Utilizing Magnetic Resonance Imaging (MRI) to Assess the Effects of Angling-Induced Barotrauma on Rockfish (Sebastes)
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1245 RAPID COMMUNICATION / COMMUNICATION RAPIDE Utilizing magnetic resonance imaging (MRI) to assess the effects of angling-induced barotrauma on rockfish (Sebastes) Bonnie L. Rogers, Christopher G. Lowe, Esteban Ferna´ ndez-Juricic, and Lawrence R. Frank Abstract: The physical consequences of barotrauma on the economically important rockfish (Sebastes) were evaluated with a novel method using T2-weighted magnetic resonance imaging (MRI) in combination with image segmentation and analysis. For this pilot study, two fishes were captured on hook-and-line from 100 m, euthanized, and scanned in a 3 Tesla human MRI scanner. Analyses were made on each fish, one exhibiting swim bladder overinflation and exophthalmia and the other showing low to moderate swim bladder overinflation. Air space volumes in the body were quantified using image segmentation techniques that allow definition of individual anatomical regions in the three-dimensional MRIs. The individ- ual exhibiting the most severe signs of barotrauma revealed the first observation of a gas-filled orbital space behind the eyes, which was not observable by gross dissection. Severe exophthalmia resulted in extreme stretching of the optic nerves, which was clearly validated with dissections and not seen in the other individual. Expanding gas from swim blad- der overinflation must leak from the swim bladder, rupture the peritoneum, and enter the cranium. This MRI method of evaluating rockfish following rapid decompression is useful for quantifying the magnitude of internal barotrauma associ- ated with decompression and complementing studies on the effects of capture and discard mortality of rockfishes. Re´sume´ : Nous avons e´value´ les conse´quences physiques des barotraumatismes sur les se´bastes du genre Sebastes, des poissons d’importance e´conomique, a` l’aide d’une me´thode ine´dite utilisant l’imagerie par re´sonance magne´tique (MRI) ponde´re´eenT2 en combinaison avec la segmentation et l’analyse des images. Dans cette e´tude pilote, nous avons capture´ deux poissons a` la ligne a` une profondeur de 100 m et les avons euthanasie´s et analyse´s dans un scanner a` 3 teslas pour humains. Les analyses faites chez les deux poissons montrent, chez l’un, une inflation excessive de la vessie natatoire et une exophtalmie et, chez l’autre, une inflation excessive faible a` mode´re´e de la vessie natatoire. Nous avons mesure´ les volumes des espaces ae´riens dans le corps par des techniques de segmentation des images qui permettent une de´finition des diverses re´gions anatomiques en MRI 3D. L’individu qui pre´sente les symptoˆmes les plus importants de barotrauma- tisme posse`de un espace orbital rempli de gaz derrie`re les yeux qui n’est pas visible a` la dissection grossie`re. L’exoph- talmie se´ve`re cause un e´tirement extro˜me des nerfs optiques qui se ve´rifie clairement a` la dissection et qui n’est pas visible chez l’autre individu. Le gaz en expansion provenant de l’inflation excessive de la vessie natatoire doit s’e´chapper de la vessie, fendre le pe´ritoine et s’infiltrer dans le craˆne. Notre me´thode d’e´valuation des se´bastes par MRI apre`s une de´- compression rapide est utile pour mesurer l’importance du barotraumatisme interne associe´ a` la de´compression et elle sert de comple´ment aux e´tudes des effets de la capture et de la mortalite´ relie´ea` la remise a` l’eau chez les se´bastes. [Traduit par la Re´daction] groundfish. Many rockfish populations are declining as a Introduction result of high discard mortality and overexploitation (Davis 2002; Parker et al. 2006). High discard mortality of In the eastern Pacific, rockfish (Sebastes spp.) comprise physoclistic fishes has been attributed to barotrauma on the one of the most important and economically valuable fish resulting in swim bladder overinflation and rupture, Received 18 December 2007. Accepted 22 April 2008. Published on the NRC Research Press Web site at cjfas.nrc.ca on 15 May 2008. J20329 B.L. Rogers,1 C.G. Lowe, and E. Ferna´ndez-Juricic. Department of Biological Sciences, California State University Long Beach, 1250 Bellflower Boulevard, Long Beach, CA 90840, USA. L.R. Frank. Center for Scientific Computation in Imaging and Center for Functional MRI, University of California San Diego, La Jolla, CA 92037, USA. 1Corresponding author (e-mail: [email protected]). Can. J. Fish. Aquat. Sci. 65: 1245–1249 (2008) doi:10.1139/F08-102 # 2008 NRC Canada 1246 Can. J. Fish. Aquat. Sci. Vol. 65, 2008 exophthalmia (eye protrusion), and prolapsed stomach and swim bladder evident by an extremely swollen abdomen. (or) cloacae (Wilson and Burns 1996; Nichol and Chilton The greenblotched rockfish (fish with no visible trauma 2006; Hannah and Matteson 2007). The number and degree (fish NVT); 22 cm SL) showed little external signs of baro- of visible trauma symptoms in rockfish (Sebastes spp.) are trauma, no visible eye trauma, and an overinflated swim highly variable among individuals and species and not a bladder visible by a moderately swollen abdomen, although good predictor of survival (Jarvis and Lowe 2008). Knowl- this fish was also rapidly decompressed. Four hours after edge of internal damage associated with barotrauma is being caught, both fish were euthanized with an overdose limited, and more empirical evidence quantifying morpho- of tricaine methanesulfonate (MS 222) and placed one at a logical and physical damage is needed. Thus far, methods time inside a magnetic resonance imaging (MRI) unit for quantifying damage in rockfish have been limited purely to whole-body scanning. Photographs taken of the fish just visual inspection. after capture and again after MRI imagining indicated little In red snapper (Lutjanus campechnus), two-dimensional change in physical appearance. X rays revealed a pattern of increasing organ displacement Images were acquired on a 3 Tesla (3T) GE SIGNA clin- following decompression (Rummer and Bennett 2005), but ical imaging system using an eight-channel head array coil volumes were estimated by measuring organ dimensions, (MRI Devices, Waukesha, Wisconsin). Two data image sets which is subject to error. Magnetic resonance imaging were acquired for each specimen, a T1-weighted sequence (MRI), on the other hand, provides a fully three-dimensional and a T2-weighted sequence. Total imaging time was 2 h –1 (3D) digital image from which any fish organ dimension can for both fish (30 minÁscan ). The T1-weighted sequence of be measured accurately (in cubic millimetres). Although a fast RF-spoiled gradient recalled echo (FSPGR) produces X rays create contrast based on bone density, MRI has an high contrast between different tissue types, whereas the T2- advantage of visualizing and depicting soft tissues based weighted sequence was a steady-state free precession (SSFP) on water content. For example, T1-weighted MRI scanning method that produces high contrast between tissues and on the shortfin mako shark (Isurus oxyrinchus) and the fluids. The T2-weighted images produced better contrast salmon shark (Lamna ditropis) allowed quantification of near the eye regions and were the only scans used for the red muscle that provided precise and unique detail about image analyses in this study. the position, volume, and types of red muscle (Perry et al. Three-dimensional image segmentation and subsequent 2007). MRI methodology can reveal internal tissue and volume measurement of anatomical regions were performed organ placement, and displacement, immediately following using ITK-SNAP v.1.4.1 (Insight Segmentation and Regis- trauma and specifically display which areas are composed tration Toolkit) (www.itksnap.org/). Segmentation was per- of tissue or gas. formed in a semi-automated fashion whereby regions of The goals of this study using MRI were (i) to characterize interest were determined by an expert user who defined the morphological changes of the internal organs and ocular- points for the full 3D segmentation. A region-growing algo- orbital regions in rapidly decompressed rockfish using MRI rithm was then used to fill out the full organ volume com- techniques and (ii) to determine whether this technique posed of individual known volume units called voxels. MRI could be used as a diagnostic tool for assessing and charac- image segmentation is always limited by a well-known terizing internal injury resulting from fishing-related baro- effect called ‘‘partial voluming’’ that reduces the contrast at trauma. Because of the high cost of MRI operations, only tissue boundaries because multiple tissue types inhabit indi- two rockfish were surveyed as part of this ‘‘proof of con- vidual voxels. cept’’ study. We quantified the relative magnitude of differ- Both fish were dissected 2 h after scanning to reveal the ences between a fish exhibiting many signs of internal organ orientation and the orbital eye regions. All external barotrauma and one showing few signs. signs of barotrauma remained virtually unchanged from the time of capture to dissection, probably because fish were Materials and methods euthanized with MS-222 and placed in an ice bath during transport to prevent any further major physiological proc- Two rockfish, one honeycomb (Sebastes umbrosus) and esses from occurring after capture. The swim bladder and one greenblotched (Sebastes rosenblatti), were caught using internal organs were made visible by cutting away a section hook-and-line off the coast of Long Beach, California (Pot- of skin and muscle on the left side of each fish (Fig. 1). The ter’s Reef) from a depth of 100 m at seafloor, euthanized, eye orbits and optic nerves were made visible by cutting and transported in an ice-bath slurry to the MRI facility. away the frontal cranial bone and clearing away all sur- The two species have very similar body morphologies, be- rounding tissue. Optic nerve lengths were measured using long to overlapping and homologous ecotypes, and inhabit digital calipers.