GPMB September 2009 Colloquium:
Abstracts of Poster Presentations
THE EFFECT OF SHELL HASH ON THE BURROWING RATE AND ABUNDANCE OF DONAX VARIABILIS
Bricker, T. (GPMB, College of Charleston) and Bergquist, D. (SCDNR, College of Charleston)
Beach nourishment projects have the potential to change the physical properties such as grain size and sediment composition of the affected shoreface. These changes to the sediment can affect the ability of infauna to penetrate the sediment. Infauna of the swash zone, such as Donax variabilis, have adapted burrowing behavior to avoid predation and exclusion by waves. Grain size has been shown to affect the burrowing rates of Donax species, but the effects of shell content remain unknown. This project examines the change in burrowing rate of D. variabilis in response to changes in shell content across a range of grain sizes. Laboratory and field trials will be carried out in a full factorial design; testing four different grain sizes (very coarse, coarse, medium, fine) against four different percentages of shell (0, 25, 50, 100%). Beaker experiments containing a known grain size of sieved play sand and percentage of crushed shell will show how burrowing rates change with varying shell content and grain size. Similar experiments will be carried out at Folly Beach, SC. In addition to burrowing rate, abundance of D. variabilis in varying shell contentwill also be examined through historical data mining of past nourishment projects and through sampling of small test plots of the various grain and shell sizes on the beach. The beach ecosystem is ecologically and economically important. Studies on the impacts of nourishment on infauna can be used to evaluate the sustainability of current nourishment practices.
DIAMONDBACK TERRAPINS (MALACLEMYS TERRAPIN) OF THE ASHLEY RIVER: ESTIMATED POPULATION SIZE, SEX RATIO, AND DISTRIBUTION
Broyles, E. (GPMB, College of Charleston), Owens, D. (College of Charleston), Roumillat, W. (SCDNR) and Levesque, E. (SCDNR)
We examined the structure of the diamondback terrapin (Malaclemys terrapin centrata) populations in the Ashley River near Charleston, South Carolina. This is an area of high boat use including the trapping of blue crabs, sport fishing, recreational boating, and industrial shipping. Little is known about current terrapin population status or distribution in this heavily used estuarine area. In conjunction with an extensive trammel netting study of estuarine fishes, mark and recapture methods were used to estimate terrapin population parameters. Terrapins were marked using passive integrated transponder (PIT) tags during 17 sampling days over 15 months that resulted in 732 terrapins tagged and 50 recaptures. Mark and recapture data were statistically analyzed using the program MARK to determine population estimates, and sex ratios were determined to be significantly male biased at 1:1.71 (p<.0001). The distribution of terrapins over a thirteen year period (1995-2008) was analyzed using the SCDNR trammel netting data in which terrapins were counted as a bycatch species. Catch per unit effort of terrapins at each of the 26 sampling sites along the Ashley have been recorded since 1995. Movements of localized terrapin densities were evaluated using ArcGIS. This information on population size, sex ratios, and distributional changes will be used to determine if regulation on habitat degradation, as well as possible protection, is needed for this species.
A COEVOLUTIONARY ARMS-RACE BETWEEN CORAL REEF HERBIVORES AND THEIR SEAWEED PREY: ARE TROPICAL URCHINS MORE RESISTANT TO LIPOPHILIC SECONDARY METABOLITES THAN TEMPERATE URCHINS?
Craft, J. (GPMB, College of Charleston), Paul, V. (Smithsonian Marine Station) and Sotka, E. (College of Charleston)
In contrast to temperate seaweeds, tropical seaweeds produce a greater diversity and higher concentrations of lipophilic secondary metabolites. This geographic variation in plant chemical defenses is likely due to a higher intensity of herbivory in tropical regions relative to temperate regions, a greater feeding tolerance by tropical herbivores relative to temperate herbivores, or both. However, few studies have tested the notion that tropical marine herbivores evolved greater feeding tolerance for tropical plant defenses. Here, we test this coevolutionary prediction by assessing feeding tolerance for ecologically-important urchins from tropical versus temperate regions. Lipophilic extracts were prepared from nine species of taxonomically diverse, chemically-rich tropical seaweed. In a series of pairwise feeding choice assays, extract-coated and control artificial-foods were offered to four tropical (Arbacia punctulata, Diadema antillarum, Echinometra lucunter, E. viridis) and two temperate (Strongylocentrotus droebachiensis and S. purpuratus) echinoid species. If there is a coevolutionary arms-race between tropical algae and these urchins, then tropical urchins should feed on extract-coated algae more readily than temperate urchins. Tropical urchins consumed greater amounts of extract-laden foods than did their temperate counterparts (ANOVA p-value < 0.001), suggesting they are more resistant to lipophilic chemical defenses produced by tropical seaweeds. This study provides support for the coevolutionary arms-race among coral reef herbivores and their seaweed prey and will help guide examinations into the biochemical adaptations urchins utilize to tolerate secondary metabolites of tropical seaweeds.
A QUANTITATIVE ASSESSMENT OF BOTTLENOSE DOLPHIN, TURSIOPS TRUNCATUS, INTERACTIONS WITH CRAB POTS AS MEANS OF UNDERSTANDING METHODS OF ENTANGLEMENT WITH FISHING GEAR
Duquette, A. (GPMB, College of Charleston), Kracker, L. (NOAA/NOS), McFee, W. (NOAA/NOS, Coastal Marine Mammal Stranding Assessments Project), Powell, J. (NOAA/NOS, Coastal Marine Mammal Stranding Assessments Project), Roth, S. (NOAA/NOS)
Entanglement in commercial fishery equipment is the number one anthropogenic cause of marine mammal death worldwide. Scientists turned to acoustic methods for monitoring and deterring marine mammals from fishing gear. However, effective avoidance solutions have not been discovered. One set back is the lack of knowledge about the methods of entanglement. Until those methods are understood, sufficient management is near impossible. The Atlantic blue crab fishery is the number one cause of fishery related incidents for bottlenose dolphins (Tursiops truncatus) in South Carolina. It is hypothesized that the dolphins are attracted to the pots by the bait and possibly the other marine life drawn to the pots. Hydrophones deployed in baited and unbaited crab pots will record dolphin sounds and allow for a quantification of the interaction. If dolphins are attracted by the bait, then they will approach and spend more time around the baited pots than the unbaited pots. Visual land observations will ground truth the acoustic data and provide insight into the behaviors bottlenose dolphins exhibit around crab pots. Any significant differences between time spent around baited pots versus unbaited pots will be determined using an ANOVA test in R statistical software. External factors that may be affecting dolphin behavior such as tide or time of day will be tested for significance via ANCOVA tests in R. Results of this research will provide more information on the dolphin behaviors and time spent at or around crab pots. With this new information, more effective avoidance measures can prevent entanglement.
RECRUITMENT OF STOCKED JUVENILE RED DRUM TO THE ADULT POPULATION IN SOUTH CAROLINA
Gerhard, J. (GPMB, College of Charleston) and Denson, M. (SCDNR)
Red drum (Sciaenops ocellatus) is one of the most significant recreational fishery species along the southeast U.S. and Gulf of Mexico coasts. Red drum stock enhancement research has been conducted in Texas, Florida, and South Carolina, and the contribution of stocked fish to the wild population in South Carolina has been measured immediately following stocking through age two. Contribution of stocked fish recruitment to the adult population has not yet been examined and was the focus of this research. Small juveniles (~ 30 mm TL) were stocked in the Ashley and Wando Rivers of Charleston Harbor in 1999 - 2002. Five years after initial stocking (2005 - 2008), adults were sampled offshore of four major South Carolina estuaries via targeted and random stratified longline sampling, and tissue samples were collected for genetic analysis to determine if stocked fish contribute to the adult population. Microsatellite-deduced genotypes of sampled fish were compared to those of broodstock to determine that some small stocked juveniles become part of the adult stock of South Carolina. The contribution of these stocked fish was calculated via several different methods. The overall contribution statewide was calculated, as well as that from each sampled estuary and each year of sampling. Age and length information gathered from 102 sacrificed adults further narrowed down these contributions by evaluating the contribution to specific age classes when fish were stocked. The overall percentage of the adult population identified as stocked fish from three stocked year classes ranged from 2.61% to 54.12%.
TRANSGENERATIONAL PLASTICITY AND HABITAT HETEROGENEITY IN MIMULUS GUTTATUS: AN ANALYSIS OF MATERNAL AND PATERNAL EFFECTS IN ASSOCIATION WITH EDAPHIC VARIATION
Henson, C. (MES, College of Charleston)
Habitat heterogeneity occurs at different spatial scales and affects landscapes, within and among populations; the ability to persist in the face of a variable landscape is of particular importance to sessile organisms. Phenotypic plasticity, environmentally dependent phenotypic expression, is one of the major mechanisms which allows for response to microhabitat heterogeneity in plant systems (Bradshaw 1965; Schlichting 1986; Schlichting and Pigliucci 1998). In this study, I am assessing whether transgenerational plasticity, a subset of phenotypic plasticity, in the form of maternal and paternal effects impacts four populations of Mimulus guttatus in their response to variable edaphic conditions. Three generations are being utilized in this experiment to establish a total of eight different edaphic pathways across the three generations. A full sibling crossing design was utilized to produce 28 maternal lines which were grown in both serpentine and nonserpentine soil environments. Dams from each line in each environment were crossed with sires from both environments to allow for the assessment of maternal and paternal effects in the offspring generation. Currently, the first two populations have been completed, and the third generation will be planted in the fall of 2009.
CHARACTERIZATION OF THE BACTERIAL PROPERTIES THAT IMPAIR RESPIRATION IN THE ATLANTIC BLUE CRAB, CALLINECTES SAPIDUS
Johnson, N. (GPMB, College of Charleston), Burnett, L. (College of Charleston) and Burnett, K. (College of Charleston)
In the blue crab Callinectes sapidus, injection with the bacterial pathogen Vibrio campbellii causes a decrease in oxygen consumption. Histological and physiological evidence suggests that the physical obstruction of hemolymph flow through the gills, caused by aggregations of bacteria and hemocytes in the gill vasculature, may underlie the decrease in aerobic function associated with bacterial infection. Alternatively, the observed decline in oxygen uptake could be a reflection of hormonal changes known to be induced by bacterial lipopolysaccharide (LPS), a major constituent of the Gram-negative bacterial cell wall. To determine whether generic particles of various sizes and surface charges are sufficient to stimulate immune and metabolic responses in C. sapidus, the effects of different inert microspheres on circulating hemocytes, hemolymph glucose, and hemolymph lactate were evaluated. None of the microspheres resulted in the dramatic, prolonged (up to 4 h) decrease in circulating hemocytes seen with injection of V. campbellii. Hemolymph glucose and lactate levels did not change in response to injected bacteria or microspheres. In future studies, C. sapidus will be injected with a variety of other bacteria and purified LPS to further elucidate the microbial elements necessary and/or sufficient to elicit changes in circulating hemocytes, hemolymph glucose, hemolymph lactate, and oxygen uptake. In exploring the range of particles capable of inducing changes in these variables, we hope to gain further understanding of the mechanisms that are responsible for inducing this decrease in aerobic metabolism, and reveal the extent to which this phenomenon may affect naturally infected crabs.
NEKTON USE OF AN INTERTIDAL RESTORED OYSTER REEF
Joyce, R. (GPMB, College of Charleston)
Over the past few decades, oyster reef restoration has become increasingly popular. The majority of restoration projects have focused on fisheries enhancement of the eastern oyster, Crassostrea virginica. However, within the past few years, restoration projects have begun to include ecosystem health as well. Currently, little is known about how the nekton community changes through time on newly formed restored oyster reefs. Also, since many of the reefs are intertidal, how nekton utilize these habitats on successive tides is not well understood. This project aims to answer two main questions. First, how will the community structure change through time on a newly restored oyster reef? Second, do organisms show site fidelity, or do they constantly move between sites? These questions will be tested by sampling on restored oyster reefs that were created by placing cement oyster “castles” in a row even with the mean low water line in a South Carolina estuary during the summer. Sampling using stop nets will begin the following spring and continue through late fall. These stop nets will be set during high tide. After most of the water has receded, nekton will be collected, identified, counted, and released. Some species will be tagged using a colored elastomer to determine site fidelity. It is expected that there will be greater species diversity and abundance on the restored reefs compared to sand or mud bottom habitat. Also, site fidelity may be present for some species, while others may be opportunistic or transient.
BASELINE DATA ON VITALITY, SUBSTRATE COMPOSITION, AND FISH DIVERSITY ON SAGHARUGHOMBE REEF, SOLOMON ISLANDS BETWEEN 2000 AND 2008
Olds, M.K. (College of Charleston), Dustan, P. (College of Charleston) and Alling, A. (Planetary Coral Reef Foundation)
Coral reef ecosystems have persisted on Earth for millions of years through mass extinctions and ice ages. They are considered to be among the most valuable planetary resources providing millions of people with direct and indirect environmental and economic services. In recent decades live coral coverage and coral vitality has significantly declined. Each reef faces a unique suite of threats depending on several environmental and anthropogenic factors. Understanding present threats depends on the collection of concrete baseline data of substrate composition, coral vitality, and reef fish diversity, enabling observations in changes over time. This study is an analysis of a long-term monitoring project that took place on Sagharughombe Reef, near Ghizo Island in the Western Province of the Solomon Islands (S 08.117° E 156.912°). This reef was visited in 2000, 2002, 2006, and 2008. This timeline includes the severe bleaching event of the great barrier reef in 2002 and the 01 April 2007 magnitude 8.1 earthquake that occurred 45 kilometers SSE from the town of Gizo, Solomon Islands (S 8.418° E 156.978°) and resulted in a series of tsunami waves. The study aims to document the state of Sagharughombe Reef between 2000 and 2008 and to assess the effects of the earthquake and tsunami.
THE EFFECTS OF A SYNTHETIC PYRETHROID PESTICIDE ON TWO ESTUARINE FISH SPECIES
Parent, L.M. (GPMB, College of Charleston), DeLorenzo, M. (NOAA/CCEHBR), and Fulton, M. (NOAA/CCEHBR)
Pesticides can enter into coastal waters via surface water runoff and aerial drift, posing a risk to non-target aquatic species. Permethrin is a synthetic pyrethroid insecticide used in agricultural, turf grass, commercial, and residential settings to control a wide range of insect pests. Permethrin application in the United States is increasing as it is being used to replace other insecticides such as organophosphates and carbamates. This study examined the effects of permethrin on two species of fish found abundantly in South Carolina estuaries, the mummichog (Fundulus heteroclitus), and the red drum (Sciaenops ocellatus). The 96h median lethal concentration of permethrin was determined to be 22 µg/L for juvenile S. ocellatus and 54 µg/L for adult F. heteroclitus. Sublethal cellular stress effects of permethrin were also assessed. Permethrin exposure did not result in significantly lower liver somatic index values. Lipid peroxidation activity of the liver, however, was significantly higher in permethrin-treated fish compared to control animals after 24h. The effects of permethrin on fish cellular stress enzymes and survival occurred at levels greater than those typically measured in the environment. Based on previous testing, the two fish species were less sensitive to permethrin than invertebrates such as shrimp. This study will further understanding of the effects of permethrin on estuarine organisms and may prove useful to the future management and regulation of pyrethroid insecticide application near estuarine habitats.
USING ANISAKIDS AS MARKERS TO IDENTIFY POTENTIAL PYGMY SPERM (KOGIA BREVICEPS) AND DWARF SPERM WHALE (KOGIA SIMA) STOCKS IN THE WESTERN ATLANTIC AND GULF OF MEXICO
Rein, R. (GPMB, College of Charleston), de Buron, I. (College of Charleston), McFee, W. (NOAA/NOS), Greig, T. (NOAA/NOS) and Arnott, S. (DNR)
Misidentification of pygmy (Kogia breviceps) and dwarf (K. sima) sperm whales is common, and although they are some of the most commonly stranded mammals on US coasts, their population structures are not well understood. It is currently thought that there are two stocks of K. breviceps and one of K. sima, however, genetic studies currently being undertaken in another laboratory are not yet conclusive. Parasites have been used to determine stock structure of their hosts species, and we will be using this approach to complement the dwarf and pygmy sperm whale genetic studies. Common parasites found in these whales include anisakids, a type of nematode, which have been successfully used as a biological tag. Anisakid specimens will be obtained from the stomachs of freshly stranded pygmy and dwarf sperm whales from the Gulf of Mexico and the northern Atlantic as well as from laboratory collections. The anterior and posterior ends of the worms will be clipped and fixed in 10% buffered formalin for morphological studies, and the middle part will be frozen at -20°C for molecular studies. Anisakids will be identified at the lowest taxon possible using scanning electron microscopy. Genetic variability of each of the anisakid species’ populations will be determined via gene sequencing and examined for the population structure among host and between regions. Results will be used to examine whether these two sperm whale species host different communities of anisakids and if their populations are composed of separate stocks in the area studied.
STABLE ISOTOPE ANALYSIS OF THE SANDBAR SHARK (CARCHARINUS PLUMBEUS): A NON-INVASIVE METHOD FOR COMPARISON OF DIET AND TROPHIC RELATIONSHIPS BETWEEN GENDERS, LOCATIONS, AND AGE CLASSES
Shiffman, D. (GPMB, College of Charleston)
Sharks, because of their role as apex predators, are ecologically and economically important animals. However, many species of sharks are declining in population at alarming rates. Federal management plans exist for many U.S. shark species, including the sandbar shark, but a management plan is only as good as the data it is based on. A recent National Marine Fisheries Service SEDAR (Southeast Data Assessment and Review) for sandbar sharks called for increased data on diet and trophic dynamics. The traditional method of stomach content analysis involves cutting open captured sharks’ stomachs to analyze the contents. This technique only shows what a shark has eaten recently, and also involves sacrificing the very animals we are trying to conserve. The use of stable isotope analysis (δ13C and δ 15N) can answer the same trophic questions as stomach content analysis, and does not require sacrificing the animal. Sandbar sharks in Virginia and South Carolina waters were captured, measured, tagged, sampled for stable isotope analysis, and released. Stable isotope analysis will be performed to determine the diet and trophic level of sandbar sharks, and to see if there are any changes in the diet or trophic level based on age class, gender, or location. We expect to find ontogenetic and location-based changes, and to contribute to non-invasive scientific sampling for threatened and endangered species.
SYSTEMATIC STUDY OF THE BREGMACEROS SP. 5 COMPLEX
Simpson, R.G. (GPMB, College of Charleston) and Harold, A.S. (College of Charleston)
The gadiform fishes of the genus Bregmaceros are small, mesopelagic planktivores that are commonly caught as bycatch in midwater trawls. Currently, taxonomists recognize 13 valid species, with several more undescribed forms. This study focuses on the more phylogenetically basal group of Bregmaceros, including B. sp.5. This undescribed form incorporates a potential complex of species found throughout the Atlantic, Pacific, and Indian Oceans. Another closely related species pair, B. sp.9 and 10, is distributed in the Indian Ocean. Using a combination of meristic values, multivariate techniques, pigmentation pattern, and various internal/external characters, these undescribed species are diagnosed and described.
AN ANALYSIS OF HABITAT MOVEMENT AND SURVIVAL RATES FOR AMERICAN OYSTERCATCHERS IN SOUTH CAROLINA
Snipes, K. (MES, College of Charleston)
The American Oystercatcher has been identified as an “extremely high priority” shorebird by the working group for the Southeastern Coastal Plain as part of the U.S. Shorebird Conservation Plan. This shorebird is especially important in South Carolina because a large proportion of oystercatchers winter here, approximately 1/3 of the overall population. This state also has one of the largest breeding populations of oystercatchers, second only to Virginia. Understanding habitat use as well as population dynamics are two very important tools in the conservation of a species. The banding and resighting of oystercatchers has been an ongoing research method that conservation agencies have been participating in all along the Atlantic coast. Incorporating banding and resighting data of oystercatchers aids in the calculation of their space use and survival rates. Although overall population estimates have been completed for the eastern race of the American Oystercatcher, individual’s habitat dispersal and movement have not been evaluated. Survival rates for oystercatchers in South Carolina have not been determined at this time either. Calculating oystercatcher survival rates can result in a better estimation of population status and trends. The results from my analysis can assist biologists in understanding this species’ behavior as well as contribute to a more accurate oystercatcher census for South Carolina.
DETERMINATION OF THE AMOUNT OF OYSTER SHELL AVAILABLE FOR RECYCLING ACROSS THE STATE OF SOUTH CAROLINA THROUGH THE USE OF SURVEYS
Stone, B. (MES, College of Charleston)
Oyster reefs create habitat for numerous fish and invertebrate species as well as provide an important commercially harvested renewable resource. Recycled oyster shell can be used to restore and enhance current oyster reefs off the coast of South Carolina. In recent years it has become harder to acquire enough oyster shell for oyster habitat restoration. Most of the shell is believed to end up as landscaping, driveways, or placed in a landfill. Although data exists for oysters harvested commercially within the state, it has been unknown what quantities of oysters are being imported into South Carolina. Although the South Carolina Department of Natural Resources (SCDNR) shell recycling program has achieved an increase in recycled shell, the quantity available still falls far short of the management needs and the state may purchase as much as 60,000 bushels of shell annually. SCDNR is interested in reducing the amount of shell purchased for restoration projects, but currently lacks the information to implement a sound expansion plan. A survey was created and conducted over the phone, by mail, and in person to determine how many oysters were being imported into the state and where the large sources of shell stock might be located across the state. This information was used to determine how much shell should be available for recycling programs. Current calculations reveal that there are about 6,718,600 pounds or 121,977 bushels of Gulf oysters being sold in South Carolina each year. It was also found that more than 50% of all the oysters sold in the state of South Carolina are sold to restaurants where they most likely end up in the trash. Targeting restaurants for their shell would make it possible for oyster restoration projects to continue without the need to purchase shell from out-of-state.
COMMUNITY AND TROPHIC STRUCTURE ANALYSIS OF THE SNAPPER GROUPER COMPLEX OFF THE SOUTHEASTERN U.S. ATLANTIC COAST
Stratton, M. (GPMB, College of Charleston)
Community/trophodynamic metrics have been utilized to detect temporal changes in the community/trophic structure of fish communities in response to environmental and fishing effects. Six community/trophodynamic metrics will be used to characterize changes in the community/trophic structure of the snapper grouper complex, a group of 73 reef-associated fish species managed by the South Atlantic Fishery Management Council: mean length, mean maximum length, mean weight, mean maximum weight, the slope of the biomass spectra, and mean trophic level. Metrics will be calculated from species-size-abundance data compiled during 1990-2009 from an annual fishery-independent sampling program (Marine Resources Monitoring, Assessment and Prediction, MARMAP) using the chevron trap as an abundance index. Stable nitrogen isotope analysis of fish white muscle tissue will be used as an index of mean trophic level (size-structure incorporated). Two isotope methods will be employed: bulk isotope analysis (BIA) and compound-specific isotope analysis (CSIA) of specific amino acids. Both methods provide trophic level estimations of fish specimens, while CSIA can also provide trophic information about basal food web organisms from consumer tissue samples. The accuracy of using CSIA to calculate trophic level in fish specimens will be evaluated in experimental feeding trials to ensure its proper use in the trophodynamic aspect of the proposed study. In addition to characterizing past trends, the ability to use the MARMAP chevron trap survey to detect future trends in the six community/trophodynamic metrics will be investigated. Strong predictive metrics would provide potential decision-making tools for fishery managers and support the implementation of ecosystem-based fishery management.
SPAWNING BY THE SOUTHERN AND GULF FLOUNDERS (PARALICHTHYS LETHOSTIGMA AND P. ALBIGUTTA) OFF THE SOUTH CAROLINA COAST
Tucker, C. R. (GPMB, College of Charleston)
The southern flounder (Paralichthys lethostigma) and the gulf flounder (P. albigutta) support an important recreational and commercial fishery in South Carolina (Wenner and Archambault 2005). Much of the life history for these species has been well documented (e.g. Gilbert 1986; Wenner and Archambault 2005; Schreiber 2006). During fall months as the temperature drops, the adults leave the estuaries to spawn in offshore waters (Gilbert 1986). This study seeks to fill in gaps in the life history of southern flounder and gulf flounder during the winter spawning months. SCUBA dives are made during the spawning season (October – March) in areas flounder are expected to spawn. At each site, the habitat is characterized and specimens are collected for histological gonad analysis and aging. To investigate transport of eggs and larvae by currents after spawning, drift bottles will be released from spawning sites (or potential spawning sites) during the 2009/10 spawning season. Since 2007, 85 flounder have been collected by the South Carolina Department of Natural Resources. In addition, 188 racks consisting of flounder carcasses turned in by recreational divers to the South Carolina Department of Natural Resources Flounder Research Project have also been examined. Histologically determined spawning stage indicates gulf flounder spawn at the same reefs they inhabit year round. Southern flounder spawning sites remain elusive, and sampling will include deeper locations further from shore during the 2009/10 season.