf of Mexico. Photo-induced toxicity following co-exposure to ultraviolet (UV) radiation is 1 mechanism by which polycyclic aromatic hydrocarbons (PAHs) from oil spills may exert toxicity. Red drum and speckled seatrout are both important fishery resources in the Gulf of Mexico. They spawn near-shore and produce positively buoyant embryos that hatch into larvae in approximately 24 h. The goal of the present study was to determine whether exposure to UV as natural sunlight enhances the toxicity of crude oil to early lifestage red drum and speckled seatrout. Larval fish were exposed to several dilutions of high-energy water-accommodated fractions (HEWAFs) from 2 different oils collected in the field under chain of custody during the 2010 spill and 3 gradations of natural sunlight in a factorial design. Co-exposure to natural sunlight and oil significantly reduced larval survival compared with exposure to oil alone. Although both species were sensitive at PAH concentrations reported during the Deepwater Horizon spill, speckled seatrout demonstrated a greater sensitivity to photo-induced toxicity than red drum. These data demonstrate that even advanced weathering of slicks does not ameliorate the potential for photo-induced toxicity of oil to these species. Environ Toxicol Chem 2017;36:780-785. © 2016 SETAC.

Acoustic monitoring indicates a correlation between calling and spawning in captive spotted seatrout (Cynoscion nebulosus).

BACKGROUND: Fish sound production is widespread throughout many families. Territorial displays and courtship are the most common reasons for fish sound production. Yet, there is still some questions on how acoustic signaling and reproduction are correlated in many sound-producing species. In the present study, our aim was to determine if a quantitative relationship exists between calling and egg deposition in captive spotted seatrout (Cynoscion nebulosus). This type of data is essential if passive acoustics is to be used to identify spawning aggregations over large spatial scales and monitor reproductive activity over annual and decadal timeframes. METHODS: Acoustic recorders (i.e., DSG-Oceans) were placed in three laboratory tanks to record underwater sound over an entire, simulated reproductive season. We enumerated the number of calls, calculated the received sound pressure level, and counted the number of eggs every morning in each tank. RESULTS: Spotted seatrout produced three distinct call types characterized as "drums," "grunts," and "staccatos." Spotted seatrout calling increased as the light cycle shifted from 13.5 to 14.5 h of light, and the temperature increased to 27.7 °C. Calling decreased once the temperature fell below 27.7 °C, and the light cycle shifted to 12 h of light. These temperature and light patterns followed the natural reproductive season observed in wild spotted seatrout in the Southeast United States. Spotted seatrout exhibited daily rhythms in calling. Acoustic signaling began once the lights turned off, and calling reached maximum activity approximately 3 h later. Eggs were released only on evenings in which spotted seatrout were calling. In all tanks, spotted seatrout were more likely to spawn when male fish called more frequently. A positive relationship between SPL and the number of eggs collected was found in Tanks 1 and 3. DISCUSSION: Our findings indicate that acoustic metrics can predict spawning potential. These findings are important because plankton tows may not accurately reflect spawning locations since egg capture is likely affected by predator activity and water currents. Instead, passive acoustics could be used to monitor spotted seatrout reproduction. Future studies can use this captive study as a model to record the estuarine soundscape precisely over long time periods to better understand how human-made stressors (e.g., climate change, noise pollution, and chemical pollutants) may affect spawning patterns.

Vitrification of Sperm from Marine Fishes: Effect on Motility and Membrane Integrity.

Our goal was to develop a standardized approach for sperm vitrification of marine fishes that can be applied generally in aquatic species. The objectives were to: 1) estimate acute toxicity of cryoprotectants over a range of concentrations; 2) evaluate the properties of vitrification solutions (VS); 3) evaluate different thawing solutions, and 4) evaluate sperm quality after thawing by examination of motility and membrane integrity. Sperm were collected from red snapper (Lutjanus campechanus), spotted seatrout (Cynoscion nebulosus), and red drum (Sciaenops ocellatus). A total of 29 combinations of cryoprotectants were evaluated for toxicity and glass formation. Samples were loaded onto 10-µL polystyrene loops and plunged into liquid nitrogen. There was a significant difference (P < 0.05) in post-thaw motility among VS and among species when using the same VS. The sperm in VS of 15% DMSO + 15% ethylene glycol + 10% glycerol + 1% X-1000™ + 1% Z-1000™ had an average post-thaw motility of 58% and membrane integrity of 19% for spotted seatrout, 38% and 9% for red snapper, and 30% and 19% for red drum. Adaptations by marine fish to high osmotic pressures could explain the survival in the high cryoprotectant concentrations. Vitrification offers an alternative to conventional cryopreservation.