Radiography and computed tomography of the heart and lower respiratory tract in toco toucans (Ramphastos toco).

This study aimed to evaluate normal features of the heart and lower respiratory tract in toco toucans by means of radiography and helical computed tomography (CT) scanner. Fifteen healthy adult toco toucans (Ramphastos toco), 10 females and 5 males, average body mass of 650 g were studied. CT examination as well as right lateral and ventrodorsal radiographic examinations of the coelomic cavity were performed under chemical restraint. Heart, lungs, air sacs, trachea and syrinx were analysed. The mean values of heart length, heart width and thoracic cavity in radiographs were, respectively, 23.76 mm, 25.94 mm and 48.87 mm. In both X-rays and CT scans, the lung parenchyma had honeycomb-like pattern. The topographic areas of the anterior and posterior air sacs were visualized as dark and air-filled spaces in X-rays. On CT evaluation, the air sacs occupied a larger area in the coelomic cavity compared to X-ray. In the lateral radiographic view, the cervical part of the trachea was positioned more ventrally in the transition from cervical to thoracic regions showing a V-shaped appearance. In all CT planes was visible division of the trachea into the right and left main bronchi at the level of 3rd thoracic vertebra. The syrinx was difficult to visualize in X-rays, but on CT it was easily identified in axial slice. In conclusion, the normal features of toco toucan's heart and lower respiratory tract that were determined on X-rays and CT scans are useful to compare with sick toco toucans, as well as other bird species.

Auditory chain reaction: Effects of sound pressure and particle motion on auditory structures in fishes.

Despite the diversity in fish auditory structures, it remains elusive how otolith morphology and swim bladder-inner ear (= otophysic) connections affect otolith motion and inner ear stimulation. A recent study visualized sound-induced otolith motion; but tank acoustics revealed a complex mixture of sound pressure and particle motion. To separate sound pressure and sound-induced particle motion, we constructed a transparent standing wave tube-like tank equipped with an inertial shaker at each end while using X-ray phase contrast imaging. Driving the shakers in phase resulted in maximised sound pressure at the tank centre, whereas particle motion was maximised when shakers were driven out of phase (180°). We studied the effects of two types of otophysic connections-i.e. the Weberian apparatus (Carassius auratus) and anterior swim bladder extensions contacting the inner ears (Etroplus canarensis)-on otolith motion when fish were subjected to a 200 Hz stimulus. Saccular otolith motion was more pronounced when the swim bladder walls oscillated under the maximised sound pressure condition. The otolith motion patterns mainly matched the orientation patterns of ciliary bundles on the sensory epithelia. Our setup enabled the characterization of the interplay between the auditory structures and provided first experimental evidence of how different types of otophysic connections affect otolith motion.

Sound Generating Structures of the Humpback Dolphin Sousa plumbea (Cuvier, 1829) and the Directionality in Dolphin Sounds.

The macroscopic morphology of structures involved in sound generation in the Indian Ocean humpback dolphin (Sousa plumbea) were described for the first time using computed tomography imaging and standard gross dissection techniques. The Indian Ocean humpback dolphin may represent a useful comparative model to the bottlenose dolphin (Tursiops sp.) to provide insights into the functional anatomy of the sound production in dolphins, since these coastal dolphins exhibit similar body size and share similarities on acoustic behavior. The general arrangement of sound generating structures, that is, air sacs and muscles, was similar in both the bottlenose dolphin and the Indian Ocean humpback dolphin. The main difference between the two species existed in a small left posterior branch of the melon in the Indian Ocean humpback dolphin, which was not found in the bottlenose dolphin and might reflect an adaptation of directionality for high frequency communication sounds as seen in some other delphinids (e.g., Lagenorhynchus sp., Grampus griseus). Thus, this may be the main reason for the asymmetry of the sound production structures in dolphins. Additionally, the longer rostrum in Indian Ocean humpback dolphins might suggest a more directional echolocation beam compared to the Lahille's bottlenose dolphin. Anat Rec, 302:849-860, 2019. © 2018 Wiley Periodicals, Inc.

Finite element simulation of broadband biosonar signal propagation in the near- and far-field of an echolocating Atlantic bottlenose dolphin (Tursiops truncatus).

Bottlenose dolphins project broadband echolocation signals for detecting and locating prey and predators, and for spatial orientation. There are many unknowns concerning the specifics of biosonar signal production and propagation in the head of dolphins and this manuscript represents an effort to address this topic. A two-dimensional finite element model was constructed using high resolution CT scan data. The model simulated the acoustic processes in the vertical plane of the biosonar signal emitted from the phonic lips and propagated into the water through the animal's head. The acoustic field on the animal's forehead and the farfield transmission beam pattern of the echolocating dolphin were determined. The simulation results and prior acoustic measurements were qualitatively extremely consistent. The role of the main structures on the sound propagation pathway such as the air sacs, melon, and connective tissue was investigated. Furthermore, an investigation of the driving force at the phonic lips for dolphins that emit broadband echolocation signals and porpoises that emit narrowband echolocation signals suggested that the driving force is different for the two types of biosonar. Finally, the results provide a visual understanding of the sound transmission in dolphin's biosonar.

Anisotropic shrinkage of insect air sacs revealed in vivo by X-ray microtomography.

Air sacs are thought to be the bellows for insect respiration. However, their exact mechanism of action as a bellows remains unclear. A direct way to investigate this problem is in vivo observation of the changes in their three-dimensional structures. Therefore, four-dimensional X-ray phase contrast microtomography is employed to solve this puzzle. Quantitative analysis of three-dimensional image series reveals that the compression of the air sac during respiration in bell crickets exhibits obvious anisotropic characteristics both longitudinally and transversely. Volumetric changes of the tracheal trunks in the prothorax further strengthen the evidence of this finding. As a result, we conclude that the shrinkage and expansion of the insect air sac is anisotropic, contrary to the hypothesis of isotropy, thereby providing new knowledge for further research on the insect respiratory system.

Penguin lungs and air sacs: implications for baroprotection, oxygen stores and buoyancy.

The anatomy and volume of the penguin respiratory system contribute significantly to pulmonary baroprotection, the body O2 store, buoyancy and hence the overall diving physiology of penguins. Therefore, three-dimensional reconstructions from computerized tomographic (CT) scans of live penguins were utilized to measure lung volumes, air sac volumes, tracheobronchial volumes and total body volumes at different inflation pressures in three species with different dive capacities [Adélie (Pygoscelis adeliae), king (Aptenodytes patagonicus) and emperor (A. forsteri) penguins]. Lung volumes scaled to body mass according to published avian allometrics. Air sac volumes at 30 cm H2O (2.94 kPa) inflation pressure, the assumed maximum volume possible prior to deep dives, were two to three times allometric air sac predictions and also two to three times previously determined end-of-dive total air volumes. Although it is unknown whether penguins inhale to such high volumes prior to dives, these values were supported by (a) body density/buoyancy calculations, (b) prior air volume measurements in free-diving ducks and (c) previous suggestions that penguins may exhale air prior to the final portions of deep dives. Based upon air capillary volumes, parabronchial volumes and tracheobronchial volumes estimated from the measured lung/airway volumes and the only available morphometry study of a penguin lung, the presumed maximum air sac volumes resulted in air sac volume to air capillary/parabronchial/tracheobronchial volume ratios that were not large enough to prevent barotrauma to the non-collapsing, rigid air capillaries during the deepest dives of all three species, and during many routine dives of king and emperor penguins. We conclude that volume reduction of airways and lung air spaces, via compression, constriction or blood engorgement, must occur to provide pulmonary baroprotection at depth. It is also possible that relative air capillary and parabronchial volumes are smaller in these deeper-diving species than in the spheniscid penguin of the morphometry study. If penguins do inhale to this maximum air sac volume prior to their deepest dives, the magnitude and distribution of the body O2 store would change considerably. In emperor penguins, total body O2 would increase by 75%, and the respiratory fraction would increase from 33% to 61%. We emphasize that the maximum pre-dive respiratory air volume is still unknown in penguins. However, even lesser increases in air sac volume prior to a dive would still significantly increase the O2 store. More refined evaluations of the respiratory O2 store and baroprotective mechanisms in penguins await further investigation of species-specific lung morphometry, start-of-dive air volumes and body buoyancy, and the possibility of air exhalation during dives.

Air space proportion in pterosaur limb bones using computed tomography and its implications for previous estimates of pneumaticity.

Air Space Proportion (ASP) is a measure of how much air is present within a bone, which allows for a quantifiable comparison of pneumaticity between specimens and species. Measured from zero to one, higher ASP means more air and less bone. Conventionally, it is estimated from measurements of the internal and external bone diameter, or by analyzing cross-sections. To date, the only pterosaur ASP study has been carried out by visual inspection of sectioned bones within matrix. Here, computed tomography (CT) scans are used to calculate ASP in a small sample of pterosaur wing bones (mainly phalanges) and to assess how the values change throughout the bone. These results show higher ASPs than previous pterosaur pneumaticity studies, and more significantly, higher ASP values in the heads of wing bones than the shaft. This suggests that pneumaticity has been underestimated previously in pterosaurs, birds, and other archosaurs when shaft cross-sections are used to estimate ASP. Furthermore, ASP in pterosaurs is higher than those found in birds and most sauropod dinosaurs, giving them among the highest ASP values of animals studied so far, supporting the view that pterosaurs were some of the most pneumatized animals to have lived. The high degree of pneumaticity found in pterosaurs is proposed to be a response to the wing bone bending stiffness requirements of flight rather than a means to reduce mass, as is often suggested. Mass reduction may be a secondary result of pneumaticity that subsequently aids flight.

A unique swim bladder-inner ear connection in a teleost fish revealed by a combined high-resolution microtomographic and three-dimensional histological study.

BACKGROUND: In most modern bony fishes (teleosts) hearing improvement is often correlated with a close morphological relationship between the swim bladder or other gas-filled cavities and the saccule or more rarely with the utricle. A connection of an accessory hearing structure to the third end organ, the lagena, has not yet been reported. A recent study in the Asian cichlid Etroplus maculatus provided the first evidence that a swim bladder may come close to the lagena. Our study was designed to uncover the swim bladder-inner ear relationship in this species. We used a new approach by applying a combination of two high-resolution techniques, namely microtomographic (microCT) imaging and histological serial semithin sectioning, providing the basis for subsequent three-dimensional reconstructions. Prior to the morphological study, we additionally measured auditory evoked potentials at four frequencies (0.5, 1, 2, 3 kHz) to test the hearing abilities of the fish. RESULTS: E. maculatus revealed a complex swim bladder-inner ear connection in which a bipartite swim bladder extension contacts the upper as well as the lower parts of each inner ear, a condition not observed in any other teleost species studied so far. The gas-filled part of the extension is connected to the lagena via a thin bony lamella and is firmly attached to this bony lamella with connective material. The second part of the extension, a pad-like structure, approaches the posterior and horizontal semicircular canals and a recessus located posterior to the utricle. CONCLUSIONS: Our study is the first detailed report of a link between the swim bladder and the lagena in a teleost species. We suggest that the lagena has an auditory function in this species because the most intimate contact exists between the swim bladder and this end organ. The specialized attachment of the saccule to the cranial bone and the close proximity of the swim bladder extension to the recessus located posterior to the utricle indicate that the saccule and the utricle also receive parallel inputs from the swim bladder extension. We further showed that a combination of non-destructive microCT imaging with histological analyses on the same specimen provides a powerful tool to decipher and interpret fine structures and to compensate for methodological artifacts.

A novel hearing specialization in the New Zealand bigeye, Pempheris adspersa.

The New Zealand bigeye, Pempheris adspersa, is a nocturnal planktivore and has recently been found to be an active sound producer. The rostral end of the swim bladder lies adjacent to Baudelot's ligament which spans between the bulla and the cleithrum bone of the pectoral girdle. The aim of this study was to use the auditory evoked potential technique to physiologically test the possibility that this structure provides an enhanced sensitivity to sound pressure in the bigeye. At 100 Hz, bigeye had hearing sensitivity similar to that of goldfish (species with a mechanical connection between the swim bladder and the inner ear mediated by the Weberian ossicles) and were much more sensitive than other teleosts without ancillary hearing structures. Severing Baudelot's ligament bilaterally resulted in a marked decrease in hearing sensitivity, as did swim bladder puncture or lateral line blockage. These results show that bigeye have an enhanced sensitivity to sound pressure and provide experimental evidence that the functional basis of this sensitivity represents a novel hearing specialization in fish involving the swim bladder, Baudelot's ligament and the lateral line.

Intermolt development reduces oxygen delivery capacity and jumping performance in the American locust (Schistocerca americana).

Among animals, insects have the highest mass-specific metabolic rates; yet, during intermolt development the tracheal respiratory system cannot meet the increased oxygen demand of older stage insects. Using locomotory performance indices, whole body respirometry, and X-ray imaging to visualize the respiratory system, we tested the hypothesis that due to the rigid exoskeleton, an increase in body mass during the intermolt period compresses the air-filled tracheal system, thereby, reducing oxygen delivery capacity in late stage insects. Specifically, we measured air sac ventilation frequency, size, and compressibility in both the abdomen and femur of early, middle, and late stage sixth instar Schistocerca americana grasshoppers. Our results show that late stage grasshoppers have a reduced air sac ventilation frequency in the femur and decreased convective capacities in the abdomen and femur. We also used X-ray images of the abdomen and femur to calculate the total proportion of tissue dedicated to respiratory structure during the intermolt period. We found that late stage grasshoppers had a lower proportion of their body dedicated to respiratory structures, especially air sacs, which convectively ventilate the tracheal system. These intermolt changes make oxygen delivery more challenging to the tissues, especially critical ones such as the jumping muscle. Indeed, late stage grasshoppers showed reduced jump frequencies compared to early stage grasshoppers, as well as decreased mass-specific CO(2) emission rates at 3 kPa PO(2). Our findings provide a mechanism to explain how body mass changes during the intermolt period reduce oxygen delivery capacity and alter an insect's life history.

Basis of acoustic discrimination of Chinook salmon from other salmons by echolocating Orcinus orca.

The "resident" ecotype of killer whales (Orcinus orca) in the waters of British Columbia and Washington State have a strong preference for Chinook salmon even in months when Chinook comprise less than about 10% of the salmon population. The foraging behavior of killer whales suggests that they depend on echolocation to detect and recognize their prey. In order to determine possible cues in echoes from salmon species, a series of backscatter measurements were made at the Applied Physics Laboratory (Univ. of Wash.) Facility on Lake Union, on three different salmon species using simulated killer whale echolocation signals. The fish were attached to a monofilament net panel and rotated while echoes were collected, digitized and stored on a laptop computer. Three transducer depths were used; same depth, 22° and 45° above the horizontal plane of the fish. Echoes were collected from five Chinook, three coho and one sockeye salmon. Radiograph images of all specimens were obtained to examine the swimbladder shape and orientation. The results show that echo structure from similar length but different species of salmon were different and probably recognizable by foraging killer whales.

The use of computed tomography for assessment of the swim bladder in koi carp (Cyprinus carpio).

Seven normal koi (Cyprinus carpio) and seven koi with negative buoyancy were examined using computed tomography (CT) to assess the swim bladder. The volume of the swim bladder was calculated in all animals. In the healthy koi there was a statistical correlation (r = 0.996) between body mass and swim bladder volume with volume (ml) being related to body mass according to the formula 4.9 +/- 0.054 x BM (g). In all koi with buoyancy problems, the gas volume of the swim bladder was reduced. Additionally, fluid was found within the swim bladder in three of the abnormal koi. CT proved to be a quick noninvasive technique for the examination of the swim bladder in koi.

Effect of body position on respiratory system volumes in anesthetized red-tailed hawks (Buteo jamaicensis) as measured via computed tomography.

OBJECTIVE: To determine the effects of body position on lung and air-sac volumes in anesthetized and spontaneously breathing red-tailed hawks (Buteo jamaicensis). ANIMALS: 6 adult red-tailed hawks (sex unknown). PROCEDURES: A crossover study design was used for quantitative estimation of lung and air-sac volumes in anesthetized hawks in 3 body positions: dorsal, right lateral, and sternal recumbency. Lung volume, lung density, and air-sac volume were calculated from helical computed tomographic (CT) images by use of software designed for volumetric analysis of CT data. Effects of body position were compared by use of repeated-measures ANOVA and a paired Student t test. RESULTS: Results for all pairs of body positions were significantly different from each other. Mean +/- SD lung density was lowest when hawks were in sternal recumbency (-677 +/- 28 CT units), followed by right lateral (-647 +/- 23 CT units) and dorsal (-630 +/- 19 CT units) recumbency. Mean lung volume was largest in sternal recumbency (28.6 +/- 1.5 mL), followed by right lateral (27.6 +/- 1.7 mL) and dorsal (27.0 +/- 1.5 mL) recumbency. Mean partial air-sac volume was largest in sternal recumbency (27.0 +/- 19.3 mL), followed by right lateral (21.9 +/- 16.1 mL) and dorsal (19.3 +/- 16.9 mL) recumbency. CONCLUSIONS AND CLINICAL RELEVANCE: In anesthetized red-tailed hawks, positioning in sternal recumbency resulted in the greatest lung and air-sac volumes and lowest lung density, compared with positioning in right lateral and dorsal recumbency. Additional studies are necessary to determine the physiologic effects of body position on the avian respiratory system.

The laterophysic connection and swim bladder of butterflyfishes in the genus Chaetodon (Perciformes: Chaetodontidae).

The laterophysic connection (LC) is an association between bilaterally paired, anterior swim bladder extensions (horns) and medial openings in the supracleithral lateral line canals that diagnoses butterflyfishes in the genus Chaetodon. It has been hypothesized that the LC makes the lateral line system sensitive to sound pressure stimuli that are transmitted by the swim bladder horns and converted to fluid flow into the lateral line system via a laterophysic tympanum. The purpose of this study was to define variation in the morphology of the LC, swim bladder and swim bladder horns among 41 Chaetodon species from all 11 Chaetodon subgenera and a species from each of four non-Chaetodon genera using gross dissection, histological analysis as well as 2D or 3D CT (computed tomographic) imaging of live, anesthetized fishes. Our results demonstrate that the lateral line system appears rather unspecialized with well-ossified narrow canals in all species examined. Two LC types (direct and indirect), defined by whether or not the paired anterior swim bladder horns are in direct contact with a medial opening in the supracleithral lateral line canal, are found among species examined. Two variants on a direct LC and four variants of an indirect LC are defined by combinations of soft tissue anatomy (horn length [long/short] and width [wide/narrow], number of swim bladder chambers [one/two], and presence/absence of mucoid connective tissue in the medial opening in the supracleithrum). The combination of features defining each LC variant is predicted to have functional consequences for the bioacoustics of the system. These findings are consistent with the recent discovery that Chaetodon produce sounds during social interactions. The data presented here provide the comparative morphological context for the functional analysis of this novel swim bladder-lateral line connection.

Radiodiagnostic method for studying the dynamics of Anguillicola crassus (Nematoda: Dracunculoidea) infection and pathological status of the swimbladder in Lake Balaton eels.

Swimbladder changes resulting from Anguillicola crassus infection of the European eel Anguilla anguilla have been the subject of several studies reported in the literature. These investigations, however, studied exclusively the status of infection at a given point in time and did not deal with changes in swimbladder infection in eels suffering from anguillicolosis over a period of time. In this study, A. crassus-induced pathological changes were monitored in 78 eels naturally infected in Lake Balaton and subsequently kept in the laboratory, thus excluding the possibility of further infection. During the 3 mo study, the status of the swimbladder was checked by radiographic examination on 4 occasions. At the end of the study the eels were dissected and the gross pathological changes in the swimbladders were compared with the radiographic findings. As compared to their starting condition, by the end of the study the pathological status of the swimbladder had deteriorated in 55% and remained the same in 37% of the cases. Tendency to improvement (1%) and variable findings (7%) were recorded in a low percentage of cases only. With the help of the radiographs presented, the dynamics of A. crassus infection and of changes in the swimbladder of individual eel specimens can be monitored easily.

Broadband acoustic backscatter and high-resolution morphology of fish: measurement and modeling.

Broadband acoustic backscattering measurements, advanced high-resolution imaging of fish morphology using CT scans and phase-contrast x rays (in addition to traditional x rays), and associated scattering modeling using the images have been conducted involving alewife (Alosa pseudoharengus), a swimbladder-bearing fish. A greater-than-octave bandwidth (40-95 kHz) signal was used to insonify live, individual, adult alewife that were tethered while being rotated in 1-deg increments over all angles in two planes of rotation (lateral and dorsal/ventral). These data, in addition to providing the orientation dependence of the scattering over a continuous band of frequencies, were also used (after pulse compression) to identify dominant scattering features of the fish (including the skull and swimbladder). The x-ray and CT scan images of the swimbladder were digitized and incorporated into two scattering models: (1) Kirchhoff-ray mode (KRM) model [Clay and Horne, J. Acoust. Soc. Am. 96, 1661-1668 (1994)] and (2) conformal-mapping-based Fourier matching method (FMM), which has recently been extended to finite-length bodies [Reeder and Stanton, J. Acoust. Soc. Am. 116. 729-746 (2004)]. Comparisons between the scattering predictions and data demonstrate the utility of the CT scan imagery for use in scattering models, as it provided a means for rapidly and noninvasively measuring the fish morphology in three dimensions and at high resolution. In addition to further validation of the KRM model, the potential of the new FMM formulation was demonstrated, which is a versatile approach, valid over a wide range of shapes, all frequencies and all angles of orientation.

Swim bladder and posterior lateral line nerve of the nurseryfish, Kurtus gulliveri (Perciformes: Kurtidae).

The morphology of the swim bladder and inner ear of the nurseryfish, Kurtus gulliveri, appear adapted for enhanced pressure wave reception. The saccule is enlarged and surrounded by very thin bone and two large fontanelles that would present reduced resistance to pressure waves. The swim bladder is elaborate, with six dorsolaterally projecting pairs of lobes that are tightly encased in ribs and an additional caudally projecting pair of lobes encased in the first hemal spine. The ribs and musculature surrounding the swim bladder laterally are very thin, so that four or five "rib windows" are readily apparent on back-lit specimens. This swim bladder-rib configuration would also present reduced resistance to pressure waves to enhance function as a peripheral auditory structure. However, high-resolution X-ray computed tomography and dissection reveal no anterior projections of the swim bladder that could serve as a mechanical coupling to the inner ear. The posterior lateral line nerve is well developed and lies directly over the tips of the ribs encasing the swim bladder lobes. This nerve is not, however, associated with a lateral line canal and a lateral line canal is absent on most of the body. We hypothesize that the posterior lateral line nerve transmits mechanosensory information from the swim bladder.

Comparative study of X-ray computerised tomography and conventional X-ray methods in diagnosis of swimbladder infection in eels caused by Anguillicola crassus.

To date, swimbladder lesions due to Anguillicola crassus infection of the European eel Anguilla anguilla have so far been studied only by conventional X-ray methods. This is the first study to report the use of computerised tomography (CT) for studying lesions induced by anguillicolosis. Of 50 eels caught by electrofishery from Lake Balaton, Hungary, in autumn 2002 and pre-selected by a conventional X-ray method, 22 specimens were examined with a Siemens Somatom Plus S40 spiral CT scanner. Tomograms, radiographs and photographs of 5 of these, showing anguillicolosis-induced swimbladder lesions of varying severity, are presented. Computerised tomograms provide information on the inner structure, air content and wall thickness of the swimbladder as well as on the number of worms it contains. When the swimbladder is not severely affected or not completely filled with worms, computerised tomography provides adequate data on the shape of the swimbladder, thickness of the swimbladder wall and the location of worms in the lumen. However, in more severe cases, i.e. when the swimbladder is tightly packed with worms or contains no air as a result of wall-thickening, this method fails to determine the number and location of helminths or the thickness of the swimbladder wall.

Radiodiagnostic examination of the swimbladder of some fish species.

Radiodiagnostic methods have not been used previously for studying the anatomy and diseases of the swimbladder of freshwater fish species. In this study, the radiographic anatomy of the swimbladder and species-related differences in swimbladder structure were studied on plain radiographs taken of 12 Hungarian fish species of major economic importance. Changes observed by radiography were also studied by conventional parasitological methods. The radiodiagnostic method reported here appears to be a useful complement to diagnostic examinations that have been based merely on dissection so far. It enables evaluation of the pathological lesions in live condition, without causing damage to the fish.