Gross morphology of the brain and some sense organs of subterranean pencil catfishes of the genus Ituglanis Costa and Bockmann, 1993 (Siluriformes, Trichomycteridae), with a discussion on sensory compensation versus preadaptation in subterranean fishes.

Subterranean organisms provide excellent opportunities to investigate morphological evolution, especially of sensory organs and structures and their processing areas in the central nervous system. We describe the gross morphology of the brain and some cephalic sensory organs (olfactory organ, eye, semicircular canals of the inner ear) and the swim bladder (a non-sensory accessory structure) of subterranean species of pencil catfishes of the genus Ituglanis Costa and Bockmann, 1993 (Siluriformes, Trichomycteridae) and compare them with an epigean species of the genus, Ituglanis goya Datovo, Aquino and Langeani, 2016. We compared qualitatively the size of the different brain regions and sense organs of the subterranean species with those of the epigean one, searching for modifications possibly associated with living in the subterranean environment. Our findings suggest that species of Ituglanis exhibit sensory characteristics that are preadaptive for the subterranean life, as only slight modifications were observed in the brains and sense organs of the subterranean species of the genus when compared with the epigean one. Because most subterranean fish species belong to lineages putatively preadapted for subterranean life, our results, discussed in the context of available information on the brain and sense organs of other subterranean species, help identify general trends for the evolution of the brain and sensory organs of subterranean fishes in general.

Sound production and mechanism in the cryptic cusk-eel Parophidion vassali.

This study investigates the sounds and the anatomy of the sound-producing organ in the male and female sand-dwelling cusk-eel Parophidion vassali. Although both sexes have similar external phenotype, they can be distinguished by their sonic apparatus and sounds. As in many Ophioidei, Parophidion vassali presents a panel of highly derived characters. Fish possess three pairs of sonic muscles, and males have mineralized swimbladder caps on which inserts the ventral sonic muscle, a neural arch that pivots, a stretchable swimbladder fenestra, an osseous swimbladder plate and a rounded pressure-release membrane in the caudal swimbladder. Females, however, do not possess anterior swimbladder caps, a swimbladder fenestra and the caudal rounded membrane. Males possess the unusual ability to produce sounds starting with a set of low amplitude pulses followed by a second set with higher amplitudes clearly dividing each sound unit into two parts. Females do not vary their sound amplitude in this way: they produce shorter sounds and pulse periods but with a higher peak frequency. Morphology and sound features support the sound-producing mechanism is based on a rebound system (i.e. quick backward snap of the anterior swimbladder). Based on features of the sounds from tank recordings, we have putatively identified the sound of male Parophidion vassali at sea. As these species are ecologically cryptic, we hope this work will allow assessment and clarify the distribution of their populations.

Slight volume changes in the duck lung do not imply a fundamental change in the structure of the parenchyma.

Slight changes in lung volume have previously been reported in ducks. We studied the functional structure of the lung of the domestic duck using classical anatomical techniques as well as ultrasound monitoring to unravel the causes of such changes. Later dorsal and medioventral secondary bronchi were superficially positioned and covered with a thin transparent and collapsible membrane, internally lined with a cuboidal to squamous epithelium. The lung parenchyma was rigid, with atria well supported by septa containing smooth muscles, interparabronchial septa reinforced by collagen fibres, and blood capillaries supported by epithelial plates. On ultrasound monitoring, an outward and inward movement of the lung surface during inspiration and expiration, respectively, was evident at the region where the airways were covered by the thin membranes. The movements plausibly facilitated air movement in the lung just like the air sacs. We conclude that volume changes in the duck lung occur due to a slight morphological adaptation rather than a change in the archetypical design of the avian lung parenchyma.

A sciaenid swim bladder with long skinny fingers produces sound with an unusual frequency spectrum.

Swim bladders in sciaenid fishes function in hearing in some and sound production in almost all species. Sciaenid swim bladders vary from simple carrot-shaped to two-chambered to possessing various diverticula. Diverticula that terminate close to the ears improve hearing. Other unusual diverticula heading in a caudal direction have not been studied. The fresh-water Asian species Boesemania microlepis has an unusual swim bladder with a slightly restricted anterior region and 6 long-slender caudally-directed diverticula bilaterally. We hypothesized that these diverticula modify sound spectra. Evening advertisement calls consist of a series of multicycle tonal pulses, but the fundamental frequency and first several harmonics are missing or attenuated, and peak frequencies are high, varying between < 1-2 kHz. The fundamental frequency is reflected in the pulse repetition rate and in ripples on the frequency spectrum but not in the number of cycles within a pulse. We suggest that diverticula function as Helmholz absorbers turning the swim bladder into a high-pass filter responsible for the absence of low frequencies typically present in sciaenid calls. Further, we hypothesize that the multicycle pulses are driven by the stretched aponeuroses (flat tendons that connect the sonic muscles to the swim bladder) in this and other sciaenids.

Histological development and integration of the Zebrafish Weberian apparatus.

BACKGROUND: The Weberian apparatus enhances hearing in otophysan fishes, including Zebrafish (Danio rerio). Several studies have examined aspects of morphological development of the Weberian apparatus and hearing ability in Zebrafish. A comprehensive developmental description including both hard and soft tissues is lacking. This information is critical for both interpretation of genetic developmental analyses and to better understand the role of morphogenesis and integration on changes in hearing ability. RESULTS: Histological development of hard and soft tissues of the Weberian apparatus, including ossicles, ear, swim bladder, and ligaments are described from early larval stages (3.8 mm notochord length) through adult. Results show a strong relationship in developmental timing and maturation across all regions. All required auditory elements are present and morphologically integrated early, by 6.5 mm SL. Dynamic ossification patterns and changes in shape continue throughout the examined developmental period. CONCLUSIONS: This study provides the first comprehensive histological description of Weberian apparatus development in Zebrafish. Morphological integration was found early, before increases in hearing ability were detected in functional studies (>10 mm total length), suggesting morphological integration precedes functional integration. Further research is needed to examine the nature of the functional delay, and how maturation of the Weberian apparatus influences functionality.

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.

Histology and structural integration of the major morphologies of the Cypriniform Weberian apparatus.

The Weberian apparatus, a diagnostic feature of otophysan fishes, is a novel hearing adaptation integrating several developmental and morphological systems (ear-vertebral column-swim bladder). Otophysan fishes are one of the largest and most successful freshwater clades, with over 10,000 species across most continents. The largest otophysan order, Cypriniformes, dominates the freshwaters of Asia, Europe, North America, and Africa. Spanning such a wide variety of environments, the Weberian apparatus undergoes morphological modifications to maintain functionality. Within Cypriniformes, we propose three distinct morphological classes of the Weberian apparatus based on the level of skeletal expansion around the swim bladder: simple (typical of most Cyprinidae), anterior plate (found in families such as Gyrinocheilidae, Catostomidae, and Botiidae), and encapsulated (either single-capsule as found, e.g., in Gobionidae and Cobitidae, or double-capsule as found, e.g., in Nemacheilidae and Balitoridae). Little ontological or comparative data exists regarding the construction or integration of these different morphologies, and less is known about the tissue level integration and variation within these morphologies. We used paraffin histology to document the hard and soft tissue anatomy of the Weberian apparatus in six species representing all morphological classes. We found sites of similarity across the morphologies including size and structure of the saccule, aspects of ossicle ossification, and swim bladder tunica composition, indicating potential sites of developmental and functional constraint. In contrast, we found differences across both auditory and nonauditory features in otic chamber size, ossification within ossicles and other vertebral elements, and composition of ligaments, indicating likely sites of adaptability. Some of these changes are likely evolutionary (taxonomic), but may be influenced by the environmental niche occupied by the clade. These results show a clear need for increased ontological and comparative study of the complete cypriniform Weberian apparatus, particularly histologically, as well as increased auditory studies across morphological types.

Gross morphological features of the air sacs of the hooded crow (Corvus cornix).

Air sacs are considered to be one of the controlling factors of bird behaviour and habits in addition to their roles in ventilation, regulating body temperature, swimming and flight. As a scavenger and an omnivorous flight bird, air sacs of the hooded crow were the focus of this study. Eight healthy, adult hooded crows were used to examine the morphological characteristics of the air sacs, which were examined grossly and with latex and cast preparations. In general, the morphological overview of the hooded crow air sacs is similar to other avian species. We observed nine air sacs; four paired sacs (cervical, cranial thoracic, caudal thoracic and abdominal air sacs) and one unpaired sac; the clavicular air sac. The cervical air sac communicated to the lung through the medioventral bronchus and had three diverticula; intermuscular, subscapular and subcutaneous. The clavicular air sac communicated with lung through the medioventral bronchus and had subscapular, axillary, humeral, subpectoral and sternal diverticula. The cranial and caudal thoracic air sacs were communicated with lung through the lateroventral bronchi and the both sacs did not have any diverticula. The abdominal air sacs were posterior to the caudal thoracic air sacs. The left abdominal sac was the largest air sac. The right and left abdominal sacs gave off branches to diverticula that pneumatized synsacrum. The abdominal air sacs gave off femoral diverticula behind the hip joint as well as perirenal diverticula.

Target Strength and swimbladder morphology of Mueller's pearlside (Maurolicus muelleri).

In the last few years, there has been increasing interest in the commercial exploitation of mesopelagic fish and a trawl-acoustic methodology has been recommended to make estimates of abundance of these resources. This study provides relevant information on the scattering properties of a key mesopelagic fish species in the Bay of Biscay, Mueller's pearlside (Maurolicus muelleri), necessary to convert the acoustic density into numerical abundance. The target strength (TS) of pearlside was estimated for the first time at five frequencies commonly used in acoustic surveys. A high-density filter was applied to reduce the bias derived from overlapping echoes erroneously assigned to single targets. Its relationship with fish length (b20) was also determined (-65.9 ± 2, -69.2 ± 3, -69.2 ± 2, -69.5 ± 2.5 and -71.5 ± 2.5 dB at 18, 38, 70, 120 and 200 kHz, respectively). Biomass estimates of pearlside in the Bay of Biscay during the four years of study (2014-2017) are given using the 38 kHz frequency. Morphological measurements of the swimbladder were obtained from soft X-ray images and used in the backscattering simulation of a gas-filled ellipsoid. Pearlside is a physoclist species, which means that they can compensate the swimbadder volume against pressure changes. However, the best fit between the model and the experimental data showed that they lose that capacity during the trawling process, when the swimbladder volume is affected by Boyle's law.

STANDING COMPUTED TOMOGRAPHY IN NONANESTHETIZED LITTLE PENGUINS (EUDYPTULA MINOR) TO ASSESS RESPIRATORY SYSTEM ANATOMY AND MONITOR DISEASE.

Multidetector computed tomography (MDCT) scans were performed in clinically healthy, nonanesthetized, standing little penguins (Eudyptula minor) to determine reference ranges for air-sac and lung volumes, as well as lung density. Five of 15 clinically healthy birds were diagnosed with pulmonary granulomas on initial MDCT scans. Granulomas were not readily apparent on radiographs, even in cases where the entire normal pulmonary parenchymal architecture was effaced on the MDCT scan. Serial MDCT scans after antifungal and antimycobacterial therapies demonstrated a response to treatment. MDCT scanning in nonanesthetized little penguins proved to be a well-tolerated, non-invasive imaging modality for respiratory diseases that are otherwise difficult to diagnose, including aspergillosis and mycobacteriosis.

Disturbance calls of five migratory Characiformes species and advertisement choruses in Amazon spawning sites.

Species-specific disturbance calls of five commercially-important characiform species are described, the Curimatidae commonly called branquinhas: Potamorhina latior, Potamorhina altamazonica and Psectrogaster amazonica; Prochilodontidae: jaraquí Semaprochilodus insignis and curimatã Prochilodus nigricans. All species have a two-chambered swimbladder and the sonic mechanism, present exclusively in males, utilises hypertrophied red muscles between ribs that adhere to the anterior chamber. The number of muscles is unusually plastic across species and varies from 1 to 4 pairs suggesting considerable evolution in an otherwise conservative system. Advertisement calls are produced in river confluences in the Madeira Basin during the high-water mating season (January-February). Disturbance calls and sampling allowed recognition of underwater advertisement choruses from P. latior, S. insignis and P. nigricans. The advertisement calls of the first two species have largely similar characteristics and they mate in partially overlapping areas in the Guaporé River. However, P. latior sounds have a lower dominant frequency and it prefers to call from river confluences whereas S. insignis shoals occur mostly in the main river channel adjacent to the confluence. These results help identify and differentiate underwater sounds and evaluate breeding areas during the courtship of commercially important characids likely to be affected by two hydroelectric dams.

Ultraviolet light illuminates the avian nature of the Berlin Archaeopteryx skeleton.

The question of whether the iconic avialan Archaeopteryx was capable of active flapping flight or only passive gliding is still unresolved. This study contributes to this debate by reporting on two key aspects of this fossil that are visible under ultraviolet (UV) light. In contrast to previous studies, we show that most of the vertebral column of the Berlin Archaeopteryx possesses intraosseous pneumaticity, and that pneumatic structures also extend beyond the anterior thoracic vertebrae in other specimens of Archaeopteryx. With a minimum Pneumaticity Index (PI) of 0.39, Archaeopteryx had a much more lightweight skeleton than has been previously reported, comprising an air sac-driven respiratory system with the potential for a bird-like, high-performance metabolism. The neural spines of the 16th to 22nd presacral vertebrae in the Berlin Archaeopteryx are bridged by interspinal ossifications, and form a rigid notarium-like structure similar to the condition seen in modern birds. This reinforced vertebral column, combined with the extensive development of air sacs, suggests that Archaeopteryx was capable of flapping its wings for cursorial and/or aerial locomotion.

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.

Relative size variation of the otoliths, swim bladder, and Weberian apparatus structures in piranhas and pacus (Characiformes: Serrasalmidae) with different ecologies and its implications for the detection of sound stimuli.

The Weberian apparatus of otophysan fishes confers acute hearing that is hypothesized to allow these fishes to assess the environment and to find food resources. The otophysan family Serrasalmidae (piranhas and pacus) includes species known to feed on falling fruits and seeds (frugivore/granivores) that splash in rivers, herbivorous species associated with torrents and rapids (rheophiles), and carnivores that feed aggressively within shoals. Relevant sound stimuli may vary among these ecological groups and hearing may be tuned to different cues among species. In this context, we examined size variation of the Weberian ossicles, swim bladder chambers, and otoliths of 20 serrasalmid species from three broad feeding ecologies: frugivore/granivores, rheophiles, and carnivores. We performed 3D-reconstructions of high resolution tomographic data (µCT) from 54 museum specimens to estimate the size of these elements. We then tested for an ecology effect on covariation of auditory structure size and body size and accounted for phylogeny with phylogenetic generalized least squares analyses. Among ecological groups, we observed differences in relative sizes of otoliths associated with sound pressure and particle motion detection, and variation in Weberian ossicle size that may impact sound transmission. Rheophiles, which live in noisy environments, possess the strongest modifications of these structures.

X-ray computed tomography study of the flight-adapted tracheal system in the blowfly Calliphora vicina, analysing the ventilation mechanism and flow-directing valves.

Following the discovery of flight motor-driven unidirectional gas exchange with rising PO2  in the blowfly, X-ray computed tomography (CT) was used to visualize the organization of the tracheal system in the anterior body with emphasis on the arrangement of the pathways for airflow. The fly's head is preferentially supplied by cephalic tracheae originating from the ventral orifice of the mesothoracic spiracle (Sp1). The respiratory airflow during flight is a by-product of cyclic deformations of the thoracic box by the flight muscles. The air sacs below the tergal integument (scutum and scutellum) facilitate the respiratory airflow: the shortening of the thorax turns the scutellum and the wings downward and the scutum upward with a volume increase in the scutal air sacs. The resulting negative pressure sucks air from Sp1 through special tracheae towards the scutal air sacs. The airflow is directed by two valves that open alternately: (1) the hinged filter flaps of the metathoracic spiracles (Sp2) are passively pushed open during the upstroke by the increased tracheal pressure, thereby enabling expiration; (2) a newly described tracheal valve-like septum behind the regular spiracular valve lids of Sp1 opens passively and air is sucked in through Sp1 during the downstroke and prevents expiration by closing during the upstroke. This stabilizes the unidirectional airflow. The tracheal volume of the head, thorax and abdomen and their mass were determined. Despite the different anatomy of birds and flies, the unidirectional airflow reveals a comparable efficiency of the temporal throughput in flies and hummingbirds.

Anatomy of the lower respiratory tract in domestic birds, with emphasis on respiration.

This manuscript describes the anatomy of the lower respiratory tract in domestic bird species including the chicken and pigeon. The here described anatomical structures play a major role avian respiration, which is fundamentally different from respiration in mammals. During inspiration and expiration, a continuous caudocranial airflow is present within the tertiary bronchi of the Paleopulmo, while the Neopulmo, which is only present in phylogenetically recent species, is characterized by tidal respiration. Various anatomical structures and aerodynamic mechanisms have been described in an attempt to explain the proposed mechanism of respiration. The air sac system that is essential for avian respiration usually comprises an unpaired clavicular air sac and paired cervical, cranial and caudal thoracic, and abdominal air sacs. The latter are by far the larger and are interwoven with the abdominal organs.

Ontogeny and morphometrics of the gills and swim bladder of air-breathing striped catfish Pangasianodon hypophthalmus.

The air-breathing fish Pangasianodon hypophthalmus has been shown to have highly plastic branchial surfaces whose area (SA) increases with temperature and aquatic hypoxia. This modulation occurs through development of inter-lamellar cell mass (ILCM). Paradoxically, in conditions where this fish has been shown capable of covering its entire aerobic scope from the water phase, it has been shown to have a very small branchial SA. To address this paradox, we measured the SA, harmonic mean diffusion distance (τh) and calculated the anatomic diffusion factor (ADF) of the branchial and swim bladder surfaces in fish ranging from 3 to 1900 g at 27°C in normoxia. Since the lamellae were distinguishable from the ILCM, we measured the actual SA as well as the potential SA if ILCM were lost. As a result of low τh, P. hypophthalmus has a high capacity for branchial oxygen uptake with or without ILCM. Actual and potential gill ADF were 361 and 1002 cm2 µm-1 kg-1, respectively, for a 100 g fish and the ADF of the swim bladder was found to be 308 cm2 µm-1 kg-1 By swimming fish to exhaustion at different temperatures, we show that modulation of this SA is rapid, indicating that the apparent paradox between previous studies is eliminated. Regression analysis of log-log plots of respiratory SA in relation to body mass shows that the gill scales with mass similarly to the SA in active water-breathing fish, whereas the swim bladder scales with mass more like the mammalian lung does. This fish presents a combination of respiratory surfaces not previously seen in air-breathing fish.

Swim bladder morphology changes with female reproductive state in the mouth-brooding African cichlid Astatotilapia burtoni.

Mouth brooding is an extreme form of parental care in which the brooding parent carries the developing young in their buccal cavity for the duration of development. Brooding fish need to compensate for the brood weight on the anterior portion of their body. For fishes with a compartmentalized swim bladder, gas distribution between the chambers may aid in regulating buoyancy during brooding. To test this hypothesis, we took radiographs of Astatotilapia burtoni to compare the swim bladder morphology of gravid, mouth-brooding and recovering females. Following spawning, females carry developing fish in their buccal cavity for ∼2 weeks, resulting in a larger and rounder anterior swim bladder compartment. Comparatively, the swim bladder of gravid females is long and cylindrical. Using small beads to mimic brood weight and its effects on female buoyancy, swim bladder changes were induced that resembled those observed during brooding. Immediately after releasing their fry, brooding females swim at a positive angle of attack but correct their swimming posture to normal within 5 min, suggesting a rapid change in swim bladder gas distribution. These data provide new insights into how swim bladder morphology and swimming behavior change during mouth brooding, and suggest a compartmentalized swim bladder may be a morphological adaptation for mouth brooding.

Intra- and Intersexual swim bladder dimorphisms in the plainfin midshipman fish (Porichthys notatus): Implications of swim bladder proximity to the inner ear for sound pressure detection.

The plainfin midshipman fish, Porichthys notatus, is a nocturnal marine teleost that uses social acoustic signals for communication during the breeding season. Nesting type I males produce multiharmonic advertisement calls by contracting their swim bladder sonic muscles to attract females for courtship and spawning while subsequently attracting cuckholding type II males. Here, we report intra- and intersexual dimorphisms of the swim bladder in a vocal teleost fish and detail the swim bladder dimorphisms in the three sexual phenotypes (females, type I and II males) of plainfin midshipman fish. Micro-computerized tomography revealed that females and type II males have prominent, horn-like rostral swim bladder extensions that project toward the inner ear end organs (saccule, lagena, and utricle). The rostral swim bladder extensions were longer, and the distance between these swim bladder extensions and each inner-ear end organ type was significantly shorter in both females and type II males compared to that in type I males. Our results revealed that the normalized swim bladder length of females and type II males was longer than that in type I males while there was no difference in normalized swim bladder width among the three sexual phenotypes. We predict that these intrasexual and intersexual differences in swim bladder morphology among midshipman sexual phenotypes will afford greater sound pressure sensitivity and higher frequency detection in females and type II males and facilitate the detection and localization of conspecifics in shallow water environments, like those in which midshipman breed and nest.

Biosonar signal propagation in the harbor porpoise's (Phocoena phocoena) head: The role of various structures in the formation of the vertical beam.

Harbor porpoises (Phocoena phocoena) use narrow band echolocation signals for detecting and locating prey and for spatial orientation. In this study, acoustic impedance values of tissues in the porpoise's head were calculated from computer tomography (CT) scan and the corresponding Hounsfield Units. A two-dimensional finite element model of the acoustic impedance was constructed based on CT scan data to simulate the acoustic propagation through the animal's head. The far field transmission beam pattern in the vertical plane and the waveforms of the receiving points around the forehead were compared with prior measurement results, the simulation results were qualitatively consistent with the measurement results. The role of the main structures in the head such as the air sacs, melon and skull in the acoustic propagation was investigated. The results showed that air sacs and skull are the major components to form the vertical beam. Additionally, both beam patterns and sound pressure of the sound waves through four positions deep inside the melon were demonstrated to show the role of the melon in the biosonar sound propagation processes in the vertical plane.